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. 2019 May 31;38(14):e101260. doi: 10.15252/embj.2018101260

Figure 5. iNKT cells provide indirect help for B‐cell class switching to IgG1 in vitro .

Figure 5

  • A
    Pooled spleen and LN cells from a Vα14 TN mouse were cocultured with wild‐type B cells with or without 1 μg α‐GalCer and with or without agnostic anti‐CD40. IgM was measured by ELISA of culture supernatants 4 days later.
  • B
    Pooled spleen and LN cells from a Vα14 TN mouse were cocultured with anti‐CD40‐activated wild‐type B cells with or without 1 μg α‐GalCer and with blocking antibody to CD1d (1 μg/ml, clone 1B1) or isotype control as indicated. IgM, IgG1, and IgA were measured by ELISA of 4‐day culture supernatants.
  • C–G
    Spleen, mLN, or PP cells from Vα14 TN iNKT mice and PP cells from an IL‐4−/− mouse were cocultured with anti‐CD40‐activated wild‐type or CD1d−/− B cells. 1 μg α‐GalCer was added to the cultures to specifically activate iNKT cells. Blocking antibodies to IL‐4 or isotype were added as indicated. IL‐4 (C) and IgG1 (D, E) were measured by ELISA of culture supernatants. IgG1+ and IgA+ class‐switched B cells were enumerated by flow cytometry after 4 days of coculture (F, G). Representative of two independent experiments. np = not performed.
Data information: Mann–Whitney test. Error bars are SD of triplicate samples. *P < 0.05; **P < 0.01; ***P < 0.001.