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IMUNITATEA = APĂRARE

Toţi ne îmbolnăvim uneori..., apoi ne
însănătoşim…

● Ce se întâmplă în organism în timpul unei
infecţii / afecţiuni ?
● De ce ne însănătoşim ?

SISTEMUL IMUN

Colecţie de CELULE şi MOLECULE care
protejează organismul împotriva infecţiilor,
transformărilor maligne şi celulelor proprii
transformate non-self

IMUNITATE
INNASCUTA / nespecifica DOBINDITA / specifica
(innate immunity
immunity)) (adaptive immunity)

 raspuns independent de antigen  raspuns dependent de antigen
 receptori pt. patogeni:  receptori pt. patogeni:
 codificati in genom  generati (rearanjare genica)
 specificitate joasa  specificitate inalta
 raspuns imediat  raspuns lent
 lipsa memorie  memorie imunologica
(expansiune clonala Ly)
 prezenta la toate organismele  prezenta doar la vertebrate
pluricelulare

SISTEM INTEGRAT

Răspunsuri imune
Barriers Skin & Mucous membranes
Invasion rapidly regenerating surfaces,
peristaltic movement, mucociliary
& infection escalator, vomiting, flow of urine/tears,
coughing

Innate immunity Cellular and humoral defences
+ lysosyme, sebaceous/mucous secretions,
stomach acid, commensal
organisms,complement proteins,
phagocytosis, NK cells

+ Inflammation
Cellular and humoral defences
Adaptive immunity Antibodies, cytokines, T helper cells,
cytotoxic T cells

Imunitatea
- piele
innăscută
1. Bariere anatomice - mucoase

• Functia de prevenire a intrarii
microorganismelor patogene
• motilitate: muco-ciliara, peristaltica
Factori fizici / mecanici • mucus
• fluxul fluidelor prin organism

• pH
Factori chimici • molecule antimicrobiene

• limfocite T intraepiteliale ()
Factori biologici • celule B-1

Imunitatea
- receptori
innăscută 2. Componente moleculare
- molecule secretate

Functia de recunoastere
Functii efectorii

Mol. anorganice: HCl, NO, H2O2
Peptide antibacteriene: defensine, cathelicidine, histatine
Proteine antibacteriene: lizozim, lactoferina, transferina
Lectine: colectine, ficoline, receptori pentru Manoza
Complement
Citokine: IFN-/, IL-1, TNF-, CSF
Chemokine: IL-8, MIP, MCP
Receptori:: TLR
Receptori

Componente celulare Functia de eliminare a microorganismelor patogene neutrofile monocite/macrofage Fagocitoza celule dendritice bazofile mastocite Inflamatie eozinofile celule NK Citotoxicitate .Imunitatea innăscută 3.

Immune responses Barriers Skin & Mucous membranes rapidly regenerating surfaces. vomiting. phagocytosis. flow of urine/tears. NK cells + Inflammation Cellular and humoral defences Antibodies. commensal organisms.complement proteins. stomach acid. sebaceous/mucous secretions. coughing Innate immunity Cellular and humoral defences + lysosyme. cytokines. mucociliary & infection escalator. Adaptive immunity cytotoxic T cells . T helper cells. Invasion peristaltic movement.

expunere la agenţi corozivi) ● biologică (ex. arsuri) ● chimică ( ex. microorganisme) ● imunologică (ex. montat să restabilească statusul normal  Iniţiată de lezarea ţesuturilor: ● mecanică/fizică (ex. precum şi excretaţi de celulele secretorii . Inflamaţia  Componentă majoră a răspunsului imun nespecific/ specific  Răspuns protectiv la injurie. hipersenzitivitate)  Implică fagocite şi mediatorii secretaţi de fagocite.

Inflamatia .Functii  Distrugerea si indepărtarea substantelor / particulelor dăunătoare / străine  Izolarea zonei infectate/inflamate  Stimularea răspunsului imun specific  Vindecare .

fagocite noi din torentul circulator sunt atrase la locul infectiei  Fagocitele ingurgiteaza particulele straine similar cu amoebele . Răspunsul inflamator celular (1)  Neutrofilele si monocitele circulante au o mobilitate foarte mare. PMN si Mo au capacitatea sa se strecoare prin spatiile inguste interendoteliale. recrutate chemotactic (chemokine) la locul infectiei ● Daca infectia se raspandeste.

Leukocyte Adhesion • Phagocytic neutrophils respond to an epithelial chemokine IL-8 • Cells migrate from the blood into the tissue underlying the infection .

Diapedesis .

Caracteristicile Inflamaţiei  Edemaţierea zonei (tumor)  Eritem (rubor)  Creşterea temperaturii (calor)  Durere (dolor)  Pierderea funcţionalităţii (functio lesa)  Reacţii / interferenţe inflamatorii locale: ● Activarea coagulării ● Căile formatoare ale Kininelor ● Fibrinoliza .

Răspunsul inflamator local Eliberarea de histamină şi prostaglandine determină vasodilataţie locală însoţită de: o increased blood flow  redness and warmth o increased capillary permeability o edema (swelling) due to fluids seeping from capillaries o more WBCs to site o phagocytes move out of vessels into extracellular fluid (ECF) o phagocytes engulf and destroy bacteria .

Loss of Function . Inflammation Heat .Swelling .Pain .Redness .

Allergy .Allergy Myeloid Basophil ?Protection of progenitor mucosal surfaces? .Allergy Protection of Mast cell mucosal surfaces . Lymphocyte Adaptive Cells Of The immunity Immune System Macrophage Phagocytosis Common Monocyte Ag presentation lymphoid progenitor Neutrophil Phagocytic PMN Anti-bacterial Pluripotent haemopoietic Anti-parasite stem cell Eosinophil Immunity .

MQ (1)  Leucocite Polimorfonucleare (PMN) ● Granulocite: o Neutrofile.PMN. eozinofile. migreaza din sange in tesuturi  se diferentiaza: o celule Kupffer in ficat o macrofage alveolare in plaman o celule mezangiale in rinichi o macrofage splenice (sinusale) in pulpa alba . mastocite ● Sunt fagocite cu viaţă scurtă.Sistemul Reticuloendotelial (SRE) / Reticulohistiocitar (SRH) o macrofage peritoneale plutind liber in fluidul peritoneal (seroase) o celule microgliale in SNC o celule Langerhans in piele .Fagocite profesionale . basofile. continând lizozomi ● Produc apă oxigenată şi radicali superoxid ● Proteine bactericide – lactoferina ● PMNs joacă un rol major in protectia împotriva infectiilor ● Defecte genetice/dobândite – infectii cronice sau recurente  Macrofagele – fagocite tisulare derivate din monocitele ciculante.

 Mastocitul: ● elibereaza amine biogene (histamina. leukotriene.  PMN: ● fagociteaza agresorul . Inflamatia locala  In tesutul interstitial exista o populatie rezidenta de leucocite. serotonina) ● secreta prostaglandine. citokine si TNF-a.

Main role is to get to site of infection rapidly and ingest microorganisms. Neutrophil Chemotaxis Neutrophils chasing yeast Neutrophil . Neutrophils Also called polymorphonucelar leukocytes 4 to 10 million per ml of blood Infection activates cytokines that Neutrophil stimulate the bone marrow to produce up to 20 million neutrophils per ml of blood. After taking up microorganisms the neutrophil will die.

Kupffer cells Lung .5 to 1 million monocytes per ml of blood Migrate into the tissues and differentiate into Macrophages Blood monocyte Phagocytose microorganisms Present antigens to T cells The name of monocyte-derived cells depends upon the tissue they reside in: Liver .Osteoclasts Tissue macrophage Human Macrophage ingesting yeast . Monocytes 0.Microglial cells Bone .Alveolar macrophages CNS .

in spatiul interstitial trec PMN: ● se acumuleaza in 30-60 minute de la aparitia agentului declansator ● fagociteaza “intrusul” si tesuturile lezate ● elibereaza enzime lizozomale  Daca inflamatia persista: ● dupa 12-18 ore interstitiul este infiltrat cu mononucleare (macrofage si limfocite) ● Macrofagele o suplimenteaza activitatea PMN o prezinta Ag. Fagocite (2)  Initial. Ly T  Daca inflamatia continua. raspunsul inflamator este suplimentat si accentuat cu elemente ale imunitatii dobandite  Ac si LyT (Ac activeaza si sistemul complementului) Vindecare .

fagocite mai putin eficiente)  Celule dendritice in splina si ganglioni  Celule interdigitate in timus  Celule Langerhans in piele . Fagocite (3)  Celulele seriei macrofagice au doua functii majore: ● Ingereaza si digera microorganisme si particule straine ● Prezinta Antigene (Ag) . il proceseaza si il prezinta Ly T * Alte celule prezentatoare de Ag (APC) (precursori hematopoietici.componenta imunogena o Preia Ag.

Fagozom . initiat de legarea fagocitului la patogen prin intermediul unor receptori •Agentul patogen este inconjurat de pseudopode membranare si apoi internalizat: .Corp rezidual .Fagocitoza (cont) •Proces activ.Fagolizozom .Vacuola digestiva .

initiat de legarea la patogen prin intermediul unor receptori  Agentul patogen este inconjurat de pseudopode membranare si apoi internalizat .Fagocitoza  Proces activ.

Bacteria. General scheme of an immune response Most microorganisms do not cause disease in humans Pathogens Disease . Worms etc Both humans and pathogens are made of How does the human proteins immune system carbohydrates distinguish & lipids pathogens from human tissues and harmless microorganisms? . Fungi. Viruses.causing organisms Protozoa.

chlatrin-independenta ● Rata & eficienta mare de internalizare  Receptori fagocitari . CD64).Fagocitoza  Caracteristici ● Definitie: preluarea unor particule de dimensiune mare (in principal microorganisme).pattern recognition receptors (PRRs) or molecules (PRMs) o Externi  FcR (CD16. peptidoglicani. Scavenger receptor (CD36).Toll like receptors . dsRNA .nonself) o Pathogen-associated molecular patterns (PAMPs): LPS. datorita capacitatii de a distinge intre tipurile de carbohidrati produsi de mamifere si bacterii (self . lipoarabinomanani. CR3 (CD11b). Mannose receptor o Interni  TLRs .substante prezente relativ constant (fara variabilitate antigenica) la un grup mare de patogeni ● Actin-dependenta.

fiind mediata de: •Ac sau •C3b .Fagocitoza (cont) opsonin dependenta.

non on--self microbian): microbian): PRR Pattern Recognition Receptor PAMP Pathogen Associated Molecular Pattern (Medzhitov & Janeway. non- non- self Recunoastere imuna self origine antigen / context biologic Ipoteze Receptori (self vs. 1997) .

Endocytic PRRs Bacterie glicoproteina proteina bact. LPS. LTA man C3b receptori manoza receptori scavenger receptori opsonine (lectina C) C) (CD36. SRB- SRB-1) (CR1 CR1)) FAGOCIT . a) ENDOCITOZA . bact. CD68.

PAMP •lipopolizaharid •peptidoglican •acid lipoteichoic •lipoproteine  invariabile •manoza  inalt conservate •ADN  specifice microbilor •ARN dc (patogeni + non-patogeni) •flagelina  comune pentru o clasa de microbi •pilina  vitale pentru microorganisme •zimozan Gram- Gram -negative Gram- Gram -positive .

Mannose-binding receptor ● Recunoaste reziduuri de manoza cu o orientare spatiala certa/unica microorganismelor ● Se gaseste doar pe MQ (nu şi pe Mo sau PMN)  2.PRR: leaga carbohidrati bacterieni  1. Glucan Receptor ● Prezent pe toate fagocitele .Receptori fagocitari de suprafata .

leaga peretele celulei bacteriene & LPS ● Fagociteaza celule apoptotice o factor nou MFG-E8 (eliberat din MQ activate se leaga la celulele apoptotice via fosfatidilserina) . LPS) o carbohidrati o Lp cu densitate mica (LDL) acetilate ● Se gasesc pe toate fagocitele ● Specific MQ.CD36 ● Recunoaste liganzi: o polianionici (ds-RNA.PRR: leagă alţi compuşi bacterieni  3.Receptori fagocitari de suprafaţă . Scavenger Receptor .

b) TRANSMITERE SEMNAL .Signaling PRRs TLR Toll--like receptors Toll NOD nucleotide--binding nucleotide oligomerization domain RIG-1 RIG- retinoic acid- acid-inducible gene- gene-1 .

.molecula identificata ca fiind esentiala pentru patern-ul embrionic de Drosophila  Conservat pe parcursul evolutiei la insecte & oameni  TLRs la mamifere este omolog cu domeniul citoplasmatic al receptorului IL-1 si IL-18 (Toll-IL1-R sau TIR domain)  Domeniul extracelular este diferit  Au fost raportati 10 TLRs (1-10)  Exprimati diferentiat pe celulele imune (nivel scazut)  Expresia este “modulata” (inductibila) ca raspuns la anumiti stimuli  Exprimati si pe alte tipuri celulare (e. celule endoteliale) . Receptorii fagocitari interni Toll-like (TLRs)  TLRs sunt proteine transmembranare  “Toll” .g.

cruzi) F Protein (RSV) Lipoarabinomannan Phosphatidylinositol dimannoside (M.CD14) TLR2 +(TLR1/TLR6) TLR4 TLR5 TLR9 Lipoproteins.Toll- Toll -like receptors (TLR) proteine transmembranare tip I (Drosophila  om)  conservate filogenetic (Drosophila  domeniu extracelular bogat in leucina  domeniu intracelular TIR (similar IL- IL-1R)  TLR mamifere (11- (11-om.cruzi) (T. 13- 13-soarece):  recunoastere PAMPs  asociere cu alti TLR sau alte proteine (MD2. lipopeptides LPS Flagellin CpG DNA Peptidoglycan Taxol Zymosan LPS Leptospira interrogans HSP60 LPS P. tuberculosis) .gingivalis Fibronectin GPI (T.

TNF- TNF- (NF (NF--kB)  activeaza imunitatea dobindita   expresia molec.Functiile imunitatii innascute PAMP Stimulare PRR  stimuleaza fagocitoza  induce activitate microbicida  induce citokine inflamatorii:  IL- IL -1. molec. CD80/CD86) CD80/CD86 .II. (MHC cls. co-stim. IL- IL-6.

cathepsina B 1O . prezentare .enzime proteolitice • eliminare a microorganismelor patogene Rol • prelucrare Atg pt.lizozim 2  OCl .lactoferina OH .Fagocitoza Mecanism dependent de oxigen independent de oxigen O2– .defensine H2 O2 .

