Activarea limfocitelor T

ORGANIZARE & FUNCTIONARE SI

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1. Scopul activarii LT
2. Declansarea RIA-
recunoastere specifica APC
3. Activarea LT: molec & cel
Activarea LT -Intrebari

1. UNDE ?
2.CINE ?
3.CUM ?
4.CE
REZULTA?

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 Activarea
LT naïve :

CCR7

1. UNDE? CCL19 /
CCL21:
HEV
CCR7
CD197

1. OLS:
Gg LIMFATICI (ZONA T-DEP)- PARACORTEX Gg (HEV*_CC7 Liganzi:
CCL19/CCL21) 5
Activarea LT naïve :
2. CINE?

2.
―APC: CD mature* – CCR, CAM, [DAg-MHC], co-stim (CD80/86, CD40)
―LT naïve* - in repaos => 2 situatii
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 CUM se activeaza LTnaive ?
aspecte
I. INTER-CELULARE : sinapsa imunologica
(global)
II. INTER-CELULARE : recunoasterea Ag
(molecular -receptorial)
III.INTRA-CELULARE: transducerea semnal
activator
(molecular- sub-celular: :
― citosolic
― nuclear
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 Activarea LT naive:

2. CUM? APC 2 1

I. INTERCELULAR IL-4
CD80 IFNγ
COOPERARE CELULARA
CD28
A.Activare anterograda:
APC → LT: 2 1 LT
―cale principala (1)
―cale “accesorie” (2)

B. Activare retrograda: 3. CE ?
LT→APC:
― CK: (IL4 & IFNγ)

― Maturizare functionala
3. CE REZULTA? ― Expansiune clonala 8
 Activarea LTnaive:
4. CUM ? nivel receptorial
A. Ag-INDEPENDENTA
Rec de adeziune celulara (LT-APC):
―CD2 – LFA-3 / CD58 (I)
―LFA1-ICAM (II)

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 Activarea LT:
4. CUM ?
II. INTER-RECEPTORIAL
A. Ag-INDEPENDENTA
Rec de adeziune celulara:
―CD2 – CD58 (LFA3) (I)
―LFA1-ICAM (II)

Etapele stabilirii sinapsei imunologice:

SINAPSA IMUNOLOGICA:
STRUCTURA:
• SMAC-central= 15nm (TCR, CD4, CD28, PKC-Θ Padapt)
• SMAC- periferal=40nm (CAM)

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 REMEMBER! SINAPSA IMUNOLOGICA:

Rol:
Contacte stranse=>
Stabilizarea contactelor
(spatiu→timp→spatiu) =>

1. Recunoastere Ag
TCR –[ DA-MHCII]
CD4- MHC II
2. R activatori accesori
(CD28-CD80/ CD40-CD40L)
3. Activarea
retrograda/anterograda
LT→IL⁺APC
4. turn-over molec semnaliz

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 Activarea LT:
4. CUM ?
B. Ag-DEPENDENTA:

II. INTER-RECEPTORIAL
1. Recunoastere Ag (extra-celulara)
― TCR
― CD4/CD8
2. Transmiterea semnal activator (intracelular)
― (TCR-) CD3-ξξ (ITAM)
― CD4/CD8 (ITAM)

III. INTRA-CELULARA:

1. Faza citosolica: cai de semnalizare intracelulare - “cascada fosforilarilor”: PTK

2. Faza nucleara: + transcriptei genelor 12
 III. Etapa INTRACELULARA-
transducerea semnalului

1. Faza
intracitosolica:
― ITAM
― PTK: Lck, Fyn, etc
― P adaptor: ZAP-70

2.Faza (intra-) nucleara:

―Fact activatori nucleari
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III. Etapa INTRACELULARA
1. Faza intracitosolica

SLP-76 (SH2domain-cont Linker for Presentation )

LAT (Linker for Activation of T cell

ZAP-70
1.Activat: Fyn & Lck (fosforilare)
2.Ataseaza de ζζ→+PTK →
3.Recrut Padaptor (LAT) →+PLC γ(fosfolipaza Cγ) →IP3 fosfatidil inozitoli
4.Modulare intensitate semnal

Reglare semnal: *Prag de activare~ prag activ ZAP-70

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III. Etapa INTRACELULARA

1. Faza intracitosolica
Cai de semnalizare
intracelulara:

PKC ( protein kinaza C- izoforma θ) 15

 Calea PLCγ1
ZAP-70-℗→LAT→
+PLCγ1℗:
→PIP2 (FLp mb)→PIP3:
Ø 1. IP3: IP3 -IP3R (ER)
Ø →↑Ca2+
→+Ca-calmodulina→
++ Calcineurina:
→Activare NFAT
2. PDK1→+℗Akt→
→Inactiv P pro-apopt Bcl-2

→DAG: +PKC →PKCθ →→

→Activare Nfkβ

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Calea Ras-MAP
kinazelor

ZAP-70→adaptori:
LAT→ Prot G (Grb2):
Ras-GDP + SOS →
Ras-GTP →MAP-kin →
ERK-1,2: F. nucleara→
Activare AP-1

MAP: Mitogen Activated Protein; Prot G: GEF: Guanine exchenge factors;

SOS= factor de transfer G;
 Schema simplificata a cuplarii
recunoasterii Ag & transducerea semnalului
• +PLCγ1→ [Ca intracelular] ↑ (via IP3)
IP3 → leg IP3R (canale Ca in ER )→ descarcarea depozitelor Ca++ in
citosol →cresterea [Ca++] intracelular →
activ calcineurina (fosfataza )→ activ factor transcriptie NFAT.

• +Protein Kinazei C via DAG

DAG (asociat mb cel) → recruteaza memberii ai caii PTK→
PKC, serine/threonine PTK →→ final activ factor transcriptie NFkB

• Cascada MAP Kinazelor (Mitogen Activator Proteins)

Proteinele SGP ( Small G-protein, ex: Ras & Rac Kin) → +MAPk → →→
final ℗ AP-1 (Fos & P-Jun )

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III. Etapa INTRACELULARA
2. Faza nucleara a semnalizarii

IL-2,4,
TNF-α
IL2, 2R + diviziuni mitotice

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Activarea LT: co-stimulrea prin

1. CD28-CD80

• Sinapsa imunologica: APC-LT naïve →CD80-CD28:

• CD28:
― Enzima LcK
― Substrat : CD28: P-Tyr (ITAM) → + IP3 =>
=> + MAP-kin => + NFKb: (CD28 response element-
promotor gena IL2) => IL-2
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 Activarea LT*: co-stimulrea
(bucla de amplificare)
1. CD40-CD40L

CD28-CD80=>
• expresia CD40L (LT activate) =>
• legaturi CD 40 (APC) =>
– APC: ↑ expresia CD80
– LT activate: + sinteza IL

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Modularea intensitatii transmiterii
semnalului
Mecanisme de amplificare
1. Fosforilare progresiva a ITAM:
afinitatea Ligand pt TCR este mare ~ ↑nr de ITAM-℗
2. Stimuli prin co-receptori (CD4/CD8)
3. Stimuli prin receptori co-stimulatori (CD28, CD40L)
Mecanism de inhibare:
1. Receptori inhibitori: CTLA-4
2. Fosfataze: defosforilare
• ITIM

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Consecintele
Activarii
LT

DAG

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sfarsit

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