Immune Netw.  2013 Oct;13(5):205-212. 10.4110/in.2013.13.5.205.

Dectin-1 Stimulation Selectively Reinforces LPS-driven IgG1 Production by Mouse B Cells

• Seo BS
• Lee SH
• Lee JE
• Yoo YC
• Lee J
• Park SR

• KMID: 1493615
• DOI: http://doi.org/10.4110/in.2013.13.5.205

Abstract

Dectin-1, which specifically recognizes beta-glucan of fungal cell walls, is a non-Toll-like receptor (TLR) pattern recognition receptor and a representative of C-type lectin receptors (CLRs). The importance of Dectin-1 in innate immune cells, such as dendritic cells and macrophages, has previously been well studied. However, the function of Dectin-1 in B cells is very poorly understood. To determine the role of Dectin-1 in B cell activation, we first investigated whether mouse B cells express Dectin-1 and then assessed the effect of Dectin-1 stimulation on B cell proliferation and antibody production. Mouse B cells express mRNAs encoding CLRs, including Dectin-1, and surface Dectin-1 was expressed in B cells of C57BL/6 rather than BALB/c strain. Dectin-1 agonists, heat-killed Candida albicans (HKCA) and heat-killed Saccharomyces cerevisiae (HKSC), alone induced B cell proliferation but not antibody production. Interestingly, HKSC, HKCA, and depleted zymosan (a selective Dectin-1 agonist) selectively enhanced LPS-driven IgG1 production. Taken together, these results suggest that, during fungal infection, beta-glucan-stimulated Dectin-1 may cooperate with TLR4 to specifically enhance IgG1 production by mouse B cells.

Keyword

Dectin-1; B cell; IgG1; C-type lectin receptor; Fungal immunity

MeSH Terms

Animals
Antibody Formation
B-Lymphocytes*
Candida albicans
Cell Proliferation
Cell Wall
Dendritic Cells
Immunoglobulin G*
Lectins, C-Type
Macrophages
Mice*
RNA, Messenger
Saccharomyces cerevisiae
Sprains and Strains
Zymosan
Immunoglobulin G
Lectins, C-Type
RNA, Messenger
Zymosan

Figure

• 

  Figure 1 Expression of Dectin-1 in mouse B cells. (A) mRNA expression of Dectin-1, Dectin-2, Mincle and mannose receptor. Total RNAs were isolated from purified mouse B cells and the indicated cell lines (L10A6.2 and RAW264.7), and levels of Dectin-1, Dectin-2, Mincle and mannose receptor (MR) mRNA were measured by RT-PCR. (B) Expression of surface Dectin-1 in mouse B cells. Isolated total mouse spleen cells from BALB/c and C57BL/6 and the indicated cell lines were stained with anti-mouse Dectin-1 FITC and anti-B220 PE, and populations of Dectin-1+ cells were determined by flow cytometric analysis.

• 

  Figure 2 Effects of Dectin-1 agonists, HKCA and HKSC, on B cell proliferation and antibody production. (A) Total spleen cells isolated from C57BL/6 were stimulated HKCA (1×107 cells/ml), HKSC (1×107 cells/ml), LPS (1 µg/ml) and anti-IgM (5 µg/ml). (B) Purified B cells from spleen of C57BL/6 were stimulated as in (A). After 2 and 3 days of culture, cell proliferation (OD) was measured by EZ-Cytox cell viability assay. Data are averages of duplicate samples with ranges (bars). After 7 days of culture, supernatants were harvested, and levels of Igs production were determined by isotype-specific ELISA. Data represent means±SEM of triplicate samples. *p<0.05.

• 

  Figure 3 Effect of the selective Dectin-1 agonist, depleted zymosan, on B cell proliferation and antibody production. Isolated total spleen cells and purified spleen B cells from C57BL/6 were stimulated with depleted zymosan (dzn, 100 µg/ml), LPS (1 µg/ml) and anti-IgM (10 µg/ml). (A) After 2 days of culture, cell proliferation was measured by EZ-Cytox cell viability assay. Data are averages of duplicate samples with ranges (bars). (B) After 7 days of culture, supernatants were harvested, and levels of Igs production were determined by isotype-specific ELISA. Data represent means±SEM of triplicate samples. *p<0.05.

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