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Distinct kinetics of memory B-cell and plasma-cell responses in peripheral blood following a blood-stage Plasmodium chabaudi infection in mice.

Nduati EW, Ng DH, Ndungu FM, Gardner P, Urban BC, Langhorne J - PLoS ONE (2010)

Bottom Line: We detected memory B cells, defined as isotype-switched IgD(-) IgM(-) CD19(+) B cells, and low numbers of Plasmodium chabaudi Merozoite Surface Protein-1 (MSP1)-specific memory B cells, in PBMC at all time points sampled for up to 90 days following primary or secondary infection.CD138(+) plasma cells in PBMC at these times expressed CD19, B220 and MHC class II, suggesting that they were not dislodged bone-marrow long-lived plasma cells, but newly differentiated migratory plasmablasts migrating to the bone marrow; thus reflective of an ongoing or developing immune response.Studies should therefore include multiple sampling points, and at times of infection/immunisation when the B-cell phenotypes of interest are likely to be found in peripheral blood.

View Article: PubMed Central - PubMed

Affiliation: KEMRI/Wellcome Trust Collaborative Research Programme, Centre for Geographical Medicine Research Coast, Kilifi, Kenya.

ABSTRACT
B cell and plasma cell responses take place in lymphoid organs, but because of the inaccessibility of these organs, analyses of human responses are largely performed using peripheral blood mononuclear cells (PBMC). To determine whether PBMC are a useful source of memory B cells and plasma cells in malaria, and whether they reflect Plasmodium-specific B cell responses in spleen or bone marrow, we have investigated these components of the humoral response in PBMC using a model of Plasmodium chabaudi blood-stage infections in C57BL/6 mice. We detected memory B cells, defined as isotype-switched IgD(-) IgM(-) CD19(+) B cells, and low numbers of Plasmodium chabaudi Merozoite Surface Protein-1 (MSP1)-specific memory B cells, in PBMC at all time points sampled for up to 90 days following primary or secondary infection. By contrast, we only detected CD138(+) plasma cells and MSP1-specific antibody-secreting cells within a narrow time frame following primary (days 10 to 25) or secondary (day 10) infection. CD138(+) plasma cells in PBMC at these times expressed CD19, B220 and MHC class II, suggesting that they were not dislodged bone-marrow long-lived plasma cells, but newly differentiated migratory plasmablasts migrating to the bone marrow; thus reflective of an ongoing or developing immune response. Our data indicates that PBMC can be a useful source for malaria-specific memory B cells and plasma cells, but extrapolation of the results to human malaria infections suggests that timing of sampling, particularly for plasma cells, may be critical. Studies should therefore include multiple sampling points, and at times of infection/immunisation when the B-cell phenotypes of interest are likely to be found in peripheral blood.

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Antibody-secreting cells in peripheral blood are migrating plasmablasts.A) Gating strategy for the identification of plasmablasts and long-lived plasma cells (LLPC) in peripheral blood, spleen and bone marrow on day 10 of infection. B) Histograms showing relative expression of CXCR4, CD19, MHC class II and CXCR5, on spleen (top two panels), blood (middle panel) and bone marrow (bottom two panels). Differences between plasma cells and plasmablasts were determined by the mean fluorescence intensity of these markers on the gated populations (red histograms) showing the gradual acquisition of CXCR4 and loss of CXCR5, CD19, MHC class II on splenic B220+ cells (top panel), B220+ CD138+ cells in spleen, blood and bone marrow (middle three panels) and B220− CD138+ LLPC in bone marrow (bottom panel). The blue histograms represent the fluorescence profile with the isotype control antibodies. C) A representative experiment out of 3 experiments showing kinetics of the appearance of plasma cells and plasmablasts during peak of infection based on their expression of B220 and CD138. The values and error bars shown are the means and the standard errors of the mean (SEM) of 5 mice.
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pone-0015007-g006: Antibody-secreting cells in peripheral blood are migrating plasmablasts.A) Gating strategy for the identification of plasmablasts and long-lived plasma cells (LLPC) in peripheral blood, spleen and bone marrow on day 10 of infection. B) Histograms showing relative expression of CXCR4, CD19, MHC class II and CXCR5, on spleen (top two panels), blood (middle panel) and bone marrow (bottom two panels). Differences between plasma cells and plasmablasts were determined by the mean fluorescence intensity of these markers on the gated populations (red histograms) showing the gradual acquisition of CXCR4 and loss of CXCR5, CD19, MHC class II on splenic B220+ cells (top panel), B220+ CD138+ cells in spleen, blood and bone marrow (middle three panels) and B220− CD138+ LLPC in bone marrow (bottom panel). The blue histograms represent the fluorescence profile with the isotype control antibodies. C) A representative experiment out of 3 experiments showing kinetics of the appearance of plasma cells and plasmablasts during peak of infection based on their expression of B220 and CD138. The values and error bars shown are the means and the standard errors of the mean (SEM) of 5 mice.

Mentions: Newly differentiated migratory plasmablasts and LLPC can be distinguished by the co-expression of the cell surface molecules B220 and CD138 (Figure 6A), in conjunction with the relative expression of other surface molecules like CD19 and MHC class II. Plasmablasts are CD138+ B220+ and express higher levels of MHC class II and CD19, whereas mature bone marrow plasma cells have downregulated CD19, B220 and MHC class II expression. In addition, the chemokine receptor CXCR4 is upregulated as B cells differentiate through plasmablasts and is maintained on bone marrow LLPC, whereas expression of CXCR5 is down regulated as B cells differentiate into plasmablasts [19], .


