Characterization of Mouse B Lymphoma Cells (CH12F3-2A) for the Study of IgA Isotype Switching
Immune Network
; : 216-223, 2004.
Article
Dans Ko
| WPRIM
| ID: wpr-13655
Responsable en Bibliothèque :
WPRO
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.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Plasmocytes
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Immunoglobuline A
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Test ELISA
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Lymphocytes B
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Transfection
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Lignée cellulaire
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Commutation de classe des immunoglobulines
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Facteur de croissance transformant bêta-1
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Luciferases
/
Lymphomes
Limites du sujet:
Animals
langue:
Ko
Texte intégral:
Immune Network
Année:
2004
Type:
Article