American Society of Hematology

Figure 5.

$Figure 5. Chemotaxis of IgG plasma cells in response to CXCL12 is limited to B220-positive plasma cells. (A) Chemotaxis assays in response to CXCL12 (100 ng/mL) were performed with CD5/Mac-1/IgM-depleted cervical lymph node cells. Overall, 7.53% ± 2.04% (n = 4) of input cells migrated toward CXCL12, whereas only 0.81% ± 0.49% (n = 4) migrated toward media alone (Basal). Input and migrated cells were analyzed by flow cytometry for the expression of B220 and syndecan-1 to identify plasma cell subsets. A representative experiment of 4 is shown. (B) Cervical lymph node (LN), spleen (SPL), and bone marrow (BM) cells were separated into B220-positive (+) and B220-negative (-) fractions as in Figure 1B prior to the CXCL12 (100 ng/mL) chemotaxis assays. The number of IgG plasma cells in the input and migrated populations for each fraction was determined by ELISPOT (n = 6), and the percentage migrated was calculated (mean ± SD). The (*) indicates statistical difference between B220-positive and B220-negative within each tissue. The (#) indicates statistical difference between the BM B220-positive fraction and B220-positive fractions from LN or SPL. A representative experiment of 3 is shown.$

Chemotaxis of IgG plasma cells in response to CXCL12 is limited to B220-positive plasma cells. (A) Chemotaxis assays in response to CXCL12 (100 ng/mL) were performed with CD5/Mac-1/IgM-depleted cervical lymph node cells. Overall, 7.53% ± 2.04% (n = 4) of input cells migrated toward CXCL12, whereas only 0.81% ± 0.49% (n = 4) migrated toward media alone (Basal). Input and migrated cells were analyzed by flow cytometry for the expression of B220 and syndecan-1 to identify plasma cell subsets. A representative experiment of 4 is shown. (B) Cervical lymph node (LN), spleen (SPL), and bone marrow (BM) cells were separated into B220-positive (+) and B220-negative (-) fractions as in Figure 1B prior to the CXCL12 (100 ng/mL) chemotaxis assays. The number of IgG plasma cells in the input and migrated populations for each fraction was determined by ELISPOT (n = 6), and the percentage migrated was calculated (mean ± SD). The (^*) indicates statistical difference between B220-positive and B220-negative within each tissue. The (#) indicates statistical difference between the BM B220-positive fraction and B220-positive fractions from LN or SPL. A representative experiment of 3 is shown.

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