Technical Report No. 18-88
Cloning of in vivo-derived thioguanine-resistant human B cells
Hakoda M, Hirai Y, Kusunoki Y, Akiyama MEditor’s note: A publication based on this report was published in Mutat Res 210:29-34, 1989.
Summary
In vivo-derived thioguanine-resistant (TGr) B cells have been cloned from peripheral blood mononuclear cells (PBMC) of four healthy adults. This was done by using Epstein-Barr virus transformation of B cells enriched from a large number of PBMC obtained with a blood cell separator. The cloned TGr B cells lacked hypoxanthine guanine phosphoribosyltransferase enzyme activity. The frequency of in vivo TGr B cells was estimated to be 8.6-13.1 x 10-6 for the four individuals by comparing the cloning efficiency of nonselected cells and TG-selected cells. This frequency is somewhat higher but comparable to the in vivo frequency of TGr T cells. Because the cloned TGr B cells can be easily expanded in vitro, this procedure provides a large amount of material for the precise characterization of in vivo mutations in humans.
In vivo-derived thioguanine-resistant (TGr) B cells have been cloned from peripheral blood mononuclear cells (PBMC) of four healthy adults. This was done by using Epstein-Barr virus transformation of B cells enriched from a large number of PBMC obtained with a blood cell separator. The cloned TGr B cells lacked hypoxanthine guanine phosphoribosyltransferase enzyme activity. The frequency of in vivo TGr B cells was estimated to be 8.6-13.1 x 10-6 for the four individuals by comparing the cloning efficiency of nonselected cells and TG-selected cells. This frequency is somewhat higher but comparable to the in vivo frequency of TGr T cells. Because the cloned TGr B cells can be easily expanded in vitro, this procedure provides a large amount of material for the precise characterization of in vivo mutations in humans.