The Role of TCR gamma-delta T cells in Celiac Disease Columbia University, College of Physicians and Surgeons By: Nertila Ujkaj Littin Kandoth Sandra.

The Role ofTCR gamma-delta T

cells in Celiac Disease

Columbia University, College of Physicians and Surgeons

By: Nertila UjkajLittin Kandoth

Sandra

Mentor: Dr. Manavalan

Celiac disease (CD) Celiac disease (CD) •a digestive disease that damages the a digestive disease that damages the small intestine and interferes with small intestine and interferes with absorption of nutrients from food.absorption of nutrients from food.•when people with CD consume when people with CD consume gluten, their immune system responds gluten, their immune system responds by damaging the small intestineby damaging the small intestine •a genetic disorder a genetic disorder •symptoms: diarrhea, weight loss, symptoms: diarrhea, weight loss, malnutrition, etc…malnutrition, etc…•diagnosis is done by intestinal biopsy diagnosis is done by intestinal biopsy histological grading and presence of histological grading and presence of serum auto- antibodies.serum auto- antibodies.•CD is considered an autoimmune CD is considered an autoimmune disorder because the body's own disorder because the body's own immune system causes the damageimmune system causes the damage

The Gut LumenThe Gut Lumen

Antigen Antigen Presenting Presenting

CellCell B cellB cellB cellB cellLPLP CD4CD4++ T cellT cell

Anti-gliadin and Anti-gliadin and anti-TTG2 anti-TTG2 antibodiesantibodies

HLA-DQ2 HLA-DQ2 or DQ8or DQ8

IELIELT cellT cell

GlutenGluten

MICMICA/BA/BNKG2DNKG2D

CD94/NKG2CCD94/NKG2C

IL-15IL-15

IELIELT cellT cell

IFN- & Cytotoxic granules

?

IMMUNOPATHOGENESIS OF CELIAC DISEASE

Gliadin Gliadin PeptidePeptide

INTRAEPITHELIAL SPACE

LAMINA PROPRIA

ObjectiveObjective• To determine if the TCR gamma-delta IELs in To determine if the TCR gamma-delta IELs in

patients with CD are Regulatory/Suppressor T patients with CD are Regulatory/Suppressor T cells cells – Regulatory T cells (also known as Regulatory T cells (also known as

suppressor T cells) are a specialized suppressor T cells) are a specialized subpopulation of T cells that act to suppress subpopulation of T cells that act to suppress activation of the immune system and activation of the immune system and thereby maintain immune system thereby maintain immune system homeostasis and tolerance to self.homeostasis and tolerance to self.

– They are characterized by having some of They are characterized by having some of the following markersthe following markers• Intracellular FOXP3 Intracellular FOXP3 •TGF-betaTGF-beta• IL-10IL-10•CTLA 4CTLA 4•GITRGITR

Methods• Isolation of Lymphocytes from the Duodenal biopsies of 30

patients with Celiac Disease

• Immunostaining for the following extracellular and intracellular markers:– T-Cell markers

• CD3• CD8• TCR alpha-beta• TCR-gamma-delta

– Markers for Regulatory/Suppressor cells:• Intra-cellular FOXP3 • Intracellular TGF-beta• GITR• CTLA-4• Intra-cellular IL-10

• Flow cytometry- technique for counting, examining and sorting microscopic particles (such as cells) suspended in a stream of fluid

http://www.flow-cytometry.de/img/fcm.gif

A. Immunohistochemistry

EPITHELIAL CELLS

B. Flow Cytometry

INTRAEPITHELIAL LYMPHOCYTES (IEL)

Results

Results

Means (error bars: 95% CI for mean)

45

40

35

30

25

20

15

10

5

0

% o

f FO

XP

3 ex

pres

sing

cel

ls

TCR alpha-beta TCR gamma-delta

Means (error bars: 95% CI for mean)

50

40

30

20

10

0

% In

trac

ellu

lar T

GF-

beta

exp

ress

ing

cells

TCR alpha-beta TCR gamma-delta

Results

Antigen Antigen Presenting Presenting

CellCell B cellB cellB cellB cellLPLP CD4CD4++ T cellT cell

Anti-gliadin and Anti-gliadin and anti-TTG2 anti-TTG2 antibodiesantibodies

HLA-DQ2/8HLA-DQ2/8

IELIELT cellT cell

TTGTTG

GliadinGliadin

MICMICA/BA/BNKG2DNKG2D

CD94/NKG2CCD94/NKG2C

IL-15IL-15

IFN- & Cytotoxic granules

TGFTGF

IELIELT cellT cellFOXP3FOXP3

Conclusion• The TCR gamma-delta T cells have

FOXP3 and secrete TGF-beta and are probably Regulator/ Suppressor T cells

Future studies• To prove that TCR gamma delta T cells

can suppress the immune response• To determine if CD patients who are on

therapy have more suppressor TCR gamma-delta T cells than Active CD patients

References• Green, P.H. and B. Jabri, Celiac disease. Annu Rev Med, 2006. 57: p. 207-21.

• Shan, L., et al., Structural basis for gluten intolerance in celiac sprue. Science, 2002. 297(5590): p. 2275-9.

• Meresse, B., et al., Coordinated induction by IL15 of a TCR-independent NKG2D signaling pathway converts CTL into lymphokine-activated killer cells in celiac disease. Immunity, 2004. 21(3): p. 357-66.

• Chen, Y., et al., Protection of the intestinal mucosa by intraepithelial gamma delta T cells. Proc Natl Acad Sci U S A, 2002. 99(22): p. 14338-43.

• Peng, S.L., et al., Propagation and regulation of systemic autoimmunity by gammadelta T cells. J Immunol, 1996. 157(12): p. 5689-98.

• Roncador, G., et al., Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. Eur J Immunol, 2005. 35(6): p. 1681-91.

• Manavalan, J.S., et al., Alloantigen specific CD8+CD28- FOXP3+ T suppressor cells induce ILT3+ ILT4+ tolerogenic endothelial cells, inhibiting alloreactivity. Int Immunol, 2004. 16(8): p. 1055-68.

Acknowledgements•Dr. Manavalan•Dr. Sat Bhattacharya•Harlem Children Society•Allison•Shannon•Harold•Celiac Disease Center of Columbia University

Thank You!!!