Prof.

Ileana Constantinescu MD, PhD
Immunology
Fundeni Clinical Institute
ileana.constantinescu@imunogenetica.ro
www.imunogenetica.ro

Course 1: Definition of general aspects of immune response









Immunology
Innate immunity
Adaptive (Acquired) Immunity
Clonal selection
Humoral and Cellular Immunity
Lymphocyte migration into lymphoid tissues
Germinal centers
Follicular dendritic cells
Mucosal immune system
Course 1: Immunology, Prof. Ileana Constantinescu

2

Immunology
Immunology: that branch of biomedical science concerned with the response of the
organism to antigenic chalenge, the recognition of self and not self, and all the biological
(in vivo), serological (in vitro) and physical chemical aspects of immune phenomena.
Immunology means the study of the structure and function of the immune system
(basic immunology); immunization, organ transplantation, blood braking and
immunopathology (clinical immunology); laboratory testing of cellular and humoral
immune function (laboratory immunology); and the use of antigen-antibody reactions in
older laboratory tests (serology and immunochemistry).

Immunological memory
Course 1: Immunology, Prof. Ileana Constantinescu

3

some smaller parasites. Prof. Course 1: Immunology.) that have gained access to our body through the skin or the lining of our internal organs. etc. Ileana Constantinescu 4 . bacteria. pollen. some bacteria) and from extracellular (most bacteria. chemicals) but also from ourselves in potentially uncontrolled growth such as malignancy or autoimmune diseases.• The innate and adaptive immune systems protect us from potentially infectious agents (viruses. large parasites) pathogens and allergens (animal hair. Such systems have evolved to protect us from not only intracellular (viruses.

Ileana Constantinescu 5 . Natural killer lymphocytes kill some tumor and virally infected cells. Prof. Large parasites are killed by eosinophils. Extracellular pathogens are immediately taken up and degraded by neutrophils and mononuclear phagocytes. which can also attach to the cell membranes of many pathogens. Preexisting soluble mediators of innate immunity are natural antibodies and complement components.INNATE IMMUNITY • Innate immunity is a nonspectific cellular and humoral response that operates as the first line of defense against pathogens. Course 1: Immunology.

The innate and adaptive systems Course 1: Immunology. Prof. Ileana Constantinescu 6 .

Ileana Constantinescu 7 .Innate immunity also includes a cellular component. Pathology abstraction in the phagocytes phagolysosome. Course 1: Immunology. Prof.

the main mediators of innate immunity are lysosomal enzymes and interferons as well as other cytokines. in contrast to B cells. all of which interact in a cascade fashion to attract phagocytic cells (chemotaxis). multicellular organisms have developed a group of largely circulating cells with very specific receptors for immunogenic peptide fragments of both intracellular and extracellular pathogens. T cells are central to an adaptive immune response. Ileana Constantinescu 8 . Course 1: Immunology. the receptors on the surface of T cells can only recognize processed antigenic epitopes presented to them in the context of surface MHC. in order to develop into mature plasma cells. they usually require some help in the form of cytokines from T cells. in multicellular organisms with an adaptive system. The ability of T cells to see only foreign antigen in the context of self MHC is called MHC restriction and is a critical safeguard for the adaptive immune system to only eliminate pathogens and not normal cells. • Thus. When these receptors are activated. Complement consists of over 20 proteins. monocytes/macrophages) and/or natural killer cells as well as the complement system. However. However. Thus. If an organism enters the body by breaking through the barriers of the skin or mucosa lining the inner surfaces of the body. • It should be noted that innate host defenses are found in all multicellular organisms. While surface immunoglobin on B cells can directly respond to an immunogenic piece (epitope) of a pathogen. complement proteins. etc. coat bacteria (opsonize) so that phagocytic cells recognize and then engulf them. many of the mediators of innate immunity can recruit adaptive immune cells. Antigen-presenting cells either process or present already processed antigenic peptides to surface immunoglobin receptors on B cells and immunoglobin-like receptors of T cells. Helper T (Th) cells not only help B cells become plasma cells.) or stimulate other cells (cytokines). they can also help cytotoxic T cells develop as well as increase macrophage activity. and acute phase proteins. eosinophils. the cells can secrete various soluble mediators that attack pathogens directly (antibodies.• The main characteristics and components of innate immunity occur in the inflammatory response discussed. Prof. it comes in contact with phagocytic (neutrophils. perforin granules. • In order to handle certain pathogens. most of which do not have an adaptive immune system. and interact with antibody to lyse the cell membrane of the invading organism.

Prof. Ileana Constantinescu 9 .Interactions between the innate and adaptive immune systems Course 1: Immunology.

Prof. Ileana Constantinescu 10 .The cells central to adaptive immune response Course 1: Immunology.

when these memory T and B cells encounter that same antigen again their response to that antigen is much faster and more robust. • Thus.• B cells develop with their appropriate receptors for a cognate antigen's epitope T and without ever seeing that antigen before. Course 1: Immunology. Ileana Constantinescu 11 . Prof. they will remember such a first encounter. once encountering that antigen and generating a full immune response to it.

Prof. Ileana Constantinescu 12 .Course 1: Immunology.

Ileana Constantinescu 13 . Prof.Course 1: Immunology.

Course 1: Immunology. and production of cytokines and chemokines to attract and activate other components of the immune system. increasing their size. Ileana Constantinescu . Prof. phagocytic activity.Activation of phagocytic cells through binding of PRRs. Binding of PRRs to PAMPs on microbial cells stimulate phagocytes to become activated. production of antimicrobial substances.

NK cells can influence the development of T cells in the initiation of adaptive immune responses. and receptors that assess the MHC I molecules present on target cells. Once engaged. If KIRs are sufficiently engaged. Prof. killing of the target cell proceeds. Killer inhibitory receptors (KIRs) "examine" the target for sufficient levels of "self" MHC I molecules. These include Fc receptors. In addition. the killing program is terminated. If not. complement receptors.Natural killer (NK) cells. (B) NK cells utilize killer activation receptors (KARs) to recognize cells undergoing stress. Course 1: Immunology. KARs activate the NK cell to kill the target. (A) NK cells use a variety of receptors to identify target cells to be killed. Ileana Constantinescu .

Ileana Constantinescu 16 . Thus. antigen selects and generates specific clones that bind and respond to it.CLONAL SELECTION • When antigen binds to either surface immunoglobulin on B cells or its specific receptor on T cells. Course 1: Immunology. • This process is called clonal selection and is an extremely important part of the adaptive immune response. these cells are induced to proliferate rapidly. Prof.

Prof.Clonal selection Course 1: Immunology. Ileana Constantinescu 17 .

which kill virus-infected cells or abnormal host cells. Ileana Constantinescu 18 . Prof. • It should be realized that in many cases a normal humoral response will not proceed unless there is also a companion Th cell response. this is because Th cells secrete various cytokines that are necessary for full B cell maturation and antibody class switching.HUMORAL AND CELLULAR IMMUNITY • Humoral immunity is mediated by the antibody secreted by terminally differentiated B cells. the plasma cells. Course 1: Immunology. Cellmediated immunity involves T cells that recognize antigen in an MHC-restricted fashion and either secrete cytokines (Th cells) or become cytotoxic cells (Tc cells).

Ileana Constantinescu 19 . Prof.Introduction to the interactions and functions of the major components of the immune system Course 1: Immunology.

Overview of the process by witch cell-mediated immunity and antibody-mediated immunity Course 1: Immunology. Ileana Constantinescu 20 . Prof.

Induction of cell mediated immunity and antibody against a viral infection Course 1: Immunology. Ileana Constantinescu 21 . Prof.

