Characterization of Mouse B Lymphoma Cells (CH12F3-2A) for the Study of IgA Isotype Switching.
- Author:
Young Saeng JANG
1
;
Seo Hyeun CHOI
;
Seok Rae PARK
;
Hyun A KIM
;
Jae Bong PARK
;
Pyeung Hyeun KIM
Author Information
- Publication Type:Original Article
- Keywords: LPS; TGF-beta1; IgA; isotype switching; CH12F3-2A; Smad
- MeSH: Animals; B-Lymphocytes; Cell Line; Enzyme-Linked Immunosorbent Assay; Immunoglobulin A*; Immunoglobulin Class Switching*; Luciferases; Lymphoma*; Membranes; Mice*; Plasma Cells; Transfection; Transforming Growth Factor beta1
- From:Immune Network 2004;4(4):216-223
- CountryRepublic of Korea
- Language:Korean
- 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.
