The role of Follicular Helper CD4+ T cells in Breast ... · The role of Follicular Helper CD4+ T cells in Breast cancer immune response Mireille LANGOUO MD. PhD student BSMO Annual

The role of Follicular Helper CD4+ T

cells in Breast cancer immune response

Mireille LANGOUO

MD. PhD student

BSMO Annual Congress 2019

Tumor-infiltrating lymphocytes (TIL) in BC

Adapted from Salgado et al, 2014; Loi et al, 2013

H&E slide of BC tumor

High TIL improve outcome of TNBC and HER2+ BC

Predict response to Anthracycline in TNBC and Trastuzumab in HER2+ BC

TGFb

TNFα

INFγ

TGFb

TIL composition in TME of BC

TGFb

Tumor suppression Tumor progression

Adapted from Salgado et al, 2014

TGFb

TNFα

INFγ

TGFb

TIL organization in TME of BC

TGFb

CXCL13

TLS: Tertiary Lymphoid Structure

TLS have similar architecture as SLO

/ IHCSLO (eg: Lymph node)

B cell follicles

Adapted from Buisseret, Garaud et al, Oncoimmunology 2017

Breast tissue

SLO – Secondary Lymphoid Organs

TLS – Tertiary Lymphoid Structures

BC – Breast Cancer

60% of primary breast

tumors have at least

one TLS

Follicular helper T cells (Tfh)

/ IHC

Tfh help B cells for immunoglobulin production and differentiation into memory cells

Adapted from David M Tarlinton et al., Nature Immunology 2011

AIMS

Understand how Tfh TIL contribute to TLS

functionnality and anti-tumor immunity in BC

Materials and methods

Untreated BC tissue

BC

Fresh BC tissue

Cell pellet

TIL

Tumor

Supernatant (SN)

FFPE block of matched fresh tumor

IHC and IF imaging

CXCR5+ TIL in BC tumor

Fresh BC tissue

BC

Pink:(CD4+CXCR5+) Tfh

Yellow: CD20+CXCR5+

Pink: CD8+CXCR5+

Yellow: CD20+CXCR5+

(T cells)(B cells)

94%

16%

12%

CD4 (T cells) CXCR5 CD20 (B cells) CD4 CXCR5/CD20

CD8 (T cells) CXCR5 CD20 (B cells) CD8/CXCR5/CD20

1

Zone 1

Zone 1

Tfh TIL characterization in BC

Fresh BC tissue

BC

Tfh TIL

ICOS+PD-1high

Tfh TIL CD4/CXCR5/CD20

CD4 PD-1 CD20 merge

CD4/PD-1/CD20

Tfh PD-1+

Non act Tfh TIL

Act Tfh TIL

Bcl6

CX

Cl1

3

IL-1

0

IL-2

1

IFN

g

MA

F

1 0 -3

1 0 -2

1 0 -1

1 0 0

1 0 1

T fh IC O S-P D -1

lo / in t

T fh IC O S+

P D -1h i

*

*

*

Re

lati

ve

RN

A E

xp

re

ss

ion

Fresh BC tissue

BC

Ig G

P D -1lo w

P D -1in t

T fh X 1 3 T fh

0 .0

0 .5

1 .0

1 .5

2 .0

2 .5

IgG

(µ

g/m

l)

n o n -T fh T IL T IL T IL

Ig A

P D -1lo w

P D -1in t

T fh X 1 3 T fh

0 .0

0 .5

1 .0

1 .5

2 .0

2 .5

IgA

(µ

g/m

l)

n o n -T fh T IL T IL T IL

Tfh TIL function in BC

Non act Tfh TILAct Tfh TIL

Fresh BC tissue

BC

CCL4

FASL

GZM

A

GZM

BIF

Ng

0

1

2

3

4

5CD8

+ICOS

-PD-1

-/int

CD8+

ICOS+

PD-1hi

Rela

tive R

NA

Exp

ressio

n

*

*

*

*

CXCR5+

PD

-1

ICOS+

PD-1hi

ICOS-PD-1int

ICOS-PD-1low

Tfh TIL function in BC

B cell zone T cell zone

CD20/CD4/CD8/PD-1

CD8/CD4/PD-1

Gate

: C

D8

+T

cells

Tumor microenvironment

Lymphatic systemT cells Priming

Local Immunoglobulin factory

Recruitment of naïve T cells Local T cell priming

Potentiation of CD8+ TIL function

BC tumor

TNFα

INFγ

TGFb

Conclusion and perspectives

TLS

Conclusion and perspectives K

i67/C

D2

0IH

C

Tumor with active TLS

Tumor area

Tumor area

Tumor with non active TLS

TN BC TUMOR

Tumor microenvironment

Lymphatic systemT cells Priming

Local Immunoglobulin factory

Recruitment of naïve T cells Local T cell priming

Potentiation of CD8+ TIL function

BC tumor

TNFα

INFγ

TGFb

Conclusion and perspectives

Memory response for long term survival after removal of tumor

TLS

MSI

TMB

PD-1 PDL-1 status

Improve the selection of patients

Improve number of responders patients

Immune biomarkers landscape

Conclusion and perspectives

Treg/CD8

Immune organization

TIL density

Circulating DNA

Tumor metabolism

Tumor inflammation

signature

Improve combo therapy choice

Institut Jules Bordet, BRUXELLES

Molecular Immunology Unit

Dr Karen Willard-Gallo

Dr Gregory Noël

Dr Soizic Garaud

Dr Pushpamali De Silva

Anaïs Boisson

Hugues Duvillier

Celine Naveaux

Laurence van Schoonwinkel

Dr Noemie Thomas

Hélène Strainchamps

Medical Oncology Department

Prof Martine Piccart

Institut Jules Bordet, BRUXELLES

Anatomical Pathology Department

Dr Roland De Wind

Prof Denis Larsimont

Dr Ligia Craciun

Sint Augustinus, GZA ANTWERP

Anatomical Pathology Department

Dr Gert Van den Eynden

Acknowledgments

Funding:

Many others

… and our patients!