E-selectin (ELAM-1) on endothelial cells binds L-selectin (LAM-1) on lymphocytes. it displays a different density of adhesion molecules specific for the tissue in which it was initially activated. thus allowing it to repeatedly return to that tissue. Ileana Constantinescu 22 . causing a loose association or tethering. Lymphocytes respond to chemotactic signals from inflamed tissues or secondary lymphoid organs initially via surface glycoproteins called selectins. memory cells tend to recirculate back to the tissue where they were activated. • Once a naive lymphocyte is activated. integrins. Migrating lymphocytes finally diapedese between endothelial cells into an organ or tissue by up-regulating production of proteinases. The endolhelial cell then directs the lymphocyte toward its basal lamina while simultaneously down-regulating ICAM expression. Firm adhesion is mediated by lymphocyte integrin proteins such as VLA-4 or LFA-1 interacting with their respective ligand VCAM or ICAM (member of the immunoglobin superfamily) on the surface of endothelial cells.LYMPHOCYTE MIGRATION INTO LYMPHOID TISSUES • Lymphocyte migration into lymphoid tissues involves a family of adhesion molecules called selectins. and members of the immunoglobin superfamily. Dr.Immunology . thus strengthening the adhesion. Course 1 .Prof. once activated. However. • This adhesive interaction causes an activation of the lymphocyte to increase expression of integrin molecules. • Both naive and memory lymphocytes continue to recirculate between blood and secondary lymphoid tissue.

dividing every 6 to 7 h. three B blasts colonize each primary follicle and are then called centroblasts. which express high levels of class II major histocompatibility complex (MHC). they also have a high propensity to recycle back into centroblasts for further proliferation and even higher affinity maturation. there is extensive fine tuning of centrocytes before they are allowed to produce antibody for export.and Th2-directed antibody class isotype production and eventual generation of plasmablasts or memory cells. Centroblasts. which are found in the base or dark zone of the follicle.GERMINAL CENTERS • Germinal centers (GCs) develop within primary follicles of lymph nodes and spleen during T cell-dependent immune responses. deletion of low affinity centrocytes. These centrocytes interact with Th2 cells and can become either plasmablasts or memory B cells. where they upregulate their somatically mutated surface lg (receptors). adhesion. down-regulate their surface lg and undergo a clonal expansion phase. Thus. Centrocytes with low affinity somatically mutated surface lg (receptors) for FDC-Ag do not upregulate bcl-2 and die via apoptosis. GCs are the sites of centroblast clonal expansion and VH region-directed somatic hypermutation.Immunology . they interact with follicular dendritic cells (FDCs). The centrocytes that express somatically mutated surface lg receptors with high ( ) affinity for FDCbound Ag up-regulate bcl-2 expression and are positively selected (which is opposite to positive selection of thymocytes). On the average. which can contain unprocessed Ag in the form of immune (Ag-Ab) complexes on their surface for months. • Centrocytes are the progeny of centroblasts that migrate apically into the basal light zone. Here. Ileana Constantinescu 23 . depending on the co-stimulatory signals they receive from FDC and Th1 or Th2 cells. and co-stimulatory molecules.Prof. these B blasts produce low affinity unmutated lg. However. Ag is presented to them by dendritic cells (DCs). which is capable of forming Ag-Ig complexes on follicular dendritic cells within primary follicles. respectively. Many of these cells die within 10 to 12 clays. Th1. • B cells in lymph nodes and spleen first encounter Ag in the T cell-rich areas of the paracortex and periarterial lymphoid sheath (PALS). During proliferation. After activation. selection of high affinity antibody-producing centrocytes. Dr. Course 1 . they activate a site-specific hypermutation mechanism that introduces random point mutations into their lg VH region genes. and within 2 days.

Ileana Constantinescu 24 . Dr.Immunology .GC zones Course 1 .Prof.

which drive genomic V(D)J rearrangements are also induced to be reexpressed in preapoptotic centrocytes of the basal light zone. Thus. however. along with the bcl-2-regulated apoptosis. interleukin-4. The development of GCs requires cluster formation between LFA-1 on B cells and ICAM-1 on FDCs as well as VLA-4 on activated B cells and VCAP on FDCs. In addition. Here Ab class switching and maturation into plasmablast or memory cells occur. Once centrocytes with high affinity surface Ig and high expression of bcl-2 have passed the discriminatory environment of the basal light zone. GCs in spleen and lymph nodes peakat 2 weeks and begin to wane after 3 weeks.Immunology .Prof. provides an additional mechanism for removing potentially autoreactive antibody-producing cells. Dr. This induction of an immature state in GC centrocytes may be necessary for further light chain receptor editing in an effort to save these cells. return to the circulation. Some activated B cells or plasmablasts can leave the secondary lymphoid organs. and -10 must be elicited from Th1 or Th2 cells in order for antibody class switching and full centrocyte development to plasmablasts or memory cells to occur. Plasmablasts migrate into the medullary cords of lymphnodes and red pulp cord of the spleen. they migrate to the apical light zone. Ileana Constantinescu . where they terminally differentiate intomature plasma cells and secrete their antibodies into the circulation . and reappear in the bone marrow and the gut. and the development of a full dark and light zone GC take 14 days. Memory B cells are found in the areas surrounding GCs (follicular mantles) a well as the marginal zone of the spleen.• • • • • • The recombination-activating genes RAG1 and RAG2. This editing of VL genes. in GC. 25 Course 1 . -5. there is an opportunity for antigen-dependent secondary V(D)J rearrangements to occur in order to fine tune the specificity of the peripheral antibody repertoire. Initial proliferation of centroblasts gives rise to the first foci in primary folicles within 2 days. GCs remain constitutively in the gut.

FOLLICULAR DENDRITIC CELLS
• FDCs have long processes and Fc, C, TNF receptors, as well as ICAM-1, and VCAM-1
but no MHC-II. They contain Ag-Ab complexes on their surface for months to years.
• Centrocytes that continue to maintain high levels of bcl-2 expression can receive
membrane-bound immune complexes (iccosomes) from FDC and induce the
expression of T cell-reactive surface molecules such as B7.2. This resultant interaction
determines whether there is differentiation into a mature GC centrocyte or
centroblast recycling. However, the formation of FDC clusters is critical dependent on
TNFα and LTα, since these cytokines are necessary for FDC cluster formation and
consequent primary as well as secondary follicles indicating that they have TNF-R1s.
• The fact that FDC can contain Ag for prolonged periods of time is the reason why we
do not need to be frequently immunized with certain antigens, since FDCs provide a
continuous depot of Ag even without an existing GC. It has recently been recognized
that the human immunodeficiency virus, upon disappearing from the blood, can be
sequestered on the dendritic arms of FDC for a number of years. Eventually, the FDC
can no longer hold the virus in check, the virus is released into peripheral blood, and
the patient enters the lethal stages of AIDS.
Course 1 - Immunology - Prof. Dr. Ileana Constantinescu

26

MUCOSAL IMMUNE SYSTEM


The mucosal immune system (MIS) consists of lymphoid tissues within and directly beneath the epithelial lining of
the respiratory, genitourinary, and gastrointestinal tracts as well as beneath the ductal system of the salivary,
lacrimal, and mammary glands. The surface area of mucosal surfaces is over 100 times greater than that of skin, and
the MIS contains up to 75% of all the B cells of the body. The primary product of the MIS is IgA.
Within the gastrointestinal tract, soluble antigen is taken up by villus epithelium and particulate antigens are
primarily taken up in the ileal portion of the small intestine by specialized surface lining microfold (M) cells. M cells
internalize Ag and transport it to underlying lamina propria macrophages, which process it and present it to
surrounding collections of lymphoid cells forming ileal Peyer's patches (PPs).
PPs contain follicles, similar to lymph nodes and spleen, with high endothelial venules and significant collections of
T cells in between. T and B blasts activated here go to the nearest mesenteric lymph node for further maturation,
alpha H chain class switching, or J chain formation or deletion. They then enter the thoracic duct and bloodstream,
homing back to the same or distant mucosal sites. Thus, even though much of the processing and reactivity to
antigen occurs in the ileum, protection through IgA occurs at many mucosal surfaces.
Serum IgA is monmeric and represents only 11% of all serum immunoglobulin. Secretory IgA represents over 95% of
all Ig found in secretions and is primarily dimeric with two monomerlic units covalently joined by a J chain. Dimeric
IgA binds to a polymeric immunoglobulin receptor (pIGR) on the basal surface of mucosal epithelial cells. This IgApIGR complex is endocytosed and transported to the apical (luminal) surface of the epithelial cell.
During this transport process, a small piece of the pIGR is cleaved with the remaining component now called the
secretory component. Thus, IgA is secreted as dimeric IgA bound to a secretory component.
Secretory IgA does not activate the complement system but coats bacteria and some viruses (polio, coxsackie, rota,
and herpes), thus preventing their adherence to mucosal lining epithelium. Also, some viruses within surface
epithelia can be neutralized by pIGR-internalized IgA. However, it should be noted that oral immunization with
soluble Ag does not always generate an immune response and can instead generate unresponsiveness.

