Figure 7
Figure 7. Avidity of the Ag-BCR interaction and TLR9 signaling strength modulate the extent of PC formation in vivo. (A,B) CFSE-labeled MD4 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates containing CpG and either HEL (left panels), HELK (middle panels), or HELKD (right panels) at high or low densities. (A) After 4 days, flow cytometry was used to measure: CFSE dilution (top panels) and CD138 up-regulation (bottom panels) in HEL-binding cells in the spleen of recipient mice. (B) Flow cytometry was used to quantify the percentage of live HEL binding B cells that have undergone proliferation (left panel) and the percentage of MD4 PCs (HEL intracellularhi, CD138+; right panel) as a proportion of total splenocytes. (C,D) CFSE-labeled MD4 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates containing HEL and various densities of CpG. The density of CpG is represented from highest to lowest moving from left to right. (C) After 4 days, flow cytometry was used to measure: CFSE dilution (top panels) and CD138 up-regulation (bottom panels) in HEL-binding cells in the spleen of recipient mice. (D) Flow cytometry was used to quantify the percentage of live HEL binding B cells that have undergone proliferation (top panel), and the percentage of MD4 PCs (HEL intracellularhi, CD138+; bottom panel) as a proportion of total splenocytes. (E) HyHEL10 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates coated with HEL alone, CpG alone, or HEL-CpG. After 7 days, ELISAs were used to measure the levels of HEL-specific Igs in the serum of recipient mice. IgM and IgG (left panel) and IgG subtypes IgG1, IgG2b, and IgG2c (right panel). (F) HyHEL10 B cells were stimulated with 1 μL particulate HEL alone, particulate CpG alone, or particulate HEL-CpG for 7 days. ELISAs were used to measure the levels of HEL-specific Igs secreted into the culture medium as described in panel E. Values represent the mean (± SD) from triplicate samples.

Avidity of the Ag-BCR interaction and TLR9 signaling strength modulate the extent of PC formation in vivo. (A,B) CFSE-labeled MD4 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates containing CpG and either HEL (left panels), HELK (middle panels), or HELKD (right panels) at high or low densities. (A) After 4 days, flow cytometry was used to measure: CFSE dilution (top panels) and CD138 up-regulation (bottom panels) in HEL-binding cells in the spleen of recipient mice. (B) Flow cytometry was used to quantify the percentage of live HEL binding B cells that have undergone proliferation (left panel) and the percentage of MD4 PCs (HEL intracellularhi, CD138+; right panel) as a proportion of total splenocytes. (C,D) CFSE-labeled MD4 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates containing HEL and various densities of CpG. The density of CpG is represented from highest to lowest moving from left to right. (C) After 4 days, flow cytometry was used to measure: CFSE dilution (top panels) and CD138 up-regulation (bottom panels) in HEL-binding cells in the spleen of recipient mice. (D) Flow cytometry was used to quantify the percentage of live HEL binding B cells that have undergone proliferation (top panel), and the percentage of MD4 PCs (HEL intracellularhi, CD138+; bottom panel) as a proportion of total splenocytes. (E) HyHEL10 B cells were adoptively transferred into C57BL/6 mice and challenged with 10 μL particulates coated with HEL alone, CpG alone, or HEL-CpG. After 7 days, ELISAs were used to measure the levels of HEL-specific Igs in the serum of recipient mice. IgM and IgG (left panel) and IgG subtypes IgG1, IgG2b, and IgG2c (right panel). (F) HyHEL10 B cells were stimulated with 1 μL particulate HEL alone, particulate CpG alone, or particulate HEL-CpG for 7 days. ELISAs were used to measure the levels of HEL-specific Igs secreted into the culture medium as described in panel E. Values represent the mean (± SD) from triplicate samples.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal