AUTHOR=Boswell Kristin L. , Watkins Timothy A. , Cale Evan M. , Samsel Jakob , Andrews Sarah F. , Ambrozak David R. , Driscoll Jefferson I. , Messina Michael A. , Narpala Sandeep , Hopp Christine S. , Cagigi Alberto , Casazza Joseph P. , Yamamoto Takuya , Zhou Tongqing , Schief William R. , Crompton Peter D. , Ledgerwood Julie E. , Connors Mark , Gama Lucio , Kwong Peter D. , McDermott Adrian , Mascola John R. , Koup Richard A. TITLE=Application of B cell immortalization for the isolation of antibodies and B cell clones from vaccine and infection settings JOURNAL=Frontiers in Immunology VOLUME=13 YEAR=2022 URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.1087018 DOI=10.3389/fimmu.2022.1087018 ISSN=1664-3224 ABSTRACT=

The isolation and characterization of neutralizing antibodies from infection and vaccine settings informs future vaccine design, and methodologies that streamline the isolation of antibodies and the generation of B cell clones are of great interest. Retroviral transduction to express Bcl-6 and Bcl-xL and transform primary B cells has been shown to promote long-term B cell survival and antibody secretion in vitro, and can be used to isolate antibodies from memory B cells. However, application of this methodology to B cell subsets from different tissues and B cells from chronically infected individuals has not been well characterized. Here, we characterize Bcl-6/Bcl-xL B cell immortalization across multiple tissue types and B cell subsets in healthy and HIV-1 infected individuals, as well as individuals recovering from malaria. In healthy individuals, naïve and memory B cell subsets from PBMCs and tonsil tissue transformed with similar efficiencies, and displayed similar characteristics with respect to their longevity and immunoglobulin secretion. In HIV-1-viremic individuals or in individuals with recent malaria infections, the exhausted CD27-CD21- memory B cells transformed with lower efficiency, but the transformed B cells expanded and secreted IgG with similar efficiency. Importantly, we show that this methodology can be used to isolate broadly neutralizing antibodies from HIV-infected individuals. Overall, we demonstrate that Bcl-6/Bcl-xL B cell immortalization can be used to isolate antibodies and generate B cell clones from different B cell populations, albeit with varying efficiencies.