Yonsei Med J.  2011 Sep;52(5):851-855. 10.3349/ymj.2011.52.5.851.

IL-10 is Predominantly Produced by CD19(low)CD5(+) Regulatory B Cell Subpopulation: Characterisation of CD19 (high) and CD19(low) Subpopulations of CD5(+) B cells

• Lee JH
• Noh J
• Noh G
• Choi WS
• Lee SS

• KMID: 1108080
• DOI: http://doi.org/10.3349/ymj.2011.52.5.851

Abstract

IL-10 production by CD19(+)CD5(+) B cells was investigated, by determining the expression levels of CD19, a classical B cell marker. Peripheral mononuclear cells were stained with fluorescence-conjugated anti-CD5, anti-CD19, anti-IL-10, and Annexin V. Interestingly, IL-10-producing B cells were found to be localised within the CD19(low)CD5(+) B cell subset. Apoptotic changes were also observed mainly in CD19(low) cells among B cells. Thus, CD5(+) B cells should be classified as CD19(high) and CD19(low) cells, and the immunological significance of CD19 for the IL-10 production by CD5(+) B cells requires further studies.

Keyword

Regulatory B cell; CD5; CD19; IL-10; CD5(+) B

MeSH Terms

Antigens, CD19/metabolism
Antigens, CD5/metabolism
Apoptosis/immunology
B-Lymphocyte Subsets/cytology/*immunology
Cell Separation
Flow Cytometry
Humans
Interleukin-10/*biosynthesis

Figure

• 

  Fig. 1 CD19(high) and CD19(low) subpopulations of B cells. Fresh PBMCs from normal human subjects were stained for CD19 and CD5. (A) Histogram of CD19 expression in CD5(-) cells and CD5(+) B cells, (B) CD19(high) and CD19(low) B cells by CD5 and CD19 expression, (C) CD19(high) and CD19(low) B cells by SSC, (D) CD19(high) and CD19(low) B cells by FSC. CD19H, CD19 (high) B cells; CD19L, CD19(low) B cells.

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  Fig. 2 Apoptosis of CD19(high) and CD19(low) B cells. CD19, APC-conjugated CD19; Annexin V, FITC-conjugated Annexin V. APC, allophycocyanin.

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  Fig. 3 Demarcation of B cell subpopulations according to the expression of CD5 and CD19. (A) Resolving CD19(high) and CD19(low) cells by the expression of CD5, (B) Zebra plotting of CD19(high) and CD19(low) cells by the expression of CD19 and CD5, (C) Demarcation of B cell subpopulations according to the expression of CD5 and CD19. *1 arrow, CD19(low)CD5(-) B cells; *2 arrow CD19(low)CD5(+) B cells; CD5(-), CD5(-) cells; CD5(+), CD5(+) cells; CD19(high), CD19(high) B cells; CD19(low), CD19(low) B cells.

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  Fig. 4 Representative IL-10 production and apoptosis profiles of B cell populations subdivided by CD19 and CD5 expression (A). (B) CD19(high)CD5(-) B cells (section a); (C) CD19(high)CD5(+) B cells (section b); (D) CD19(+)CD5(-) B cells; (E) CD19(low)CD5(-) B cells (section c); (F) CD19(low)CD5(+) B cells (section d). The percentage of each population is indicated in the plotting box. X-axis, PE-conjugated IL-10 expression; Y-axis, FITC-conjugated Annexin V; Arrow, IL-10-producing CD19(+) B cells.

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  Fig. 5 IL-10 production from CD19(low)CD5(+) B cells among B cells. (A) Resolving CD19(high) and CD19(low) cells by the expression of CD19 and IL-10 (upper box, zebra plotting; lower box, density plotting of CD19(+) B cells for IL-10 production, (B) IL-10-producing cell frequency (%) in B cell populations divided on the basis of CD19 and CD5 expression. *1 arrow, IL-10-producing CD19(low) B cells; *2 and *3 arrow, non-IL-10-producign CD19(high) and CD19(low). B cells; IL-10(+) total, total IL-10-producing B cells; Annexin V(+)/IL-10(+), IL-10-producing B cells undergoing apoptosis; Annexin V(-)/IL-10(+), IL-10-producing B cells without apoptosis.

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