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Figure S1 IgG4 and IgE are not strongly induced by IL-10, IL-10-secreting Tr1 cells or CD4+CD25+ Treg cells without IL-4 and sCD40L. PBMC from healthy individuals were incubated with IL-10 (50 ng/ml), IL-10-secreting Tr1 cells or CD4+CD25+ Treg cells. (A) Quantitative RT-PCR determined after 5 days of incubation (mean values ± SE; n = 4). (B) The IgE and IgG4 concentrations in the supernatants  measured after 12 days of incubation (mean values ± SE; n = 4).

Figure S2 Regulation of IgE and IgG4 by IL-4- and IFN-@-secreting cells. Der p 1 or bee venom phospholipase A2-specific IL-4-secreting and IFN-@-secreting T cells were used as controls. IL-4-secreting T cells significantly increased IgG4 and IgE production, whereas IFN-@-secreting T cells did not affect IgE and IgG4 production.

Figure S3 Role of IL-10 in suppression of IgE and induction of IgG4 by Treg cells. B cells from healthy individuals are incubated with CD4+CD25+ Treg cells or with CD4 + CD25- T cells. Quantitative RT-PCR determined after 5 days of incubation. (A) IgE germline transcripts and IgG4 productive transcripts (one representative experiment). (B) IL-10-secreting Tr1 cells were cultured in the upper compartment of the transwells and PBMC were cultured in the lower compartment. IL-10-producing Tr1 cells induced IgG4 production without any requirement of cell-to-cell contact (data represent 2 independent experiments). (C) In PBMC, IL-10 or IL-10RA-chain was blocked by mAbs in the presence of IL-10. IgG4, which was significantly induced by IL-10 was strongly inhibited either by blocking the cytokine itself or its receptor (data represent 3 independent experiments) *P < 0.05. IC: isotype control.

Figure S4 IgA is elicited by stimulation of TLR7 or 9 but not by IL-10, IL-10-secreting Tr1 cells or CD4+CD25+ Treg cells. (A) PBMC or pure B cells from healthy individuals are incubated with IL-10 (50 ng/ml), IL-10-secreting Tr1 cells or CD4+CD25+ Treg cells. Quantitative RT-PCR determined after 5 days of incubation (mean values ± SE; n = 3-5). (B) PBMC from healthy individuals are stimulated with TLR2, 3, 4, 5, 7, 8 or 9 ligands. Quantitative RTPCR determined after 5 days of incubation (n = 3). IL-4 and sCD40L, which upregulated IgE and IgG4 suppressed TLR7 and TLR9 induced IgA.

Figure S5 IgA was determined after stimulation of PBMC with type B CpG2006 and type A CpG10 and CpG2216 (data represent 3 independent experiments). CpG2006 was found to be the most potent inducer of IgA. In contrast, induction of IgA by A type CpGs was very weak. IgA mRNA was detected on day 5.

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