Abstract
Neutralizing antibody (NAb) assays for human immunodeficiency virus (HIV) are used to study the immune response in infected individuals, to examine monoclonal antibodies and viral diversity, and to judge the potential value of candidate vaccine immunogens in preclinical and clinical trials. An important aspect of these efforts is an ability to achieve and document equivalent assay performance across multiple laboratories. Recent advances in assay technology have led to major improvements in how HIV NAbs are measured. Stable cell lines containing HIV Tat-regulated reporter genes are now available that permit rapid, sensitive and reproducible measurements of virus neutralization after a single round of infection in a high throughput format.Moreover, these assays may be used with molecularly cloned Env-pseudotyped viruses for greater reagent stability and traceability.A luciferase (Luc) reporter gene assay performed in TZM-bl (JC53bl-13) cells was recently optimized and many of its performance parameters have been validated. This assay has become the main endpoint neutralization assay used by the NIH-sponsored HIV Vaccine Trials Network and by a growing number of laboratories worldwide.
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Acknowledgments
The author thanks Dr. Marcella Sarzotti-Kelsoe for guidance in optimizing and validating the assay, and Kelli Greene for her critical review of the document. He also thanks Drs. George Shaw and John Kappes for sharing their reagents. This work was supported by NIH grants AI30034 and AI46705.
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Montefiori, D.C. (2009). Measuring HIV Neutralization in a Luciferase Reporter Gene Assay. In: Prasad, V.R., Kalpana, G.V. (eds) HIV Protocols. Methods In Molecular Biology™, vol 485. Humana Press. https://doi.org/10.1007/978-1-59745-170-3_26
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DOI: https://doi.org/10.1007/978-1-59745-170-3_26
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