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Effect of immunomodulating agent on intracellular killing and phagocytosis of polymorpho nuclear leukocytes (PMNL) Singh Vinod1, Das Siddartha Kumar2 1Department of Applied Microbilogy and Laboratory Medicine, College of Life sciences, Cancer Hospital and Research Institute, Gwalior - 474001, India. 2Clinical Immunology and Rheumatology Unit, Department of Medicine, King George's Medical College, Lucknow - 22600, India. Abstract The in vitro effect of immunomodulating agent methotrexate on phagocytosis and intracellular killing of E. coli by circulating human polymorphonuclear leukocytes (PMNL) had been assessed. PMNL were isolated from the whole blood of ten human healthy volunteers by density gradient centrifugation and incubated with 10−3 M methotrexate. E. coli was opsonized in human serum and added to the prepared PMNL. Phagocytosis was determined by serial dilutions plating of nonphagocytized bacteria. After the treatment with lysostaphin and trypsin intracellular killing of bacteria was determined at set intervals by serial dilutions and plating of viable bacteria released by PMNL lysis. Methotrexate treated PMNL phagocytized 56% and control PMNL 67% of bacteria in 15 minutes (p>0.5) and 96% of ingested bacteria were killed in 15 minutes by both populations. In these experiments the in vitro incubation of circulating PMNL with methotrexate produced no significant alterations in PMNL phagocytosis or intracellular killing of E. coli. Top Key words Immunomodulating Agent, Phagocytosis, PMNL. Top | | |
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