Immune Netw.  2009 Aug;9(4):133-137. 10.4110/in.2009.9.4.133.

Further Characterization of Activin A-induced IgA Response in Murine B Lymphocytes

• Lee HJ
• Kim PH

• KMID: 1474585
• DOI: http://doi.org/10.4110/in.2009.9.4.133

Abstract

We have recently shown that activin A, a member of TGF-beta superfamily, stimulates mouse B cells to express IgA isotype but other isotypes. In the present study, we further characterized effects of activin A on B cell growth and IgA expression. We found that activin A did not have effect on LPS-stimulated cell viability. In parallel, CFSE staining analysis revealed that activin A did not alter cell division. An increase of IgA secretion by activin A was completely abrogated by anti-activin A Ab but not by anti-TGFbeta1 Ab. In the same conditions, no other isotypes are significantly affected by each antibody treatment. Finally, activin A, as similar to TGF-beta1, increased IgA secretion by mesenteric lymph node cells. These results suggest that activin A can specifically stimulate IgA response, independent of TGF-beta in the gut.

Keyword

activin A; TGF-beta; IgA; mesenteric lymph node; CFSE

MeSH Terms

Activins
Animals
B-Lymphocytes
Cell Division
Cell Survival
Fluoresceins
Immunoglobulin A
Lymph Nodes
Mice
Succinimides
Transforming Growth Factor beta
Transforming Growth Factor beta1
Activins
Fluoresceins
Immunoglobulin A
Succinimides
Transforming Growth Factor beta
Transforming Growth Factor beta1

Figure

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  Figure 1 Effect of activin A and TGF-β1 on the growth kinetics of splenic B cells. Normal spleen B cells (1×106) stimulated with LPS were cultured in the presence of TGF-β1 (0.2 ng/ml) and activin A (10 ng/ml). Numbers of viable cells were enumerated by trypan blue exclusion. Data are means±SEM (bars) of triplicate culture wells.

• 

  Figure 2 Effect of activin A on B cell proliferation. CFSE-labeled splenic B cells (1×106) were cultured as in Fig. 1. After 48 h, 72 h, and 96 h incubation, B cell proliferation was assessed by analyzing the dilution of CFSE in the same number of viable cells.

• 

  Figure 3 Effect of anti-activin A Ab on IgA secretion by B lymphoma cells. B lymphoma cells were stimulated with LPS (12.5µg/ml) and activin A (10 ng/ml). Titrated anti-activin A Ab and anti-TGF-β Ab were pre-incubated for 1 h at room temperature before addition to culture. Supernatants were harvested after 3 days of culture. IgA secretion was determined by ELISA. Data are means of triplicate cultures±SEM.

• 

  Figure 4 Effect of anti-activin A Ab on Ig secretion by normal spleen B cells. Mouse spleen B cells were stimulated with LPS (12.5 g/ml) and activin A (10 ng/ml). As in Fig. 3, anti-activin A Ab (0.5µg/ml) and anti-TGF-β Ab (5µg/ml) were added to cultures. Supernatants were harvested after 7 days of culture. Secretion of Ig isotypes was determined by ELISA. Data are means of triplicate cultures±SEM.

• 

  Figure 5 Activin A increases IgA secretion by mesenteric lymph node cells. Mesenteric lymph node (1×106) cells were treated 10 ng/ml of activin A, and 0.2 ng/ml of TGF-β1. Supernatants were harvested after 7 days of culture. IgA, IgG1, and IgG2b secretion was determined by ELISA. Data are means of triplicate cultures±SEM.

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