1. M.Boothapandi Research Scholar Department of Zoology University of Madras Chennai Prof. R. Ramanibai Research Supervisor & Guide Department of Zoology University of Madras Chennai

2. Immunomodulators are drugs which either suppress or stimulate the immune system – (Immunosuppressants and Immunostimulants) INTRODUCTION Immunostimulants Chemical agents - levomisole, Cimetidine, Isoprinosine(ISO) Microbiological agents- BCG ( bacille Calmette-Guerin,) Indication- Tumor, Immunodeficiency, Infectional diseases Immunosuppressant Chemical agents- Cyclophosphamide, Azathioprine Hormone- steroids Microbiological agents- Cyclosporine A(CsA) Indication- Organ transplantation, Hypersensitivity diseases, Autoimmune diseases, Infectious diseases

4. Antiproliferative activity in human lung cancer (Kameswaran and Ramanibai, 2008) Indirubin- Anticancer drug (Han, 1994) Indigotin- Hepatoprotective activity (Singh et al., 2001) Root and leaves-To treat epilepsy and hydrophobia (Tyagi et al., 2010) Medicinal uses of I.tinctoria

5. OBJECTIVES 1.To evaluate the effect of ethanolic and aqueous extract of I.tinctoria on in vitro Macrophage functions. 2.To estimate the effect of ethanolic and aqueous extract of I.tinctoria on in vitro lymphocyte proliferation

6. Kingdom :Plantae Division : Magnoliophyta Class : Magnoliopsida Order : Fabales Family : Fabaceae Genus : Indigofera Species : tinctoria linn. Systematic position of the plant

7. MATERIALS AND METHODS 1.Extract preparation – Aqueous extract and ethanolic extract were prepared in respective solvents 2.Macrophage isolation from peritoneal fluid and lymphocytes isolated from Peripheral blood (Cheng et al., 2008) of wistar albino rat. 3.Nitric Oxide (NO) production (Green et al., 1982) 4.Arginase activity (Munder et al., 1998) 5.Pinocytic activity (Plytycz et al., 1992) 6.Lysosomal activity (Suzuki et al., 1990) 7.Phagocytic assay on nitroblue tetrazolium (NBT) reduction Test (Rainard, 1986) 8.Lymphocytes proliferation (Mossmann, 1983)

8. RESULTS

9. Field Emission- Scaning Electron microscope (FE-SEM) image of isolated (A) peripheral blood lyphocytes and (B) peritoneal macrophage (PEM) from wistar albino rat A B

10. Effect of I.tinctoria extracts on PEM Nitric Oxide (NO) Production and Arginase activity

11. Effect of I.tinctoria extracts on PEM Pinocytic Activity

12. Effect of I.tinctoria extracts on PEM Lysosomal Activity

13. Effect of I.tinctoria extracts on PEM Phagocytosis

14. Effect of I.tinctoria extracts on invitro proliferation

15. Effect of I.tinctoria extracts on invitro proinflammatory cytokine production

16. SUMMARY  In innate immune system both aqueous ethanolic and aqueous extracts were enhanced the Macrophage NO production and simultaneously decrease the activity of Arginase.  The other predominant activity of PEM like Pinocytic and lysosomal activity also substantially enhanced by both ethanolic and aqueous extracts  The principal role of Macrophage phagocytosis also notably enhanced.  On the other hand the it enhance the adaptive immune system through the proliferation of lymphocytes.

17. OUTCOME  The present findings revealed that both the ethanolic and aqueous extracts of I. tinctoria did not cause direct toxic effects on isolated immune cells and suggested that they could potentially enhance innate as well as adaptive immune responses.  These strong immunomodulatory activities of I. tinctoria noted here in vitro help to explain some of the medicinal benefits attributed to the plant in traditional medicine and could be used to provide a future basis for the development of this plant as an immuno-regulating substance.

18. Future Plan of the work....  To find out the active compounds responsible for the immunomodulatory potential of I.tinctoria and find out the immunomodulatory effect through invivo studies on Wistar albino rat model.

19. ACKNOWLEDGEMENT We acknowledge to UGC- UPE – PHASE II New Delhi (No. 2013/PFEP/C3/280) for financial support.

20. THANK YOU