Cell Proteins Interacting with the Human Immunodeficiency Virus in Immunoblotting can be Detected by R5- or X4- Tropic Human Immunodeficiency Virus Particles

Zeina Soayfane; Bilal Houshaymi; Mamdouh H Kedees; Laurent Belec; Nadine Nasreddine

doi:10.4103/ijabmr.IJABMR_398_18

Cell Proteins Interacting with the Human Immunodeficiency Virus in Immunoblotting can be Detected by R5- or X4- Tropic Human Immunodeficiency Virus Particles

Int J Appl Basic Med Res. 2020 Apr-Jun;10(2):81-85. doi: 10.4103/ijabmr.IJABMR_398_18. Epub 2020 Apr 2.

Authors

Zeina Soayfane¹, Bilal Houshaymi², Mamdouh H Kedees³, Laurent Belec⁴, Nadine Nasreddine²

Affiliations

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, and University of Paris Descartes, Paris, France.

Abstract

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-1_JRCSF and the X4-tropic HIV-1_NDK.

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.

Keywords: Free human immunodeficiency virus particles; gp160; human immunodeficiency virus; western blot.