Course 12 - Immunology - Prof. Dr. Ileana Constantinescu

27

Ag and cell traffic in gut
Course 1 - Immunology - Prof. Dr. Ileana Constantinescu

28

E. Roitt. ed. In Janeway CA Jr. 12th Edition. C. In Cellular and molecular immunology. Evolution of the immune system. and Bradley. Ivan M. R. Immunology Today 18. April 2011. Immunobiol-ogy: The immune system in health and disease.A. Humana Press Inc. American Society for Histocompatibility and Immunogenetics. Ileana Constantinescu 29 .  Dennis R. Danovitch. 111 :S442.Immunology . Seamus J. Berkower IJ. http://www. 5th ed. and Trowsdale. Histocompatibility Testing. General properties of immune responses.org Baxter-Lowe. 5th ed. In Paul WE. Fundamental immunology. 2005 by Lippincott Williams & Wilkins. Jr. ed. Cecka.. 6th ed.. and Allocation of Kidney Transplants in Handbook of Kindey Transplantation. Histocompatibility Testing (19). 2004:665-682. In Paul WE. Philadephia: WB Saunders Co. Cross-Matching. J. 1 in Transplantation Immunology Methods and Protocols edited by Philip Horniclx. Philadephia: Garland Publishing. Overview of the immune response.References • • • • • • • • • • • Roitt's Essential Immunology. Lichtman AH. Philadelphia: Lippincott Williams & Wilkins. in Lange Medical Immunology. Immunogenicity and antigen structure..D. 5th ed. Burton. eds. Philadelphia: Lippincott Williams & Wilkins.M. J. Martin. Delves. J... Fourth Edition. Course 1. Berzofsky JA. Callaghan. J Allergy Clin Immunol 2003. 2006.Prof. Tenth Edition.ashi-hla. Chaplin DD. and Reed F. 2003. 1997. Fundamental immunology. Travers P. Current status of Renal Transplantation Cpt. Campbell. J. 2001. Gabriel M. 2003. Includes Desktop Edition. Janeway CA.W. A. Peter J. 43. Paul WE. B. and Colombe. 2003. Walport M. The immune system: An introduction. L. Wiley-Blackwell Abbas AK. Marlene Rose. Shlomchik P.

Ileana Constantinescu 30 .. Handbook of Kindey Transplantation Fourth Edition. C. U: In the queue for a cadaver donor kidney transplant: new rules and concepts in the Eurotransplant International Foundation. Kidnei Int... 333-338. 62. Garland Publishing. W. Cherikh.ac. http://www.J. Lippincott Williams & Wilkins. 2002.. S. Human Immunology 56: 6-16. ASHI Quarterly. 2000. B. • Janeway. G. S. 2000.. 2005 • De Meester.. B. Gore. Bradley.. • Danovitch. Bresnahan.M.Prof.F...A. Microarrays: New tools for transplantation research. 1997.H.A. The role of HLA in renal transplantation.. Tolleris. A. in Histocompatibilitz Testing.3.. Pediatric Nephrology. Posttransplant renal function in the first year predicts long-term kindey transplant survival. W.Immunology .R. M.. Frei. Third Quarter. J.. Tissue Antigens. B.• Cecka.ebi. 104. • European Bioinformatics Institute. Travers P: Antigen recognition by T lymphocytes. Imperial Collage Press. 2003. • Hariharan. Course 1. Selection of Platelet Donors and Provision of HLA Matched Platelets. Persijn..S. Luminex. C. 311-318. F. New York and London. J. McBride.. G. 1988. J.G. Vendor Forum.. Claas. M. 2003. C. • Chua. Sarwal. C. Navarrete. W.M. 379 • Herczyk.A.. 18:319-327. Ed 3 Immunobiology 41-46. and Johnson..S. Matchability in kidney transplantation. Nephrology Dialysis Transplantation 15. 1997. M. P. • Harrison. 32:121-129.uk/imgt/hla • Gilks.M.

Reviews in Immunogenetisc. 2001. 2003. C. Hurley.D. L. Bios Scientific Publishers. 3. J. K. 2001. The Journal of Molecular Diagnostics.A. G. in Clinical Diagnosis and Management by Laboratory Methods. S. Katovich.. 1Ş105-123. Human Immunology 2001. P: Polymorphism and evolution of class I and class II genes and molecules. http://www. M. Human Leucocyte Antigen Gene Polymorphism and the Histocompatibility Laboratory. J.E. Transplantation 1997.. J. Twentieth Edition. 40. • Williams..L.M. D. S. • Wujciak. N. Immunogenetics 41. Academic Press..M. A. and Leone... J. • Moore. W.• Jerius. J. D.. • Rhodes.H. R. 3.213. S.: HLA antibody screening: Comparison of a solid phase enzyme-linked immunoassay with antiglobulin augmented lymphocytotoxicity.uk.. 2000...1-17. Parham. 62:3.. 64:1617./HGP/Chr6/MHC/shtml • Thelan. Luminex 100 User Manual Version 1. 1996..G. J. R. M. Journal of Urology.4:252 • Luminex Corporation..1995. 1998-2000. Matas. L. Rodey. Clin Lab Diagn Lab Immunol. Kremers. HLA Beyond Tears. J. 1:21-31.C. et. American Journal of Transplantation.: Detection of HLA class I specific antibodies by the QuikScreen Enzyme-Linked Immunosorbent Assay. • Lucas.. John Bernard Henry. T. 2000. Double renal transplants from marginal donors: 2-year results.A. Transplantation Proceedings. Barber. A. • Marsh. Eds). Evaluation of the GTI Quik-ID Assay and Analysis of Antibody Patterns.. P. D. Trowsdale. J. Characterization of HLA Class I Specific Antibodies by ELISA Using Solubilized Antigen Targets: I. 715-721. Inc. Ileana Constantinescu 31 . De Novo..Prof. Hartzman. 2000. 1999. A-M. M. P.. Taylor. Ploeger. Lucas. • Little. • Zachary. Matchabililty as an important factor for kidney allocation according to the HLA Match.sanger.B. S..A. G. Mart. Barber L. 163. Zachary. American Society of Histocompatibility and Immunogenetics Laboratory Manual. 3.P. In HLA and MHC: Genes. Human Leucocyte Antigen : The Major Histocompatibility Complex of Man. Improved scoring system to assess adult donors for cadaver renal transplantation. A. Wade.... Laffell. M.ac. • Nyberg. al. Reviews in Immunogenetics.: Genetics and molecular genetics of the MHC.... N. • Sanger Centre. Meyers.. • Trowsdale. D. Saunders. 927-946. T. 1999.P.D. Delaney. • Johnson. T.. edn. A. 1997.A. DeGoey.7 Rev B. D. 423-425..Immunology . Opelz..J. 228-235 Course 1. The HLA FactsBook. 27: 1403-1405...A.S: ASHY. • Trowsdale. J. Oxford. K. • Rodey Glenn E.. A.R.. Molecules and Function (Browning. Paparounis. And McMichael.. Parham.S. Murillo. 1997. Lenxa.

Cant Andrew J.Tsongalis Gregory J. 2006.Jorde Lynn B. . .Wood Peter : Understanding Immunology.Forsythe John L. Coleman William B :Molecular Diagnostics a training and study guide. Editura Universitara "Carol Davila". Mosby Elsevier.Ileana Constantinescu : Imunologia transplantului.Inc.Carey John C.R :Transplantation. Introduction to human histocompatibility. second edition. Jackson Graham :Practical Hematopoietic Stem Cell Transplantation. Prof. .. Bamshad Michael J : Medical Genetics. 2009. . Galloway Angela. 2009. AACC Press. . Leor Jonathan. De Novo. Fourth Edition. Blackwell Publishing Ltd.Battler Alexander. .CO.2010 Curs 1: Imunologie.Verlag London Limited. second edition. Bucuresti. 2000. Eds. Ileana Constantinescu 32 . Saunders Elsevier.2002.Rodey Glenn E : HLA beyond tears. 2007 . Stem Cell and Gene-Based Therapy.Durango.Springer . Frontiers in Regenerative Medicine.

destination and structure of cells and tissues of the immune system • • • • • • • • • • • • • • • • Bone Marrow Stem Cells Erythropoiesis Granulolcytopoiesis Eosinophils Basophils Neutrophls Lymphopoiesis Natural Killer Cells Monocytes Dendritic Cells Thymus Lymph Nodes Spleen Mucosal Associated Lymphoid Tissue (MALT) Circulation and Recirculation Course 2: Immunology.Course 2: Origin. Ileana Constantinescu 33 . Prof.

Prof. • It also provides a microenvironment necessary for B lymphocyte maturation and formation of pre-T cells (lymphopoiesis).BONE MARROW • The bone marrow constitutes almost 5% of total body weight and is responsible for the formation of all blood (hemopoiesis) in adults. Course 2: Immunology. Ileana Constantinescu 34 .

Prof. granulocytes.STEM CELLS • All of these cells originate from undifferentiated pluripotential hemopoietic stem cells (PHSC). monocytes. When SCs divide. SCs have limited proliferative capacity and exhibit the potential to differentiate into all cells of the myeloid (erythrocytes. the two daughter cells (DCs) can either continue as SCs or proceed into any of the differentiation pathways Course 2: Immunology.1 % of all cells in adult marrow. which can be found first in the mammalian embryo within the liver then spleen. and platelets) or lymphoid (T and B cells) lineage. Ileana Constantinescu 35 . • SCs are pluripotential CD34+ and represent less than 0.

hematopoiesis Course 2: Immunology. Ileana Constantinescu 36 . Prof.Cells of the immune system .

