Division of Genetic and Cell BiologyChromatin Dynamics lab

University of San Raffaele, Milan, Italy

Rohini R NairMarie Curie-COFUND fellow

Myeloid-Derived Suppressor Cells as A Potential Therapuetic Target For Women With Early Pregnancy Loss

Inflammation

Acute Inflammation

Chronic Inflammation

Inflammatory State

Quiescent State

Inflammatory state during pregnancy

I G D

I – Implantation, G – Gestation, D - Delivery

No implantation 30%

delivery 30%

Miscarriage 10%

Biochemical miscarriage 30%

Prognosis of pregnancy

Teklenburg et al., 2010

S100A8 & S100A9

Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) of S100A8 & S100A9 in endometrial deciduas and chorionic villi

In EndometriumMaternal tissue

In Chorionic villiFetal tissue

Role of S100A8 & S100A9 pregnancy loss

S100A8 S100A9

Quantitative real time PCR of S100A8 & S100A9 in endometrial deciduas and chorionic villi

Western blot analysis of S100A8 & S100A9 in endometrial deciduas and chorionic villi

S100A8

S100A9

S100A8 level remain high during 7th day of menstrual cycle compared to 21st day of menstrual cycle. Serum from five healthy volunteers in proliferative (7th day LMP) and secretory phase (21st day LMP) of menstrual cycle were collected. S100A8 levels were analyzed through ELISA assay. Bar diagram representing S100A8 level (pg/ml) in proliferative (7th day of LMP) and secretory phase (21st day of LMP) of menstrual cycle (A). S100A8 levels did not change in 1st, 2nd and 3rd trimester of pregnancy. Bar diagram representing 1st (n=10), 2nd (n=7) and 3rd (n=5) trimester of pregnancy (B). S100A8 levels were found to be elevated in early pregnancy loss patients compared to healthy controls. Bar diagram representing controls (n=10) and EPL cases (n=10) * represents p<0.01 (C).

S100A8 cytokine levels in peripheral blood

Myeloid derived Suppressor Cells

21st day of LMP7th day of LMP

26.48

0.07

11.1023.05 11.20

0.14CD

11b

CD33

%

CD

11b+

CD

33+

cell

s in

blo

od

15

10

5

0

1st Trimester 2nd Trimester 3rd Trimester21st day

%

CD

11b+

CD

33+

cell

s in

blo

od

7th day 21st day

15

10

5

0

1st Trimester 2nd Trimester 3rd Trimester

17.518.5

0.87

1217

0.30

45.83

0.30

CD

11b

CD33

***

***

Level of Myeloid derived suppressor cells (MDSC) in menstrual cycle and pregnancy

P=0.02

Mean Fluorescence Intensity of Cd11b

Tolerance of Embryo at

Implantation stage

Uterine epithelium

7th Day

21st Day

1st trimester3rd trimester

Loss of Tolerance i.e.

Parturition

Treg

Treg

Treg

Treg

MDSC

MDSC

MDSC

MDSC

MDSC

MDSC

MDSC

MDSCMDSC

MDSC

MDSC

MDSC

%

CD

11b+

CD

33+

cell

s in

blo

od

Controls EPL

15

10

5

0

Abs

orba

nce

(570

nm)

Undepleted CD33 depleted

15

10

5

0

20

25

3

2

1

0

4

5

IL-2

(p

g/m

l)

Undepleted CD33 depleted

Controls EPL

0.4

0.2

0.0

0.6

und

eple

ted

/ CD

33

depl

eted

IL

-2 (

pg/m

l)

0.8

Controls EPL

0.4

0.2

0.0

0.6

und

eple

ted

/ CD

33

depl

eted

A

bsor

banc

e (5

70nm

)

Pregnant Females in 1st trimester

17.518.5

0.87

Early Pregnancy Loss patients

4.3220.77

0.23C

D11

b

CD33

**

* *

*

Level and suppressive function of MDSC in early pregnancy loss

Controls EPL

CD33+ cells in endometrium

Ave

rage

No

of

CD

33+ c

ells

Controls EPL

15

10

5

0

MDSC level in Implantation Site

**

Reduced CD11b+CD33+ MDSC correlate negatively with the increases S100A8 level in peripheral blood of Recurrent Early Pregnancy loss

(RPL) patients.

Myeloid-Derived Suppressor Cells can be used as a Potential Therapuetic Target For Women With Early

Miscarriage

Patients and volunteers for providing blood samples and tissue samples.

This work was financially supported by the Department of Biotechnology, New Delhi, India.

I would also acknowledge the help from DST-Confocal facility in BHU for confocal microscopy and Interdisciplinary School of Life Science for BD FACS calibur.

Acknowledgement

Thank You

Chromatin Dynamic Group

Reproductive Genetic Group