Abstract
It has been realized that nitric oxide (NO) plays several regulatory roles in vivo. Despite very beneficial effects, NO has also been implicated as a deleterious agent in various pathophysiological conditions. In this study, we estimated immune response by using CD4/CD8 for NHCEC with added nitrate or nitrite ions. The growth medium of NHCEC was prepared by combining one part of Dullbecco-modified Eagle's medium and one part of Ham's nutrient mixture. The medium was further supplemented with cholera toxin, insulin, transferrin, epidermal growth factor, hydrocortisone, penicillin, streptomycin, gentamicin, HEPES and fetal bovine serum 10%FBS. From this three cultures were made. To the first, nitric acid was added. To the second, nitrous acid was added. The third was a control. The NHCEC's suspension was placed into centrifuged tubes and FITC-attached anti-CD4 antibodies and R-PE-attached anti-CD8 antibodies were added. The tubes were incubated for 30 minutes on ice in a dark room. The samples were washed once in phosphate buffered saline (PBS). The cell sediment was resuspended in paraformaldehyde in PBS. And then an immune response to NHCEC was estimated by comparing the fluorescence area ratio CD4/CD8. We could find the effect of nitrate ion and nitrite ion by CD4/CD8. This study is a first step in the investigation immune response using the fluorescent antibody method.
