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2016
INTRODUCERE N
IMUNOLOGIE
INTRODUCERE N
IMUNOLOGIE
NOIUNI ELEMENTARE
INFLAMAIA IMUNITATEA/ RSPUNSUL IMUN
Definiie Tipurile de imunitate
Factori declanatori Rspunsul imun nnscut
Etapele procesului inflamator
Fagocitoz
Consecine
PROCESUL INFECIOS Sistem complement
Colonizare Cale clasic
Infecie inaparent Cale altern
Infecie latent Cale lectinic
Boal subclinic
Rspunsul imun dobndit
Stare de purttor
Umoral
Celular
Contextul

Toate organismele (omul, animalele, plantele, fungii,


bacteriile) dein mecanisme de aprare.
PROCESUL INFLAMATOR Definiie

Proces fiziopatologic complex


reacie tisular nespecific,
de aprare local,
ce include
fenomene alterative,
reactive,
vasculo-exudative,
proliferative,
Reparatorii.
Scop = a limita aciunea i a neutraliza agentul agresor.
Reechilibrarea/ aducerea la normal i asanarea focarului inflamator,
conduc la vindecarea structurii anatomice afectate.
PROCESUL INFLAMATOR Ageni declanatori
Ageni fizici
ex. Traumatisme
Microorganisme
ex. Prioni, Virusuri, Bacterii, Fungi, Parazii
Ageni chimici exogeni/endogeni
ex. Arsur chimic
ex. Reflux gastro-esofagian
Semnele inflamaiei Celsus
Rubor
Calor
Tumor
Dolor
Functio laesa
INFLAMAIA- FORME ANATOMO-CLINICE
INFLAMAIE ALTERATIV

Miozit Clostridium perfringens


INFLAMAIA- FORME ANATOMO-CLINICE
INFLAMAIE EXSUDATIV SEROAS
INFLAMAIE EXSUDATIV HEMORAGIC

Bacillus anthracis
Antrax cutanat Pneumonie hemoragic
Mycobacterium tuberculosis- Pleurit
INFLAMAIA- FORME ANATOMO-CLINICE
INFLAMAIE EXSUDATIV FIBROAS INFLAMAIE EXSUDATIV
PURULENT

Staphylococcus aureus
Chlamydia trachomatis Infertilitate
INFLAMAIA- FORME ANATOMO-CLINICE
INFLAMAIE PROLIFERATIV
GRANULOMATOAS Lupus vulgaris Mycobacterium tuberculosis
Flora normal Peptide i proteine Activarea S. Fagocitoza patogenilor Vindecarea infeciei cu
antimicrobiene complement n esutul limfoid RIU, RIC Ac specifici,
Activarea fagocitelor Celulele dendritice Imunitate dobndit activarea macrofagelor
Activare S. complement migreaz n ggl. limfatici prin APC de limf. Th, activarea
Distrugerea Fagocitoza limf. T citotoxice
microorganismelor Activare celule NK
Activarea limf. Producie de citokine,
chemokine
ETAPA DE DECLANARE
1. Agentul patogen depete bariera anatomic... esut
interstiial, esut parenchimatos
2. Rspunsul inflamator acut
Sunt implicai
Factori chemotactici- chemokine
Factori de comunicare intercelular- citokine
Factori enzimatici: Proteaze, Kinaze
Sistemul complement- opsonizare agent patogen
ETAPA DE DECLANARE
2. Rspunsul inflamator acut
Modificri microcirculaie (! Celulele endoteliale):
Vasodilaaie local Rubor/ Eritem
Crete permeabilitatea vascular...
Exsudare de proteine plasmatice Tumor/Edem
Infiltrat inflamator-migrarea leucocitelor prin marginaie, diapedez
Sistem mononuclear fagocitar monocite-macrofage
PMN- neutrofile
Celulele endoteliale
Proteine vasoactive
1.VASODILATAIEcretere flux sangvin, migrare leucocite
Endothelium derived relaxing factor EDRF
NO
Sistemul kinin/bradikinin vasodilataie, crete debitul sangvin local
...edemul perilezional comprim celulele endoteliale care ncep s produc

