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- Title
Detection of HIV-1 Neutralizing Antibodies in a Human CD4<sup>+</sup>/CXCR4<sup>+</sup>/CCR5<sup>+</sup> T-Lymphoblastoid Cell Assay System.
- Authors
McLinden, Robert J.; LaBranche, Celia C.; Chenine, Agnès-Laurence; Polonis, Victoria R.; Eller, Michael A.; Wieczorek, Lindsay; Ochsenbauer, Christina; Kappes, John C.; Perfetto, Stephen; Montefiori, David C.; Michael, Nelson L.; Kim, Jerome H.
- Abstract
Sensitive assays are needed to meaningfully assess low levels of neutralizing antibodies (NAbs) that may be important for protection against the acquisition of HIV-1 infection in vaccine recipients. The current assay of choice uses a non-lymphoid cell line (TZM-bl) that may lack sensitivity owing to over expression of CD4 and CCR5. We used transfection of a human CD4+/CXCR4+/α4β7+ T-lymphoblastoid cell line (A3.01) with a CMV IE promoter-driven CCR5neo vector to stably express CCR5. The resulting line, designated A3R5, is permissive to a wide range of CCR5-tropic circulating strains of HIV-1, including HIV-1 molecular clones containing a Tat-inducible Renilla luciferase reporter gene and expressing multiple Env subtypes. Flow cytometric analysis found CCR5 surface expression on A3R5 cells to be markedly less than TZM-bl but similar to CD3.8 stimulated PBMC. More importantly, neutralization mediated by a diverse panel of monoclonal antibodies, HIV-1 positive polyclonal sera and sCD4 was consistently greater in A3R5 compared to TZM-bl cells. The A3R5 cell line provides a novel approach to guide the development and qualification of promising new HIV-1 vaccine immunogens.
- Subjects
IMMUNOGLOBULINS; CD4 antigen; CXCR4 receptors; LYMPHOBLASTOID cell lines; BIOLOGICAL assay; ANTIVIRAL agents
- Publication
PLoS ONE, 2013, Vol 8, Issue 11, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0077756