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Methods To better understand the requirements for activation of B cells producing bNAbs, we generated a series of cell lines and transgenic mice expressing such antibodies or selected germline-reverted versions as B cell surface receptors. The bNAb cell lines included those that recognize the CD4 binding site (b12, VRC01, PGV04, PGV19, NIH45-46), the MPER of gp41 (4E10), and additional glycan-dependent sites on the trimer (PG9, PG16, PGT145, 2G12, PGT128, PGT135, PGT121). Different Env-containing antigens and virions were tested for the ability to stimulate bNAb cell lines. Mouse strains expressing germline or mutated forms of 4E10 and b12 bNAb Ig genes were generated by gene targeting to the physiological loci. These “knock-in” mice were studied for their B cell development, and responses to HIV immunogens.

Engineered mice and B cell lines expressing broadly neutralizing antibodies and their unmutated precursors: tools for HIV vaccinology