2.VASOCONSTRICIE
Endoteline, ET1 vasoconstricie brutal, intens
Celulele fagocitare
Diapedez
Cuplarea Fagocit/ factor chemotactic
Receptor C5a- Agent patogen opsonizat cu C5a
Receptor pt Crystal Induced Chemotactic Factor CCF
Receptor pentru LTB4
Modificarea de form a fagocitului (rotund...triunghiular) i motilitatea
Rearanjarea citoscheletului
Polarizarea fa spate a microtubulilor
Filamentele de actin- acumulare n faa i n spatele celulei
Fagocitarea agentului determinant opsonizat
Fagolizozomi, cuplare cu lizozomi
Inactivare complet
Prezentarea superantigenului cascade de reacii imune
Neutrofilele
Molecule de adeziune celular
ICAM-1 (intercellular adhesion molecule),
ELAM-1 (endothelial cell leucocyte adhesion molecule),
LECAM-1 (leucocyte endothelial cell adhesion molecule),
Adezine (altele ...),

ISCOM (immunostimulating complex),


GMP-140 (platelet alpha-granule membrane protein),
PAF (platelet-activating factor),
MAC (membrane atack complex),
MACIF (MAC inhibitory factor),
... ... ...
Neutrofilele
Reactani de faz acut (glicoproteine sintetizate n ficat)
pot duce la necrobioz, remaniere tisular
proteina C reactiv (PCR)
fibrinogenul,
a2-macroglobulina,
a1-antitripsina,
a1-antichimotripsina,
haptoglobina,
hemopexina,
ceruloplasmina,
componenta C3,
amiloidul P seric
VSH
Nr. crescut de leucocite (PMN)
Celulele prezentatoare de antigen
Eliberare citokine, chemokine
Expunerea Ag HLA clasa a II-a
Rezultate etap declanare

Rspunsul inflamator acut


Vasodilaaie local (eritem)
Exsudare de proteine plasmatice (edem)
Infiltrat inflamator- acumulare de PMN
ETAPA EFECTOARE
SUBETAP MOLECULAR
Activarea sistemului complement- MAC
Activarea sistemului de coagulare/fibrinoliz
Activarea cii de metabolizare a acidului arahidonic
COX
PGI (prostaciclin)- vasodilataie, antiagregant trombocitar, anti aderare PMN
PGE2 (prostaglandin), TXA2 (tromboxan A2)- vasoconstrictor, proagregant trombocitar,
permabilitatea vascular
LOX
LTD4, LTG4 (leucotriene)- vasoconstricie sistemic, vasodilataie local, bronhoconstricie
ETAPA EFECTOARE
SUBETAP MOLECULAR (continuare)
Activarea sistemului kinin, bradikinin
Eliberare citokine
Eliberare interferoni
Actina- mobilitatea celular, filamente de actin circulatorii-oc
septic
SUBETAP VASCULAR
SUBETAP EXSUDATIV bariera fibrino-imuno-leucocitar
IMUNITATEA- RSPUNSUL IMUN
IMUNITATEcapacitatea de aprare specific a organismului fa de agresori externi
(virusuri, bacterii, fungi, protozoare, toxine), ct i fa de propriile molecule i unele
celule degradate sau modificate.
Sarcina fundamental a imunitii distincia dintre moleculele i celulele proprii (self)
i cele strine (non-self)
Fr imunitate

Bubble boy- SCID/ SISC


Sindromul imunodeficienei
severe combinate (Lim B,
Lim T)
Pneumonie
Meningit
Sepsis

David Vetter
1971-1984
Fr imunitate

AIDS/ SIDA- sindromul imunodeficienei dobndite


Terapii imunosupresoare- cancere

Accidentul nuclear Cernobl 1986


Pacient neoplazie imunosupresat
Celulele sistemului imun

preluat din Microbiology and Immunology on-line, University of South California, School of Medicine
http://pathmicro.med.sc.edu/book/immunol-sta.htm
CELUL TUMORAL LIMFOCITE T LIMFOCIT T HIV
esutul limfoid
Distribuia organelor limfoide

preluat din Microbiology and Immunology on-line, University of South California, School of Medicine http://pathmicro.med.sc.edu/book/immunol-sta.htm
Neutrofil care prsete mduva osoas Neutrofil care fagociteaz Candida albicans
Circulaia limfocitelor n organele limfoide periferice

preluat din Microbiology and Immunology on-line, University of South California, School of Medicine
http://pathmicro.med.sc.edu/book/immunol-sta.htm
SISTEM IMUN

Imunitate Imunitate
nnscut dobndit
(prima linie de aprare) (a II-a linie de aprare)