Cells and growth factors in hemopoiesis Course 2: Immunology. Ileana Constantinescu 37 . Prof.

The precursor cell gradually loses its ability to self-renew and then divide as it approaches its mature phenotype. Ileana Constantinescu 38 . Some of these growth factors such as stem cell factor (SCF) and interleukin (IL-7) are secreted by marrow stromal cells. • SCs not only depend on a specific microenvironment for their maturation. Prof. but most others are produced elsewhere and transported to the marrow by the blood stream (endocrine). it becomes a dividing multipotential colony forming unit-spleen (CFU-S) or CFU (colony forming unit-lymphoid )(CFU-Ly) progenitor cell and then a progressively maturing unipotential precursor cell for any of the mature cells of peripheral blood.• If a DC (daughter cell) proceeds into the differentiation pathway. Course 2: Immunology. they also depend on numerous glycoprotein growth factors that act at different stages to control the cell type as well as rate of cell formation.

ERYTHROPOIESIS • Erythropoiesis occurs when a CFU-S cell gives rise to burst-forming units erythrocyte (BFU-E) and then colony-forming units-erythrocyte (CFU-E) all under the influence of erythropoietin (EPO) from the kidney cortex as well as IL-3 and granulocyte-macrophage colony-stimulating factor (GMCSF). Prof. Ileana Constantinescu 39 . Course 2: Immunology. • CFU-E form the precursor of erythrocytes.5 X 10 11 erythrocytes are produced each day. • These maturation events occur around macrophages. the proerythroblast. During this sequence. the nucleus is condensed and then lost as the cell becomes 6-7 micro. 2. which phagocytose the extruded nuclei as well as provide some growth factors. Under normal conditions. Proerythro blast —► basophilic erythroblast —► polychromatophilic erythroblast —> ortho-chromatophilic erythroblast —» reticulocyte —» erythrocyte is the sequence of events in maturation.

Course 2: Immunology. Prof. • Mature cells are named because of staining of their specific granules. -BASO or CFU-NM -+ CFU-N -* myeloblast -* promyelocyte —> myelocyte -* metamyelocyte -H» Stab -* mature cell.GRANULOCYTOPOIESIS • Granulocytopoiesis is the generation of three types of granulocytes (neutrophils. Ileana Constantinescu 40 . eosinophils. basophils) from CFU-S. All derive from CFU-S (spleen) —> CFU-Eo.

IL-3. and IL-5 for their maturation. Prof. Their specific granules contain a number of factors including histaminase. phagocytosing antigenantibody complexes. acid phosphatase. They are important in destroying parasites. and major basic protein. They represent 2 to 4% of peripheral blood white cells. Ileana Constantinescu 41 . Course 2: Immunology.Eosinophils • Eosinophils require GM-CSF. and combating histamine levels during allergic reactions.

Ileana Constantinescu 42 . They represent <1% of peripheral blood white cells. IL-3. and chemotactic factors for eosinophils and neutrophils.Basophils • Basophils require GM-CSF. Course 2: Immunology. and IL-4 for their maturation. heparin. Prof. Their specific granules contain histamine. • They are similar to mast cells in that they participate in IgE-mediated immediate hypersensitivity responses.

the CFU-S gives rise to a CFUNM cell. Ileana Constantinescu 43 . lysozyme. Course 2: Immunology. • Their specific granules contain alkaline phosphatase. IL-3. lactoferrin. can differentiate into a CFU-Neut or CFU-M cell. depending on the level of granulocyte or macrophage colony-stimulating factor in the environment. Prof. Unlike eosinophils and basophils. and granulocyte colonystimulating factor (G-CSF) for their maturation. 60% to 70% of all white cells are neutrophils.Neutrophils • Neutrophils require GM-CSF. They are the earliest phagocytic cells to appear in a bacterial infection and are a prominent constituent of cellular immune response. phagocytic and type IV collagenase. which.

• Thus. gut-associated lymphoid tissue. etc. and lymph nodes. Prof.LYMPHOPOIESIS • Lymphopoiesis is the ability of CFU-Ly to form either CFU-LyB or CFU-LyT in the bone marrow. • CFU-LyB cells develop in the presence of IL-7 and IL-3 in the marrow to immunocompetent B cells with slgM. spleen. the bone marrow and thymus are considered primary lymphoid tissue. Course 2: Immunology. Ileana Constantinescu 44 . CFU-Ly T cells become pre-T cells. • Further maturation of these cells occurs in germinal centers of secondary lymphoid tissue. which leave the bone marrow and find their way to the subcapsular area of the thymic cortex where they proliferate and mature in the deeper thymic cortex into either CD3 + 4+ or CD3 + 8+ cells. are considered secondary lymphoid tissue.

NATURAL KILLER CELLS
• Natural killer (NK) cells also develop within the
marrow and then are found in peripheral
blood, liver sinusoids (pit cells), and spleen
sinusoids but not thoracic lymph. duct.
• NK cells appear early in bacterial infections,
can secrete interferon-y, and spontaneously
kill some viral infected cells and tumor cells.

They respond to IL-2 and develop
independently of the thymus.
Course 2: Immunology, Prof. Ileana Constantinescu

45

MONOCYTES
• Monocytes develop from CFU-M progenitors under the influence of GM-CSF,
IL-3, and G-CSF. They represent 3 to 8% of leukocytes in peripheral blood.
• Monocytes have numerous azurophilic granules (lysosomes) in their
cytoplasm and can readily leave the circulation to give rise to macrophages
within almost every organ. They are thus the source of the mononuclear
phagocytic system throughout the body.
• They are even more phagocytic than neutrophils or eosinophils in that they
can degrade larger bacteria within phagosomes via the formation of
hydrogen peroxide, hypochlorous acid, and superoxide.
• They can also fuse to form foreign body giant cells in response to a large
antigenic load. Macrophages express class II major histocompatibility
complex and can function as antigen-processing and -presenting cells. They
secrete the pro-inflammatory cytokines IL-1, IL-6, and tumor necrosis factor a
(TNFα) upon activation.
Course 2: Immunology, Prof. Ileana Constantinescu

46

DENDRITIC CELLS
• Dendritic cells are bone marrow-derived cells specialized
for presenting antigens to either CD4 + or CD8+ T cells in
order to initiate a primary immune response.
• These cells appear to be generated from CD34+ cells and
require GM-CSF, TNF alpha, and possibily TGF beta for
full maturation. Dendritic cells are found in all lymphoid
tissues where they have specific names. Because of their
highly efficient function in antigen presentation, they
have come to be called "nature's adjuvant“.

Course 2: Immunology, Prof. Ileana Constantinescu

47

Prof. Ileana Constantinescu 48 .Course 2: Immunology.

and dendritic cells have developed in the marrow and CD3+4+ and CD3+ 8+ cells have matured in the thymus. Course 2: Immunology. • After slg+ B cells. with the medulla containing mainly mature cells. • Most of the deletion of cells occurs in the cortex. pre-T cells. they leave these primary lymphoid organs and migrate into peripheral secondary lymphoid tissue: lymph nodes spleen and mucosal associated lymphoid tissue (MALT). Ileana Constantinescu 49 . • The stroma of the thymus is composed of epithelial cells that originate from the third pharyngeal pouch in the embryo. The parenchyma is made up of thymocytes that are either positively and then negatively selected for maturation (<1%) or deleted (>99%) by apoptosis. Prof.THYMUS • The thymus is a primary lymphoid organ that is required for the generation of immunocompetent CD3+4+ and CD3 + 8+ cells necessary for an adaptive immune response . NK cells. monocytes.

LYMPH NODES • Lymph nodes (LNs) are encapsulated structures that are strategically placed throughout the body to receive and filter antigens and cells from peripheral interstitial fluid and lymph. Their indented hilus contains blood and lymphatic vessels. Course 2: Immunology. Prof. lymph nodes in the body. 1-10 mm in diameter. Ileana Constantinescu 50 . • All LNs eventually drain into the thoracic duct system and back to the peripheral blood. • There are approximately 550 kidney-shaped.

Ileana Constantinescu 51 .Course 2: Immunology. Prof.

Prof. Ileana Constantinescu 52 .Structure of a lymph node Course 2: Immunology.

