Immune Netw.
2004 Dec;4(4):216-223. 10.4110/in.2004.4.4.216.
Characterization of Mouse B Lymphoma Cells (CH12F3-2A) for the Study of IgA Isotype Switching
- Affiliations
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- 1Department of Microbiology, College of Natural Sciences, Kangwon National University, phkim@kangwon.ac.kr
- 2Vascular System Research Center, Kangwon National University, Korea.
- 3Department of Biochemistry, College of Medicine, Hallym University, Chunchon, S. Korea.
- KMID: 1515488
- DOI: http://doi.org/10.4110/in.2004.4.4.216
Abstract
- BACKGROUND
It is well known that IgA isotype switching is induced by TGF-beta1. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of TGF-beta1. 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-beta1, then levels of germ-line (GL) transcripts were measured by RT-PCR, and GLalpha 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-beta1, regardless of the presence of LPS, increased level of GLalpha transcripts but not GLgamma2b transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and TGF-beta1. Both mIgA and IgA secretion in the presence of TGF-beta1 were further increased by over-expression of Smad3/4. Finally, GLalpha promoter activity was increased by TGF-beta1. 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.
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Cited by 1 articles
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SUMO Proteins are not Involved in TGF-β1-induced, Smad3/4-mediated Germline α Transcription, but PIASy Suppresses it in CH12F3-2A B Cells
Sang-Hoon Lee, Pyeung-Hyeun Kim, Sang-Muk Oh, Jung-Hwan Park, Yung-Choon Yoo, Junglim Lee, Seok-Rae Park
Immune Netw. 2014;14(6):321-327. doi: 10.4110/in.2014.14.6.321.
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