Measuring HIV neutralization in a luciferase reporter gene assay

DC Montefiori - HIV protocols, 2009 - Springer
HIV protocols, 2009Springer
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 …
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.
Springer