The immune system is made up of a diverse set of cells that have specific roles for destroying foreign agents that enter the body and connect to various other systems and functions to support overall health, Researchers and physicians are interested in immune stimulation to enable the immune system to fight illness, disease, and infection on its own. Immune stimulation is defined by an increase in proinflammatory cells and cell types relating to the immune system. This project seeks to examine immune stimulation and the intersection between oncology and immunology to understand if a natural herb, echinacea, can increase immune cells. Two reputable cancer foundations, Memorial Sloan Kettering Cancer Center and Susan G. Komen Breast Cancer Foundation, write that echinacea improves a low white blood cell count related to chemoradiotherapy. There is little to no data supporting that echinacea increases all immune cells. In my research, hemocytes, insect immune cells analogous to mammalian immune cells, are counted after echinacea exposure to quantify the immunostimulatory properties of the herb. The purpose is to understand if echinacea at concentrations safe for human supplementation increases immune cell count in healthy organisms. It is expected that echinacea will increase the number of hemocytes because of the speculated immunostimulatory properties that have been researched in vitro.

D.melanogaster are used in this study because they are widely studied and contain a small amount of hemolymph that can be extracted and used to count hemocytes.

To introduce echinacea, Gaia Herbs' “Echinacea Supreme" tincture, containing various species and parts of the echinacea genus, alcohol, and water, is introduced to the D. melanogaster in the media until the third instar larvae stage when data is collected. The concentration introduced represents 5% of the daily recommended dosage from Gaia Herbs after accounting for food intake and mass of D.melanogaster, Higher concentrations of echinacea caused mortality. Equal amounts of ethanol in the tincture are introduced to the media of a control culture to standardize culture conditions across all D. melanogaster populations.

To collect hemolymph, larvae are pierced with a lancet, placed in a pierced PCR tube inside an Eppendorf tube, and centrifuged. The hemolymph containing hemocytes drains into the Eppendorf tube. Then 10uL of the hemolymph solution is placed on a glass slide, incubated, fixed in methanol, and stained with Giemsa's blood stain. The immune cells are then counted using a microscope at 40X magnification.

The control group had 30 hemocytes/10 uL of hemolymph solution, the ethanol group had 24 hemocytes/10 L of hemolymph solution, and the echinacea group had 25 hemocytes/10 uL of hemolymph solution. The control group averaged five trials, ethanol averaged three, and echinacea averaged four. One way ANOVA determined that the decrease was insignificant (0.36<P<0.98). No immunostimulatory properties were found in the echinacea group. The data collected showed essentially the same results for the echinacea and ethanol groups, meaning echinacea in the tincture used had no effect on the hemocyte count. This data accepts the null hypothesis.