Componente Componente Componente Componente


celulare umorale celulare umorale

adaptat dupa Microbiology and Immunology on-line, University of South California, School of Medicine
http://pathmicro.med.sc.edu/book/immunol-sta.htm
TIPURILE DE IMUNITATE

I.EREDITAR/ NNSCUT (natural, de specie)


II.Dobndit (achiziionat, adaptativ)
1. Activ
-Natural (postinfecioas)
-Artificial (n urma vaccinrii)
2.Pasiv
-Natural (transplacentar, prin laptele matern)
-Artificial (administrarea imunoglobuline)
IMUNITATE
NNSCUT/ nespecific DOBNDIT
/ specific
(innate immunity) (adaptive immunity)

Dependena fa RI independent RI dependent


de antigen
Receptori pentru Codificai n genom Generai (rearanjare genic)
patogeni specificitate joas specificitate nalt
Viteza de rspuns Imediat (minute...ore) Lent (exp 1...sptmni, exp 2...zile)
Memorie Absent Prezent- Expansiune clonal
limfocitar
Toate organismele pluricelulare Prezent doar la vertebrate
Celule efectoare Fagocite, celule NK, proteine, Limfocite B, Limfocite T (helper,
complement, bariere naturale citotoxice, reglatoare)

SISTEM INTEGRAT
RSPUNSUL IMUNITAR NNSCUT

Mecanism universal de aprare fa de infecii


Prima linie de aprare a organismului
Precede rspunsul imun adaptativ
prezent la toate organismele pluricelulare

receptori i efectori ancestrali


rspunde la o mare varietate de patogeni

Distincie perfect self-nonself


Defecte foarte rare (... n general letale)
Levy O. Innate immunity of
the newborn: basic mechanisms and
clinical correlates. Nat Rev
Immunol. 2007 May;7(5):379-90.
RInnscut

ROL I FUNCII
originea i contextul antigenului
Recunoatere non-self antigen
Funcii efectoare
prevenire intrare
microorganisme
eliminare
patogene
Rol instructiv asupra imunitii specifice

Iniierea i tipul rspunsului


RInnscut

ELEMENTE CONSTITUTIVE

1. Bariere anatomice
2. Componente moleculare
3. Componente celulare
RI nnscut

1. Bariere anatomice

Pielea
Conductul auditiv extern
Mucoase
Ocular
Tract respirator
Tract gastro-intestinal
Tract genito-urinar
RI nnscut

1. Bariere anatomice - piele


- mucoase
Funcia de prevenire a intrrii
microorganismelor patogene

motilitate: muco-ciliar, peristaltica


Factori fizici / mecanici mucus
fluxul fluidelor prin organism

Factori chimici pH
molecule antimicrobiene

Factori biologici limfocite T intraepiteliale ()


Bariera anatomic
RInnscut
2. Componente moleculare
- receptori
- molecule secretate
Funcia de recunoatere
Funcii efectorii

Receptori TLR (toll like receptor, pattern recognition molecule)


Mol. anorganice: HCl, NO, H2O2
Peptide antibacteriene: defensine, cathelicidine, histatine
Proteine antibacteriene: lizozim, lactoferina, transferina
Lectine: colectine, ficoline, receptori manoza
Complement
Citokine: IFN-gamma, IL-1, TNF-alpha, CSF
Chemokine: IL-8, MIP (macrophage inflammatory protein), MCP (macrophage chemotactic protein)
S. epidermidis PSM- phenol soluble modulin- activeaz macrofagele
RInnscut

3. Componente celulare
Funcia de eliminare a microorganismelor patogene

neutrofile
monocite/macrofage Fagocitoza
celule dendritice

bazofile
mastocite Inflamaie
eozinofile

celule NK Citotoxicitate
RInnscut

FUNCIA DE RECUNOATERE IMUN


origine antigen / context biologic

SELF vs NON self

non-self

self
RInnscut

RECEPTORI PENTRU ANTIGEN/PAMP

IMUNITATE NNSCUT IMUNITATE DOBNDIT

Originea Codificai n genom Generai prin rearanjare genic

Distribuia Distribuie non-clonala distributie clonal


receptorilor
(expansiune clonal limfocitar)

inta recunosc structuri conservate recunosc detalii de structur molecular


PAMP (pathogen associated Antigen (epitopi)
molecular pattern)