LT.Mecanismele uciderii bacteriilor .  “Explozia oxidativa” ● Activata in urma fagocitozei ● Stimulata de PRRs ● Consum crescut de O2 ● Produce substante care sunt toxice directe pentru bacterii: o Produsi derivati ai Oxigenului o Produsi derivati ai Nitrogenului  NO (monoxidul de azot)  Produs de NO sintetaza inductibila (iNOS)  Enzima este indusa de cytokine. TNF .cont.

cresterea oxidarii glucozei ● Formarea anionului superoxid. superoxid dismutaza (SOD).+ NADP+ + H+ (NADPH oxidaza) o O2. apei oxigenate si a acidului percloric: o 2O2 + NADPH  2O2. catalaza & glutation peroxidaza (H2O2 ) . doar in PMN nu si in MQ) ● Radicalii derivati de O2 sunt detoxifiati de ceruloplasmina.granulele azurofile.“Explozia oxidativa”-speciile reactive ale O2 Speciile reactive ale oxigenului se formeaza prin “Explozia oxidativa”.+ 2H+  H2O2  HO  (SOD superoxid dismutaza) (Fe2+ ) o H2O2 + Cl2  2HOCl (MPO mieloperoxidaza . transferina. care presupune: ● Cresterea consumului de O2 ● Glicogenoliza .

.

tissue damage .temperature shifts .Danger hypothesis Naive T cells Signal 1 Signal 2 (co- (co- stimulation) APC Danger signal .stress cells Damaged Normal .hypoxia cell cell .hsp .infection .

 acces cel. acces cel.febra . efect. efect.activeaza . producere Atc.activeaza Ly .activeaza Ly . Citokine Macrofag activat IL- IL-1 TNF- TNF- IL- IL-6 IL- IL -12 EFECTE LOCALE .activeaza NK endoteliul vascular endoteliul vascular .febra .producere de IL-6 .induce dif.mobilizare metaboliti . EFECTE SISTEMICE .creste .inducere proteine . permeab. vasculara . .activeaza . Th1 .febra .soc faza acuta .

IFN- sunt importante in mod special in inflamatii. ●  interferon: o Inhiba replicarea virala. etc.  Interferonii  Produsi de celulele infectate cu virusuri. TNF- si -. creste numarul NK si induce antigenele MHC-I ●  interferon: o Activeaza macrofagele si induce antigenele MHC-II o Apararea imuna impotriva infectiilor si proliferarilor maligne. creste numarul NK si induce antigenele MHC-I ●  interferon: o Inhiba replicarea virala. Mediatorii specifici  Citokinele ● Produsi celulari de natura proteica cu rol de “mesaj” pentru alte celule. ● Cresc expresia endoteliala a moleculelor de adeziune. carora “le spun cum sa se comporte” ● IL-1. . actioneaza ca si mesageri de scurta durata care protejeaza celulele invecinate de infectia virala. activarea si agregarea PMNs.

Immune Leukocyte Fibroblast Inductori MAJORI Virus Virus. mitogens. T cells. natural Macrophages epithelial cells killer cells •Dupa expunere corespunzatoare cele mai multe celule sunt apte sa produca cel putin un tip de IFN I. B cells Fibroblasts. IL-1 si TNF. LPS Antigens. rol crucial intre mecanismele de aparare ne- specifica a gazdei impotriva a numerosi patogeni. Interferonii Proprietati IFN- IFN- IFN- NOMENCLATURA Type I. Type II. •Sinteza IFN- este inalt reglata numai in anumite tipuri de celule si este indusa de stimuli specifici IFN- factorul major de activare macrofagica. . TNF- + IL-12 Nr de GENE 26 1 1 SURSA CELULARA T cells. Type I. •Tipul I de IFN poate fi indus de asemenea de LPS (endotoxina bacteriana).

Functii efectorii: Rolul interferonului interferonului Celula infectata viral Celula neinfectata IFN receptor IFN- IFN-/ 2-5 (A) PKR Sintetaza degradare mRNA  translatie mRNA Inhibare sinteza proteica .

Pathogens Epithelial barrier Activated epithelial cells Epithelial cytokines Permeabilised endothelium Cell & fluid migration Interaction with other cells of the innate and adaptive Opsonisation immune systems Phagocytosis .

Monocytes/macrophages - Hours Hours to days NK cells - Hours to days Short- lived Long-lived & connect with adaptive immune system . Timing of innate immunity after infection Barriers . Epithelial activation .Complement - Seconds Minutes Minutes Cytokines/chemokines - Minutes to days Neutrophils .

Eosinophil Eosinophil Basophil Neutrophil Eosinophil Neutrophil Lymphocyt e Basophil Monocyte .

.Eosinophils attacking a schistosome larva in the presence of serum (IgE) from an infected patient.

increase neutrophil chemotaxis. Heparin. neutrophil and monocyte chemotaxis. bronchoconstriction. IL-6. IL-5. anticoagulation. Tryptase. Chymase. increased fibroblast proliferation & platelet activation . Prostaglandin D2 Platelet Activating Factor. contraction of smooth muscle. IL-8. increased vasopermeability. increase eosinophil. TNF- Causing: Vasodilation. Mast Cells Resting Mast cell Degranulated mast cell Mediators released include: Leukotriene C4 & D4. Histamine IL-4.

non-B cells No classical antigen receptors Part of the innate immune system Recognise and kill abnormal cells such as tumour cells Directly induce apoptosis in virus infected cells by pumping proteases through pores that they make in target cells Similar cytolytic mechanisms to cytotoxic T lymphocytes (CTL) Involved in antibody-dependent cellular cytotoxicity (ADCC) .Natural Killer (NK) cells Non-T.

Missing self MHC NKR cls I Celula  Celula Absenta NK  tinta citotoxicitatii NCR Ligand activator NKR Celula  Celula tinta Citotoxicitate NK  NCR Ligand activator .

IL-12 Growth factors.Interactions between phagocytes and other innate immune components: Natural Killer cells IFN NK Cytokines – TNF. angiogenic factors. proteinases - REPAIR & REMODELLING Activated macrophage Interferon  (IFN) receptor .

Sistemul proteinelor plasmatice - raspunsul imun nespecific umoral .

dar si de fagocite.Secreted Pattern Recognition Molecules (sPRM) Produse mai ales de ficat. cu rol in: •Activarea Complementului •Opsonizarea celulelor microbiene Proteine de faza acuta .

Cascada coagularii .

vascular permeability. Complement System  Nonspecific defense system ● The combination of antibodies with antigens does not cause destruction of the antigens or pathogen. ● C5-C9: attack (complement fixation). increases phagocytosis (C3b.  Antibodies serve to identify the targets for immunological attack. ● C4. r. increases integrin avidity (C5a) o As an opsonin. C3: activation.  The complement proteins are designated C-1 to C-9. Become activated by the attachment of antibodies to antigens. mast cell degranulation (C3a. s: recognization. C5a) o Leukocyte chemotaxin.  Complement proteins can be subdivided into 3 components: ● C1q. ● These proteins are in an inactive state. C2. C3bi) . Antibody- induced activation of the complement.  Activated via classic (C1) or alternative (C3) pathways to generate MAC (C5 – C9) that punch holes in microbe membranes  In acute inflammation o Vasodilation.

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b C4b C3b + + C2b Bb .Activation of C’ System .cont.

faze .Imunitatea Innăscută .

Răspunsul inflamator sistemic (1)  Induction of fever – ● Caused by many bacterial products (endotoxins from G(-) bacteria) ● Endogenous pyrogens from monocytes and macrophages (IL-1 and certain interferons)  Increased WBC production / releasing  Increased synthesis of hydrocortisone and adrenocorticotropic hormone (ACTH)  Production of acute phase proteins  C-reactive protein binds to membranes of certain microorganisms to activate the complement system .

Răspunsul inflamator sistemic (2) .

Răspunsul inflamator sistemic (3) Febra are efecte pozitive şi negative asupra infecţiei şi funcţiei organismului POSITIVE NEGATIVE  indicate a reaction to  extreme heat  enzyme infection denaturation and interruption  stimulate phagocytosis of normal biochemical  slow bacterial growth reactions ● increases body temperature > 39° C (103°F) is dangerous beyond the tolerance of some > 41°C (105°F) could be fatal and bacteria requires medical attention ● decreases blood iron levels .

menit sa tina in loc infectia pina la interventia imunitatii dobindite  “Instruieste” sistemul imunitatii dobindite pentru a raspunde la infectii Decizia majora de a raspunde sau nu unui antigen este luata de imunitatea innascuta. prin receptorii codificati in genom ! .Imunitate innascuta  Nu este doar un sistem de aparare simplu.

stomach acid. Immune responses Barriers Skin & Mucous membranes rapidly regenerating surfaces. Invasion peristaltic movement. mucociliary & infection escalator.complement proteins. commensal organisms. cytokines. cytotoxic T cells . flow of urine/tears. coughing Innate immunity Cellular and humoral defences + lysosyme. T helper cells. sebaceous/mucous secretions. vomiting. NK cells + Inflammation Cellular and humoral defences Adaptive immunity Antibodies. phagocytosis.

Imunitatea innăscută ROL și FUNCTII originea si contextul Recunoastere Ag Functii efectoare • prevenire intrare microorganisme • eliminare patogene Rol “instructiv” asupra imunitatii specifice initierea si tipul raspunsului .

135- 135-145) . Nature Reviews Immunology. 1. INNATE ADAPTIVE IMMUNITY IMMUNITY cellular immune response PAMP pathogen TLR Endocytic IL-12 Th1 PRR IFN- CD80/86 CD28 DC TCR Naive Th1 MHC-II T cells IFN- (adapted after Medzhitov R. 2001.

proteins.000 D) cell receptors (TCRs) found on T lymphocytes n gradul de diferenţă structurală faţă de componentele endogene nimmunogen: substance that induces an immune n distanţa filogenetică între antigen şi gazdă accesibilitatea epitopului pentru recunoaştere imună response (able to trigger) .000+). corpusculare – virusuri. (e. Minodora Dobreanu Antigens Haptens Antigen (Ag) – the molecule an antibody (Ab) binds to n Small organic molecules can become antigens • usually a foreign substance if they bind to proteins.Anticorpi n REACTIA ANTIGEN-ANTICORP n Aplicatii ale reactiei Ag-Ac Dr. Allergen n hapten: low molecule weight substance that combines with specific antibodies but is incapable of eliciting an immune response by itself • examples in the case of allergy could be pollen.. poate să producă paralizia sitemului imun al gazdei. immunogens. or a chemical in soap Antigens. amino acids) cat dander. nu se cunoaşte cu precizie mecanismul. bacterii. molecular form important in cantităţi excesive. several different antibodies – By binding haptens to proteins in the laboratory. n Dimensiunea moleculei (>10. toleranţa imună determining immunogenicity apare şi dacă un antigen este administrat în doze similare pe căi diferite: enteral.g.g.Not every antigen can n n calea de administrare a antigenului (intravenos pentru Ag trigger an immune response. • each antigen has different sites that antibodies n Become antigenic determinant sites on the can bind to. ducând la manifestarea unei imunotoleranţe.g. fosfolipide. lipids. n ANTIGEN n ANTICORP Antigene .. adăugarea sărurilor de poor immunogens aluminiu sau de calciu). molecule n doza de antigen: pătrunderea într-un organism a unui antigen în complexity. subcutanat sau intravenos.. n frecvenţa de administrare a antigenului (administrări multiple îl fac carbohydrates) = good immunogens mai imunogen) imunogenicitatea Ag solubile este crescută prin adăugarea •repeating polymers (e. sugars. •nonrepeating polymers (e. acizi nucleici) •molecular size (MW of 10. new antigens are created for research or diagnostic purposes. epitopes: Antigene nantigen: a substance that interacts with either Factorii care determină / influenţează potenţialul imunogen al unui antigen sunt: antibodies (immunoglobulins) or antigen-specific T. so that one antigen can be bound by proteins. nucleic acids) = n adjuvanţilor (de exemplu la vaccinuri. dar se pare că sunt angrenate şi macrofagele 1 . subcutan pentru Ag solubile – proteine.

fluide tisulare și ca receptori membranari pe limfocitul B (rol în contactul cu Ag pentru diferențierea Ly B în plasmocit).Glicoproteine prezente in ser. groove. 2007 IgE 2 . extended surface. IgG2. The panels below show space- filling or computer-generated models indicating where contact between the peptide antigen and antibody occurs. IgG4 IgM IgA : IgA1. center. încărcarea cu carbohidrați) IgG : IgG1. Recunoasterea Ag – scopul raspunsului imun Imunoglobulinele o imunoglobulinele o receptorii pentru Ag de pe celulele sistemului imun Diversitate si heterogenitate Definitie . IgG3.limfocit B . 6th Ed. IgA2 IgD Abdul K. Celular and Molecular Immunology. modificate de diferiti factori) -antigene incomplete (haptena) Modalitate de obtinere Grad de inrudire genetica -antigene naturale -antigene autologe -antigene artificiale (antigene naturale modificate) -antigene alogenice -sintetice (homopolimeri. heteropolimeri liniari sau -antigene xenogenice ramificati) Origine Tip de celula implicata in raspunsul imun -antigene animale -antigene timodependente (antigene T-dependente) -antigene vegetale limfocit T . pocket. Abbas.sintetiza anticorpi : -antigene bacteriene majoritatea antigenelor -antigene virale -antigene timoindependente (antigene T-independente) -heteroantigene (origini multiple) direct limfocitului B): antigene cu epitopi repetitivi How Antibody Binds to Antigen n ANTIGEN n ANTICORP n REACTIA ANTIGEN-ANTICORP n Aplicatii ale reactiei Ag-Ac The top part of this figure shows how different shaped antigens can fit into the binding site of antibodies: left. Clase și subclase (Izotopuri si Allotipuri) (secventa de Aa. right. Antigene Antigene Criterii de clasificare Criterii de clasificare Sursa Capacitatea de a induce un raspuns imun -externa -antigene complete -interna (normale. mărime.

