Imunologie

Aparare Ne-specifica
Celule
 General Introduction
• Primitive pattern recognition receptors (PPRR)
are conserved across evolution
• They recognize common motifs on pathogen
• Innate immune system is triggered as soon as
physical and chemical barriers are penetrated by
pathogens

Why do you think it is Innate immune system

that kicks in first?
 GENERAL FEATURES
• Complement activation
• Phagocytosis
• Cell-mediated cytotoxicity

• Cytokines and chemokines also contribute

to the enhancement of innate immunity
 Role of complement system
opsonization by phagocyte
(fig.2-1)
• Alternative pathway
• Spontaneous hydrolysis of C3 protein to produce
C3b
• C3b deposits on microbial surface and activates
cascade
• Various proteins are generated
• C3b on the bacteria recognizes CR1 receptors
on phagocytes and the opsonization takes place
 Alternative pathway: MAC
• Generation of membrane attack complex
(MAC)
• Insertion of MAC into bacterial cell
membrane induces lysis of the cells or
bacteria
 Alternative pathway: C5a,
C4a, and C3a
• Some proteins are anaphylatoxins (C5a, C4a,
C3a)
• They bind to cognate receptors on mast cells and
basophils which have receptors for anaphylatoxins
• Induction of degranulation takes place
• Release of histamine and inflammatory mediators
(This is a sign why we feel sick. So what do we do
about it? We take antihistamine such as Advil)
 PHAGOCYTOSIS

• Neutrophils
• Macrophages
 Phagocytic cells
• Neutrophils mature in bone marrow
• Small % is stored in bone marrow
• Majority released into circulation
• Number of inflammatory cytokines
enhance release of neutrophils from bone
marrow
• *cytokines enhance activity of Innate and
Adaptive Immunity
 Phagocytic cells cont.
• Macrophages are more effective than
monocytes
• Longer half-life than neutrophils
• Serve as antigen presenting cells to CD4+
T-cell activation
 Recruitment of phagocytes
into infected tissues
• Neutrophils and monocytes are attracted to site of
infection by chemotactic molecules and
chemokines
• They marginate along and attach to lumenal surface
• Secrete enzymes that break basement membrane
• Diapedesis takes place (squeezing through the
endothelial wall)
• Monocytes differentiate into macrophages in tissues
fig. 2-2
 Direct recognition of
pathogens
• Occurs via primitive receptors (PPRR)
• Receptors on cell recognize bacterium
directly
• Phagocytosis takes place
 Indirect recognition of
pathogens
• Uses cell surface receptors
• Recognize molecules bound to pathogen
cell surface
• Molecules that attached to pathogen are
called opsonins
 Complement activation
opsonins
• Bacterium carries C3b (opsonin)
• It is recognized by CR1 receptor which is
present on phagocyte
• Phagocytosis takes place
• must have receptor!
 B-cell activation opsonins
• Bacteria has IgG (opsonin)
• It is recognized by FcR receptor on
phagocyte
• Phagocytosis takes place
• must have receptor!
*IgG is synthesized by B-cells, specifically
by plasma cell
 Pathogen

Pathogen

Anti-pathogen antibody (IgG)