These cells are localized into: • SUPERFICIAL CORTEX: containing primary and secondary follicles with germinal centers containing dark and light zones. Prof. lymph and blood.• Thus. which permeate a stroma of largely type III and I collagen. Course 2: Immunology. forming lymphatic sinuses supporting a parenchyma of immunocompetent T and B cells as well as macrophages and other antigen-presenting cells. Ileana Constantinescu 53 . The type III collagen is arranged in a chicken wire fashion. • MEDULLA: with medullary cords. • DEEP CORTEX: containing high endothelial post-capillary venules. LNs have two circulations.

They then decrease their endothelial height. Course 2: Immunology. Any cells that enter through afferent lymphatics can also enter the parenchyma of the LN. where it leaves the hilus via efferent lymphatics. They can remain in the LN for an undetermined period or enter a lymph sinus and exit via an efferent lymphatic. • Most B cells migrate superficially into follicles. We will develop further the structure of the LN during an immune response. It then percolates through the cortex into the medullary sinuses. • The hilar artery supplies the medulla and deep cortex but then empties into a capillary net on either side of the superficial cortical follicles. Prof. These capillaries then drain into post capillary venules (PCVs) with a very high cuboidal endothelium in the deep cortex.• Lymph enters a Lymph nod (LN) at its surface as numerous afferent lymphatics pierce its type I collagenous capsule and empty into the subcapsular sinus. and leave the LN as hilar veins. Ileana Constantinescu 54 . with most of the T cells remaining in the deep cortex. pass through the medulla. It is at the PCVs (post capillary senules) in the deep cortex where circulating T and B cells adhere to PCV endothelium and migrate between these cells (diapedesis) into the node.

which can respond to blood-borne antigens surrounded by red pulp (RP). • Similar to LN. Ileana Constantinescu 55 .SPLEEN • The spleen is an encapsulated organ with lymphoid tissue called white pulp (WP). More lymphocytes circulate through it each day than any other secondary lymphoid organ. Prof. the spleen has a stroma of type III and I (capsule and trabecular) collagen that supports the parenchyma of WP and a blood sinusoidal system with cords of hemopoietic and lymphoid tissue called RP separated from each other by a marginal zone. Course 2: Immunology.

Structure of spleen white pulp with perarteriolar lymphoid sheath (PALS) Course 2: Immunology. Prof. Ileana Constantinescu 56 .

respectively. The periphery of WP contains B cellrich lymphoid follicles just like the periphery of LN. The marginal zone and sinus function as a part of B and T cell entry points into the WP similar to high endothelial venules of LN. Some RP cords contain granulocytes. B and T cells can migrate out of the WP into an RP cord through bridging channels (BCs) in the marginal zone. Between the periphery of the WP and the marginal zone is a blood filled marginal sinus. but the marginal sinus has a low endothelium. Course 2: Immunology.• The part of the WP around a central arteriole is called the periarteriolar lymphoid sheath (PALS) and contains primarily T cells. The sinusoidal macrophages destroy old platelets and erythrocytes. Prof. • The marginal zone contains macrophages and some memory B cells. which receives capillary branches from the central arteriole. The central arterioles empty directly into the RP cord or sinuses and then into trabecular veins. Ileana Constantinescu 57 . It is thus analogous to the deep cortex of LN. T and B cells leave the splenic circulation here and then migrate to the PALS or follicles. • The RP contains blood sinusoids with associated macrophages that line cellular cords. but others connect with WP follicles and contain plasma cells.

or aggregates of secondary nodules that can become encapsulated. Thus. diffuse aggregates. solitary primary nodules. and submucosa of the gastrointestinal. most intraepithelial lymphocytes (IEL) are CD8+ αβ (60%) or γδ(40%) T cells. Course 2: Immunology. Prof. • While most of the diffuse and aggregated lymphoid tissue of the MALT is responsible for producing IgA. which transport luminal antigen to under lying antigen reactive lymphocytes. It appears that the γδ cells may react to bacterial antigens. The epithelial lining over Peyer's patches (PP) contains specialized antigen transporting cells called M cells. After re-entering the circulation. and genitourinary tracts contain wandering cells. • It should also be realized that most of the cells activated in the MALT stay in or return to the MALT. All of these are collectively called mucosal associated lymphoid tissue (MALT). respiratory. Ileana Constantinescu 58 .MUCOSAL ASSOCIATED LYMPHOID TISSUE (MALT) • The epithelial lining. they home back to the MALT. lamina propria. cells stimulated in PP (Peyper’s patches) or tonsils enter mesenteric or cervical lymph nodes for further maturation.

Prof.Course 2: Immunology. Ileana Constantinescu 59 .

Approximately 1-2% recirculate every hour. blood —> lymph —> blood. spleen. NK cells are only found in blood. Ileana Constantinescu 60 . and liver.CIRCULATION AND RECIRCULATION Both T and B cells continuously circulate and recirculate. Prof. Course 2: Immunology. However.

Ileana Constantinescu 61 .Prof.Course 3: The Antigen • • • • • • • • Definition Immunogenicity Specificity Antigen determinants Superantigens Structure and functions Immunization Vaccination Course 3 .Immunology .

Immunology . it the target of a humoral or cell mediated immune response.Prof. Original antigenic sin: tendency to react to immunization by producing antibodies to a determinant on the stimulating antigen that resembles a determinant on an antigen encountered previously. See antigenic drift. antigenic shift. In the interval between infections. Course 3 .• • • • • • • • • • Definition Antigen: substance that can elicit an immune response and that can react specifically with the corresponding antibodies or T cell receptors. the influenza virus has undergone antigenic change. Ileana Constantinescu 62 . Called also self antigen. Antigenic drift: minor change in a surface antigen of a pathogenic microorganism. Autoantigen: an antigen that. as in autoimmune disease. Influenza A virus antigenic drift is the result of mutation in the hemagglobulin and/or neuraminidase genes. despite being a normal tissue constituent. The term was first used to describe the response to reinfection with influenza when the host markes a secondary immune response to the strain of influenza of a previous infection. Antigenic shift: abrupt major change in a surface antigen of a pathogenic microorganism. An antigen may contain determinants. Infection virus type A: the major antigenic variants are called subtypes antigenic variation. Ex. A protein many have sequential and/or conformational determinants. The antigenic variants emerge by selection of mutants that are less susceptible to neutralization by the prevailing antibodies in the infected host. Antigenic determinant or epitope: portion of an antigen that marks contact with a particular antibody or T cell receptor. Antibodies that cross-react with the original antigen are produced preferentially.

• An antigen binds to a specific antibody to provide a mechanism by which the antigen is recognized and inactivated. • The binding process of antigen to specific antibody forms the basis of a test called an enzyme-linked immunosorbent assay (ELISA) that can be used to measure the amount of antigen in a biological fluid. Ileana Constantinescu 63 .Immunology . an antigen complexes with a specific antibody so that the complex can attach itself to specialized immune cells that either internalize the complex to destroy it or release biologic mediators such as histamine to induce an allergic/inflammatory response.     Course 3 . the greater the amount of antigen present in the assayed sample).Prof. The color intensity of the solution is indicative of the amount of antigen in the fluid sample (the darker the color. This antigen binds to the coating antibody and is detected by the addition of another antibody that specifically recognizes another specific part of the antigen that is bound to the coating antibody. In this manner. the binding of an antigen to a specific antibody provides the physician with a means to measure the concentration of a specific antigen in biological fluids. Thus. • As shown in a sample of biological fluid such as blood serum is placed into a solid support vesicle that has been coated with an antibody that specifically recognizes a specific part of some antigen found in blood. ANTIGEN: An antigen is a molecule that binds to a specific protein structure called antibody. • This detecting antibody is linked to an enzyme that specifically reacts with a substrate that is then added to the reaction mixture to yield a colored solution.

Ileana Constantinescu 64 .Schematic of the enzyme-linked immunosorbent assay (ELISA).Immunology .Prof. Course 3 .