Specificitate Joas nalt


Microorganismele- PAMP- pathogen associated molecular pattern
PRR- pattern recognition receptors
Extracelulari lizozim, PCR etc.
Citoplasmatici NOD
Membranari
Care conduc la endocitoz/fagocitoz
-Receptori Manoz
-Receptori scavenger ALT, LPZ
-Receptori C3b-opsonine
Care transmit semnalul intracelular
-TLR- toll like receptors
----cascada semnal intracelular NFkB---producie citokine
tiai c?
NFkB
Nuclear factor kappa-light-chain-enhancer of activated B cells
a protein complex
that controls transcription of
DNA,
cytokine production and
cell survival.
NF-B
in almost all animal cell types
is involved in cellular responses to stimuli
stress,
cytokines,
free radicals,
heavy metals,
ultraviolet irradiation,
oxidized LDL,
and bacterial or viral antigens.
key role in regulating the immune response to infection
NFkB
Nuclear factor kappa-light-chain-enhancer of activated B cells

Incorrect regulation of NF-B linked to:


cancer,
inflammatory and autoimmune diseases,
septic shock,
viral infection, and
improper immune development.

NF-B has also been implicated in


synaptic plasticity and
memory.
PAMP
pathogen associated molecular pattern

invariabile lipopolizaharid
comune pentru o clas de microbi peptidoglican
nalt conservate acid lipoteichoic
specifice microbilor (patogeni + nepatogeni) lipoproteine
PAMP/MAMP manoza
vitale pentru microorganisme ADN
ARN
flagelina
pilina
zimozan
Gram-pozitive Gram-negative
Pattern Recognition Receptors - PRR
a) Extracelulari

Lizozim--- peptidoglican
Psoriazina--- Escherichia coli (poate fi indus de UV - raze solare)
MBL (mannose binding lectine)--- activeaz Sistemul complement
PCR (proteina C reactiv)--- recunoate microorganismele i celulele
umane deteriorate
LPZ binding protein--- bacterii Gram negative
Pattern Recognition Receptors - PRR
b) Membranari ENDOCITOZA Endocytic PRRs

Bacterie

glicoproteina proteina bact.


bact. LPS, LTA
man C3b

receptori manoza receptori scavenger receptori opsonine


(lectina C) (CD36, CD68, SRB-1) (CR1)

FAGOCIT
Pattern Recognition Receptors - PRR
b) Membranari TRANSMITERE SEMNAL
Signalling PRRs

TLR Toll-like receptors

C) Citoplasmatici
NOD nucleotide-binding
oligomerization domain
RIG-1
retinoic acid-inducible gene-1
tiai c?
"Das ist ja toll!"
Toll-like receptors (TLR)
proteine transmembranare tip I
conservate filogenetic (Drosophila ... om)
domeniu extracelular bogat n leucin
domeniu intracelular TIR (similar IL-1R)
TLR mamifere (11-om, 13-oarece)
recunoatere PAMPs
asociere cu ali TLR sau alte proteine
(MD2,CD14)
TLR
Infecie

PAMP
Toll

Mamifere
NFkB

Peptide antimicrobiene
iNOS (oarece)
Rol instructiv Molecule costimulatoare
asupra imunitii specifice Citokine proinflamatorii
TLR FUNCII

TLR4 TLR2 TLR6 TLR9 TLR5


flagelina
Lipo
LPS proteine PGN ADN
(Gram-) (Gram+) (+TLR2)

rspuns direct apoptoz oc rspuns


anti-microbian septic adaptativ

(adaptat dupa Donata Vercelli: Innate Immunity in Diseases of Lung, Heart and Blood)
TLR- INTE FARMACOLOGICE

Adjuvani vaccinare
agoniti
Boli inflamatorii (sepsis, astm, artrit reumatoid, LES
etc)
antagoniti
inhibitori/supresori
TLR & ci semnalizare intracelular
Vaccinare creterea eficienei vaccinurilor existente
noi vaccinuri profilactice (boli infecioase)
vaccinuri terapeutice (cancer, Alzheimer)

Vaccinuri ce conin liganzi TLR


antigen: BCG [TLR2 & TLR4]
adjuvant: HiB OMBC conjugate [TLR2]
(outer membrane complex Neisseria)
Adjuvani liganzi TLR (agoniti)
Pam3Cys [TLR1 + TLR2]
MPL A, CFA, BCG, analogi LPS [TLR4]
Imiquimod, Resiquimod [TLR7]
CpG ODN [TLR9]
Immune Response to Toll-Like Receptor 9-Agonist Adjuvanted
Pneumococcal Vaccination in HIV Infected Adults

This study is currently recruiting participants.