FcR
Phagocyte
Pathogen
Opsonin recognition
Followed by ingestion
 Cytokine mediated opsonins
• Phagocyte containing CRP (Compliment-
reactive protein or heat shock protein)
binding site
• Recognizes bacterium carrying CRP
(opsonin)
• Phagocytosis takes place
• must have binding recognition site!
 PHAGOSOME
• Phagocytic vacuole
• Contains lysosomal enzymes, reactive
oxygen intermediates, reactive nitrogen
intermediates
 Phagosomes and
Phagolysosomes
• Binding of microorganisms triggers:
-NADPH oxidase assembly
-formation of phagosome
• Phagosomes fuse with lysosomes and form
phagolysosome
• Lysosomal granules (lactoferrin [binds iron],
lysosyme [destroys muramic acid in bacterial cell
wall], defensins [permeabilize bacterial and fungal
membranes]) are released in phagolysosome
• Antigen is degraded and released by exosytosis
Fig. 2-5
 NADPH oxidase and reactive
oxygen intermediates
• Increase in oxygen consumption due to
phagocytosis
• Activation of NADPH oxidase
• Uses oxygen
• Generates superoxide anion (ROI)
Fig. 2-6
(you will be tested on this concept)
 Nitric oxide and reactive
nitrogen intermediates
• NO is a lipid and water soluble gas which is
cytotoxic to microbes
• Releases RNIs
• Important for elimination of pathogens resistant
to ROIs
• In phagocytes, synthesis of NO requires
induction of nitric oxide synthase (got to know
this enzyme)
• Catalyzes conversion of L-arginine to L-citruline
and NO in presence of oxygen
 Regulation of nitric oxide
synthase
• 2 signals needed for NO synthase activation:
• 1) Induction of NOS occurs in response to phagocytosis
of Mycobacteria, leishmania, or tumor necrosis factor
• 2) cytokine INF signal
• Downregulation occurs in response to cytokines IL-10,
IL-4, and transforming growth factor beta (TGF), which
are Th2 cytokines
• TGF is the most effective inhibitor of NO synthesis
• Th1 cells are responsible for these signals (intracellular
bacteria)
(there is an antagonistic activity of Th1 and Th2 cells)
 Chronic granulomatous
disease
• Inherited immunodeficiency disorder
• Individuals are susceptible to normally nonpathogenic and
pathogenic organisms
• Characterized by absence or decreased NADPH oxidase
activity in neutrophils
• In phagosome NADPH oxidase complex assembly is
diminished or absent
• Degradation of microbe is diminished
• Must rely on phagolysosome and lysosomal enzymes
• Nitrozole tetrazolium (NBT) test is used to diagnose CGD
• NBT is a yellow dye that becomes insoluble and turns
purple in presence of NADPH oxidase activity
 CGD
• Thus, CGD inhibits NADPH oxidase complex
assembly
• That leads to inability to form superoxide,
hydrogen peroxide, hydroxyl radicals, hydroxyl
anions
• The phagosome takes a different rout here
through lysosomal enzymes: lactoferrin,
lysozyme, defensins

Fig. 2.8-very important figure

 NATURAL KILLER CELLS
• Destruct host cells in order to eliminate virus
• Express receptors that recognize viral proteins
embedded in cell membrane
• NK cells also express killer inhibitory
receptors (KIRs), whose ligand is class I MHC
molecule
• Upon interaction with class I MHC, NK cell
receives a signal that inhibits killing of cell
expressing class I MHC
 Receptor-mediated
cytotoxicity
• Virus invades cell
• Viral proteins expressed on cell surface
• NK cell receptor binds to viral protein
• Polarization of granules
• Granules release perforin (this protein
punches or “perforates” holes in
membrane of NK cells and CD8 cells)
• Cell’s osmotic lysis
 Release of perforin
from granules FcR

Virus NK cell

Antibodies are generated that

recognize viral envelope proteins

Any
cell

Viral envelope proteins Osmotic lysis

on the cell surface

Release of perforin
from granules

NK receptor Osmotic lysis

 Antibody-mediated cellular
cytotoxicity (ADCC)
• Virus invades cell
• Viral proteins expressed on cell surface
• Antibodies generated that recognize viral
envelope proteins
• FcR receptors on NK cells recognize antibody
bound to viral proteins
• Polarization of granules from NK cells
• Release of perforin
• Cell’s osmotic lysis
*B-cell has to be activated to produce antibody
 NK cell in a nutshell
• The bottom line is that Natural Killer cells
destroy cells that engulfed virus by releasing
perforin from granules
• There are two mechanisms:
-ADCC (antibody directed cell cytotoxicity)
-Receptor mediated cytotoxicity
• Activity of NK cells is enhanced by IL-2 and
IL-12
• In case of ADCC, B cell must be activated in
order to produce antibodies
 Eosinophils
• Derived from bone marrow
• Exist both in circulation and tissues
• Major role in immunity to parasites (helminths)
• Requires antibodies (IgE) which binds to helminth
• Recognition is indirect
• Have FcR that recognizes Fc region of IgE
• Triggers degranulation of eosinophils
• Release of major basic protein (MBP) and eosinophil
cationic protein (ECP)
• IL-5 is a peptide secreted by T-cells and is a precursor of
eosinophils
• Schema experimentala de cultivare a celulelor stem. Celulele stromale ale maduvei osoase formeaza o matrice
pe care celulele hematopoietice prolifereaza. Celulele singulare pot fi apoi trasnferate pe un gel semisolid de agar
pentru a le oferi posibilitatea sa creasca in colonii.
• Celule din maduva osoasa in cultura de lunga durata.
• Macrofagele sunt de 5-10 ori mai mari decat monocitele,
contin mai multe organite celulare, in special lisosomi.
• Pseudopode lungi, care ajung in contact cu bacteria, un prim pas spre
fagocitoza.
• Fagocitoza si procesarea antigenelor exogene. Materialul digerat este
exocitat; unele peptide sunt prezentate de catre MHC II.
• Limfocit normal (in timus)
• Timocite in apoptoza
• Timocit normal
• Timocit in apoptoza.