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Anteprima

ESTRATTO DOCUMENTO

DENDRITIC CELL BIOLOGY

DCs are the most potent antigen-

presenting cells that play a central

role in coordinating human

immunity

They comprise different subsets at

different stage of maturation

They have potential advantages as

cellular adjuvants for

immunotherapy strategies

Antigen cross-presentation and T cell immune response

TAAs Immune attack against

the tumor

Tumor Antigens Tumor cells CD8

migration

Dendritic cell CD4 Lymphocyte

CD4

CD8

Activated/Mature DC

DC migration Lymph node

RCC : from where we started….

 RCC Cell Lines Characterization to obtain the most immunogenic RCC line:

60 RCC lines: 10% (5-6 RCC lines).

IFNg Elispot Assay

350

cells 300

T

numbers/100.000 250

200

150

100

Spot 50

0 TU

T21

T0 U

U YS

LYS

II

LYS I

I II

II I

I II T

T Ab

Ab b

Ab Ab Ab

NA b L

b +

+ A

NA

A A +

DC +

T21

DC +

T0 +

+ + +

+ + DC

U

U YS

U

S U

NA

YS YS T

+

T

+ Y T NA T

L +

L + T21

T0 L + L +

C T21

DC T0

C T0 C T21

NA D

D D

NA

NA +

NA

+ NA

+ + NA

T21

T0 T0 T21

NA NA NA

NA

ELTHEM cell line

A B

(A) Real Time-PCR of tumor markers (OFA, CE,

hTERT, Ruas) and interleukin-6

(B) Microsatellite instability (MSI) and loss of

heterozygosity (LOH)

(C) Immunocytochemistry (CAM 5.2, the

mitochondrial markers, the vimentin,

cytokeratin AE1/AE3, cytokeratin 19, EGF-R, the

EMA, CD10 and Ki 67)

C

ELTHEM cell line

A (A) Mixed Lymphocytes Tumor

cell Cultures Cytotoxicity test of

+

CD8 T lymphocytes against

autologous ELTHEM clone (60%

lysis, E / T = 60:1).

(B) ELISPOT test for IFNg

B release. The block of the

response of class I test shows

the 91% specificity.

ELTHEM proteomic profile

A B

MW

150 KDa C

10 3 10

pH

(A) Representative 2-DE map of proteins extracted from ELTHEM clone

(B) Biological function and (C) localization of identified protein.

Therapeutic vaccines based on the use of autologous

dendritic cells as natural cellular adjuvants

Peripheral Blood Cytokines

Dendritic

Cell

Re-infusion RCC Antigen

Patent N. MI2005 A002018 (21.10.05).

: Linea cellulare di carcinoma renale e suo uso". Brevetto Industriale

Inventors: E. Ranieri, L. Gesualdo, W. Herr, M. Battaglia.

International Patent : PCT/EP2006/06763, 20.10.06

DC MONITORING Immunity

DC phenotyping In induction

RCC tissue and lymph nodes cancer, infective

disease

RCC PATIENT DC subsets

analysis by IHC

Tolerogenicity

Induction

DC Phenotyping in

Peripheral Blood Transplant,

autoimmunity,

allergy

Human Dendritic cells

DC comprise two subsets functionally and phenotypically different:

Myeloid DCs

+ +

• BDCA-1 /BDCA-3

• potent stimulators of T lymphocyte

• IL-6, IL-12p70, IL-10, TNF-a production

Plasmacytoid DCs

+ +

• BDCA-2 /BDCA-4

• generate a Th2 response

• impressive producers of IFN-a in viral

infection

Dendritic cells analysis in RCC peripheral blood

35000

blood 30000 P<0,01

peripheral 25000 BDCA1

20000 BDCA2

15000

of BDCA3

cells/mL 10000

5000

N° 0 Normal RCC

A significant decrease (p<0,01) in the percentage and in the absolute number

of myeloid DC and plasmacytoid DC subsets are observed in RCC pts compared

with healthy controls Gigante M. et al, Mol Immunol. 2009

Dendritic cells analysis in RCC tissue

Plasmacytoid DC

Myeloid DC

RCC

Healthy

A significant increase of myeloid DC and plasmacytoid DC infiltrate in RCC biopsies

compared to normal kidneys (p<0.001) Gigante M., Ranieri E et al, Mol Immunol. 2009

Dendritic Cells distribution in RCC lymph nodes

+ +

A significant decrease of mature DC (CD11c , CD83 ) infiltrate in RCC lymph

nodes compared to normal (p< 0.001) Normal

RCC

Green:: CD11c-FITC

Red: CD83-TRITC 16

cells/hpf 14

12 *

10

CD11c+/CD83+ 8

6

4

2

0 PATIENTS CONTROLS

Gigante M. Ranieri E et al, Mol Immunol. 2009

Mechanism of Action

DC subsets Immature DC Immature DC

Peripheral

Peripheral blood Lymphoid tissue

tissues/Cancer

IFN-a-conditioned DC

 IFN-a is an important adjuvant for the development of

DC-based vaccines with high clinical efficacy.

 IFN-a potently enhances both T cell and antibody

responses and promote immunological memory by direct

action on DC.

IFN-a-conditioned DC preferentially stimulate Type-1 and

limit Treg-type in vitro T cell responses in RCC patients

IFNa DC :

 promoted significantly stronger Tc1 

80

effector T cell responses than cytokine 70

cocktail-matured DC as revealed by 60 p = 0.022

IFN-g ELISPOT assay 

50 IFN-DC

40

 mDC

were more efficient in inducing the  aDC1

30

+

generation of suppressor CD8 T cells 20

than mDC 10

0

 induced a lower expansion of Treg MAGE-6 EphA2 SEB

cells than mDC

 IFN-a-conditioned DC may be candidates for use in novel

therapeutic vaccines in the setting of RCC

Gigante M., Ranieri E. J. Immunother 2008


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DETTAGLI
Corso di laurea: Corso di laurea magistrale in medicina e chirurgia (a ciclo unico - 6 anni)
SSD:
Università: Foggia - Unifg
A.A.: 2012-2013

I contenuti di questa pagina costituiscono rielaborazioni personali del Publisher kalamaj di informazioni apprese con la frequenza delle lezioni di Medicina di Laboratorio - Patologia Clinica e studio autonomo di eventuali libri di riferimento in preparazione dell'esame finale o della tesi. Non devono intendersi come materiale ufficiale dell'università Foggia - Unifg o del prof Ranieri Elena.

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