NITRIC OXIDE SYNTHESIS FROM ACTIVATED MONOCYTES INDUCED BY FRACTIONATED SOLUBLE (CYTOPLASMIC) ANTIGEN OF B. MELITENSIS 16M IN CELL CULTURE.

Abstract

The present research was designed to investigate the production of nitric oxide by monocytes in cell culture induced by different fractionated soluble antigen of Brucella melitensis 16M. For this purpose soluble antigen of B. melitensis was prepared by treating the culture with ultrasonic waves in an ultrasonicator. This antigen was then subjected to gel filtration chromatography to fractionate the antigen according to its approximate molecular weight. Nitrite and other reactive intermediates have been postulated to participate in arginine dependent tumor cytotoxicity by activated macrophages, supernatant were collected after 48 hr. of culture assayed for the presence of these molecules in a standard Griess reaction. This investigation revealed that fractionated antigen SAG3 produced maximum nitrite followed by SAG1, SAG2, SAG5, SAG4 and SAG6 respectively. Therefore, it was observed that NO play an important role in killing of B. melitensis and it had been concluded that much of the antimicrobial activity of macrophages against B. melitensis is due to nitric oxide and substances derived from it.