– Fab regions are variable. n 2 long H chains are joined to 2 shorter L chains. Several Flavours of Ab’s: Antibody Structure 3 . • Provide specificity for a binding to an antigen.structura Antibody Structure n 100 million trillion antibody molecules that contain 4 polypeptide chains (Idiotypes). – Fc region of different antibodies are constant.structura Situsuri de legare specifica pentru antigen Regiunea Fab C1q Regiunea Fc Imunoglobuline . n B cells have antibodies that serve as receptors for antigens. Bifunctionalitatea moleculei de imunoglobulina Imunoglobuline .

placenta.000-60.0 0.56 - So how can your body produce Ab to so many antigens. most of which are not Ab Fluid nasal 70-846 8-304 0 Salivă 194-206 42 64 n so there cannot be one gene for one antibody to code for Fluid Intestinal 166 4 8 these – we wouldn’t have enough antibodies! Urină 0. s mast mostly in & function fixes B-cell crosses cells to secretions complement.03 12 0.1-1.0003 2 human adult serum level (mg/ml) Half-life in 5 3 25 2 6 serum (days) Number of 5 1 1 1 1-3 four-chain monomers Special Early Found Activates Stimulate Found properties appearance.500 10. Fluid Cervical 3-133 1-285 5-118 even those it’s never seen? Fluid Vaginal 35 52 - Tonegawa Theory Fundamental Immunology (5th Edition) Lippincott.000 genes which code for all the proteins you need in your entire body.971) 4 . mainly on complement.500 Lacrimi 80-400 0-16 0-18 n but you only have about 30. Williams & Wilkins Chapter 31 (p. Major antibody classes Properties of antibody classes Properties IgM IgD IgG IgE IgA Heavy m     chain mean 1 0. secrete activates traces in bind to histamine macrophage serum macrophage s s s and granulocytes Distributia Ig in diverse lichide biologice The Number Dilemma Concentratiile imunoglobulinelor in secretiile umane externe (mg/l) Secreţie IgA IgG IgM n You have about a trillion different antibodies able to react with millions of different types of Ag Ser 2.047 1.06-0. surfaces.

D.1011 5 .10 x104 Total potențial cu diversitate joncțională 109 . and J for the heavy chain.4 x107 H x Ll 5 . V each B cell gets a unique and J are joined to C for the light chain combination of segments! Mecanisme care contribuie la generarea diversității anticorpilor H Lk Ll Gene din linia germinativă n ANTIGEN Gene V 250-1000 250 2 Gene J 4 4 3 n ANTICORP Gene D 12 0 0 Diversitate n REACTIA ANTIGEN-ANTICORP combinatorie V x J (x D) 10. there are many thousands of possible V-D-J combinations so that Antibody Genes each B cell gets a unique combination of segments! Additional diversity occurs because there are two types of light chains.crz 14 • variable segments (V) – many different versions • diversity segments (D) – several different versions • joining segments (J) – a few different versions • constant segments (C) – a few different versions that are nearly identical A unique recombination A unique recombination occurs in each B cell occurs in each B cell n each B cell combines these gene segments to make an Ab chain like shuffling a Unique combination of segments becomes joined by deck of cards somatic gene rearrangement . V and J for the light chain n since there are multiple types of each gene • each B cell combines these gene segments to make an segment. there are many Ab chain like shuffling a deck of cards thousands of possible V-D- J combinations so that . m. Genes for antibodies aren’t like most other genes - they come in pieces (“gene-lets”):  k .V.000 1000 6 n Aplicatii ale reactiei Ag-Ac Asocierea lanțului H si L H x Lk 1 . .crz 2  l .V.000-40.crz 22  . and J are joined to C for the heavy chain. . Since there are multiple types of each gene segment.  . D.

macrophages or complement • Ac îmbracă țintele celulare care urmează a fi lizate Receptor for • agregarea FcR de pe NK – activare. neutrofile.activarea caii clasice a complementului (IgG-C2. IgE. IgM) •complexele imune (Ag-Ac) se leaga de C1q . IgA) act as a “trash tag”.receptori pentru antigen de pe limfocitul B •ontogenia B . IgM. the Antibody binds to • eozinofilele au FcR si leaga Ac (IgE. Question: What types of immune reactions are IgG. it will bind there and Citotoxicitatea celulară mediată de Ac (IgG. TNF. marking it for destruction by “killer” • Celule care lizează celule: NK. eozinofile cells. IgE and IgD involved in? Functii efectoare Funcții efectoare (1) . IgA. paraziti se complexeaza cu Ac si astfel nu se leaga de tinte celulare •Activarea complementului (IgG. virusuri.legarea incrucisata a Ig de membrana de catre Ag multivalente Neutralizarea Ag de catre Ac circulanti •toxine.Cm3) Ac se leaga la bacterii si maresc capacitatea de fagocitoza •Opsonizarea (IgG) . IgM+IgD. medicamente. IgM .IgM. orice fel de Ig de suprafata •activarea B .creste capacitatea fagocitantă Cresterea fagocitozei implica intensificarea legarii de suprafata a •fagocitele au FcR si leaga particule opsonizate cu Ac bacteriilor opsonizate si activarea fagocitului (procese mediate de FcR) IgM creste indirect fagocitoza – C3b Funcții efectoare (2) How an Antibody Works When an Ab finds its Ag on an invader. bacterii. IgA) care au complexat Ag target antigen of antibody on NK NK cell is signaled by apoptosis and/or cell recognizes a to kill the target membrane damage parazitare – secretie de enzime bound antibody cell 6 .Opsonizare Imunoglobuline de membrana . IFN constant region The target cell dies After binding.

and treatment of certain n REACȚIA ANTIGEN-ANTICORP cancers n Monoclonal antibodies are pure antibody preparations n Aplicații ale reacției Ag-Ac – Specific for a single antigenic determinant – Produced from descendents of a single cell n Hybridomas – cell hybrids made from a fusion of a tumor cell and a B cell – Have desirable properties of both parent cells – indefinite proliferation as well as the ability to produce a single type of antibody 7 . leukotriene. agregarea FcR (IgA) in prezenta alergenului duce la eliberarea de mediatori ai inflamatiei •celulele epiteliale au FcR. șoarece) – invers ca IgA Inhibarea răspunsului imun (IgG) •LyB au FcR și leagă Ac din complexele imune → se inhibă activarea B Antibody functions Antibody functions Monoclonal Antibodies n ANTIGEN n Commercially prepared antibodies are used: n ANTICORP – To provide passive immunity – In research. Funcții efectoare (3) Functii efectoare (4) Hipersensibilitatea imediata (IgE) Imunitatea de la nivelul mucoaselor •mastocitele si bazofilele au FcR pentru IgE monomeric. hipersensibilitatea imediata unde neutralizează Ag Imunitatea neonatala la mamifere (IgG matern) •IgG matern trece prin bariera placentară în circuitul sanguin fetal (om) •IgG din lapte e captat de FcR din intestinul nou-născutului și intră în sânge (porc. clinical testing. citokine) – secrețiile mucoaselor ca IgAs. captează IgA din sânge și îl trec în (amine vasoactive. prostaglandine.

Elispot Mutiplexare 8 .Aplicațiile reacției Antigen-Anticorp Tehnici de imunodetecție Western Blotting Imunoprecipitarea Imunohistochimia / Imunocitochimia Citometria in flux Radioimunoanaliza (RIA) ELISA .

SELF .Procesare / Prezentare Antigene .non SELF Limfocite T si B MHC .

Cells of the specific immune system T cell B cell •Involved with cell mediated immunity •Involved with humoral •Two types: immunity • helper T cells (CD4) •Secrete antibodies • cytotoxic T cells (CD8) •Generally eliminate •Generally eliminate intracellular pathogens extracellular pathogens .

 the specific antibody produced by a plasma cell is also secreted in soluble form and circulates in the blood . and then die. Others become memory B cells.some then become plasma cells that all produce the same antibody (huge numbers of antibodies -2000/second). .when a B cell encounters a matching antigen. B cells  Lymphocytes that react directly with antigens  produced and mature in bone marrow  require stimulation from Helper T Cells  each B cell produces and wears a unique antibody on its surface  clonal selection . it begins to divide rapidly.

T cells  T cells display TCR as their antigen recognition protein  When stimulated they become Cytotoxic or Helper T cells  Secrete cytokines that recruit other cells of the Immune System Types of T cells:  helper T cells – start the immune response ● Inflammatory T cells (TH1) .Activate B cells to make antibody  cytotoxic T cells (CTLs) ● Kill virally infected cells ● Kill cells containing cytosolic bacteria ● Kill tumor cells  suppressor T cells – suppress the activities of other T cells. helping to end the immune response .Activate macrophages to kill intracellular bacteria ● Helper T cells (TH2) .

 The intracellular part signals through the CD3 complex. These molecules are involved in signaling the T cell when it engages antigen .TCR Structure The TCR plus CD3 is referred to as the TCR complex  The TCR is composed of two protein chain: or  ● Crz ● Crz–  The extracellular part binds to MHC+peptide. The TCR is always associated with CD3 () and 2  (zeta) chain.

.  Creates a highly diverse repertoire of TCRs.TCR Gene Rearrangement  Similar to Ig gene rearrangement: ● Heavy () chain has V+D+J(+C) segments.  The mechanisms are the same as in B cells. ● Light () chain has V+J(+C) segments.  TCRs can be either  or .

because if they did attack “self”. CDR3  mature in the thymus. T cells  there are millions of different T cells – the difference is in their receptors (surface markers)  each T cell has a unique receptor that will recognize a different foreign substance: V . VCDR1. CDR2.critical. autoimmune disease could result . where they learn „to tell” the difference between self and “non-self” .

•Helper T cells (Th) recognize peptide associated with MHC class II molecules •Cytotoxic T cells (Tc) recognize peptide associated with MHC class I molecules . MHC & T cells  T cells have a requirement to recognise both the ANTIGEN and the MHC molecule  This is because the molecular structure of the MHC-Antigen complex is arranged so that some of the polymorphic amino acids of the MHC molecule are in direct contact with the TCR ● Therefore T cell recognition of antigen is said to be MHC „restricted‟.

binds MHC class-II. TCR Structure – accessory molecules Accessory molecules which also bind MHC: •CD4 on helper T cells. binds MHC class-I. . •CD8 on killer T cells.

CD4 and CD8 Molecules  Transmembrane molecules  Referred to as co- receptor or accessory molecules  Member of the Ig superfamily  T cells are either ● CD4  CD4(+) CD8(-) ● CD8  CD4(-) CD8(+) .

MHC Clasa I si Clasa II  Clasa I  Clasa II ● Distribuţie tisulară ● Distribuţie tisulară o Toate celulele o Celule prezentatoare nucleate de Antigen (APC) o Hematiile la unele  Macrofage specii  Celule dendritice  Limfocite B  Celule stromale timice ● Functie ● Functie o Prezinta antigene o Prezinta antigene Ly Tc (CD8+) Ly Th (CD4+) .HLA.

.

Proteinele MHC Clasa I si Clasa II  Clasa I  Clasa II ● Lanţul Alfa ● Lanţul Alfa o 3 domenii externe o 2 domenii externe o 1 transmembranar o 1 transmembranar o 1 rest C citoplasmatic o 1 rest C citoplasmatic o Codificat in MHC o Codificat in MHC (crz 6p) (crz 6p) ● Beta-2 Microglobulina ● Lanţul Beta o 1 domeniu extern o 2 domenii externe o nu e ancorat in o 1 transmembranar membrana o 1 rest C citoplasmatic o fara rest citoplasmatic o Codificat in MHC o codificat in crz 15 (crz 6p) .

Proteinele MHC I si MHC II .

beta 2 (2-mg) regiuni  Leagă strâns oligopeptide ● ancorat .pe toate  MHC II .alfa cu 2 ● liber . pe celulele nucleate din imunocite: limfocitele B si organism monocite (APC) * Exceptii (He.alfa cu 3  Alcătuit din 2 lanţuri: regiuni ● ancorat .Expresie  MHC I .beta cu 2 mici (aproximativ 9 Aa) regiuni  Sunt extrem de polimorfe  Acceptă oligopeptide cu cel puţin 15 Aa  Polimorfism limitat .limitat. MHC = CMH . celule ** Exceptii : germinale) ● limfocitele T activate  Alcătuit din 2 lanţuri: ● Celule epiteliale / timus ● ancorat .

 Moleculele MHC clasa II sunt exprimate nativ doar pe un subset de celule hematopoietice si de celulele timice stromale. Distributia tisulara a exprimarii MHC  Expresia moleculelor MHC difera de la tesut la tesut.  Moleculele MHC clasa I sunt exprimate pe toate celulele nucleate. foarte bine exprimate pe celulele hematopoietice. . alte celule exprimându-le doar in urma expunerii la citokine inflamatorii (interferon-).

● procesul este realizat orice celula poate de celule interactiona cu limfocite “profesionale” (de ex. Celule cu virus dendritice. bacterii) sau ● deoarece MHC Clasa I Ag care nu necesită este prezent pe toate procesare celulele din organism. daca este infectata Macrofage. MHC & Antigenele  MHC Clasa I  MHC Clasa II-a ● prezinta peptide ● prezinta antigene endogene exogene care au fost ● originea peptidelor: self fagocitate si procesate sau de origine virala (de ex. LyB) . T.

.

. care contine cele mai polimorfe reziduuri. Structura moleculara a MHC clasa I  Doua structuri in  helix ale zonelor 1 si 2 ale lantului  formeaza “peptide-binding groove” al moleculei  TCRs se leaga la “vârful” moleculei.

MHC clasa I 2-microglobulina .

1) formează peptide- binding groove al moleculei  TCRs se leaga la “vârful” moleculei.Structura moleculară a MHC clasa a II-a  Cele doua domenii polimorfe (1. care conţine cele mai polimorfe reziduuri. .

MHC clasa a II-a DR chain .

Aceste situsuri corespund aminoacizilor care vin in contact cu antigenul. Variabilitate similara se observa in cazul moleculelor MHC clasa II.Polimorfismul MHC clasa I Variatia allelica survine la nivelul unor situsuri specifice ale moleculei MHC. .

stabilizindu-l . • Aminoacizii pozitionati de-a lungul santului interactioneaza (prin legaturi de hidrogen si atractii de tip ionic) cu amino acizii peptidului. • Regiunile 1 si 2 ale MHC clasa I si 1 / 1 ale MHC clasa II formeaza “imaginile in oglinda” cu rolul de a alcatui santul de legare al peptidului antigenic.„80) a dovedit existenta unui sant alcatuit din foi -pliate anti- paralele (la baza santului) si -helixuri (pe laturile santului). Polimorfismul MHC clasa I • Structura cristalina (clarificata târziu .

Expression of HLA is co-dominant Father Mother Kids .

Polymorphism and polygeny of MHC Two different properties make it difficult for pathogens to evade immune responses Definitions: Alleles: different forms of one gene Allotypes: different forms of one protein (isoforms) Polymorphic: alternative forms of one gene = Many alleles Oligomorphic: a few forms of one gene = Few alleles Monomorphic: no polymorphism Homozygous: same allele on both inherited chromosomes Heterozygous: different allele on both inherited chromosomes .

ncbi. The MHC gene region http://www.nlm.gov/mhc/MHC.fcgi?cmd=init&user_id=0&probe_id=0&source_id=0&locus_id=0&locus_group=0&proto_id=0&banner=1&kit_id=0&graphview=0 .nih.

