Detection and activation of HIV broadly neutralizing antibody precursor B cells using anti-idiotypes

Bancroft, Tara; DeBuysscher, Blair L.; Weidle, Connor; Schwartz, Allison; Wall, Abigail; Gray, Matthew D.; Feng, Junli; Steach, Holly R.; Fitzpatrick, Kristin S.; Gewe, Mesfin M.; Skog, Patrick D.; Doyle-Cooper, Colleen; Ota, Takayuki; Strong, Roland K.; Nemazee, David; Pancera, Marie; Stamatatos, Leonidas; McGuire, Andrew T.; Taylor, Justin J.

Human PBMC collection and storage Blood was obtained by venipuncture from healthy, HIVseronegative adult volunteers enrolled in the General Quality Control study in Seattle, WA, which was approved by the Fred Hutchinson Cancer Research Center institutional review board. Informed consent was obtained from all study participants before enrollment into the parent protocols. PBMCs were isolated from whole blood using Accuspin System Histopaque-1077 (Sigma-Aldrich) resuspended in 10% dimethylsulfoxide in heat-inactivated fetal bovine serum and cryopreserved in liquid nitrogen before use. . Briefly, the D Q52 -J H cluster was replaced in a C57BL/6derived C2 embryonic stem cell line with the iglb12 heavy chain VDJ exon or inferred germline VRC01 heavy chain VDJ exon. Mice derived from these modified embryonic cells carrying the transgenic heavy chain were interbred and homozygous transgenic heavy chain +/+ male or female mice were used for experiments. Control transgenic animals expressing the iglVRC01 heavy chain were generated in an identical manner .