.Prof. hepatitis B surface a. but anti-HBc antibodies appear during the acute infection.g. that is. CD2. (HBeAg). only the portion of the protein or polysaccharide molecule known as the antigenic determinant (q.Definitions and comments • antigen (an'ti-jon) [antibody + Gr. any of a number of cell surface markers expressed by leukocytes and used to distinguish cell lineages. they do not protect against reinfection. such as bacteria and tissue cells. Antigens may be soluble substances. under appropriate conditions. isoantigen. a core protein antigen of hepatitis B virus present in the blood in some infected individuals. Anti-HBe antibodies appear transiently during convalescence. however. Abbreviated Ag. • blood-group a's. Originally called Australia (An) antigen because it was first found in an Australian aborigine. one occurring in some but not all individuals of the same species. • 65 Course 3 . of inducing a specific immune response and of reacting with the products of that response. CD3.Immunology . also formerly called hepatitis-associated a.. • hepatitis B e a. such markers can be identified by specific monoclonal antibodies and are numbered CD1. Ileana Constantinescu . a coat protein antigen of the hepatitis B virus present on complete virions (Dane particles) and smaller spherical and filamentous particles circulating in the blood of individuals with active or chronic infections. • hepatitis B core a. • allogeneic a. erythrocyte surface antigens whose antigenic differences determine blood groups. developmental stages. (HBcAg). and functional subsets. the antigen is not present in the blood of infected individuals. such as toxins and foreign proteins.v. Anti-HBs antibodies appear in the blood in late convalescence and are protective against reinfection. a core protein antigen of the hepatitis B virus present inside complete virions (Dane particles) and in free core particles in the nuclei of infected cells. with specific antibody or specifically sensitized T-lymphocytes. gennan to produce] any substance which is capable. (HBsAg). they do not protect against reinfection.) combines with antibody or a specific receptor on a lymphocyte. • CD a. histocompatibility antigens and human blood group antigens. (HAA) and serum-hepatitis (SH) a.. e. etc. or particulate. or both.

Course 3 .Immunology . Ileana Constantinescu 66 .Prof.

Immunology .Course 3 .Prof. Ileana Constantinescu 67 .

systems of allelic alloantigens that can cause transplant rejection. 2.. antigens of low-frequency blood groups. a region on the short arm of chromosome 6 containing several genetic loci. 1.Definitions and comments • heterologous a. Called also transplantation antigens. HLA antigens found only on the gene product of a single allele. -DR. platelet a. 1. major. private a's. each having multiple alleles. -C. -DP.. about 15-30 such systems have been found in mice. Ileana Constantinescu .. 1. 3. • histocompatibility a's. so cal because they occur in only a few kindreds. HLA antigens occurring on the products of several allelic gene. a tumor antigen expressed only on a particular type of chemically induced tumor public a's public a's. Forssman antigen. so called because they occur in the general population at high frequencies. I table at platelet group. isoantigen. systems of allelic alloantigens that can stimulate an immune response that leads to transplant rejection when the donor and recipient are mismatched. • • • •   homologous a.. HLA a's. the antigen that induces the formation of an antibody. but with a long delay (up to 100 days). histocompatibility antigens governed by genes of the HLA complex (the human major histocompatibility complex). e. heterophile a. HLA-A. -B. antigens of high-frequency blood groups. HLA antigens in humans and H-2 antigens in mice • histocompatibility a's. 2. any of a group of cross-reacting antigens occurring in several species and having a species distribution that does not correspond to phylogenetic relationships. -DQ. 2. 68 Course 3 . (Human Leukocyte Antigens). • histocompatibility a's. • heterophil a. those in the major histocompatibility complex. any of several isoantigens expressed by platelets..Immunology . an antigen that reacts with an antibody that is not the one (the homologous antigen) that induced its formation.g.Prof. Loci are designated by letters. Called also heterogenetic a. minor..

Ileana Constantinescu 69 . stomach.v. See lipopolysaccharide. mol. a glycoprotein. q. a serine endopeptidase secreted by epithelial cells of the prostate gland. alpha-fetoproten carcinoembryonic antigen. Cf. and pancreatic oncofetal antigen.]. any antigen that occurs exclusively in a particul organ and serves to distinguish it from other organs. (PSA).the lipopolysaccharide-protein somatic ant gens of gram-negative bacteria. • O a. pancreas. lung. wt. Ha. Measurement of PSA serum levels is sometimes used as a screening test for prostate cancer. and breast. an antigenic gene product that is expressed durin fetal development. e. (2) second-order organ specificity is attributed to an antigen characteristic of the same organ many.. • Prostate-specific a.()00. The primary use of CEA is in monitoring response to treatment of colorectal cancer. • oncofetal a. important in the serological classifcation of enteric bacilli..Definitions and comments • nuclear a's. mol. inflammatory bowel disease. it also occurs at lower levels in the serum of patients with cancer at other sites and some normal adults.. oncofetal antigens. 2()(). rectal polyps. wt.g. Course 3 . secreted into the glycocalyx coating the luminal surface of gastrointestinal epithelia. and cigarette smoking. and derepressed in some tissues that have undergone neoplastic transformation. Originally thought to be a specific antigen of the fetal digestive tract and adenocarcinoma of the colon. 800. • (CEA). (POA). [ohne Hauch. • pancreatic oncofetal a. Two types organ specificity have been proposed: (1) first-order or tissue speificity is attributed to the presence of an antigen characteristic of particular organ in a single species.0 found in fetal and neoplastic pancreatic tissue but not in that normal adults. alcoholic cirrhosis and pancreatitis. CEA is now known to occur normally in feces and pancreaticobiliary secretions and to appear in the plasma in a diverse group of neoplastic and non-neoplastic conditions including cancers of the colon.Prof. • organ-specific a.Immunology . the components of cell nuclei with which antinuclear antibodies (see under antibody) react. even unrelated species. are thus useful tumor markers. partially or completely repressed in adult tisssues. a glycoprotein. serum levels are elevate benign prostatic hyperplasia and prostate cancer.

Therefore. The smallest individual entity that can be detected by a receptor is termed an epitope. the antibody response against a microbe can consist of many different immunoglobulins recognizing different specific sites on the microbe.Prof.Immunology .Epitopes. Invasive microbes express many antigenic molecules that can be individually identified by different antigen receptors (immunoglobulins). Course 3 . An individual molecule can even consist of smaller parts that are specifically recognized by different immunoglobulins. Ileana Constantinescu 70 .

and antibodies will generally stimulate the strongest overall immune responses (they have greater immunogenicity).Immunogenicity. Antigenicity: ability of an antigen to: 1.Immunology . 71 Course 3 . Ileana Constantinescu . induce an immune response 2. combine with specific antibodies and/or cell receptors.Prof. Antigens that are more complex in structure tend to have more binding sites and express a greater variety of different epitopes. Antigens that can be recognized by the greatest range of available T and B cell receptors.

Antigenemia – the presence of antigen in the blood.Specificity • Ability of antibodies or lymphocytes to discriminate among different - ligands. - Antigenic – having the properties of an antigen - Antigenicity – the property of being able to induce a specific immune response or the degree to which a substance is able to stimulate an immune response.Immunology .Prof. - Antigenemic – exhibiting antigenemia. Called also immunogenicity. Ileana Constantinescu 72 . Course 3 .

Immunology .Binding affinity of antibody Course 3 .Prof. Ileana Constantinescu 73 .

be a target of an immune response Subsequent exposures to the same immunogen generally lead to heightened secondary responses by the adaptive immune system. both stimulate an immune response and.Immunogens. and tolerogens.Prof. and tolerogens • There are three types of antigens: immunogens. by themselves can both stimulate immune responses and serve as targets of those responses. subsequent exposure of the immunline system to the same tolerogen leads to diminished responsiveness against it. • Unlike immunogens. haptens.Immunology . • Immunogens are molecules that can. • Haptens are small molecules that. Ileana Constantinescu 74 . haptens. by themselves. Course 3 . in turn.

by themselves.Immunogens.Immunology .Prof. Haptens cannot stimulate responses on their own. Reexposure to immunogens and haptens usually generate heightened responses. Ileana Constantinescu 75 . can both stimulate and serve as targets of immune responses. but can serve as targets of responses generated if the hapten is attached to an immunogen. Reexposures to tolerogens usually result in diminished responses. Immunogens and tolerogens. haptens and tolerogens. Course 3 .

a special category of antigens. • The binding of superantigens to relatively nonpolymorphic regions of MHC and TCR molecules promotes adherence of T cells to antigen-presenting cells. as well as to MHC class II molecules outside of the peptide presentation groove.SUPERANTIGENS • Superantigens are certain bacterial and viral glycoproteins that bind TCR and MHC class II antigens outside of the conventional groove for antigenic peptide binding. irrespective of TCR specificity. • While a single peptide antigen may be recognized by or is immunogenic for a small number of T cell clones. Such cross-linking of TCR with MHC molecules leads to the activation of multiple clones of T cells.Immunology . leading to nonspecific activation of multiple T cell clones. have the capacity to stimulate multiple T cell clones. known as superantigens. Superantigens.Prof. which have been identified thus far as bacterial and viral glycoproteins. are super-stimulators of T cells because they are capable of binding to a large number of T cell receptor Vβ sequences. Ileana Constantinescu 76 . especially CD4+ cells. Course 3 .