Genele MHC  Determina capacitatea unui organism de a interactiona cu antigene  Sansa unei persoane de aceeasi etnie sa aiba gene MHC compatibile este de aprox 1:1000  Genele MHC sunt denumite HLA de la “human leukocyte antigen”  La alte specii au denumiri diferite (la soarece genele sunt denumite H-2)  Genele MHC I si MHC II sunt înşiruite pe acelaşi cromozom 6p  Genele MHC sunt poligenice şi polimorfe .

factorul properdinic B.Genele MHC  Genele MHC utilizeaza loci diferiti pe cromozomul 6p  Genele MHC sunt mostenite de la ambii parinti. combinatia genelor numindu-se haplotip  Expresia genelor MHC este co-dominanta ● Indivizii pot exprima 2x3 subclase de gene I si 2x3(4) subclase de gene II  Alte gene ale sistemului imun înnăscut sunt localizate între cele două regiuni (I şi II) în acelaşi “ciorchine” (“cluster”) . C4.C2. TNF .

Major Histocompatibility Complex .

Human MHC (HLA) encoded on cromosome 6 .

HLA-DZ  In consecinţă. HLA-E (6). HLA-B (728). moleculele MHC sunt limitate în capacitatea lor de a lega peptide  Aceasta se numeşte “restricţie MHC” . HLA-DQ(68 allele). HLA-DP(120 allele). HLA-DR(3 allele). HLA-DX. HLA-DR(503 allele). HLA-C (210). HLA-DO (16). HLA-F (1). HLA-G (15)  Genele MHC Clasa II-a sunt denumite: HLA-DP(23 allele). HLA-DM (10).Genele MHC  Gradul de polimorfism variaza in cele doua clase  (Polimorfism = numarul de allotipuri sau izoforme posibile)  Genele MHC Clasa I sunt denumite: HLA-A (414 allele). HLA-DQ(32 allele).

Figure 5-13 .

com/HIG/index.html . Human Leukocyte antigen (HLA=MHC in humans) polymorphism .alleles http://www.anthonynolan.

Figure 3-24 part 1 of 2  Heavy Chain .

 Heavy Chains .

Diversitatea imunologică generată de locusurile MHC .

HLA Diversity .2 x 3 HLA‟s per person http://www.html .edu/dept/HPS/transplant/html/tt_1.stanford.

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APC  Macrofage/monocite  Celule Dendritice (Langerhan‟s cells)  Ly B .Celulele prezentatoare de Ag .

Proprietatile APC .

RELATIONSHIP BETWEEN PATHOGEN AND MHC COMPARTMENTS: A TWO-PRONGED APPROACH TO ANTIGEN PRESENTATION  Two places for pathogens to be found within eukaryotic cells.  One place for antigens to end up in order for the TCR to recognize foreign antigen- on the surface of the cell in the context of MHC.  Two pathways for antigen presentation for peptides (MHC class I and II). . cytosol/endogenous and vesicles/exogenous.

Demarcation between MHC class I and class II Processing Pathways .

Reits . MHC Class I pathway Figure by Eric A.J.

Generation of Peptide-class I MHC Complexes • Calnexin Tapasin + Calreticulin TAP = Transporter associated with Ag Processing . Endogenous Antigen Processing .

Reits .J. MHC class II pathway Figure by Eric A.

etc. Exogenous Antigen Processing  APC engulf antigens (bacteria.) ● MQ. dendritic cells eat anything ● B cells only endocytose Ags bound by their mIg receptors  Ag peptides generated in endocytic vesicles .

Demarcation between MHC class I and class II Processing Pathways .

MHC Proteins  Ambele tipuri de proteine MHC sunt importante pentru activarea Ly T: ● Class I proteine MHC o Sunt recunoscute de Ly T CD8 o Prezenta peptide (antigene) endogene ● Class II proteine MHC o Sunt recunoscute de Ly T CD4 o Prezenta peptide (antigene) exogene .

Co-receptors on Helper and Killer T cells .

Prezentare Ag .

Staphylococcus aureus . cytokine release. systemic inflammation Enterotoxin A .Toxins that act as superantigens: • superantigen binds to regions outside the normal binding sites on TCR. MCH II • result is massive T cell activation.

Sistemul limfatic Organe si tesuturi limfoide .

Cells of the specific immune system T cell B cell •Involved with cell mediated immunity •Involved with humoral •Two types: immunity helper T cells (CD4) •Secrete antibodies cytotoxic T cells (CD8) •Generally eliminate •Generally eliminate extracellular pathogens intracellular pathogens .

Lymphatic System: Overview Function:  Drain fluid from around cells  Absorb fat from intestines  Circulate lymph  Filter lymph  Immunity .

ANATOMY OF THE IMMUNE SYSTEM  The immune system is localized in several parts of the body – immune cells develop in the primary organs .bone marrow and thymus (yellow) – immune responses occur in the secondary organs (blue) .

The bursa of Fabricius in birds .

Originea celulelor implicate în
răspunsul imun
Celula stem
Hematopoietica Progenitor mieloid
Monocit
GM CSF M CSF

IL1,3,6 G CSF

IL 3
Progenitor
Basofil Macrofag
limfoid IL 5
Celula
dendritica
IL7
B Neutrofil

NK Timus Mastocit

Eosinofil
CD8 CD4
T T

Plasmocit

Lymphocyte Maturation

ANATOMY OF THE IMMUNE SYSTEM

 Bone marrow – blood-producing tissue located inside certain
bones (5% BW)
– blood stem cells give rise to all of the different types of
blood cells

 Thymus – glandular organ near the heart – where T cells learn
their jobs

 Spleen – serves as a filter for the blood
– removes old and damaged red blood cells
– removes infectious agents and uses them to activate cells
called lymphocytes

 Lymph nodes – small organs that filter out dead cells,
antigens, and other “stuff” to present to lymphocytes

 Lymphatic vessels – collect fluid (lymph) that has “leaked” out
from the blood into the tissues and returns it to circulation

B-CELL DEVELOPMENT

B-CELL DEVELOPMENT

it is found in the inferior neck and extends into the mediastinum where it partially overlies the heart – It increases in size and is most active during childhood – It stops growing during adolescence and then gradually atrophies .Thymus  A bilobed organ in mediastinum above the heart  The size of the thymus varies with age – In infants.

T-CELL DEVELOPMENT .

T-CELL DEVELOPMENT .

Positive selection occurs in the thymic cortex T cells (CD4+CD8+) that recognise foreign antigen presented in the form of antigen/MHC complexes by antigen-presenting cells within the thymus are allowed to live MHC self- recognition molecules This is called positive selection . Steps in T cell development Step 1.

T cells are presented with self antigen/MHC complexes by antigen-presenting cells within the thymus If T cells bind and recognise these self antigens they are destroyed by apoptosis The immune system destroys T cells specific for self-antigen This is called negative selection .Steps in T cell development Step 2. Negative selection occurs in the thymic medulla.

T-CELL DEVELOPMENT The result is a T cell repertoire that recognises foreign antigen and is tolerant towards self antigen .

Know your Flow! .

Spleen  Largest lymphoid organ  In upper left quadrant of abdomen  Has a hilum and a capsule  Sinuses contain blood instead of lymph  Filters blood – Worn out RBC – Bacteria  Lymphocytes  Monocytes .

Structure of the Spleen  Surrounded by a fibrous capsule. macrophages. and huge numbers of erythrocytes and platelets  Two distinct areas of the spleen are: – White pulp – area containing mostly lymphocytes suspended on reticular fibers and involved in immune functions – Red pulp – remaining splenic tissue (MQ) concerned with disposing of worn-out RBCs and bloodborne pathogens . it has trabeculae that extend inward and contains lymphocytes.

.

Additional Spleen Functions  Stores breakdown products of RBCs for later reuse – Spleen macrophages salvage and store iron for later use by bone marrow  Site of fetal erythrocyte production (normally ceases after birth)  Stores blood platelets .

1-10 mm Filter lymph. Cancer cells.Lymph Fluid of the lymphatic system Similar to blood plasma and interstitial fluid Lymphatic Vessels Transport lymph Lymph is returned to the circulatory system at either the right or left subclavian veins Lymph Nodes 500-600. Microorganisms. Lymphocytes Monocytes .

Lymph Nodes  Lymph is filtered through lymph nodes  Found in clusters  Vary in size  Principal groupings in deep thoracic. abdomen and cervical.  Provide biological filtration  Site of cancer growth and metastasis . axillary. inguinal regions.

cortex and medulla:  Cortex contains lymph nodules  Follicular dendritic cells  Germinal centers – B cells proliferate  Lymphatic vessels enter node on convex side  Lymph passes through irregular channels called sinuses  Leaves node through one or two efferent vessels at the hilum or hilus . Lymph Node Capsule.

Flow of lymph .

MALT  MALT – mucosa-associated lymphatic tissue is composed of: – Peyer’s patches. and the appendix (digestive tract) – Lymphoid nodules in the walls of the bronchi (respiratory tract)  MALT protects the digestive and respiratory systems from foreign matter . tonsils.

Tonsils  Lymphoid tissue of tonsils contains follicles with germinal centers  Tonsil masses are not fully encapsulated  Epithelial tissue overlying tonsil masses invaginates. forming blind-ended crypts  Crypts trap and destroy bacteria and particulate matter .

form a ring of lymphatic tissue around the pharynx  Location of the tonsils – Palatine tonsils – either side of the posterior end of the oral cavity – Lingual tonsils – lie at the base of the tongue – Pharyngeal tonsil – posterior wall of the nasopharynx – Tubal tonsils – surround the openings of the auditory tubes into the pharynx . Tonsils  Simplest lymphoid organs.

Aggregates of Lymphoid Follicles  Peyer’s patches – isolated clusters of lymphoid tissue. preventing them from breaching the intestinal wall – Generate “memory” lymphocytes for long- term immunity . similar to tonsils – Found in the wall of the distal portion of the small intestine – Similar structures are found in the appendix  Peyer’s patches and the appendix: – Destroy bacteria.

adenoids. appendix .Gut associated lymphoid tissue (GALT) .tonsils. Peyer’s patches.

adenoids. Peyer’s patches.Gut associated lymphoid tissue (GALT) .tonsils. appendix .

Lymphatic vessels Resemble veins (same 3 layers) Found throughout body except: – Avascular tissues – Central nervous system – Splenic pulp – Bone marrow .

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and right side of head.Lymphatic vessels join to form lymphatic trunks Lymphatic trunks join to form : •Thoracic duct (3/4 of body) •Right lymphatic duct (drains right arm. . neck and upper torso) These empty into subclavian veins at junction with internal jugular vein.

Fluid Movement Formation of lymph: Fluid leaves capillaries by diffusion and filtration Escaped proteins If lymph flow blocked = tissue swelling or edema Specialized lymphatic capillaries in vili of small intestine transport lipids - they are called lacteals. and the fluid is called chyle. .

Edema  Accumulation of interstitial fluid Causes of Edema  Blockage of lymphatic system  Increased pressure in veins  Lack of albumin – Decreases fluid returning to blood capillaries by osmosis  Inflammation .

) . Overview of the immune response: Antibody mediated Cell (humoral) mediated (CMI) (Fig. p. 22.6. Madigan et al. 762.

Antigen Recognition Provides the key for the immune system to recognize the presence of intracellular microorganisms MHC proteins are ignored by T cells if they are complexed with self protein fragments .

Antigen Recognition If MHC proteins are complexed with endogenous or exogenous antigenic peptides. they: – Indicate the presence of intracellular infectious microorganisms – Act as antigen holders – Form the self part of the self-antiself complexes recognized by T cells .

T Cell Activation: Step One – Antigen Binding  TC cells are activated by antigen fragments complexed with class I MHC proteins  APCs produce co-stimulatory molecules that are required for TC activation  TCR that acts to recognize the self-antiself complex is linked to multiple intracellular signaling pathways  Other T cell surface proteins are involved in antigen binding (e..g. CD4 and CD8 help maintain coupling during antigen recognition) .

16 .T Cell Activation: Step One – Antigen Binding Figure 21.

it must recognize one or more co- stimulatory signals  This recognition may require binding to other surface receptors on an APC – Macrophages produce surface B7 proteins when nonspecific defenses are mobilized – B7 binding with the CD28 receptor on the surface of T cells is a crucial co- stimulatory signal  Other co-stimulatory signals include cytokines and interleukin 1 and 2 .T Cell Activation: Step Two – Co-stimulation  Before a T cell can undergo clonal expansion.

and form clones – Differentiate and perform functions according to their T cell class .T Cell Activation: Step Two – Co-stimulation  Depending on receptor type. T cells: – Become tolerant to that antigen – Are unable to divide – Do not secrete cytokines  T cells that are activated: – Enlarge. co-stimulators can cause T cells to complete their activation or abort activation  Without co-stimulation. proliferate.

including hormonelike glycoproteins released by activated T cells and macrophages  Some are co-stimulators of T cells and T cell proliferation  Interleukin 1 (IL-1) released by macrophages co- stimulates bound T cells to: – Release interleukin 2 (IL-2) – Synthesize more IL-2 receptors . Cytokines  Mediators involved in cellular immunity.

which sets up a positive feedback cycle that encourages activated T cells to divide – It is used therapeutically to enhance the body’s defenses against cancer  Other cytokines amplify and regulate immune and nonspecific responses .Cytokines  IL-2 is a key growth factor.

Helper T Cells (TH) .

require TH co-stimulation to activate B cells  Cytokines released by TH amplify nonspecific defenses .Helper T Cell  TH cells interact directly with B cells that have antigen fragments on their surfaces bound to MHC II receptors  TH cells stimulate B cells to divide more rapidly and begin antibody formation  B cells may be activated without TH cells by binding to T cell– independent antigens  Most antigens. however.

or killer T cells. Cytotoxic T Cell (Tc)  TC cells. are the only T cells that can directly attack and kill other cells  They circulate throughout the body in search of body cells that display the antigen to which they have been sensitized  Their targets include: – Virus-infected cells – Cells with intracellular bacteria or parasites – Cancer cells – Foreign cells from blood transfusions or transplants .

T-CELL FUNCTIONS .

T-CELL FUNCTIONS .

18a.Mechanisms of Tc Action Figure 21. b .

.A Cytotoxic T Cell Attacking and Killing a Virus- Infected Target Cell CELLS alive! Here. the smaller cytotoxic T cell or Tc (arrow) is attacking and killing a much larger virus-infected cell. The T cell will survive while the infected cell is destroyed.

