Immune Netw.  2015 Feb;15(1):37-43. 10.4110/in.2015.15.1.37.

Lactoferrin Combined with Retinoic Acid Stimulates B1 Cells to Express IgA Isotype and Gut-homing Molecules

• Kang SH
• Jin BR
• Kim HJ
• Seo GY
• Jang YS
• Kim SJ
• An SJ
• Park SR
• Kim WS
• Kim PH

• KMID: 2150828
• DOI: http://doi.org/10.4110/in.2015.15.1.37

Abstract

It is well established that TGF-beta1 and retinoic acid (RA) cause IgA isotype switching in mice. We recently found that lactoferrin (LF) also has an activity of IgA isotype switching in spleen B cells. The present study explored the effect of LF on the Ig production by mouse peritoneal B cells. LF, like TGF-beta1, substantially increased IgA production in peritoneal B1 cells but little in peritoneal B2 cells. In contrast, LF increased IgG2b production in peritoneal B2 cells much more strongly than in peritoneal B1 cells. LF in combination with RA further enhanced the IgA production and, interestingly, this enhancement was restricted to IgA isotype and B1 cells. Similarly, the combination of the two molecules also led to expression of gut homing molecules alpha4beta7 and CCR9 on peritoneal B1 cells, but not on peritoneal B2 cells. Thus, these results indicate that LF and RA can contribute to gut IgA response through stimulating IgA isotype switching and expression of gut-homing molecules in peritoneal B1 cells.

Keyword

Lactoferrin; Retinoic acid; Peritoneal B1 cell; IgA; Gut-homing molecule

MeSH Terms

Animals
B-Lymphocytes
Immunoglobulin A*
Immunoglobulin Class Switching
Immunoglobulin G
Lactoferrin*
Mice
Spleen
Transforming Growth Factor beta1
Tretinoin*
Immunoglobulin A
Immunoglobulin G
Lactoferrin
Transforming Growth Factor beta1
Tretinoin

Figure

• 

  Figure 1 Effect of LF, RA, and TGF-β1 on Ig secretion by mouse peritoneal B cells. Mouse whole peritoneal B cells were stimulated with LPS (12.5 µg/ml), RA (25 nM), LF (60 µg/ml), and TGF-β1 (0.2 ng/ml) for 7 days. Supernatants were collected, and Ig production was determined by isotype-specific ELISA. Data are means of triplicate samples±SEM. *p<0.05.

• 

  Figure 2 Effect of LF, RA, and TGF-β1 on Ig secretion by mouse peritoneal B1 and B2 cells. Mouse peritoneal B1 and B2 cells were stimulated with LPS (12.5 µg/ml), RA (25 nM), LF (60 µg/ml), and TGF-β1 (0.2 ng/ml) for 7 days. Supernatants were collected, and Ig production was determined by isotype-specific ELISA. Data represent the results of one of the two independent experiments and are means of triplicated samples±SEM. *p<0.05.

• 

  Figure 3 Combined effect of LF and RA on Ig secretion by mouse peritoneal B1 and B2 cells. Mouse peritoneal B1 and B2 cells were stimulated with LPS (12.5 µg/ml), RA (25 nM), and LF (30, 60, 120, 240 µg/ml in the serial dilution exp.; 60 µg/ml in the combination exp.) for 7 days. Supernatants were collected, and Ig production was determined by isotype-specific ELISA. Data represent the results of one of the two independent experiments and are means of triplicated samples±SEM. *p<0.05.

• 

  Figure 4 Measurement of membrane IgA on freshly isolated peritoneal B1 and B2 cells. Expression of mIgA and B220 was determined by FACS.

• 

  Figure 5 Effect of LF and RA on gut homing molecule expressed by peritoneal B1 and B2 cells. Mouse peritoneal B1 and B2 cells were cultured with LPS (12.5 µg/ml), RA (25 nM), and LF (60 µg/ml). After 4 days of culture, expression of surface CCR9 and α4β7 within B220 gated cell population was determined by FACS. Data represent the results of one of the two independent experiments.

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