Prof.Immunology . Ileana Constantinescu 77 .Course 3 .

such as penicilloic acid. • In order to be immunogenic.Prof. Course 3 . penicillin must first become a chemically reactive molecule. that is able to covalently attach itself to an endogenous carrier protein. it cannot activate immune cells to elicit an immune response to produce those specific antibodies.Immunology . an immunogen is an antigen. In this manner. Thus. the antigen. • By itself. Ileana Constantinescu 78 . but it is not immunogenic. but an antigen is not necessarily an immunogen. such as serum albumin or other cell surface membrane proteins.An immunogen is an antigen that activates immune cells to generate an immune response against itself. penicillin is an antigen that can bind to a specific antibody. the penicilloic acid-protein molecule becomes an immunogen that is capable of activating both T and B lymphocytes to elicit the production of antibodies by B cells that are able to specifically bind to penicillin.

Course 3 .Penicilloic acid-protein conjugate Penicilloic acid groups can be readily conjugated to proteins.Immunology . Conjugation usually takes place to the amino groups of lysine.Prof. Ileana Constantinescu 79 .

  Other  examples  of  haptens  include  nucleic  acids. This approach is used successfully to prepare vaccine against Haemophilus influenzae type b.. Course 3 .  a  specific  antibody can be produced that either binds strongly to that specific hapten alone. As a result. Ileana Constantinescu . and not  to related haptens.Prof. meningitis due to Haemophilus b. CLINICAL APPLICATION OF HAPTEN CONJUGATION Coupling of haptens to a protein carrier improves the immunogenicity of the hapten. researchers have  synthesized  small  organic  molecules  whose  structures  can  be  specifically  modified  at  specific sites. benzene rings with either nitroso or arsonate groups at different positions in  the  ring  possess  only  one  site  for  the  binding  of  an  antibody  and  are  referred  to  as  haptens.  and  small  molecular  80 weight molecules such as penicillin. which was the most common cause of meningitis in children. and weakly to related haptens. The latter finding provides a mechanistic basis for the ability of an antibody to bind other  molecules  that  closely  resemble  a  particular  antigenic  structure  (a  cross-reacting  antibody). • • • • In order to study the specificity of antigen binding by antibody molecules. Using  a  specific  hapten  coupled  to  an  immunogenic  protein  carrier.Immunology . or binds strongly to that specific hapten.  For example.  phospholipids. HAPTEN: A hapten is a small organic molecule that is not immunogenic by itself. has been virtually eliminated.

This risk has increased dramatically in recent years. may initially complex with host proteins and elicit the initial production of antibodies against the hapten. Course 3 . However. When these haptens gain entry into the host. since humans are exposed to a ever increasing number of haptens in their environment. In this manner.Immunology . The other reaction is pathological and involves the binding of antigen to specific IgE antibody that is attached to mast cells. two separate reactions can occur. One reaction is protective and involves the production of antibodies that bind to the hapten and clear it from the circulation. haptens pose a serious health risk to humans. haptens are small nonimmunogenic antigens that can be made into immunogens that can elicit either clinical or pathological immune reactions. precipitating histamine release and an allergic response. upon reexposure to nickel. Ileana Constantinescu 81 . such as nickel. an environmental hapten. they may complex with host proteins to create immunogemic haptenprotein molecules. Thus.COMMENTS • • • • Outside of the laboratory.Prof.

intermediate doses of antigen elicit strong immune responses that provide protection to the HOST against the antigen. Course 3 . In contrast. In contrast. However. Most antigens require several administrations to achieve optimal immunogenicity.Prof. but they probably involve the activation of antigen-presenting cells. frequency of antigen administration. Ileana Constantinescu 82 . Adjuvants. help to augment the immune response that is generated against a particular antigen. The frequency of antigen administration also determines the magnitude of the immune response that is generated against an antigen. soluble antigens. such as proteins. for efficient antigen uptake and processing for the presentation of degraded immunogenic peptides to T cells. the immunogenicity of soluble antigens can be augmented when they are administered in conjunction with compounds known as adjuvants. parenterally administered antigens may also induce a state of tolerance to the antigen if it should gain entry to the internal environment a second time via another route in an appropriate dose. A second administration of the antigen elicits a secondary response that is of a higher magnitude as compared with the primary response. The mechanisms responsible for the action of adjuvants are not well understood at present. A third administration of the antigen elicits an even greater immune and finally. Also. such as killed mycrobacteria or Bordetella pertussis. This latter phenomenon may be related to the finding that very low or very high doses of antigen are not able to elicit a strong immune response and may actually induce a state of immune nonresponsiveness/tolerance in the host. are more immunogenic when they are administered intravenously. nucleic acids and carbohydrates. are more immunogenic when administered subcutaneously.Immunology . and use of adjuvant. For ex. antigens given orally (parenterally) may elicit immune responses in the gut that allow for the production of antibodies that protect us from the antigen crossing the gut lining and entering the internal compartments of the body.COMMENTS • • • • • • • • Other factors that determine the degree of immunogenicity of antigen include the route of antigen administration. For example. antigen dose. particulate antigens. a primary administration of antigen elicits a primary immune response of a given magnitude. such as bacteria and viruses. such as macrophages.

Ileana Constantinescu 83 .Immunology .Course 3 .Prof.

cells can sample not only the general characteristics of their environment. but also the nature of the cells and molecules with which they come into contact. Initial protection against infection is provided by mechanical barriers (such as the skin and mucous membranes) and by chemical barriers (such as microbial molecules secreted by some skin cells) that can quite effectively bar the entry of microbes in the body. One is an ancient system of self-defense—the innate immune system (sometimes called the "nonspecific" immune system). The human immune system is complex.Immunology . cell types. We use two immune systems to combat invasion by infectious organisms. and molecules that must work together. The other is the antigen-specific adaptive immune system (sometimes called the "specific" immune system).g. a normal cell becoming cancerous). Nonself may enter the body from the outside or represent an unacceptable change within the body (e. Course 3 . Using a variety of surface receptors. Ileana Constantinescu 84 .Prof. composed of multiple organs.Summary • • • • • The immune system must be able to distinguish self—that which belongs within the body—from nonself..

but can be recognized by the immune system if bound to an immunogenic molecule. (2) are potentially reactive against self. The process in which a surface receptor transfers information into the cytoplasm and often to the nucleus is known as signal transduction. in turn. carbohydrates. be a target of an immune response. Ileana Constantinescu 85 . lipids. This process involves a series of steps that activate sequential sets of enzymes that can modify their substrate molecules to become biologically active. Subsequent exposures usually elicit heightened responses against that specific immunogen. A given antigen may provoke different responses in different individuals. cannot stimulate an immune response. The smallest identifiable part of an antigenic molecule that can be recognized by a T or B cell receptor is called an epitope or determinant. Tolerogens are molecules that. can both stimulate immune responses and serve as targets of those responses. but subsequent exposures result in diminished response to that specific tolerogen.Immunology . or (3) that will (in the case of T lymphocytes) be unable to function in the body.• • • • • • • • An antigen is a structure that can be recognized and bound by the specific receptors of T and B lymphocytes. like immunogens. both stimulate an immune response and. Antigens may be proteins. by themselves. The adaptive immune response is initially random and results in T and B cell receptors that (1) appropriately identify nonself.Prof. Haptens are small molecules that. by themselves. Course 3 . or combinations such as glycoproteins or glycolipids. Immunogens are antigenic molecules that can.

Prof. even as many others around them might be dying. for thousands of years. Dr.PREVENTION OF INFECTION: VACCINATION • Humans have known of vaccination. Ileana Constantinescu 86 . Those who survived diseases such as plague. if not its mechanisms. • Not only was their survival a blessing to the survivors but also provided society with a set of individuals able to aid others and perform critical activities in the face of subsequent epidemics. smallpox.Immunology . and other epidemic diseases were generally sale from the same diseases developing again. • Vaccination is an inoculation of non-virulent or inactivated microbes as a means of inducing specific immunity. Course 3 .

the discovery by Robert Koch of the role of specific microbes in specific diseases stimulated activity in the field. Since that time. a virus that normally infects cattle) conferred protection against smallpox. has in turn been practically eliminated and the smallpox virus now exists only in a few protected laboratories. •In 1794. At the time. Routine childhood vaccinations have eliminated much of the misery and occasional permanent crippling or fatal consequences of once commonplace diseases such as measles. a disease that once killed humans by the thousands. vaccination has revolutionized human and animal health.History of vaccination •The ancient Egyptians and Chinese performed forms of vaccination to attempt to protect themselves against diseases such as smallpox by exposing individuals to powders formed from the crusts and scales of pockmarks on infected individuals.Immunology . a potentially fatal human disease caused by a related but more virulent form of vaccinia. but those individuals found themselves protected. diphtheria. Ileana Constantinescu 87 . Dr. •Smallpox.Prof. Edward Jenner demonstrated that intentional inoculation with cowpox (a mild disease in humans caused by a form of vaccinia. Later. of course. Course 3 . Sometimes mild forms of the disease developed in individuals so treated. Jenner called his procedure vaccination. or sometimes no visible disease developed at all. and the word was later adapted to name the viral organism found lo be the causative agent. Louis Pasteur advanced the science of vaccination by developing effective vaccines against epidemic diseases of agricultural animals and eventually performed a dramatic demonstration of a rabies vaccine that saved the life of a young boy bitten by a rabid dog. there was no knowledge of the role of microbes in causing such diseases. and polio.