Other T Cells Suppressor T cells (TS) – regulatory cells that release cytokines. which suppress the activity of both T cells and B cells Gamma delta T cells (Tgd) 5 – 10% of all T cells found in the intestines that are triggered by binding to MICA receptors .

Selection of B cells by antigen (clonal selection)

Different types of B cells have
different receptor molecules.


When a pathogen (germ) “locks
on” to a receptor, that type of B
cell is selected.


The selected B cell divides
rapidly to make lots of copies
of itself. The copies make lots
of antibodies against the
pathogen.

Plasma cells secrete antibody at a high rate but can no longer respond to
antigen or helper T cells.

Clonal Selection Theory (continued)

 Some of the cells
become plasma
cells that secrete
primary response.
 Others become
memory cells that
secrete
antibodies during
the secondary
response.
 Antigens select
lymphocytes that
are already able
to make
antibodies.

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Memory & specificity – key features of the adaptive immunity .

Cinetica răspunsului imun la o infecţie virală tipică .

Raspunsul imun in tumori .

fizică (radiaţii ionizante). HSV 8 (Herpes simplex virus 8) poate determina sarcom Kaposhi o sau de origine internă (oncogene) . tetraclorură de carbon. Hodgkin sau carcinomul nazo- faringean. HBV (virusul Hepatitei B). Există virusuri oncogene:  de tip ADN: EBV (Epstein-Barr). HPV (Papiloma virus) determină carcinomul cervical uterin. etc) şi biologică (mai ales virală). chimică (benzen. respectiv limfomul cu limfocite T. Factori cancerigeni o de origine externă. poate cauza carcinom hepatocelular  de tip ARN: HTLV (human T lymphotropic virus): poate determina Leucemia.atacă limfocitele B determinând limfomul Burkitt.

este inactivă sau • Mutaţii ale acestor gene lipseşte la peste 50% din determină ignorarea din partea celulele canceroase celulei a unuia sau mai multor componenţi ai reţelei de semnale inhibitorii. ceea ce conduce la o creştere celulară necontrolată. declanşează apoptoza celulei .1. pentru a permite repararea ADN) - in caz de leziuni ireparabile. • p53 = Proteina citopl. a creşterii tu. Gene supresoare tumorale • Inhibă sau frânează creşterea •Gena supresoare p53 a celulară şi ciclul celular. monitorizează integritatea ADN în timpul diviziunii celulare (poate opri / încetini ciclul celular în faza S. prevenind dezvoltării tumorilor – crz dezvoltarea tumorilor 17p13. înlăturând “frânele” ciclului celular. Supres.

Antigene care aparţin şi celulelor normale  Deşi nu toate celulele neoplazice dispun de markeri deosebiţi de cei fiziologici. Antigene tumorale  TSA-Tissue TSA Specific Ag . care pot reapare după naştere – Antigenele de diferenţiere: diferenţiere apar numai într-o fază determinată a maturaţiei unor serii celulare. În această grupă sunt înşirate: -fetoproteina (AFP) şi antigenul carcinoembrionar (CEA). ca în cazul elementelor imature din cadrul unor leucemii (limfatice sau mielocitare). numite neoantigene neoantigene. . la majoritatea pot fi identificate structuri antigenice diferite.Antigene care aparţin doar celulelor tumorale  TAA TAA-Tissue Associated Ag . dar nu şi a celor de după naştere. Aceste structuri pot fi înşirate în următoarele 2 grupe: – Antigenele retrogenetice retrogenetice: în condiţii fiziologice ele sunt exprimate exclusiv pe suprafaţa celulelor embrionare. Din această cauză se numesc şi antigene oncofetale.

CD5. CD4 – LLC cu T: CD3. lanţuri  libere intracitoplasmatic – LLA cu T: CD3 (citoplasmatic). CD8 TdT-CD34-HLA DR-CD19-CD10-CD79b-cy Ig-FMC7-CD5-CD23-CD22-sIg-CD38 . TdT(n) – LMA: CD13.Antigenele de diferenţiere  Pentru leucemii acute: – LLA: CD10 (CALLA)-forma comună. CD33. CD21. CD7. CD19. TdT(n) .forma null – LLA cu preB: CD19. mieloperoxidaza Terminal deoxynucleotidyl transferase (nuclease)  Pentru leucemii cronice: – LLC cu B: CD19. CD23 – Sindromul Sezary: CD3.

Markeri tumorali . de ex. receptori hormonali)) sau este secretată în umorile înconjurătoare (markeri markeri umorali / solubili în lichide biologice) şi a cărei identificare / dozare este utilă in diagnosticul sau management- ul clinic al afecţiunilor canceroase. care rămâne în / pe celula producătoare (markeri markeri celulari.definiţie Markerul tumoral este substanţa produsă de către un ţesut tumoral. . p53. crz Ph.

 Do not require prior exposure for sensitization to the tumor antigens. – Release perforins and granzymes.  Stimulated by interferon. but distinct from T cells. – NK cells attach to cells that lack class- 1 MHC antigens.  Provide first line of cell- mediated defense. – NK cells destroy tumors in a nonspecific fashion. Natural Killer (NK) Cells  Lymphocytes that are related to. .

AFP la pacienţii cu ciroze hepatice alcoolice .risc înalt de carcinom hepatocelular) • diagnosticul tumorilor. chimioterapic sau radioterapic • stabilirea recurenţei / recăderii / metastazării .Markeri tumorali . concentraţia markerului este în strânsă relaţie cu masa ţesutului tumoral) • monitorizarea răspunsului la tratament chirurgical. pentru completarea simptomatologiei clinice (5 HIAA urinar în sindr. carcinoide) • stabilirea prognozei (în unele cazuri.utilitate • screening – populaţie la risc (Calcitonina in familii cu afecţiuni neoplazice endocrine multiple tip 1.

Carbohidraţi – CA 15-3. Antigene oncofetale – CEA şi AFP B. Proteine non-oncofetale – Tireoglobulina. CA 27. CA 125. TPA. Enzime – NSE. Markeri tumorali – clasificare – utilitate în practica clinică A.29. CA 72-4. Calcitonina D. Markeri genetici – p53 . Feritina. S-100. Hormoni – hCG. CA 19-9. 2-mg. TK F. Bence Jones E. PSA C.

4. 6.sensibilitate 100% . Valoarea cut off: este valoarea concentraţiei la care sensibilitatea şi specificitatea metodei sunt optime.capacitatea de a indica modificările induse de tratament. 2. 3.capacitatea de a discrimina între indivizii sănătoşi şi cei cu neoplazii.specificitate 100% .capacitatea de a identifica toţi pacienţii cu tumoră.capacitate prognostică în funcţie de concentraţie. 5. Caracteristicile unui marker tumoral ideal 1. De obicei corespunde concentraţiei la care specificitatea este de minimum 80% . în stadiu cât mai precoce.specificitate de organ – capacitatea de a localiza tumora.corelaţie între concentraţia serică a markerului şi stadiul tumorii.

•. concentraţii de peste 35 ng/ml (dacă infecţia urinară este exclusă) poate indica un carcinom al vezicii urinare •valori crescute în lcr. cervical. fixată în membrana celulei neoplazice (de unde se eliberează în sânge) S-au descris cel puţin 6 epitopi (heterogenitatea se datorează conţinutului variabil în glucide) Valori normale serice: 0-4.6 ng/ml. mamar. apar în metastaze cerebrale .Antigene oncofetale – CEA În viaţa embrionară se produce în pancreas şi în tractul gastro- intestinal. Fumători : < 10 ng/ml Timpul de înjumătăţire în ser : 1-2 săptămâni Valori crescute se întâlnesc în carcinoame ale tractului gastro- intestinal: colo-rectal. gastric şi pulmonar.în urină. pancreatic. ovarian. Este o glicoproteină complexă – 180 kD cu 45-60% carbohidraţi.

primare şi secundare Tumori primare Se (%) Locul metastazei Se (%) Carcinom pulmonar cu celule mici 60-70 Multiple 87 Carcinoame pulmonare altele 75-85 Ficat 71 decât cele cu celule mici Oase 57 Cancer mamar preoperator (T3) 40-95 Cancer mamar postoperator 75-85 Plămâni 56 Ficat 30-40 Peritoneu 33 Pancreas 30-40 Sistem limfatic 11 Carcinom colo-rectal preoper. 10-85 Carcinom colo-rectal postoper. Antigene oncofetale – CEA Sensibilitatea în tu. gât 60-80 postoperator indică o tumoră reziduală . 65-75 •Concentraţia plasmatică a markerului Vezică urinară <10 se corelează destul de slab cu masa Prostată <10 tumorală Ovar 20-30 •Se/Sp sunt modeste – nu se poate utiliza în scop de screening Tiroidă <10 • Persistenţa valorilor patologice Cap.

ovariene. Antigene oncofetale – AFP Este o glicoproteină de origine oncofetală. hepatita cronică activă . testicular. ciroza hepatică.000 : 100 :1 Timpul de înjumătăţire: 5 zile Valori crescute (peste 500 ng/ml): carcinom hepatocelular (poate fi utilizată şi pentru screening la persoane AgHBs+).5 U/ml) la adult 2-5 mg/ml la făt. În cursul vieţii intrauterine se formează în sacul vitelin. sinteza ei fiind suprimată la adult. Valori normale: < 5 ng/ml (< 3. retro­peritoneale. < 500 ng/ml la femeia gravidă Raportul normal al concentraţiei de AFP în : serul fetal : lichidul amniotic : serul matern = 10. fiind o proteină plasmatică. cu masa moleculară 70 kD (conţine 4% carbohidraţi). mediastinale şi în boli non-maligne: hepatita alcoolică.în săpt 14-20). în serul mamei concentraţia max este atinsă în săpt 35-38). moderat crescute: în neoplazii testiculare. iar mai târziu în ficat şi tract gastro-intestinal (sinteza max . hepatita acută virală.

Sensibilitatea AFP în diverse afecţiuni tumorale şi non-tumorale AFP ng/ml <10 10-215 215-7500 >7500 Persoane sănătoase ~100% Ciroza hepatică/hepatita ~80% ~15% ~5% Carcinoame extrahepatice ~65% ~30% ~5% cu metastaze hepatice Carcinoame primare ~5% ~10% ~35% ~50% hepatocelulare .

CA 15-3 Este o mucină (GP) polimorfă epitelială Sensibilitatea CA 15-3 şi CEA în (300 . Locale 46 12 Osoase 79 44 Are o sensibilitate semnificativ mai Hepatice 79 75 mare în comparaţie cu CEA în detectarea cc mamar aflat în Media: 69 41 diferite stadii .450 kD) diverse stadii ale cc mamar A fost identificat cu 2 Ac Mo diferiţi: CA15-3 > Stadiul CEA > • 115 D8 (obţinut prin imunizare cu 25 bolii 5 U/ml(%) lipide din laptele uman) şi U/ml(%) • DF3 (prin imunizare cu membrana celulelor din cc mamar). T1 25 9 T2 32 11 Valori normale: 0-22 U/ml T3+T4 35 14 Timpul de înjumătăţire: 4-6 zile Media: 30 11 Peste 40 U/ml probabilitatea Meta neoplaziei este de 80%.

Valoarea CA 15-3 în monitorizarea postoperatorie Cut-off Valori crescute înainte Valori în momentul „Lead time” (U/ml) de diagnostic clinic (%) diagnosticului (%) maxim (luni) 25 70 76 9 40 63 76 7 . CA 15-3  CA 15-3 este folosit pentru monitorizare postoperatorie.  „Lead time” este perioada de timp care trece de la prima creştere a concentraţiei markerului şi până când detectarea clinică a recăderii devine posibilă. pentru că indică recidiva cu câteva luni înainte ca diagnosticul clinic să fie posibil.

7+/-10 Hepatita cronică % 57 43 29 29 14 25.5 0.09 13.6+/-76 Ovare % 66 64 55 39 34 59+/-64 .8+/-19 Boli maligne Stomac % 14 7 7 7 7 20.5+/-30 Ficat % 36 28 21 14 14 44.2+/-53 Prostată % 100 50 30 28 20 26.4+/-13 Ciroză % 39 39 31 27 23 26.3 0. Specificitatea CA 15-3 Concentraţia CA15-3 în alte boli benigne şi maligne U/ml >22 >25 >30 >35 >40 X +/.2+/-11 Plamâni % 71 71 60 53 40 63.3+/-6 sănătoase% Hepatita acută % 63 50 38 25 0 24.2+/-124 Pancreas % 80 80 70 70 50 51.DS Persoane 9.5 1.4 5.

după tratament în std II şi III  Determinarea CA 27. Recomandări pentru utilizarea Markerilor pentru dg.29 este utilizat doar in monitorizarea pacientelor aflate în fază avansată de boală  Descreşterea CD 15-3 = succes terapeutic  CEA – dg precoce al metastazelor . Cancerului de sân  CA 15-3 nu se utilizează ca şi marker de screening de masă (Se ↓)  Determinarea receptorilor pentru Estrogen / Progesteron este recomandată pentru a aprecia raspunsul la terapie hormonală  Determinarea CD 15-3 şi CA 27.29 sunt utile pentru detecţia precoce a recurenţei la pacientele clinic asimptomatice.

hipertrofie de prostată) şi în carcinom de prostată. se recomandă dozarea free PSA  Este un marker specific de organ.  Valori normale: 0-2. iar cut-off: 3 ng/ml  Timp de înjumătăţire: 1-2 zile. care poate fi folosit şi pentru screening-ul grupelor de risc (bărbaţi cu vârste cuprinse între 50-70 ani). pentru excluderea dg de hipertrofie benignă. sintetizată de celulele epiteliale prostatice şi secretată în lichidul seminal. .  Valori crescute apar în boli benigne (prostatită.PSA  Este o glicoproteină cu masa moleculară de 36 kD (Ablin.Antigenul specific prostatic (Prostate-Specific Antigen).5 ng/ml.  In intervalul 4-10 ng/ml. 1970). mai ales în asociere cu tuşeul rectal  PSA poate fi folosit şi pentru monitorizarea terapiei şi detectarea recidivelor.

pentru excluderea dg de hipertrofie benignă. Recomandări pentru utilizarea PSA pentru dg. se face înainte de orice manipulare /manevre asupra prostatei  La valori < 4 ng/ml. se recomandă dozarea f PSA (dacă < 25% din PSA. → risc ↑) Standard PSA Probabilitate % free PSA Probabilitate cancer cancer 0-2 ng/mL 1% 0-10% 56% 2-4 ng/mL 15% 10-15% 28% 4-10 ng/mL 25% 15-20% 20% >10 ng/mL >50% 20-25% 16% >25% 8% . Cancerului de prostată  PSA se utilizează în asociere cu tuşeul rectal ! (cut-off: 3 ng/ml)  Recoltarea sângelui pentru analiză. nu se recomandă biopsia  În intervalul 4-10 ng/ml.

