TY - JOUR
T1 - Interleukin-7 contributes to human pro-B-cell development in a mouse stromal cell-dependent culture system
AU - Taguchi, Tomoko
AU - Takenouchi, Hisami
AU - Shiozawa, Yusuke
AU - Matsui, Jun
AU - Kitamura, Noriko
AU - Miyagawa, Yoshitaka
AU - Katagiri, Yoko U.
AU - Takahashi, Takao
AU - Okita, Hajime
AU - Fujimoto, Junichiro
AU - Kiyokawa, Nobutaka
N1 - Funding Information:
We thank S. Yamauchi for her excellent secretarial work. We also thank Dr. A. Manabe and Dr. K. J. Mori for gifting murine bone marrow stromal cell line, MS-5. This work is supported in part by Japan Society for the Promotion of Science. KAKENHI 17591131, 18790751, 18790750, 18790263, 18790264, the Budget for Nuclear Research of the Ministry of Education, Culture, Sports, Science and Technology, based on the screening and counseling by the Atomic Energy Commission, grant from the Japan Health Sciences Foundation for Research on Health Sciences Focusing on Drug Innovation. This work was also supported in part by Health and Labour Sciences Research Grants and Grant for Child Health and Development from the Ministry of Health, Labour and Welfare of Japan and by CREST, JST.
PY - 2007/9
Y1 - 2007/9
N2 - 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 α 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 α 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 α 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.
AB - 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 α 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 α 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 α 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.
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U2 - 10.1016/j.exphem.2007.05.019
DO - 10.1016/j.exphem.2007.05.019
M3 - Article
C2 - 17656007
AN - SCOPUS:34548140548
VL - 35
SP - 1398
EP - 1407
JO - Experimental Hematology
JF - Experimental Hematology
SN - 0301-472X
IS - 9
ER -