Distinct kinetics of memory B-cell and plasma-cell responses in peripheral blood following a blood-stage Plasmodium chabaudi infection in mice.

Nduati EW, Ng DH, Ndungu FM, Gardner P, Urban BC, Langhorne J - PLoS ONE (2010)

Antibody-secreting cells in peripheral blood are migrating plasmablasts.A) Gating strategy for the identification of plasmablasts and long-lived plasma cells (LLPC) in peripheral blood, spleen and bone marrow on day 10 of infection. B) Histograms showing relative expression of CXCR4, CD19, MHC class II and CXCR5, on spleen (top two panels), blood (middle panel) and bone marrow (bottom two panels). Differences between plasma cells and plasmablasts were determined by the mean fluorescence intensity of these markers on the gated populations (red histograms) showing the gradual acquisition of CXCR4 and loss of CXCR5, CD19, MHC class II on splenic B220+ cells (top panel), B220+ CD138+ cells in spleen, blood and bone marrow (middle three panels) and B220− CD138+ LLPC in bone marrow (bottom panel). The blue histograms represent the fluorescence profile with the isotype control antibodies. C) A representative experiment out of 3 experiments showing kinetics of the appearance of plasma cells and plasmablasts during peak of infection based on their expression of B220 and CD138. The values and error bars shown are the means and the standard errors of the mean (SEM) of 5 mice.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2990717&req=5

pone-0015007-g006: Antibody-secreting cells in peripheral blood are migrating plasmablasts.A) Gating strategy for the identification of plasmablasts and long-lived plasma cells (LLPC) in peripheral blood, spleen and bone marrow on day 10 of infection. B) Histograms showing relative expression of CXCR4, CD19, MHC class II and CXCR5, on spleen (top two panels), blood (middle panel) and bone marrow (bottom two panels). Differences between plasma cells and plasmablasts were determined by the mean fluorescence intensity of these markers on the gated populations (red histograms) showing the gradual acquisition of CXCR4 and loss of CXCR5, CD19, MHC class II on splenic B220+ cells (top panel), B220+ CD138+ cells in spleen, blood and bone marrow (middle three panels) and B220− CD138+ LLPC in bone marrow (bottom panel). The blue histograms represent the fluorescence profile with the isotype control antibodies. C) A representative experiment out of 3 experiments showing kinetics of the appearance of plasma cells and plasmablasts during peak of infection based on their expression of B220 and CD138. The values and error bars shown are the means and the standard errors of the mean (SEM) of 5 mice.
Mentions: Newly differentiated migratory plasmablasts and LLPC can be distinguished by the co-expression of the cell surface molecules B220 and CD138 (Figure 6A), in conjunction with the relative expression of other surface molecules like CD19 and MHC class II. Plasmablasts are CD138+ B220+ and express higher levels of MHC class II and CD19, whereas mature bone marrow plasma cells have downregulated CD19, B220 and MHC class II expression. In addition, the chemokine receptor CXCR4 is upregulated as B cells differentiate through plasmablasts and is maintained on bone marrow LLPC, whereas expression of CXCR5 is down regulated as B cells differentiate into plasmablasts [19], .

Bottom Line: We detected memory B cells, defined as isotype-switched IgD(-) IgM(-) CD19(+) B cells, and low numbers of Plasmodium chabaudi Merozoite Surface Protein-1 (MSP1)-specific memory B cells, in PBMC at all time points sampled for up to 90 days following primary or secondary infection.CD138(+) plasma cells in PBMC at these times expressed CD19, B220 and MHC class II, suggesting that they were not dislodged bone-marrow long-lived plasma cells, but newly differentiated migratory plasmablasts migrating to the bone marrow; thus reflective of an ongoing or developing immune response.Studies should therefore include multiple sampling points, and at times of infection/immunisation when the B-cell phenotypes of interest are likely to be found in peripheral blood.

View Article: PubMed Central - PubMed

Affiliation: KEMRI/Wellcome Trust Collaborative Research Programme, Centre for Geographical Medicine Research Coast, Kilifi, Kenya.

ABSTRACT
B cell and plasma cell responses take place in lymphoid organs, but because of the inaccessibility of these organs, analyses of human responses are largely performed using peripheral blood mononuclear cells (PBMC). To determine whether PBMC are a useful source of memory B cells and plasma cells in malaria, and whether they reflect Plasmodium-specific B cell responses in spleen or bone marrow, we have investigated these components of the humoral response in PBMC using a model of Plasmodium chabaudi blood-stage infections in C57BL/6 mice. We detected memory B cells, defined as isotype-switched IgD(-) IgM(-) CD19(+) B cells, and low numbers of Plasmodium chabaudi Merozoite Surface Protein-1 (MSP1)-specific memory B cells, in PBMC at all time points sampled for up to 90 days following primary or secondary infection. By contrast, we only detected CD138(+) plasma cells and MSP1-specific antibody-secreting cells within a narrow time frame following primary (days 10 to 25) or secondary (day 10) infection. CD138(+) plasma cells in PBMC at these times expressed CD19, B220 and MHC class II, suggesting that they were not dislodged bone-marrow long-lived plasma cells, but newly differentiated migratory plasmablasts migrating to the bone marrow; thus reflective of an ongoing or developing immune response. Our data indicates that PBMC can be a useful source for malaria-specific memory B cells and plasma cells, but extrapolation of the results to human malaria infections suggests that timing of sampling, particularly for plasma cells, may be critical. Studies should therefore include multiple sampling points, and at times of infection/immunisation when the B-cell phenotypes of interest are likely to be found in peripheral blood.

Show MeSH
Related in: MedlinePlus