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Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
Published on: September 14, 2014
Isolation and characterization of an HIV-1 envelope glycoprotein-specific B-cell from an immortalized human naïve
Zehua Sun1,2,3, Shiqiang Lu1,4, Zheng Yang5
1AIDS Institute, Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong, PR China.
Abstract:
With the recent development of single B-cell cloning techniques, an increasing number of human immunodeficiency virus type 1 (HIV-1)-specific broadly neutralizing antibodies have been isolated since 2009. However, knowledge regarding HIV-1-specific B cells in vivo is limited. In this study, an HIV-1-specific B-cell line was established using healthy PBMC donors by the highly efficient Epstein-Barr virus transformation method to generate immortalized human naïve B-cell libraries. The enrichment of HIV-1 envelope-specific B cells was observed after four rounds of cell panning with the HIV-1 envelope glycoprotein. An HIV-1 envelope-specific stable B-cell line (LCL-P4) was generated. Although this cell line acquired a lymphoblastic phenotype, no expression was observed for activation-induced cytidine deaminase, an enzyme responsible for initiating somatic hypermutation and class switch recombination in B cells. This study describes a method that enables fast isolation of HIV-1-specific B cells, and this approach may extend to isolating other B-cell-specific antigens for further experiments.
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