Interleukin-7 contributes to human pro-B-cell development in a mouse stromal cell-dependent culture system.

OBJECTIVE

The role of interleukin (IL)-7 in human B lymphopoiesis is still controversial. We used an in vitro culture system to verify involvement of IL-7 in development of human pro-B cells from hematopoietic stem cells.

MATERIALS AND METHODS

Human CD34(+) bone marrow cells were cultured for 4 weeks on MS-5 mouse stromal cells to induce pro-B cells. Expression of IL-7 receptor alpha or other B-cell differentiation marker genes on cultured human CD34(+)bone marrow cells was investigated by reverse transcription polymerase chain reaction (RT-PCR). Colony assay of human CD34(+) bone marrow cells was also performed to determine the effect of IL-7 on colony-forming ability. Neutralizing antibody or reagent that eliminates the effect of IL-7 was added to the culture system, and the number of pro-B cells induced was estimated by flow cytometry.

RESULTS

RT-PCR analysis revealed mRNA expression of IL-7 receptor alpha as well as B-cell differentiation marker genes in not only CD19(+) pro-B cells but also CD19(-) CD33(-) cells induced from CD34(+) bone marrow cells after cultivation for 4 weeks on MS-5 cells. Addition of anti-mouse IL-7 antibody, anti-human IL-7 receptor alpha antibody, or JAK3 kinase inhibitor reduced the number of pro-B cells induced, demonstrating that elimination of IL-7 reduces pro-B-cell development. Addition of anti-mouse IL-7 antibody emphasized the colony-forming ability of burst-forming unit erythroid cells.

CONCLUSIONS

IL-7 produced by MS-5 cells is required for human pro-B-cell development from CD34(+)bone marrow cells in our culture system, and IL-7 appears to play a certain role in early human B lymphopoiesis.