IgA Isotype Switching 연구를 위한 마우스 B Lymphoma Cell (CH12F3-2A)의 특성 연구
Characterization of Mouse B Lymphoma Cells (CH12F3-2A) for the Study of IgA Isotype Switching
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Abstract
Background: It is well known that IgA isotype switching is induced by TGF-β1. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of TGF-β1. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and TGF-β1, then levels of germ-line (GL) transcripts were measured by RT-PCR, and GLα promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: TGF-β1, regardless of the presence of LPS, increased level of GLα transcripts but not GLγ2b transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and TGF-β1. Both mIgA and IgA secretion in the presence of TGF-β1 were further increased by over-expression of Smad3/4. Finally, GLα promoter activity was increased by TGF-β1. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels. (Immune Network 2004;4(4):216-223)
참고문헌 (14)
- "Transforming growth factor beta 1 increases IgA isotype switching at the clonal level", 145, pp.3773~3778 (1990)
- "Transforming growth factor beta specifically enhances IgA production by lipopolysaccharide-stimulated murine B lymphocytes", 170, pp.1039~1044 (1989)
- "Transforming growth factor beta induces IgA production and acts additively with interleukin 5 for IgA production", 170, pp.1415~1420 (1989)
- "Immunoglobulin heavy-chain switching may be directed by prior induction of transcripts from constant- region genes", 85, pp.7704~7708 (1988)
- "High frequency class switching of an IgM B lymphoma clone CH12F3 to IgA cells", 8, pp.193~201 (1996)
- "Receptor-associated Mad homologues synergize as effectors of the TGF-beta response", 383, pp.168~172 (1996)
- "DPC4, a candidate tumor suppressor gene at human chromosome 18q21.1.", 271, pp.350~353 (1996)
- "Interleukin 5 and interleukin 4 produced by Peyer's patch T cells selectively enhance immunoglobulin A expression", 139, pp.2669~2674 (1987)
- "Role of Rac GTPase in the nuclear signaling by EGF", 407, pp.7~12 (1997)
- "Differential regulation of IgA production by TGF-beta and IL-5 whereas IL-5 promotes their survival and maturation into IgA-secreting cells", pp.158~172 (1992)
- "Transforming growth factor beta 1 selectivity stimulates immunoglobulin G2b secretion by lipopolysaccharide-activated murine B cells", 177, pp.1031~1037 (1993)
- "Smad3 and Smad4 mediate transforming growth factor-beta1-induced IgA expression in murine B lymphocytes", 31, pp.1706~1715 (2001)
- "p300 cooperates with Smad3", 4 and Runx3 in TGFbeta1-induced IgA isotype expression, 2003, pp.33, 3386~3392
- "Analysis of transforming growth factor-β1-induced Ig germ-line γ2b transcription and its implication for IgA isotype switching", 35 (2005)