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 FcR 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)

FcR
Phagocyte
Pathogen

Opsonin recognition
Followed by ingestion

Cytokine mediated opsonins

Phagocyte containing CRP (Complimentreactive 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)
Cells osmotic lysis

Release of perforin
from granules

FcR

NK cell

Virus

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
FcR receptors on NK cells recognize antibody
bound to viral proteins
Polarization of granules from NK cells
Release of perforin
Cells 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 FcR 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.