Rhesus macaque Bcl-6/Bcl-xL B cell immortalization: Discovery of HIV-1 neutralizing antibodies from lymph node

Authors

Jakob Samsel, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America; Institute for Biomedical Sciences, George Washington University, Washington, D.C., United States of America. Electronic address: jakob.samsel@mail.nih.gov.
Kristin L. Boswell, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.
Timothy A. Watkins, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.
David R. Ambrozak, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.
Rosemarie Mason, ImmunoTechnology Section, VRC; Humoral Immunology Section, VRC.
Takuya Yamamoto, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.
Sungyoul Ko, ILAb Co., Ltd.; College of Pharmacy, The Catholic University of Korea, South Korea.
Yongping Yang, Structural Biology Section, VRC.
Tongqing Zhou, Structural Biology Section, VRC.
Nicole A. Doria-Rose, Humoral Immunology Section, VRC.
Kathryn E. Foulds, Nonhuman Primate Immunogenicity Core, VRC.
Mario Roederer, ImmunoTechnology Section, VRC.
John R. Mascola, Humoral Immunology Section, VRC.
Peter D. Kwong, Structural Biology Section, VRC.
Lucio Gama, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.
Richard A. Koup, Immunology Laboratory, Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD, United States of America.

Document Type

Journal Article

Publication Date

2-25-2023

Journal

Journal of immunological methods

DOI

10.1016/j.jim.2023.113445

Keywords

Antibody discovery; B cell immortalization; HIV; Rhesus macaque; Vaccination

Abstract

Many HIV-1 vaccines are designed to elicit neutralizing antibodies, and pre-clinical testing is often carried out in rhesus macaques (RMs). We have therefore adapted a method of B cell immortalization for use with RM B cells. In this system, RM B cells are activated with CD40 ligand and RM IL-21 before transduction with a retroviral vector encoding Bcl-6, Bcl-xL, and green fluorescent protein. Importantly, RM B cells from lymph nodes are more effectively immortalized by this method than B cells from PBMC, a difference not seen in humans. We suggest the discrepancy between these two tissues is due to increased expression of CD40 on RM lymph node B cells. Immortalized RM B cells expand long-term, undergo minimal somatic hypermutation, express surface B cell receptor, and secrete antibodies into culture. This allows for the identification of cells based on antigen specificity and/or functional assays. Here, we show the characterization of this system and its application for the isolation of HIV-1 neutralizing antibodies from a SHIV.CH505-infected animal, both with and without antigen probe. Taken together, we show that Bcl-6/xL immortalization is a valuable and flexible tool for antibody discovery in RMs, but with important distinctions from application of the system in human cells.

APA Citation

Samsel, Jakob; Boswell, Kristin L.; Watkins, Timothy A.; Ambrozak, David R.; Mason, Rosemarie; Yamamoto, Takuya; Ko, Sungyoul; Yang, Yongping; Zhou, Tongqing; Doria-Rose, Nicole A.; Foulds, Kathryn E.; Roederer, Mario; Mascola, John R.; Kwong, Peter D.; Gama, Lucio; and Koup, Richard A., "Rhesus macaque Bcl-6/Bcl-xL B cell immortalization: Discovery of HIV-1 neutralizing antibodies from lymph node" (2023). GW Authored Works. Paper 2353.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/2353

Department

School of Medicine and Health Sciences Student Works