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Medicinal plant products are generally used as immunomodulatory (stimulatory/suppressive/adjuvant) agents for curing various ailments. These activities could be due to the presence of primary and secondary metabolites as already described in the literature. In the present study, we focused on those proteases isolated from the flowers of Caesalpinia pulcherrima and Phanera variegata against specific protein antigen i.e. Bovine serum albumin (BSA) on human diabetic blood samples using flow cytometry. For these studies, determination of protein (Nanodrop) and protease (spectropho-tometer) content from the flowers of Caesalpinia pulcherrima and Phanera variegata using phosphate buffered saline (PBS, pH 3 to 9). Out of these, the most active protease treated with diabetic human blood samples (n=6; fasting glucose level, >180) using diluted and concentrated form of protease (100 µl) and determined its blood counts in the form of lymphocytes, monocytes and granulocytes count and also analysed its forward and side scatter of the cell. The results showed that acid protease (pH 5) showed enhancement in granulocytes count but sudden decline in monocytes count as compared to control. Similarly, there is enhancement in both forward and side scatter after treating with acid protease of Caesalpinia pulcherrima and Phanera variegata. Overall, flow cytometric results revealed that acid protease (pH 5) from the flowers of Caesalpinia pulcherrima and Phanera variegata showed antidiabetic property in human blood samples.

Flow cytometry is a technology that measures simultaneously both optical and fluorescence properties of single cells or any other particle including cell organelles (i.e. nuclei, chromosome etc.), microorganisms, and latex beads. This technology also exploits bright beams of light (usually from laser) to directly disclose various aspects i.e. size, granularity (internal complexity of cell) including relative fluorescence intensity of cells by the way of light is scattered or indirectly by coupling wth fluorescent probes to cell components or suspended particle. Some components like dichroic mirror, optical filter, photomultiplier tube, detectors etc. that played a crucial role in flow cytometry. Therefore, light scattering propertiesand measurement of fluorescence through flow cytometry is widely used in life science laboratories including preclinical and clinical research for many applications such as immunological, entomological, pharmaceutical etc. This review provides information about the optical system that are used in flow cytometry and showed various biological applications.

There are number of medicinal plant products (leaves, root, stem bark, seeds etc.) including its metabolites i.e. primary (protein) and secondary (flavonoids, terpenoids, glycosides etc.) that have been used as immunomodulatory (stimulatory, suppressive or adjuvant) agents. For the last several years, researchers focused on various immunopharmacological aspects especially its therapeutic action of these medicinal plants against various diseases. The objective of our study was to identify and evaluate the action of aqueous leaves extract from these medicinal plants i.e.Azadirachta indica, Syzygium cuminiandProsopis spicigeraon the development of spleen cells (ex vivocellular proliferation assay) from mice using hepatitis B vaccine containing surface antigen (HBsAg; 20 µg/ml) and also estimated its pro-inflammatory (IFN-gamma and TNF alpha) cytokines from spleen cell culture supernatant. For these studies, mice were immunized subcutaneously with HBsAg on day 0. After three days, spleens were excised aseptically and prepared single cell suspension (106cells/ml) using variable doses of aqueous leaves extract (0.625-5 mg; 50 µl) incubated with or without HBsAg. After 48 h, cell culture supernatant was collected for the estimation of Th1 (IFN-gamma and TNF alpha) cytokines. However, cell proliferation was revealed by blue tetrazolium reduction assay (MTT) and cytokines by ELISA. The results showed these aqueous leaves extracts showed dose-dependent reduction of cell proliferation including pro-inflammatory cytokines. Overall, these medicinal plants showed immunosuppressive activity inex vivomurine model based studies.

The objective of our present study is to investigate the immunosuppressive activity of variable doses of crude flavonoids (6.25-100 mg/ml; 50 µl) extracted from the leaves of Mesua ferrea, Ficus benghalensis and Jasminum auriculatum on human whole blood using specific antigen (i.e. hepatitis B vaccine containing surface antigen, 20 µg/ml; 10 µl) is to estimate its blood counts, CD14 monocyte marker and observed its cytotoxicity. The results showed that the crude flavonoids extracted from these three medicinal plants showed maximum inhibition of blood counts, CD14 monocyte marker and cytotoxicity at higher doses. Overall, the data suggests that crude flavonoids extracted from these medicinal plants i.e. Mesua ferrea, Ficus benghalensis and Jasminum auriculatum showed immunosuppressive activity against HBsAg.