Wednesday, August 7, 2019
Critical Essay Example | Topics and Well Written Essays - 1000 words
Critical - Essay Example They established a system to investigate the membrane dynamics of the events occurring at the interface of HIV-1 infected and receptor expressing T cells. The authors selected appropriate CD4+/CXCR4+ T cell lines for the study and maintained them in an antibiotic supplemented cell growth medium and established the purity of the cell lines at a level greater than 90% by flow cytometry by indirect immunofluorescence. These CD4+ cells were labeled as target cells. Jurkat CE 6.1 cells infected with HIV-1 strain LAI were used as effector cells. After phenotyping the cells for surface Env and CD4 expression, the effector and the target cells were mixed in equal quantities on cover slips, with or without inclusion of mAb (monoclonal antibody). For specific time intervals after which they were fixed and stained. Kinetic studies were conducted in separate experiments by immunostaining of conjugates for specific mAbs. Appropriate software and methods for confocal microscopy and photography wer e employed. Inhibition of cytoskeletal rearrangement and signaling were studied in separate experiments. Cell-cell fusion assay and transmission electron microscopy were the other experiments conducted. The authors have been able to develop a novel system to study the cell to cell dissemination of HIV-1 by demonstrating a close packing and concentration of the virus particles in the plasma membranes of both effector and target cells, though they could not actually demonstrate the formation of a synapse between the two cells, which they suggest is the most likely mechanism. The role of an actin dependent mechanism in the Env-dependent recruitment of CD4, CXCR4, and LFA-1 has successfully been demonstrated. This cytoskeleton dependent receptor movement during infection of the target cells along with formation of an adhesive junction has been proposed as the likely
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