Verified by University of Aarhus, January 2008
Purpose
Pneumococcal disease is a major source of morbidity and mortality
in HIV-patients. HIV-patients are vaccine hyporesponders.A good
immune response to pneumococcal vaccination enhances vaccine
effectiveness, thereby preventing the morbidity and mortality
caused by pneumococcal disease. Even when an optimized
regimen containing both conjugated and polysaccharide
pneumococcal vaccine is used, only 13% of the immunized HIV
patients are high responders at week 96. Recent data indicate that
TLR9-agonists have excellent vaccine adjuvant potential and
are safe to use in immunocompetent as well as
immunocompromised individuals. The aim of this study is to
evaluate the qualitative and quantitive immune response to
pneumococcal vaccination with or without TLR9-agonist in HIV-
infected adults
Recunoatere imun
IPOTEZE

Receptori (self vs. nonself microbian)


(Medzhitov & Janeway, 1997)

Semnale de pericol Danger hypothesis


(self vs. self alterat)
(Polly Matzinger, 1994)
Danger hypothesis
Teoria self-non-self = orice non-self = int a sistemului imun
unele molecule non-self pot fi tolerate, dac nu reprezint un pericol ex. ftul

Naive
T cells
Signal 1 Signal 2
(costimulation)

APC

Danger
signal

- infection
- tissue damage Damaged Normal
- stress cells cell cell
- hypoxia
- temperature shifts
- hsp
Recunoaterea imun- IPOTEZE

- Receptori
(Medzhitov & Janeway, 1997)
- Semnale de pericol (danger hypothesis)
(Polly Matzinger, 1994)

- Recunoatere prin lips (missing self)


Missing self
Celulele NK distrug celulele care nu prezint CMH I

MHC
NKR cls I
Celula Celula Absenta
NK
tinta citotoxicitii
NCR Ligand
activator

NKR
Celula Celula Citotoxicitate
NK tinta

NCR Ligand
activator
Funciile imunitii nnscute

PAMP

Stimulare PRR
stimuleaza fagocitoza
induce activitate microbicid
induce citokine inflamatorii:
IL-1, IL-6, TNF-alpha (NF-kB)
activeaz imunitatea dobndit
expresia molec. costim. (MHC cls.II,
CD80/CD86)
Co-stimulare: iniierea rspunsului imun specific

PAMP
Infecie
TLR

APC

MHC / peptide costimulator

TCR CD28

T cell Activare
FAGOCITOZA

eliminarea microorganismelor patogene


prelucrare antigenului pt. prezentare
Celule fagocitare

neutrofile
eozinofile
monocite
macrofage
celule dendritice
limfocite B
MACROFAG
Mycobacterium tuberculosis
Macrofag

Carl Zeiss Microscopy Sursa: https://www.flickr.com/photos/zeissmicro/8765512496/in/photostream/


Neutrofil care prsete mduva osoas Neutrofil care fagociteaz Candida albicans
1. Ataarea
2. Ingerarea
3. Distrugerea
SISTEMUL COMPLEMENT
Definiia sistemului complement
complex multienzimatic format din circa 30 de componente
proteice, celulare sau plasmatice
C1,2,3..., factor B, factor D
clivare componente a i b (C3a/C3b, C5a/C5b...)
cascad de reacii
Definiia sistemului complement
cascad de reacii
complexul de atac membranar---por--liza membranar/ liza
microbian
Opsonizare fagocitoza de ctre macrofage/neutrofile
ndeprtarea CIC
Activarea altor rspunsuri imune...anafilatoxine
sursa: hepatic (majoritatea), monocite, macrofage, epitelii
component normal a serului
Calea altern Calea lectinic Calea clasic
Activarea sistemului complement-3 ci
Calea clasic Calea lectinic
formarea complexelor imune (Ag-Ac) lectina care leag manoza de pe suprafaa bacteriilor
(MBL, Mannose-Binding Lectin)
celule apoptotice, alte proteine asemntoare
virusuri
proteina C reactiv cuplat cu anumii liganzi
Calea altern
Calea clasic

C5 convertaza=C4b2a3b
Calea altern
C3b Bb 3b
Calea altern
C3b Bb 3b
Calea lectinic
C4b2a3b
MASP

Lectin- domeniu care recunoate 1 carbohidrat


mannose-associated serine protease 1
-