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ORIGINAL ARTICLE
Year : 2020  |  Volume : 10  |  Issue : 2  |  Page : 81-85

Cell proteins interacting with the human immunodeficiency virus in immunoblotting can be detected by R5- or X4- tropic human immunodeficiency virus particles


1 Department of Cell Biology, Faculty of Science, Lebanese University, Beirut, Lebanon
2 Department of Microbiology, Faculty of Health, Lebanese University, Beirut, Lebanon
3 Department of Cell Biology, State University of New York, New York, NY, USA
4 Virology Lab, Georges Pompidou European Hospital, University of Paris Descartes, Paris, France

Correspondence Address:
Nadine Nasreddine
Department of Microbiology, Faculty of Health, Lebanese University, Beirut
Lebanon
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijabmr.IJABMR_398_18

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Introduction: The present study reported a new immunoblot assay, with revelation by R5- or X4-whole free human immunodeficiency virus (HIV) particles or recombinant gp160. Materials and Methods: The assay was optimized to identify cell proteins interacting with HIV. Whole cell lysates were prepared from peripheral blood lymphocytes (PBLs), dendritic cells (DC), monocyte-derived macrophage (MDM), and Henrietta Lacks (Hela, wild-type or transfected with DC-specific intracellular adhesion molecule-3-Grabbing Non-Integrin, HeLa) and Human endometrial cells (HEC-1A) lines; HIV particles used were the R5-tropic HIV-1JRCSFand the X4-tropic HIV-1NDK. Results: Experiments with PBL lysates and both viruses demonstrated different bands, including a unique band at 105–117 kDa in addition to nonspecific bands. The 105–117 kDa band migrated at the same level of that observed in controls using total PBL lysate and anti-CD4 mAb for detection and thus likely corresponds to the cluster difference (CD) 4 complex. Blots using lysates of DCs, MDM, HeLa cell line, and HEC-1A cell line allowed identifying several bands that positions were similar to that seen by recombinant gp160 or whole R5- or X4-HIV particles. Conclusion: Blot of whole lysates of various HIV target cells is recognized by free HIV particles and allows identifying a wide range of HIV-interacting cell proteins. Such optimized assay could be useful to recognize new cellular HIV attachment proteins.


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