Recomandări pentru utilizarea CEA pentru dg. este utilizat pentru completarea stadializării  Dozarea CEA imediat post-operator nu este recomandabilă. Cancerului colo-rectal  CEA nu este recomandat pentru screening (Se ~). datorită eliberării unei mari cantităţi intra-operator  Poate fi utilizat pentru monitorizarea răspunsului terapeutic şi progresiei bolii .

Recomandări pentru utilizarea Markerilor pentru Cancerul pulmonar (NSE.3 ng/ml •Valori crescute: carcinom bronşic non-parvicelular – specificitate 95%.9% Persoane sănătoase 92 0% ( 7.8) CYFRA 21-1 – fragmente solubile de Citokeratina 19 (în NSCLC) •Valori normale: < 3.0) 0% ( 1.1% 13. Antigenul celulelor carcinomatoase scuamoase (SCC) •Valori normale: < 3 ng/ml Se recomanda dozarea tuturor markerilor pt a putea depista “Leading marker-ul”. CYFRA 21-1.2. CEA.0.8 +/. SCC) Sensibilitatea↓a markerilor NSE (SCLC) – CEA (NSCLC) în cancerul pulmonar Tipul bolii N NSE (> 13 ng/ml) CEA (> 5 ng/ml) Carcinom cu celule mici 81 74% 47% Alte carcinoame 108 15% 46% Boli pulmonare benigne 79 5. cu care se va monitoriza tratamentul !!! (cel mai adesea biopsia este imposibilă !) .1 +/.

5% 12.6% III 64% 46% 32% IV 85% 70% 65% . CA19-9. CA19-9. CEA şi stadiul tumorii Stadiul CA 72-4 > 8ng/ml CA 19-9 > 37 U/ml CEA> 5ng/ml I 14% 4% 1% II 28.5% 12. Recomandări pentru utilizarea Markerilor pentru Cancerul gastric Sensibilitatea şi specificitatea comparativă a CA72-4. CEA CA72-4 > 8 ng/ml CA19-9 > 37 U/ml CEA > 5 ng/ml Carcinom 78% 60% 51% Boli benigne 1% 7% 9% Corelaţia între valorile crescute ale CA72-4.

Recomandări pentru utilizarea Markerilor pentru Cancerul ovarian  Nu se recomandă utilizarea CA 125 pentru screening-ul populaţiei asimptomatice (Se ↓)  Istoric familial de cancer în sfera genitală → se recomandă dozarea la 6 luni interval + Ultra- sonografie transvaginală  CD 125 poate fi utilizat ca marker prognostic în terapia primară .

1 Neuroblastom NSE FIA 0. CA 19-9 EIA 1-0.00001 . TPA.01 Tiroidiană Tireoglobulina. CEA Pancreatică CA 19-9. fPSA Gastrică CA 72-4. TPA Pulmonară cu celule mici NSE. TPA Ovariană CA 125. CEA Germinale AFP + hCG Prostată PSA. CA 72-4 Vezica urinară TPA Metoda Limita detecţiei (fmol) Colo-rectală CEA.. Calcitonina LIA 0. Markeri tumorali de primă linie recomandaţi în neoplazii Tumori Markeri tumorali Hepatică primară AFP. TPA.01-0. CEA. TPA.

Combinarea markerilor tumorali în practica clinică  ROC (Receiver Operating Characteristics) markeri tumorali în carcinomul mamar  CEA: 42%  hCG: 21%  CA 15-3: 30%  p53: 10%  C-erB-2: 20-30% MCA = mucinous carcinoma associated antigen (cut off: 11-14 U/l) .

Combinarea markerilor tumorali în practica clinică Carcinoamele pulmonare: Calcitonina: 54%  CEA: 88%  NSE: 90%  Carcinomul cu celule în boabe de ovăz: •C-k-ras2: 20% •C-myc: 45% .

. Imunoterapia in tumori  Imunoterapia cu anticorpi antitumorali  Doar anticorpii specifici antiTSA pot fi utilizati în scop diagnostic sau terapeutic.  Terapia antitumorală cu anticorpi izolaţi: produce citoliză mediată de complement sau prin fagocitoză (ADCC):  Anticorpi antiHER2/neu (Herceptin) -în cancerul de sân  Anticorpi antiCD20 (Rituxan) -în LLC  Anticorpii 17-1A reacţionează cu un antigen asociat carcinomului colorectal  Necesitatea obţinerii unor anticorpi monoclonali ‟‟umani‟‟ sau „‟umanizaţi‟‟ este obligatorie pentru a putea fi utilizaţi timp îndelungat.  O formă mutantă a receptorului factorului de creştere epidermic (cu o del a domeniului extracelular) este antigenică şi poate fi administrată sub forma unui ‟‟vaccin‟‟ (cuplat cu celulele tu) – de bază pentru terapia cu anticorpi a tumorilor. Actualmente există puţini anticorpi specifici reali antitumorali:  Receptorii B (în LLC). anticorpi anti CD21). idiotipul anticorpilor care este receptorul limfocitar B poate dezvolta antiidiotip (de ex.

CD3 – declanşează eliberarea / formarea de perforine) Este terapia antitumorală ”de viitor”. Ricina). deoarece utilizează înalta specificitate a anticorpilor monoclonali şi capacitatea citolitică a sistemului imun celular specific. Acest tip de terapie poate ucide însă şi celule învecinate normale („‟inocent bystander‟‟) Terapia antitumorală cu anticorpi bispecifici: la TAA la o moleculă “trigger” de pe T (de ex. inhibă molecule / procese intracelulare esenţiale.Imunoterapia in tumori Terapia antitumorală cu anticorpi cuplaţi cu toxine sau radioizotopi: Anticorpii cuplaţi cu toxine (de ex. Anticorpii cuplaţi cu radioizotopi mediază citoliza prin denaturarea ADN. .

Imunitate de transplant .

Transplant • Definitii – Autogrefa: transfer de tesuturi de la – la acelasi organism (piele) – Allogrefa: transplant intre indivizi diferiti ai aceleiasi specii – Isogrefa (singenic): transplant intre gemeni identici (univitelini) – Xenogrefa: transplant de la o specie la alta .

in particular pentru locii D . Compatibilitate de Transplant • Pentru a creste sansa de supravietuire a transplantului: – Cel mai important: compatibilitate ABO – Absenta Ac citotoxici preformati impotriva Ag HLA ale donatorului – Compatibilitate HLA.

Antibodies Direct diagnostic tests. rubella virus. VZV. molecular biology tests (PCR. HSV 1 and 2. RT- PCR) – DNA. Methods Indirect diagnostic tests (serological) . CMV. HDV.RNA-virus . EBV. HCV. HTLV 1/2 . toxoplasma gondii and chlamydia. VIROLOGICAL ASSESSMENT Both donor and recipient are tested for: HBV. HIV 1/2.

HLA .

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AHG CDC .ELISA 2. Anti-HLA antibody detection and identification .sequence-specific oligonucleotide probe hybridization (medium resolution ) SSP – sequence-specific primers (high resolution) SBT – Sequence Based Typing allele SEQR (the highest available resolution) 3.ELISA .CDC . IMMUNOGENETICS 1. Cross.match . HLA Typing by molecular biology methods – PCR SSOP.

Lysis of donor lymphocytes is indicative of cytotoxic antibodies in the recipient’s serum directed against donor lymphocytes • The identity of these antibodies must then be determined in order to find a suitable donor who is negative for the corresponding HLA antigen(s). Major Histocompatibility Complex • Lymphocyte crossmatch – Used to screen recipient serum for anti-HLA antibodies • Recipient’s serum. complement and donor B lymphocytes are mixed together in a test tube. .

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Sample of cells or tissue Combine DNA with sequence- specific primer fix for each allele Amplify by PCR .

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DNA 80ng for Class I 40 ng for Class II .

Importance of DNA Quality 100 ng Genomic DNA 1% Agarose Gel .

Assign-SBT Resolves Ambiguities Sequences are arranged in “layers” …Master sequence .

also has been associated with amoebic transmission (granulomatous amebic encephalitis) . Types of Transplants • Corneal – Best graft survival rate since the cornea is avascular and the lymphatic drainage from the eye is not as well developed as in other tissues – Associated with transmission of prions- Creukfeld-Jacob disease-Transmissible spongiform encephalopathy.

Similar statistics to 1 haplotype match when the recipient receives multiple blood transfusions prior to the surgery (induces tolerance to the allograft) and is placed on immunosuppressive therapy . 5 year survival – With a 1 haplotype match = 80%. Types of Transplants • Renal – Between living donors with a 2 haplotype match = 90- 95%. 5 year survival – Cadaver transplants between unrelated donors is the most common type of transplantation.

Types of Transplants • Liver – In adults with chronic active hepatitis or cirrhosis – In children with biliary atresia – 1 year survival rate is slightly greater than 90% .

Types of Transplants • Cardiac transplantation – In adults. endocardial fibroelastosis is the usual indication – Endomyocardial biopsies are the best means of diagnosing allograft rejection – Approximately 80% of transplants survive 1 year . used in patients with chronic ischemic heart disease and congestive cardiomyopathy – In children.

leukemia and immunodeficiencies – Goal is to infuse donor marrow containing pluripotential hematopoietic stem cells that will eventually repopulate the lymphoid. myeloid. – GVH occurs in almost 2/3rds of cases – Increased incidence of CMV pneumonitis . erythroid. and megakaryocytic series in the recipient. Types of Transplants • Bone marrow transplants – Used in the treatment of aplastic anemia.

or 2 haplotype match is: – 25% .1 haplotype match – However. a 2 haplotype match is rarely achieved due to crossovers between the individual loci during meiosis when homologous chromosomes line up close to each other . 1. Transplant Rejection • The chance of a sibling in a family having another sibling with 0.0 haplotype match – 25% .2 haplotype match – 50% .

Transplant Rejection • Three types of transplant rejections – Hyperacute rejection – Acute rejection – Chronic rejection .

a blood group A recipient would have anti-B IgM antibodies and would react against a group B donor heart) • Hyperacute rejection is rare because ABO and anti-HLA cytotoxic antibody screening is performed prior to the surgery . Transplant Rejection • Hyperacute rejection: – occurs within minutes of attaching the allograft to the recipient’s blood supply – Due to the presence of an ABO mismatch or preformed cytotoxic antibodies in the host against foreign HLA antigens in the donor tissue (example.

Transplant Rejection • Acute rejection – Most common type of rejection encountered – Usually occurs within the first 3 months of the transplantation – Involves cell-mediated and antibody-mediated reactions:  Cell-mediated has the greatest role in rejection  The type II antibody-mediated hypersensitivity produces a necrotizing vasculitis with subsequent vessel damage and intravascular thrombosis .

. corticosteroids. Transplant Rejection • Acute rejection – Vessel events can occur over a period of time leading to fibrosis and vessel lumen obliteration – The cell-mediated component involves cytotoxic T cells producing extensive interstitial infiltrate in the graft with edema and damage to the tissue (Type IV hypersensitivity) – Can be reversible with immunosuppressive drugs such as cyclosporin A. and OKT3.

Transplant Rejection • Chronic rejection – Irreversible – Occur over a period of months to years – Extensive fibrosis and loss of organ structure characterize the histologic findings in the transplant – Activated macrophages release growth factors that stimulate fibroblasts to deposit collagen – There is also chronic ischemia secondary to antibody- mediated damage to the vessels .

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blocking their reaction with the graft . Transplant Rejection • Cyclosporin A inhibits CD4 helper T cell release of interleukin-2 (blocks calcineurin) which stimulates the proliferation of cytotoxic and helper T cells • Corticosteroids inhibit macrophage production of interleukin-1 and tumor necrosis factor and are cytotoxic to immature cortical derived thymocytes • OKT3 is a monoclonal antibody preparation that attaches to the CD3 antigen receptor of T cells.

jaundice. 2+ jaundice. relative eosinophilia. right upper quadrant pain. CBC/PBS: falling blood counts. he received radiotherapy and chemotherapy as well as broad-spectrum antibiotics PE VS: normal blood pressure. two months ago. HPI Prior to the transplant. moderate dehydration. he underwent an apparently uncomplicated bone marrow transplantation. Elevated direct serum bilirubin and transaminases. and dyspnea. no infectious agents on stool exam . a generalized rash. PE: cachexia. violaceous and erythematous macules as well as papules and bullae with scale formation over extremities Labs Elevated IgE level.ID/CC A 45 year old male with refractory acute myeloid leukemia is brought to the emergency room with fever. severe diarrhea.

Graft versus Host Reactions • Potential complication in bone marrow and liver transplants and in blood transfusions administered to patients with T cell immunodeficiency • Donor lymphocytes produce interleukin-2 • -->activation of NK cells (primary effector cells in acute GVH reactions)-->lymphokine-activated NK cells are called LAKs and produce extensive epithelial cell necrosis in the biliary tract (jaundice). and GI tract (diarrhea) . skin (maculopapular rash).

Graft versus Host Reactions • May progress into chronic GVH which is marked by the presence of extensive fibrosis • To lessen the risk of GVH. • Cyclosporin A is used also . donor tissue is pretreated with anti-thymocyte globulin to remove donor T cells.

Transplant complications • Immunosuppressive therapy has increased the incidence of: – Cervical cancer – Malignant lymphomas (immunoblastic) – Basal and squamous cell carcinomas of the skin • Squamous cell CA is the most common overall malignancy • Other complications include infection and bone marrow suppression .

Diagnostic Immunopathology…. Immunodeficiency Disorders .

in certain cases. What is Immunodeficiency? Under-reaction to antigen • A failing of one or more of the body’s defensive mechanisms resulting in morbidity or mortality. • Immunodeficiency may be Primary or Secondary. • Severity is variable. proteins. • Or. other homeostatic systems in the body will be disrupted by the defect. • Any part of the immune system can be deficient – cells. • The body is susceptible to infection by organisms that meet with little or no resistance. signalling mechanisms……. .

diagnosis and therapy options for primary immunodeficiencies involving: – Phagocytes (Chronic Granulomatous Disease) – The Complement cascade (HereditaryAngioedema) – Lymphocytes (X-Linked Agammaglobulinaemia) . Learning Points • Be aware of the range and classification of immunodeficiency disorders • Understand the nature of disease.

.Primary Immunodeficiency: • Arises without a pre-existing causative disorder. • Usually has a genetic basis.

g. – Infection – Renal failure. e. or protein losing enteropathy – Leukaemia or Lymphoma – Myeloma – Extremes of age – Certain Drug Therapies .Secondary Immunodeficiency: • Arises because of another pre-existing pathology or intervention.