fatigue. however. because there is no known effective treatment.Immunology . and vomiting.Prof. Approximately 2 to 3 days after the onset of the symptoms. in an 1806 letter to Edward Jenner. a rash appears. Early symptoms are similar to those of influenza and include high fever. no case of small pox has been reported since 1977. Ileana Constantinescu 88 . with death usually occurring during the second week of illness.CLINICAL APPLICATION SMALLPOX • Smallpox is a highly contagious disease spreading from person to person. In the United States. Course 3 . routine childhood vaccination for small pox ended in 1972. Treatment for smallpox is mainly supportive. The rash is initially seen on the face and oropharynx. legs. Vaccination within 4 days of exposure to smallpox can prevent a fatal outcome and can reduce the severity of the illness. wrote that. "Future nations will know by history only that the loathsome smallpox has existed and by you has been extirpated. Dr. • Thomas Jefferson. it then spreads to the upper arms. current concern about the potential use of smallpox as a weapon of bioterrorism. muscle aches. often by saliva droplets from the affected person's mouth. There is. • The mortality rate is approximately 30%." Because of a successful worldwide vaccination program. headache. and trunk.

Ileana Constantinescu 89 . Dr.Prof.• The standard vaccination schedule in USA Course 3 .Immunology .

although rare. So long as a sufficiently large population is vaccinated. the infection could spread rapidly among them.• The beneficial impact of vaccination has been so widespread dial we now face the situation in which many people no longer recognize the dangers that have been overcome. and some individuals advocate the elimination of vaccinations because they believe them to be responsible for several.Immunology . the danger exists that a pool of unprotected individuals may be created that could once again be subject lo main of the infectious diseases dial once terrorized human populations. Course 3 . As a result. This concept is called herd immunity. The inherent risk is dial if an infectious organism infects a significant number of unprotected individuals. Dr. • It is probably not necessary dial every individual in a population be vaccinated. Ileana Constantinescu 90 .Prof. A growing number of parents fail to recognize the need to vaccinate children. and the population as a whole remains essentially resistant. side effects. and ensuing mutations unanticipated by the vaccine could endanger vaccinated individuals its well. the chances of an infectious agent "finding" an unprotected individual become very small.

• It must stimulate the production of neutralizing antibodies to minimize reinfection. several characteristics must be present: • The vaccine must provide effective protection against the pathogen from which it is derived without significant danger of actually causing the disease or severe side effects.Immunology . transport. • The vaccine must stimulate development of those immune responses that are most effective against the pathogen in question (protective T-cell responses). Course 3 . • The vaccine must be economically feasible for widespread use. Ileana Constantinescu 91 . • The protection provided by the vaccine must be effective over a long period of time. and use.Essential characteristics of vaccines For vaccinations to be effective in protection of intended populations. • It must be sufficiently stable for storage. Dr.Prof.

the safer the vaccine. Living. Course 3 . These vaccines would be most appropriate for protection against diseases in which the organisms are so virulent that even killed vaccines are not used because of the risk that a few organisms may have survived the treatments intended to kill them. • DNA vaccines are those in which the naked genetic material of the organism is injected into the host. and express the gene products from the pathogen.Prof. • Attenuated vaccines are those in which the organisms included are live. • Killed vaccines include organisms that have been killed by treatment with physical or chemical agents and may include inactivated toxins (toxoids). live vaccines generate the most efficient immune responses. replicating organisms may synthesize and express molecules that are highly stimulatory to the immune systems. The intent is that host cells will pick up the DNA. The DNA has usually been modified to remove some genes that are critical to causing disease but includes those whose products will stimulate immune responses. but their ability to replicate and cause disease has been damaged by treatment with heat. chemicals. • Extracts are vaccines composed of materials derived of materials isolated from disrupted and lysed organisms. They should be incapable of replication or infection but still able to provoke immune responses. • Live vaccines are those that include organisms capable of normal infection and replication.Types of vaccines • Vaccines may be prepared from pathogenic organisms in a variety of ways. with attenuated vaccines less so. • Modern techniques of molecular biology have permitted two additional forms of vaccines to lie generated. Ileana Constantinescu 92 . the less effective it may be. or other means. but those molecules may be absent from vaccines containing only killed or attenuated organisms. Dr. Paradoxically. The antigenic stimulus remains intact longer than methods in which the vaccine is rapidly eliminated from the host and the stimulation of the immune system stops. These vaccines cause only subclinical or mild forms of the disease at worst. • Recombinant vaccines are those in which the organisms have been engineered in the laboratory by the removal of certain genes critical to their ability to actually cause the disease (the gene encoding a critical toxin) or reproduce completely. followed by killed vaccines and extracts. incorporate it.Immunology . The potency of vaccines can be elevated by administering them alter mixing diem with adjuvants. These vaccines are not used against pathogens causing severe diseases. These organisms are typically able to infect host cells and even proliferate but cannot induce the disease they are associated with. • In general.

In DTP (diphtheria-tetanuspertussis) vaccine.Prof. the causative agent of whooping cough) is an effective adjuvant.Adjuvants • Adjuvants are substances or mixtures that are given together with vaccines lo heighten the effectiveness of the vaccination. Adjuvants are mixtures of bacterial components suspended in sonic medium such as oil to slow and prolong their dispersal into the tissues. • Adjuvants prolong the period of time that a vaccine persists to stimulate the immune responses. and/or attracts phagocytic cells to the site of application and stimulates their activation so that their antigen presentation to lymphocytes is increased. Dr. Course 3 . • The latter is not used for human vaccinations in the United States and other countries because it uses material derived from Mycobacterium and can cause false-positive indications in persons being tested for tuberculosis. attracting phagocytic and other cells to the site.Immunology . • The bacterial material provokes a mild inflammation. Other adjuvants include alum and BCG (Bacillus Calmette Guerin). Some vaccine components themselves can serve as adjuvants. Ileana Constantinescu 93 . the pertussis component (from Bordetella pertussis.

Although main dangerous infectious diseases are now prevented or minimized by routine vaccination.Prof. • In many cases. new generations of new escape mutants have already been generated. many others such as malaria. as described earlier. Perhaps the most dramatic example of this is HIV. large numbers of new surface structures are constantly being generated that are new to the immune system. Often the difficulty in developing a vaccine is related to characteristics of the infectious organism. histosomiasis. if and when it does. clearance of the pathogen is difficult. Course 3 .Difficulties in vaccine development • The development of effective vaccines is not necessarily a straightforward one. the difficulty lies with the ability of the pathogen to change its antigens so that immune responses generated to that point are ineffective. Unfortunately. some pathogens such as Plasmodium enter erythrocytes that do not express MHC class 1 or 2 molecules and are essentially invisible to T cells. but because the virus mutates so rapidly during replication. And. As mentioned earlier. • These escape mutants are free to continue replicating as the immune system tries to catch up. the ability of cells to sequester themselves within certain types of host cells shelters them from the effects of antibodies. however. If the host is not generating sufficiently effective cellular responses. and the immune response often plays a hopeless game of catchup. Dr. and AIDS still lack effective vaccines.Immunology . Ileana Constantinescu 94 . The immune system can generate strong antibody responses against certain structures on the viral surface.

Philadelphia: Lippincott Williams & Wilkins. 2004:665-682. Jr. In Janeway CA Jr. Walport M. Course 3 . 5th ed. • Berzofsky JA. 6th ed.. • Chaplin DD. Immunobiology: The immune system in health and disease. Fundamental immunology. 2003. • Paul WE.Prof. Evolution of the immune system. General properties of immune responses. In Paul WE. Ileana Constantinescu 95 . Travers P. The immune system: An introduction. Immunogenicity and antigen structure. 5th ed. In Paul WE. Shlomchik P. ed. J Allergy Clin Immunol 2003. 2003. eds. ed. • Janeway CA. Berkower IJ.Immunology . Philadelphia: Lippincott Williams & Wilkins. Philadephia: WB Saunders Co. Philadephia: Garland Publishing. 5th ed.Suggested readings • Abbas AK. Overview of the immune response. Lichtman AH. Fundamental immunology. 2003. In Cellular and molecular immunology. 111 :S442.