INTRODUCERE N

IMUNOLOGIE
Dr. MARA MIHAI
drmaramihai@gmail.com

NOIUNI ELEMENTARE
INFLAMAIA
Definiie
Factori declanatori
Etapele procesului inflamator
Consecine
PROCESUL INFECIOS
Colonizare
Infecie inaparent
Infecie latent
Boal subclinic
Stare de purttor

IMUNITATEA/ RSPUNSUL IMUN

Tipurile de imunitate
Rspunsul imun nnscut
Fagocitoz
Sistem complement
Cale clasic
Cale altern
Cale lectinic
Rspunsul imun dobndit
Umoral
Celular

Contextul
Toate organismele (omul, animalele, plantele, fungii,
bacteriile) dein mecanisme de aprare.

PROCESUL INFLAMATOR Definiie

Proces fiziopatologic complex care const ntr-o reacie tisular
nespecific, de aprare local, ce include fenomene alterative,
reactive, vasculo-exudative, proliferative, reparatorii, cu
principalul scop de a limita aciunea sau de 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: 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

EXSUDATIV HEMORAGIC
INFLAMAIE EXSUDATIVINFLAMAIE
SEROAS

Bacillus anthracis
Antrax cutanat
Mycobacterium tuberculosis- Pleurit

Pneumonie hemoragic

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
antimicrobiene
Activarea fagocitelor
Activare s. complement
Distrugerea
microorganismelor
Activarea limf.

Activarea s.
complement
Celulele dendritice
migreaz n ggl. limfatici
Fagocitoza
Activare celule NK
Producie de citokine,
chemokine

Fagocitoza patogenilor
n esutul limfoid
Imunitate dobndit
prin APC

Vindecarea infeciei cu
RIU, RIC Ac specifici,
activarea macrofagelor
de limf. Th, activarea
limf. T citotoxice

ETAPA DE DECLANARE
1. Agentul patogen depete bariera anatomic... esut
intersiial, 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

Modificarea de form a fagocitului (rotund...triunghiular) i motilitatea

Fagocitarea agentului determinant opsonizat

Receptor C5a- Agent patogen opsonizat cu C5a

Receptor pt Crystal Induced Chemotactic Factor CCF
Receptor pentru LTB4

Rearanjarea citoscheletului
Polarizarea fa spate a microtubulilor
Filamentele de actin- acumulare n faa i n spatele celulei
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),
GMP-140,
adezine,
LECAM-1 (leucocyte endothelial cell adhesion molecule),
ISCOM (immunostimulating complex),
PAF,
MAC (membrane atack complex),
MACIF (MAC inhibitory factor),

Reactani de faz acut (glicoproteine sintetizate n ficat)- necrobioz, remaniere tisular

proteina C reactiv
fibrinogenul,
a2-macroglobulina, a1-antitripsina, a1-antichimotripsina, haptoglobina, hemopexina, ceruloplasmina, componenta C3, amiloidul P
seric
VSH

Celulele prezentatoare de antigen

Eliberare citokine, chemokine
Expunerea Ag HLA clasa a IIa

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

MACROFAG Escherichia coli

MACROFAG SPORI FUNGICI

Escherichia coli

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

Mduva osoas

 Neutrofil care prsete mduva osoas

Neutrofil care fagociteaz Candida albicans

Circulatia limfocitelor in 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

TIPURILE DE IMUNITATE

SISTEM IMUN

Imunitate
innascuta
(prima linie de aparare)

Componente
celulare

Componente
umorale

Imunitate
dobndita
(a II-a linie de aparare)

Componente
celulare

Componente
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 (achizitionat, adaptativ)
1. Activ
-Natural (postinfecioas)
-Artificial (n urma vaccinrii)
2.Pasiv
-Natural (transplacentar, prin laptele matern)
-Artificial (administrarea imunoglobuline)

IMUNITATE
NNSCUT/ nespecific
(innate immunity)
Dependena fa
de antigen

RI independent

RI dependent

Receptori pentru codificati n genom

patogeni
specificitate joasa
Viteza de rspuns Imediat (minute...ore)
Memorie
Absent
Toate organismele pluricelulare

Celule efectoare

DOBNDIT
/ specific

(adaptive immunity)

Fagocite, celule NK, proteine,

complement, bariere naturale

Generai (rearanjare genic)

specificitate inalta
Lent (exp 1...saptamani, exp 2...zile)
Prezent- Expansiune clonal
limfocitar
Prezent doar la vertebrate
Limfocite B, Limfocite T (helper,
citotoxice, reglatoare)

SISTEM INTEGRAT

RSPUNSUL IMUNITAR NNSCUT

Mecanism universal de aparare fata de infecii
Prima linie de aparare a organismului

Precede raspunsul imun adaptativ

prezent la toate organismele pluricelulare

receptori si efectori ancestrali

raspunde la o mare varietate de patogeni

Distincie perfecta self-nonself

Defecte foarte rare i 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 si contextul antigenului

Recunoatere non-self antigen

Funcii efectoare
prevenire intrare
eliminare

microorganisme
patogene

Rol instructiv asupra imunitatii 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

Functia de prevenire a intrrii

microorganismelor patogene

Factori fizici / mecanici

Factori chimici

Factori biologici

motilitate: muco-ciliara, peristaltica

mucus
fluxul fluidelor prin organism
pH
molecule antimicrobiene
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)