Immunodeficiency Disorders • Most immunodeficiency disorders are acquired (most common acquired disorder is AIDS) • Approximately • 50% are B cell disorders. and • 5% each for phagocytic and complement problems • Most cases are in children less than 15 years of age (80% male) . • 40% T cell disorders.

B cells. or complement • these deficiencies are grouped as: – primary: inherited or congenital • severe combined immunodeficiency (SCID) – secondary: acquired • HIV infection David Vetter 1972-1984. the original bubble boy . Under-reaction to antigen Immunodeficiencies • occur when some part of the immune system is defective or missing – T cells. phagocytes.

– Common variable immunodeficiency (CVID). differentiation failure leading to decrease in one or more of the IgG subclasses or IgA subclasses. – Adhesion molecule defects. a common but poorly defined collection of syndromes with reduced IgG. defective killing. – Chronic Granulomatous disease. oscillation of neutrophil numbers. – Specific granule defects.• Phagocyte deficiencies: – Cyclic neutropenia. and IgA/IgM. • Antibody deficiencies: – X-linked agammaglobulinaemia – Selective Ig deficiency. . affects chemotaxis.

Factor I. also low platelets and low IgM. – Hereditary Angioedema (C1-INH deficiency). gene defects affecting thymus development and hence T cells. . • Complement deficiencies: – Deficiency of individual components. or inhibitors Factor H. C3. – Wiskott-Aldrich.• T cell deficiencies: – DiGeorge’s Syndrome. C5 – C9. Now known as CATCH-22 syndromes. e. progressive reduction in T cells.g.

Primary Immunodeficiency: Phagocyte-specific Disorder .

? Chronic Granulomatous Disease (CGD) ? What is CGD? • A group of diseases in which there is a defect in a major killing mechanism of phagocytes. • CGD is the most significant of the phagocyte defects. • Patients are vulnerable to specific bacterial and fungal infections. ? . inherited and result from ? genetic abnormalities in key enzymes of the ‘oxidative burst’. • The diseases are rare.

. Clinical findings • Recurrent severe bacterial and fungal infections occurring at sites acting as environmental barriers: – Skin -Lymph nodes – Liver or spleen -Lungs – Gastrointestinal tract • Infecting organisms are specifically ‘catalase positive’ which means the faulty phagocyte cannot use the organism’s endogenously produced hydrogen peroxide to kill them. • Patients are no more susceptible to viral infection than anyone else.

gingivitis. gum disease. Aspergillus.. Aureus. abscesses. Pseudomonas. although milder forms may present in adulthood • Symptoms may include boils. • Delayed growth and malabsorption in children. lymphadenopathy. . pneumonia.and unusual organisms such as Serratia marcescens • • Patients are usually children. granuloma.• Organisms include: Staph. diarrhoea. lymphadenopathy. • Chronic inflammation at various sites – e. Candida…. GI or urinary obstruction. Salmonella. fever.g.

• Neutrophils are morphologically normal. Laboratory Diagnosis • Patients often have an elevated • Nitroblue tetrazolium test white cell count (NBT) where NBT (colourless) (neutrophil is reduced to formazan (blue) by the phox enzymes. flourescing cells are counted. . Slide or tube assay – observing the presence of blue inclusions in phagocytes. • Dihydro-rhodamine (DHR) reduction assay – fluorescent rhodamine (reduced DHR) is • Chemotaxis and detected by flow cytometry. leucocytosis). endocytosis are Flourescence in cells is measured and normal.

St. When to suspect CGD? • In a patient (particularly a male child): • with recurrent infection with catalase positive organisms • Infection with rare organisms • Infection in unusual sites e.g. aureus liver abscesses .

• Individual susceptibility varies but 1 serious infection can be expected every 3-4 years in X-CGD cases even with prophylaxis. . • Trials of Gamma IFN – may reduce number and severity of episodes. • Tissue biopsy/drainage – may need hospitalisation. • Antibiotic prophylaxis. • Steroids and NSAIDS sometimes used to treat granulomas. Therapy and Management • Prompt aggressive treatment of infection with antibiotics.

Primary Immunodeficiency: Disorder of the Complement System Hereditary Angioedema (HAE) .

A low level of C1 esterase inhibitor is the confirmatory test • Treatment with synthetic androgens which increase the synthesis of the inhibitor . also GI • Laboratory tests show a decreased level of C4 (best screen) and C2. Complement Disorders • C1 esterase inhibitor deficiency (hereditary angioedema) – Autosomal dominant disease resulting in excessive release of C2-derived kinins that increase vessel permeability resulting in edema • See involvement of the face and oropharynx sometimes with obstruction of the airway.

? What happens in HAE? ? • A deficiency of C1INH due to genetic mutations of the C1INH gene on chromosome 11p11-q21. • Patients are also susceptible to immune complex disease and SLE (failure to remove IC’s). ? . • The loss of control of the complement and contact systems results in severe subcutaneous and submucosal oedema due to fluid leakage into the extravascular space. – Secondary or Acquired forms (AAE) occur in ? some LPD’s or in association with C1INH autoantibodies.

Different variants exist: • Variant I • Variant II • 80-85% of cases • 10-15% of cases • Low levels of normal • Normal or increased levels of C1INH are found (10. abnormal C1INH are found 30% of normal) • Autosomal dominant. • Variant III genetic mutations often • Not associated with C1INH ‘missense mutations’ deficiency (X-linked) No one with homozygous HAE has ever been observed. genetic • C1INH present is mutations are usually single produced by the normal ‘point mutations’ allele • Autosomal dominant. .

• Episodes may be as frequent as weekly or as rarely as once a year or less. • Tingling or tightness may precede attacks. nausea and vomiting – often mistaken for acute abdomen. • Extreme swelling of tissues – oedema. Clinical findings • Occurs in childhood or adolescence. . feet. • Often in hands. airways. • Pain and swelling of abdomen. • Swelling of larynx – may cause fatal asphyxiation. • Oedema lasts usually 24-72 hours. face.

Triggers……… • Minor trauma • Surgery • Anxiety/stress • Mild illness – colds or ‘flu • Dental procedures • Hormones – pregnancy or OCP • Physical activity – typing. . writing. gardening etc….

C3. C3. Laboratory Diagnosis • Firstly . • Low C1INH with normal C1.measure C4 levels. C1q (Type 2 HAE) . If low: • Measure C1INH level (single radial-immunodiffusion) and function (ELISA). C1q (Type I HAE) • Normal or raised levels of dysfunctional C1INH with normal C1.

XLA (Bruton’s Disease) .Primary Immunodeficiency: Lymphocyte-specific disorder affecting antibody production. X-Linked Agammaglobulinaemia .

. • Also seen: joint problems. kidney problems. meningitis. ear (otitis media). blood (sepsis). malignancy in older patients. • Sites of infection: mucous membranes. Clinical findings • LITTLE BOYS WITH BIG INFECTIONS! • Symptoms appear at 6-9 months of age (after loss of maternal Ig) or as late as 3-5 years of age. gut (Giardia. lungs (bronchitis/pneumonia). skin. eyes. neutropenia. or enterovirus).

• If diagnosis has been delayed then permanent lung damage (bronchiectasis). • Growth retardation may be present. or hearing loss due to repeated infection may be present. .• Typical organisms involved include: Haemophilus Influenzae. • Affected boys have little or no tonsils or palpable lymph nodes. • There may be no family history of XLA. Streptococcus Pneumoniae.

• Profound reduction in circulating B lymphocytes. severe. . • Hypogammaglobulinaemia (decreased immunoglobulin levels) – absent in severe cases. Significant Laboratory Findings Serum protein electrophoresis • Microbiological evidence of early-onset. recurrent infection. • For confirmed diagnosis – mutation in the Btk gene.

Counting lymphocytes by Flow Cytometry 488 nm CD19-PE CD3-FITC CD45-Per CP .

Lymphocyte subsets by Flow Cytometry .

• Prompt treatment of any infection and use of prophylactic antibiotics. • Avoidance of live vaccines (Polio). including siblings. Therapy Options & Management • Intravenous Immunoglobulin (IvIg). . • Education of patient and family. • Genetic counselling. • Prenatal testing.

• Treatment but with chronic lung disease: – May have a shortened life span. Prognosis • No treatment: – Death at an early age. • Early diagnosis and treatment: – Normal active life. .

Canada. U.S. which – HIV 1 is the most common decrease in number isolate in the E. first identified in 1981 • infects CD4+ (helper) T cells. the activity of HIV proteins.. U. AIDS • Acquired Immunodeficiency • caused by the human immunodeficiency syndrome is the most virus (HIV . HIV 2 is primarily • decreased numbers of CD4+ T cells lead isolated in West and Central Africa-transmitted to increased susceptibility to heterosexually opportunistic infections.U.. of age thereby preventing production of the virus ..1984) and is spread by common acquired contact with body fluids immunodeficiency in the E.S.U.. – AIDS is the most common cause of death in men and • treatments include drugs that inhibit women between 25-44 years RT-ase.

represent the remainder) • Adults: – Intimate sexual contact is the most common mode of transmission.000 risk) – Health care workers most commonly contract the disease from accidental needle sticks where the risk for becoming HIV positive is 0.3% . AIDS Modes of Transmission • Subsets of individuals that become HIV positive in decreasing order of frequency – Homosexual or bisexual men – IV drug abusers – Heterosexual contacts – Individuals receiving blood or blood products (includes hemophiliacs) • Men account for approximately 88% of all people who are HIV positive (women-IV drug abusers. usually by anal intercourse between homosexuals or by vaginal intercourse in heterosexuals – Inoculation of the virus through blood or blood products is the second most common mode of transmission with the majority of cases related to the sharing of needles between intravenous drug abusers and less commonly by receiving a blood transfusion (1:40.000 to 1:200.

and assays for the p24 antigen in the serum . AIDS Modes of Transmission • Children – Aids is contracted in greater than 90% of cases from an infected mother who is usually an intravenous drug user – Transplacental (vertical) transmission accounts for approximately 30-50% of cases. therefore. – Treatment of asymptomatic pregnant women with AZT has reduced the rate of infants developing AIDS from 13-40% to less than 10% • All newborns of HIV positive women are HIV Ab positive due to transplacental passage of IgG antibodies. documentation of HIV infection in infants is often difficult during the first 2 months – Tests in this setting that are useful include the isolation of the virus in peripheral blood monocytes. the detection of viral RNA using PCR. while perinatal factors associated with blood contamination during delivery and breast feeding (most common perinatal factor) represent the remainder of cases – Pregnant women who have CD4 helper T cell counts less than 400 cells/uL or who are positive for the viral p24 antigen (a marker of disease activity) have an increased risk for transmitting HIV to their baby.

• The co-receptor is CXCR4 on T cells and  chemokine receptor CCR5 on macrophages • Macrophages are the primary reservoir for the virus and are the vehicle for carrying the virus into the CNS .AIDS • AIDS virus attacks and destroys CD4 T helper cells by first attaching to the CD4 molecule on the surface membrane of the lymphocyte with the gp120 envelope protein • A co-receptor must also be present with the CD4 molecule for fusion and entry of T cell tropic strains of HIV-1.

integrase. Reverse transcriptase catalyzes the reverse transcription of viral RNA into double-stranded DNA. fusion with the host cell membrane occurs via the gp41 molecule. AIDS • Following binding. • The HIV genomic RNA is uncoated and internalized. • The virally encoded enzyme. causes the viral DNA to be integrated into the host cell DNA .

.AIDS • Cells with CD4 molecules on their surface in which the virus can replicate without killing the cell are monocytes. tissue macrophages. because viral DNA cannot integrate efficiently into the host cell’s genome unless activation has occurred • Activation of the host cell is also required for initiation of transcription of the integrated proviral DNA into either genomic RNA or mRNA. and microglial cells (macrophages) in the CNS • Host cell activation is CRITICAL to the pathogenesis of HIV. dendritic cells in the skin and lymph nodes.

HIV Life Cycle .

HIV & Immune System .

and lymphadenopathy – CD4 counts are usually normal. AIDS Clinical Findings • Divided into acute. sore throat. but the p24 antigen is increased . latent and late phases • Acute phase – Develops within 3-6 weeks of contracting the virus – Initial infection is characterized by fatigue.

normal . AIDS Acute phase – p24 – High RNA levels – CD4+ cell counts .

– This finding is not associated with an increased likelihood of developing AIDS. a loss in lymphadenopathy or a decrease in lymph node size may be a prognostic markers of disease progression . however. AIDS: Early Symptomatic Disease • Generalized lymphadenopathy – The presence of enlarged lymph nodes (> 1 cm) in two or more extrainguinal sites for more than 3 months without an obvious cause.

. AIDS: Early Symptomatic Disease • Other non-specific findings: – Fever – Weight loss – Diarrhea – Malaise – A distinct group of patients with early AIDS develop immune thrombocytopenic purpura (ITP) due to the presence of anti-platelet antibodies.

AIDS Laboratory Findings • Antibodies against gp120 and gp160 (ELISA test) are not usually present for approximately 4-8 weeks (window period). – screening tests in blood banks may be negative during this phase • Positive ELISA screens are confirmed by the Western blot analysis which detects p24 and gp41 antibodies in order to increase overall specificity Testul Western Blot. 2: CTR negativ. Linia 1: CTR pozitiv. 3-4: teste pozitive HIV-1. 7-10: teste pozitive seriate la un pacient infectat HIV-1 . 5-6: teste pozitive HIV-2.

AIDS Asymptomatic latent phase – Virus is actively proliferating and is present in follicular dendritic cells (macrophages) in lymph nodes – The rate of disease progression correlates with HIV RNA levels – The CD4+ cell counts fall progressively during this asymptomatic period at the rate of approximately 50 cells/microliter/year .

hairy leukoplakia and aphthous ulcers are very common during this stage. but are not severe enough to be indicative of a defective cell-mediated immune response – Oral lesions: Thrush. The first two are associated with declining immunologic function (< 300 CD4+ T cells) . AIDS Symptomatic phase • Clinical Findings – Follows an average span of 10 years – CD4 count drops to below 400 cells/uL – p24 antigen appears again – Patients develop minor opportunistic infections.

causing bacterial pneumonias). and Streptococcus pneumoniae • Due to the use of trimethoprim and aerosolized pentamidine. disseminated CMV infections. Pneumocystis carinii is no longer the most common cause of death • Average life span from beginning of infections to death is 10 years . AIDS Advanced AIDS • Diagnosis of AIDS is based upon an opportunistic infection (usually Pneumocystis carinii pneumonia) and/or a CD4 count of  200 cells/uL in an HIV positive individual • Most common cause of death today is bacterial pneumonia (most common pathogen is Mycobacterium avium intracellulare –MAIC.

associated with HHV-8 . Organ Pathology in AIDS • Lungs are the most frequently involved organ in AIDS • Gastrointestinal tract • Kaposhi’s sarcoma is the most common cancer in AIDS and the most common cutaneous lesion.

