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A study on cell mediated immunity to rabies virus: lymphocyte-mediated cytotoxicity in vitro


A study on cell mediated immunity to rabies virus: lymphocyte-mediated cytotoxicity in vitro



Indian Journal of Medical Research 64(10): 1414-1420



ISSN/ISBN: 0971-5916

PMID: 1017854

An attempt was made to demonstrate lymphocyte-mediated cytotoxicity as an index of cell-mediated immunity, using sensitized rabbit lymphocytes as effector cells and rabies virus infected BHK-21 (baby hamster kidney tissue culture) cells as target cells. A carrier state of rabies virus infection was established in BHK-21 cells. At the 78th passage of the infected cell line, 90% of the cells in the monolayer showed specific fluorescence of rabies virus antigen in the cytoplasm. Rabbits were sensitized by 1 s.c. inoculation of 125,915 mouse LD50 dose of live avianized rabies virus. The spleen lymphocytes from the sensitized animals were harvested on 2nd, 4th, 6th, 8th 10th, 13th and 16th post-sensitization (PS) days and were used in lymphocyte cytotoxicity assay. To the infected BHK-21 cells, which were labeled with 51Cr (0.75 .mu.Ci/5 .times. 104 cells), sensitized lymphocytes of different PS days were added (at a ratio of 1 target cell:100 lymphocytes) and were incubated for 18 h at 37.degree. C. Control preparations were also made. The amount of 51Cr released from target cells increased from 37% when treated with sensitized lympocytes obtained on the 2nd PS day, to 93% when treated with sensitized lymphocytes obtained on the 6th PS day. Thereafter, the Cr release dropped to a level of 20-21%, when tested with lymphocytes obtained on the 13th or 16th PS day. When unsensitized lymphocytes or MEM [minimal essential media] with 10% fetal calf serum was added to 51Cr labeled infected BHK-21 cells, the release of 51Cr ranged from 22-20%. Maximum sensitization of lymphocytes was obtained on the 6th PS day, as indicated by effective killing of rabies infected BHK-21 cells. The minimum level of antibody to rabies virus was detected on the 10th PS day which persisted up to the 16th PS day.

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