1. Marie Curie-COFUND fellow
Myeloid-Derived Suppressor Cells as A Potential Therapuetic Target For Women With Early Pregnancy Loss
Rohini R Nair
Marie Curie-COFUND fellow
Division of Genetic and Cell Biology
Chromatin Dynamics lab
University of San Raffaele, Milan, Italy

2. Inflammation
Acute Inflammation
Chronic Inflammation

3. I – Implantation, G – Gestation, D - Delivery
Inflammatory State
Quiescent State
Inflammatory state during pregnancy I G D
I – Implantation, G – Gestation, D - Delivery

4. Prognosis of pregnancy
Teklenburg et al., 2010

5. S100A8 & S100A9

6. Role of S100A8 & S100A9 pregnancy loss
In Endometrium
Maternal tissue
In Chorionic villi
Fetal tissue
Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) of S100A8 & S100A9 in endometrial deciduas and chorionic villi

7. S100A8
S100A9
Quantitative real time PCR of S100A8 & S100A9 in endometrial deciduas and chorionic villi
S100A8
S100A9
Western blot analysis of S100A8 & S100A9 in endometrial deciduas and chorionic villi

8. S100A8 cytokine levels in peripheral blood
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).

9. Myeloid derived Suppressor Cells

10. Level of Myeloid derived suppressor cells (MDSC) in menstrual cycle and pregnancy
Mean Fluorescence Intensity
of Cd11b
21st day of LMP
7th day of LMP
26.48
0.07
11.10
23.05
11.20
0.14
CD11b
CD33
% CD11b+CD33+ cells in blood 15 10 5
1st Trimester
2nd Trimester
3rd Trimester
21st day
7th day
17.5
18.5
0.87 12 17
0.30 4
5.83
***

11. Tolerance of Embryo at Implantation stage
MDSC
MDSC
21st Day
MDSC
Uterine epithelium
MDSC
MDSC
MDSC
MDSC
MDSC
MDSC
MDSC
Treg
MDSC
7th Day
1st trimester
MDSC
3rd trimester
Treg
Treg
Tolerance of Embryo at Implantation stage
Treg
Loss of Tolerance i.e.
Parturition

12. Level and suppressive function of MDSC in early pregnancy loss
Pregnant Females in 1st trimester
17.5
18.5
0.87
Early Pregnancy Loss patients
4.32
20.77
0.23
CD11b
CD33
% CD11b+CD33+ cells in blood
Controls
EPL 15 10 5 **
Absorbance (570nm)
Undepleted
CD33 depleted 15 10 5 20 25 3 2 1 4 5
IL-2 (pg/ml)
Undepleted
CD33 depleted * *
Controls
EPL
0.4
0.2
0.0
0.6
undepleted / CD33 depleted
IL-2 (pg/ml)
0.8
Absorbance (570nm) *

13. MDSC level in Implantation Site
Controls
EPL
CD33+ cells in endometrium
Average No of CD33+ cells
Controls
EPL 15 10 5 **
Reduced CD11b+CD33+ MDSC correlate negatively with the increases S100A8 level in peripheral blood of Recurrent Early Pregnancy loss (RPL) patients.

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

15. Acknowledgement
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.

16. Reproductive Genetic Group
Thank You
Chromatin Dynamic Group