Organ Pathology in AIDS • Other malignancies include: – Cervical cancer (HPV) – Anal squamous carcinoma (HPV) – Non-Hodgkin’s lymphomas – Primary CNS lymphomas (HIV) .

ketoconazole therapy – Clinical diagnosis of adrenal insufficiency should be suggested by hypotension with electrolyte disturbances . Organ Pathology in AIDS • Endocrine disease – Adrenal insufficiency is very common – HIV induced abnormalities in steroid synthesis. adrenalitis from CMV.

Organ Pathology in AIDS • CNS and PNS disease – Dementia – Meningitis – Peripheral neuropathies – Spinal cord lesions – Opportunistic infections • Toxoplasmosis is the most frequent cause of encephalitis and a space occupying lesion in the CNS • CMV chorioretinitis is the leading cause of blindness in AIDS • Cryptococcus is the most common cause of meningitis in AIDS .

lymphopenia.primary renal disease • Dermatologic Disease – Seborrheic dermatitis • Seen in up to 50% of patients with HIV • Caused by Malassezia furfur • Hematologic problems – Cytopenias (neutropenia. thrombocytopenia) • Autoantibodies against platelets – Anemia of chronic disease – Autoimmune hemolytic anemia . Organ Pathology in AIDS • Renal Disease – Focal segmental glomerulosclerosis (HIV- associated nephropathy) presents with nephrotic syndrome .

hepatitis. AIDS Laboratory Abnormalities • To detect Antibodies against gp120 and gp160 – Detected by EIA (enzyme-linked immunoassays) within 4-12 weeks after contracting the virus) – False negatives in patients who are immunocompromised or who have serum collected before the antibody has developed – False positive in patients with autoimune diseases. with anti MHC II Ab . receiving recent vaccins.

5% . AIDS Laboratory Abnormalities • AIDS antibodies • Other laboratory – Duplicate negative tests are reported as negative abnormalities – Second test turning positive must be – Reversal of the CD4/CD8 verified by obtaining suppressor T cell ratio positive results in 2 out of 3 repeated tests – Absolute lymphopenia – All positive or indeterminate EIA tests – Hypergammaglobulinemia are confirmed with a secondary to polyclonal B Western blot assay which measures p24 and cell stimulation by EB gp41 antibodies to virus and CMV improve the overall specificity of the test – Decreased mitogen • A positive EIA in combination with a positive blastogenesis Western blot has a positive – Anergy to skin testing predictive value of 99.

Western Blot Results .

one during the acute infection and the other at the onset of AIDS . AIDS Laboratory Abnormalities • Other laboratory abnormalities – Decreased production of cytokines – NK cells are not activated since gamma interferon from CD4 cells is decreased – Increased p24 core protein indicates active disease and has two peaks.

AIDS Treatment and Prevention • Initial treatment: Azidothymidine (AZT) – Blocks reverse transcription • HIV reverse transcriptase is not accurate and produces frequent transcription errors – This high mutation rate causes resistance to drugs • Treatments include: – Reverse transcriptase inhibitors – Protease inhibitors (saquinavir and ritonavir) – New drugs currently being developed that block HIV’s entry to helper T cells • Other drugs include – Gamma interferon – Granulocyte-macrophage colony stimulating factor – Granulocyte colony stimulating factor .

AIDS Treatment and Prevention • Prevention in order of decreasing effectiveness – Abstinence – Monogamous relationship with a confirmed seronegative partner – Manual sex – Kissing – Intercourse with latex condoms using nonoxynol-9-a spermicide with some anti-viral activity .

Bazele celulare şi moleculare ale reactiilor imune patologice .

dobândit) . autoimunitate)  deficit (genetic. IMUNITATEA PATOLOGICA  exces (hipersensibilitate.

Hiperergie = Răspuns Imun care cauzează leziuni tisulare Se disting diferite tipuri de hipersensibilitate după: • Evoluţia în timp • Tipul de protagonişti imuni implicaţi (Ac sau Tc)  Alergiile mediate de Ac sunt de tip imediat/acut şi subacut  Cea mai importantă reacţie de hipersensibilizare de tip întârziat este cea mediată celular .Hipersensibilitate .

IgG. fagocite (Mo/MQ). Mastocite / Bazofile. PMN Celule  Tipul IV ● Mediată prin limfocite Tc . M. şocul anafilactic)  Tipul II ● Ag asociate celulelor. Eozinofile “Atopia”: Tendinţa exagerată de a dezvolta un răspuns prin Anticorpi ● IgE (forme clinice de prezentare a RIP tip I: astmul. Clasificarea Gell şi Coombs (după mecanismul de producere) a tipurilor de Hipersensibilitate  Tipul I (“Alergia”) ● Ag solubile. alergia alimentară. rinita alergică. limfocite NK (ADCC)  Tipul III ● Complexe imune Ag-Ac. sistemul C. urticaria. IgE.

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Type I hypersensitivity – sensitization to an inhaled allergen or bee sting cytokines   Mast cell Antigens (red dots) from inhaled pollen are ingested and presented by macrophages to T cells. . which is responsible for allergy symptoms. Activated T cells produce cytokines leading to the production of IgE. which binds to receptors on mast cells and causes the release of histamine. Onset is usually within minutes of contact with antigen.

because the bound drug makes them look foreign to the immune system. the red blood cells are more susceptible to lysis or phagocytosis.Type II hypersensitivity – immune-mediated destruction of red blood cells Drug (p=penicillin) modified red blood cells  induce the production of antibodies. . When these antibodies are bound to them. Onset is dependent on the presence of specific antibodies.

and and C5a). systemic. . The location of the inflammation depends on the location of the antigen . under skin.inhaled. Onset is usually within 2-6 hours. Type III hypersensitivity – immune complex formation and deposition Immune complexes Immune complexes activate complement of antigen (red dots) Inflammation and (green dots. which activate complement and the inflammatory response. and antibody form in edema occur. In sensitized individuals. C4a. and mast cells target organ organ is damaged (yellow cell) degranulate.C3a. allergen (antigen) combined with antibody leads to the formation of immune complexes.

leading to tissue injury. peaks by 24-48 hours.Type IV hypersensitivity – delayed-type or contact T cells (blue cells) that recognize Antigen (red dots) antigen are Inflammatory are processed by activated and response causes local APCs release cytokines tissue injury. Inflammation by 2-6 hours. Antigen is presented by APCs to antigen-specific memory T cells that become activated and produce chemicals that cause inflammatory cells to move into the area. .

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asparaginaza.  -lactoglobulina  VENIN HIMENOPTERE  fosfolipaza A2  MEDICAMENTE  insulina.Nomenclatura OMS a unor alergeni  INHALATORI  polen de mesteacăn (Betula verucosa): Bet v1  acarian (Dermatophagoydes pteronissinus): Der p 1-6  gândac bucătărie (Blatella germanica): Bla g 1-2  peri pisica (Felix domesticus): Fel d 1  peri cal (Equus cabalus): Equ c 1-3  Aspergillus fumigatus: Asp f 1  ALIMENTARI  ou (Gallus domesticus): Gal d 1-3  lapte vacă: caseina. lactalbumina. latex (Hev b 1) .

dar sensibilizează sistemul imun: ● mecanismul implică IL-4 secretate de limfocitele Th2 ● IL-4 stimulează B să producă IgE La contactul ulterior cu alergenul. Hipersensibilitatea imediată – tipul I Contactul iniţial cu alergenul este asimptomatic. are loc: •Legarea IgE de receptorii specifici din membrana Ma •Eliberarea de amine vasomotorii si producerea de agenti inflamatorii -> răspuns inflamator •Simptomatologia poate fi localizată sau sistemică .

bazofilul tisular 1.Gut & lung: •T cell dependent •Short lived <40 days •25x105 IgE receptors •Lower histamine content •Chondroitin sulphate •Lower tryptase Proteoglicani. TNF. LTB4. Ubiquitous in connective tissues: •Long lived >40 days •3x104 IgE receptors •High histamine content •Heparin & high tryptase 2. -5. Mucosal . -6.E4). Tromboxan GM-CSF. Lipoxine . Mastocitul . Proteaza neutra. -4. IL-3. Calea cicloxigenazei:Prostaglandine ß-Glucozaminidaza. PAF Calea lipoxigenazei: SRS-A (LTC4.D4.

leukocyte-derived histamine-releasing peptide. neuropeptide. citokine (IL-1. declansata de factori asociati cu coagularea (F XII) ● Vasodilatatie. Pl ca raspuns la injurie (traume. agregarea plachetara declanseaza eliberarea. Mediatori specifici eliberaţi  Amine vasoactive: ● Histamina: eliberata de Ma. reactii imune (IgE-FcRI Ma). cu largirea jonctiunilor ● Serotonina: prezenta in granulele dense Pl. IL-8). . Ba. permeabilitate vasculara (edem) ● Posibil efect chemotactic Leu (declanseaza expresia ECAM pe endoteliul vascular) ● Contractia musculaturii netede non-vasculare (bronsice) ● Durere ● Inactivate rapid (kininaze) ● Actioneaza mai lent decat histamina. temperaturi extreme). efect vasodilatator similar cu al histaminei  Kinine plasmatice: ● Formarea bradikininei (BK) prin clivarea HMWK. anafilatoxine (fragmente C3a. produce vasodilatatie si contractia celulelor endoteliale venulare. C5a).

.

Anafilaxia localizată şi Şocul anafilactic

Caracteristicile răspunsului imun IgE

• componentă normală a răspunsului imun secundar

• durată scurtă

• inductibil cu doze foarte mici de antigen

• modulat de citokine: întărit de IL-4, suprimat de
IFNγ

• în principal, produs ca reacţie imună la interfaţa
organism-mediu

• relativ uşor de întrerupt: doze foarte mari de
antigen, administrări repetate (imunoterapie
specifică)

Tratament
 Influentarea raportului Th1/Th2 : desensibilizare

Hipersensibilitatea subacută
 Cauzată de IgM şi IgG, poate fi transferată via plasmă
sau ser
● Debut lent (1–3 ore) după expunerea antigenului
● De durată (10–15 ore)

Reacţia citotoxică (tip II)

• Ac se leagă la Ag fixate in
membrana celulară,
stimulând reacţia
citotoxică dependentă de
Ac şi citoliza mediată prin
complement

• Exemple: reacţiile
patologice din
transfuzia de sânge cu
incompatibilitate în
sistemul ABO şi Rh
• R TSH – LATS
(hipertiroidie)
• RAC - Miastenia gravis
• RPL – hiposecreţia
lactată
• RSTH – nanism

Diagnosticul fenomenelor de tip II

Demonstreaza fixarea anticorpilor si/sau a
factorilor de complement la suprafata
celulelor purtatoare de Ag
● Tehnica: imunofluorescenta
(Anticorpi anti-Ig umane cuplate cu
o molecula fluorescenta)
● Teste functionale:

o Crossmatch (verifica existenta
Ac preformati la primitor,
prevenind rejetul superacut in
transplant)
o test Coombs direct sau indirect:
 Direct – Cauta Ig sau C3 pe hematiile
pacientului
 Indirect - Cauta Ig anti-hematii in sreul
pacientului

lezarea vasculară. PAR.“de sistem” (LED. Hipersensibilitatea subacută Reacţia imună patologică prin complexe imune (tip III):  Antigenele sunt răspândite în sânge sau în ţesuturi (adesea autoAg)  Se datorează: ● Persistenţei infecţiei / Administrării repetate a Ag (local sau general) ● Sintezei continue. DM)  Relativ comune (mai ales la vârstnici) . ineficiente de Ac  IgM/IgG formează CIC: ● La un raport corect al concentraţiei componentelor. renală. tegumentară “ inocent bystander” (prin activarea sistemului complementului şi a proteazelor Leu) ● Reacţie inflamatorie locală (fenomen Arthus) sau generală (boala serului)  Caracterizează bolile autoimune generalizate . sunt rapid epurate de MQ ● Complexele mici/solubile (Ag >>> Ac) sunt greu epurate -> Inflamaţia intensă.

... alveolite « alergice ».) .Fenomene de tip III localizate Fenomenul Arthus Implicatii clinice: afectiuni pulmonare in care alergenul este inhalat in mod repetat (plamanul de fermier.

Boala Serului  Simptome ● Febra ● Eruptii ● Adenopatie ● Artrita ● Glomerulonefrita .

. . etc.  Medicamentos ● Reactiile medicamentoase se pot manifesta clinic prin mai multe tipuri de reactii imune patologice  Autoimun ● LED o complexe: anticorpi anti-DNA si DNA depozitati in articulatii. Fenomene de tip III  Postinfectios ● Glomerulonefrita postinfectioasa (streptococica) ● hepatite ● mononucleoza ● malarie ● . piele. rinichi..

antibiotice) ● B.Hipersensibilitatea întârziată (Tip IV)  Debut lent (1–3 zile)  Mediată prin mecanisme implicând limfocite Th (pentru Ag solubile) şi Tc (pentru Ag asociate celulelor) ● Citokinele din Tc activate sunt mediatori ● Activează Tc (cu Ag procesate de MQ si prezentate prin MHC II) rejectia grefelor (HVGR)  Activează TH1 (cu Ag procesate de MQ si prezentate prin MHC I sau II) cu eliberarea de citokine (IFN) ● TB test cutanat ● Dermatitele de contact (Triggers: chimicale. bijuterii ieftine (Ni).autoimune (DZ I.Tiroidite) .

Patogeneza RIP Tip IV .

RIP tip IV mediată de Th1 – Alergen injectat Infiltrat celular!! Exemplu: testul la tuberculină .

RIP tip IV mediată de Th1 – Alergen absorbit Exemplu: Dermatitele de Contact .

Nickel RIP tip IV – contact cu metale grele Heavy metals bind to proteins and haptenylate them. Stimulates an immune response Transport to lymph node and T cell stimulation to hapten:protein .

RIP tip IV – DZ I “Insulita beta” .

allo-.auto-. toleranţă . Funcţii şi disfuncţii ale Th Ts TH1 TH2 SĂNĂTATE (imunoprotecţie: răspuns inflamator adecvat. xeno-) TH2 TH1 TH2 TH1 Alergie (toleranţă xeno Boli autoimune (toleranţa self scăzută) scăzută) Răspuns excesiv inflamator Răspuns excesiv inflamator .