Cutting Edge: IL-4, IL-21, and IFN-γ Interact To Govern T-bet and CD11c Expression in TLR-Activated B Cells - PubMed (original) (raw)

. 2016 Aug 15;197(4):1023-8.

doi: 10.4049/jimmunol.1600522. Epub 2016 Jul 18.

Arpita Myles 1, Daniel P Beiting 2, Kenneth J Roberts 3, Lucas Dawson 2, Ramin Sedaghat Herati 4, Bertram Bengsch 5, Susanne L Linderman 6, Erietta Stelekati 5, Rosanne Spolski 7, E John Wherry 5, Christopher Hunter 2, Scott E Hensley 6, Warren J Leonard 7, Michael P Cancro 8

Affiliations

Cutting Edge: IL-4, IL-21, and IFN-γ Interact To Govern T-bet and CD11c Expression in TLR-Activated B Cells

Martin S Naradikian et al. J Immunol. 2016.

Abstract

T-bet and CD11c expression in B cells is linked with IgG2c isotype switching, virus-specific immune responses, and humoral autoimmunity. However, the activation requisites and regulatory cues governing T-bet and CD11c expression in B cells remain poorly defined. In this article, we reveal a relationship among TLR engagement, IL-4, IL-21, and IFN-γ that regulates T-bet expression in B cells. We find that IL-21 or IFN-γ directly promote T-bet expression in the context of TLR engagement. Further, IL-4 antagonizes T-bet induction. Finally, IL-21, but not IFN-γ, promotes CD11c expression independent of T-bet. Using influenza virus and Heligmosomoides polygyrus infections, we show that these interactions function in vivo to determine whether T-bet(+) and CD11c(+) B cells are formed. These findings suggest that T-bet(+) B cells seen in health and disease share the common initiating features of TLR-driven activation within this circumscribed cytokine milieu.

Copyright © 2016 by The American Association of Immunologists, Inc.

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Figures

Figure 1

Figure 1. IL4 and IL21 act in a cell intrinsic manner to regulate TBET expression in vitro

Magnetically enriched CD23+ splenic B cells were cultured in vitro with various combinations of α-Ig-μ (IgM), α-CD40 (40), IL4 (4), IL21 (21), and IFNγ (γ). Mouse data are representative of 3 independent experiments. (A) WT or _Cd19_cre/+_Tbx21_f/f B cells treated for 48hrs and probed for TBET (ΔMFI=WT-mutant). (B) Tbx21 mRNA levels in WT cells treated for 20hrs. (C) WT, _Il21r_−/−, or _Stat6_−/− B cells were labeled with either CFSE (green plots) or Violet Cell Trace (VCT, purple plots), treated with ODN1826 and indicated cytokines for 48h, then stained for CD11c and TBET. (D) Magnetically enriched CD27−CD19+ human B cells were labeled with CFSE, treated for 108h, and probed for TBET on live, CFSE− cells. (E) Frequency of TBET+ B cells from each treatment across 6 healthy, adult donors.

Figure 2

Figure 2. TBET+CD11c+ cells delineate a BMEM cell subset and accumulate in _Il21_tg mice

(A–B) GC B and BMEM cells were analyzed for TBET and CD11c expression by FACS. GC B and BMEM cell gating strategies are in Supplemental Fig. 1G. All panels are representative of 3 independent experiments with ≥ 3 mice per strain. (A) TBET staining on GC B cells from C57BL/6 (B6, n=14) or BALB/c (n=23) mice with frequency enumeration. (B) TBET and CD11c staining on BMEM cells from B6 mice. (C) TBET and CD11c staining on splenic B-2 cells from WT and _Il21_tg mice. (D) Serum IgG1 or IgG2a/c (IgG2a + IgG2c) levels in WT and _Il21_tg mice were determined by ELISA. Values are means ± S.E.M. from 5 WT and 7 _Il21_tg mice.

Figure 3

Figure 3. Influenza virus infection drives TBET+CD11c+ BMEM cell formation in the absence of both IFNγ and IL4

Splenocytes were harvested from non-infected (−) or day 10 post i.n. 30 TCID50 PR8 infection (+) WT (n=21, black bars), _Ifng_−/− (n=10, white bars), or _Il4_−/−_Ifng_−/− (n=13, gray bars) mice across 3–7 experiments with ≥3 mice per group. GC B, BMEM, and TFH cell gating strategies are in Supplemental Figures 1G & 1J. (A) Enumeration of GC B cells. (B) TBET staining on GC B cells. (C) Il4 and (D) Il21 mRNA levels from sorted naïve CD62L+ CD4 T (TN, n=9) or TFH cells. (E) Proportions and (F) numbers of TBET+CD11c+ BMEM cells.

Figure 4

Figure 4. Activated B cells express T-BET independent of IFNγ in IL4 limiting conditions

Splenocytes and sera were harvested from non-infected (−) or day 14 post oral gavage (+) of 200 H. polygyrus in WT (n=20, black bars), _Il4_−/− (n=24, white bars), or _Il4_−/−_Ifng_−/− (n=11, gray bars) mice across 3–6 experiments with ≥3 mice per group. GC B, BMEM, and TFH cell gating strategies are in Supplemental Figures 1G & 1J. (A) Enumeration of GC B cells. (B) TBET staining on GC B cells. (C) Serum concentrations of IgG1 and IgG2c + IgG2b. (D) Il21 mRNA levels from sorted TFH cells. (E) Proportions and (F) numbers of TBET+CD11c+ BMEM cells.

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