RInnscut

3. Componente celulare
Functia de eliminare a microorganismelor patogene

neutrofile
monocite/macrofage
celule dendritice

Fagocitoza

bazofile
mastocite
eozinofile

Inflamaie

celule NK

Citotoxicitate

RInnscut

FUNCIA DE RECUNOATERE IMUN

origine antigen / context biologic

SELF vs NON self

self

nonself

RInnscut

RECEPTORI PENTRU ANTIGEN/PAMP

IMUNITATE NNSCUT

IMUNITATE DOBNDIT

Originea

Codificai n genom

Generai prin rearanjare genic

Distribuia
receptorilor

Distribuie non-clonala

distributie clonal

inta

recunosc structuri conservate

PAMP (pathogen associated
molecular pattern)

recunosc detalii de structur molecular

Antigen (epitopi)

Specificitate

Joas

nalt

(expansiune clonal limfocitar)

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

PAMP
pathogen associated molecular pattern

invariabile
comune pentru o clas de microbi
nalt conservate
specifice microbilor (patogeni + nepatogeni)
PAMP/MAMP
vitale pentru microorganisme

lipopolizaharid
peptidoglican
acid lipoteichoic
lipoproteine
manoza
ADN
ARN
flagelina
pilina
zimozan

Gram-pozitive

Gram-negative

Pattern Recognition Receptors - PRR

a) Extracelulari

Lizozim--- peptidoglican
Psoriazina--- Escherichia coli, poate fi indus de UVB (soare)
MBL (mannose binding lectine)--- activeaz s. 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
bact.
receptori manoza
(lectina C)

man

LPS, LTA

receptori scavenger
(CD36, CD68, SRB-1)

FAGOCIT

proteina bact.
C3b
receptori opsonine
(CR1)

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

Toll-like receptors (TLR)

proteine transmembranare tip I

conservate filogenetic (Drosophila..om)
domeniu extracelular bogat in leucina
domeniu intracelular TIR (similar IL-1R)
TLR mamifere (11-om, 13-soarece)
recunoastere PAMPs
asociere cu alti TLR sau alte proteine
(MD2,CD14)

TLR

Infectie

PAMP
Toll

Mamifere
NFkB

Rol instructiv
asupra imunitatii specifice

Peptide antimicrobiene
iNOS (soarece)
Molecule costimulatoare
Citokine proinflamatorii

TLR FUNCII
TLR4
LPS
(Gram-)

TLR2
Lipo
proteine
(Gram+)

rspuns direct
anti-microbian

TLR6

TLR9

TLR5

flagelina
PGN
(+TLR2)

apoptoz

ADN

oc
septic

rspuns
adaptativ

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

TLR- INTE FARMACOLOGICE

Adjuvanti vaccinare
agonisti
Boli inflamatorii (sepsis, astm, artrita reumatoida, LES
etc)
antagonisti
inhibitori/supresori

TLR & ci semnalizare intracelulara

Vaccinare

imbunatatire eficienta vaccinuri existente

noi vaccinuri profilactice (boli infectioase)

vaccinuri terapeutice (cancer, Alzheimer)

Vaccinuri ce contin liganzi TLR

antigen: BCG [TLR2 & TLR4]

adjuvant: HiB OMBC conjugate [TLR2]

(outer membrane complex Neisseria)

Adjuvanti liganzi TLR (agonisti)

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 HIVinfected adults

Recunoastere imuna
IPOTEZE

Receptori (self vs. nonself microbian)

(Medzhitov & Janeway, 1997)

Semnale de pericol Danger hypothesis

(self vs. self alterat)

(Polly Matzinger, 1994)

Danger hypothesis
Spre deosebire de teoria self-non-self unde orice era non-self reprezenta o int a sistemului imun,
aici unele molecule non-self pot fi tolerate, dac nu reprezint un pericol Ex: fetus
Naive
T cells

Signal 1

Signal 2
(costimulation)

APC

Danger
signal

- infection
- tissue damage
- stress cells
- hypoxia
- temperature shifts
- hsp

Damaged
cell

Normal
cell

Recunoasterea imun- IPOTEZE

- Receptori

(Medzhitov & Janeway, 1997)

- Semnale de pericol (danger hypothesis)

(Polly Matzinger,
1994)
- Recunoastere prin lipsa(missing self)

Missing self
Celulele NK distrug celulele care nu prezint CMH I

Celula
NK

NKR

Celula
tinta

NCR

Celula
NK

MHC
cls I

Ligand
activator

NKR

NCR

Absenta
citotoxicitatii

Celula
tinta

Ligand
activator

Citotoxicitate

Functiile imunitatii innascute

PAMP

Stimulare PRR

stimuleaza fagocitoza
induce activitate microbicida
induce citokine inflamatorii:

IL-1, IL-6, TNF-alpha (NF-kB)

activeaza imunitatea dobndit

expresia mol. costim. (MHC cls.II,
CD80/CD86)

Co-stimulare: initierea raspunsului imun specific

PAMP

Infectie

TLR

APC

MHC / peptide

costimulator
CD28

TCR
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

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

-Este o component normal a serului

 Calea comun C5 este clivat n C5a i C5b

...................................................... por C9

 Calea altern Calea lectinic Calea clasic

Activarea sistemului complement-3 ci

Calea clasic
formarea complexelor imune (Ag-Ac)
celule apoptotice,
virusuri
proteina C reactiv cuplat cu anumii liganzi

Calea altern

Calea lectinic
lectina care leag manoza de pe suprafaa bacteriilor
(MBL, Mannose-Binding Lectin)
alte proteine asemntoare

Calea clasic
C5 convertaza=C4b2a3b

Calea clasic

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