As described in their previous paper, the authors used ViroMag to improve infection and transduction of HIV virus in primary human CD8+ T lymphocytes
This paper demonstrate the efficiency of ViroMag from OZ Biosciences to highly improve transduction even on cells cultured in suspension. Classical protocol was optimized and adapted by Sacha JB as previously described in Nature Protocol 2010.article reference: J Virol. 2013 Mar;87(5):2628-38
Early Antigen Presentation of Protective HIV-1 KF11Gag and KK10Gag Epitopes from Incoming Viral Particles Facilitates Rapid Recognition of Infected Cells by Specific CD8+ T Cells.Kløverpris HN, Payne RP, Sacha JB, Rasaiyaah JT, Chen F, Takiguchi M, Yang OO, Towers GJ, Goulder P, Prado JG.
Abstract
CD8(+) T cells are major players in antiviral immunity against human immunodeficiency virus type 1 (HIV-1) through recognition of viral epitopes presented on the surface of infected cells. However, the early events involving HIV-1 epitope presentation
to CD8(+) T cells remain poorly understood but are nonetheless crucial
for the rapid clearance of virus-infected cells. Here, we
comprehensively studied the kinetics of antigen presentation of two protective
epitopes, KF11Gag and KK10Gag, restricted by HLA alleles B*57:01 and
B*27:05, respectively, and compared these to KY9Pol and VL9Vpr epitopes
in a single cycle of HIV-1 replication. We consistently demonstrate differences in epitope presentation kinetics, with very early presentation, within 3 h postinfection, for the protective KF11Gag, KK10Gag epitopes, and KY9Pol but only late presentation for VL9Vpr. We show that this early presentation relies on the antigen
being presented from incoming viral particles and is correlated with
rapid CD8(+) T cell activation and clearance of virus-infected cells.
Additionally, our data indicate a dose-response dependency between the
levels of CD8(+) T cell activation and the amount of virus inoculum.
These data reflect a proof of principle emphasizing the importance of
identifying early-presented viral epitopes for rapid elimination of HIV-1-infected cells.
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