Antibody-secreting plasma cells with unique CD5+IgG+CD21lo phenotype developed in humanized NOG mice

The developmental process of human antibody (Ab)-secreting B cells in humanized mice is not fully defined. To examine the characteristics of class-switched and Absecreting B cells in humanized mice, we analyzed NOD/Shi-scid, common γc-null mice reconstituted with cord blood-derived hematopoietic stem cells (CB-NOG). Most of the CD138-positive plasmablasts (PBs)/plasma cells (PCs) in the NOG spleen maintained CD5 expression. Moreover, IgG-bearing B cells, including PBs and memory cells, predominantly had a CD21-CD24hiCD5+ phenotype. We examined the expression of CD21, CD24 and CD5 on B cells developed in CB-NOG mice. We observed three types of B cells (CD24high CD21-; T1, CD24low/CD21-; T2, CD24-CD21+; T3) within CB-NOG CD5+ cells, which are similar to transitional B cells. However, the surface marker expression was different from normal transitional B cells. First, CD21 expression was significantly lower than in human peripheral blood and CB-derived mature B cells. Second, the expression of CD5 increased according to the developmental stage. The splenocytes were sorted according to the transitional subsets and examined for IL-10 expression. The CD21-CD24hiCD5+ fraction had the highest IL-10 expression among the CD5+ cells, suggesting the involvement of regulatory B cells. The same fractions were cultured in the presence of CpG and IgM. These stimulated B cells secreted a low level of IgM and IgG Abs to CpG and IgM stimulation in vitro and maintained a CD5+CD21low phenotype. These results demonstrate that IgG-bearing B cells with a CD21-CD24hiCD5+ irregular phenotype are developed in humanized NOG mice and have become the source of antibodies in CB-NOG mice. (250 words). Correspondence to: Yoshie Kametani, Ph.D, Department of Molecular Life Science, Tokai University School of Medicine, 143, Shimokasuya, Isehara, Kanagawa, 259-1193, Japan, Tel: +81-463-93-1121; Fax: +81-463-94-2976; E-mail: [email protected]