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KMID : 0390320110210010213
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Chungbuk Medical Journal
2011 Volume.21 No. 1 p.213 ~ p.225
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Function of CD5 in Human Tonsillar B cells
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Hong Jeong-Won
Kim Mei-Shan
Choi Song-Yi
Lee Dong-Wook
Song Hyung-Geun
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Abstract
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Purpose:Using the anti-CD5 mAb P9 and P17, which were produced in our laboratory, we evaluated the expression and function of CD5 in human tonsillar B cells.
Materials and Methods : Human tonsils were obtained from children undergoing tonsilectomy, For B cell purfication, tosil monoclear cells were first submitted to E-rosetting with 2-aminoethylisothio-uronium bromide (AET) treated sheep -red blood cells. Tonsilar B cell were futher separated into high density and low density B cell by centrifugation through percoll gradient. The high density and low density cells were then cultured in RPMI 10% FCS medium with or without IgM, PMA and IL-2 at 1, 2, 3 day, check the CD5 surface expression by flow cytometry. High density cells were further seperated into CD5+ and CD5- B cells using magnetic cell sorting with indirect microbeads (MACS) kit. The cells then cultured with or without PMA, anti-CD5 antibodies (P9, P17) for 3 day and checked the proliferation effect.
Results: In human tonsillar B cells, the average percent of CD5 expression is about 10%. We demonstrated that CD5 expression can be induced by coculture with PMA, IgM and IL-2 both high and low density tonsillar B cells. We also studied the proliferative response of anti-CD5 on tonsillar B cells. Ligiaton of CD5 using P17 mAb both high density human tonsillar B cell and PMA activated B cells inhibit cell proliferation.
Conclusion:CD5+ B cell is the minor population in human tonsil, and mainly expressederatio in the mantle zone. Engagement of the CD5 on resting tonsillar B cells inhibit cell proliferation.
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KEYWORD
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CD5, inhibition, tonsilar B cell, monoclonal antibody (P9 and P17)
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