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. Author manuscript; available in PMC: 2017 Sep 15.
Published in final edited form as: J Immunol. 2016 Aug 12;197(6):2119–2130. doi: 10.4049/jimmunol.1600133

Figure 2.

Figure 2

T cell requirement differs for initial development of TEC H/P (A) vs. induction of TEC H/P by splenocytes or transferred T cells (B, C). A, Naïve IFN-γ−/−CD28−/− NOD.H-2h4 males, 5–6 wk old, were given NaI water. One wk later, groups of mice were given rat IgG (Iso), anti-CD4 (GK1.5), anti-CD8 (116–13.1) or both antibodies every 10–12 days (see Methods). Thyroids were removed after 8 wk on NaI water (15 wk of age). Anti-CD4 or anti-CD8 alone had little effect on TEC H/P severity in most mice (p >0.1). Depletion of both CD4+ and CD8+ T cells significantly inhibited TEC H/P development (p < 0.0001). N=8 (Iso), 7 (anti-CD8), 9 (anti-CD4) and 9 (anti-CD4 + 8). B, splenocytes from IFN-γ−/−CD28−/−I females with severe TEC H/P were cultured and transferred to female SCID recipients (see Methods). Recipients were given NaI water and thyroids were removed 28 or 60 days later as indicated. Groups of recipients were given rat IgG (Con), anti-CD4 or anti-CD4 and anti-CD8 beginning 6 or 30 days after cell transfer as indicated. Antibody injections were repeated at 12–14 day intervals. Thyroids were removed from all treated mice at day 60. Mice given anti-CD4 or anti-CD4 and anti-CD8 at day 6 (before recipients had severe disease) developed minimal TEC H/P at day 60 (p < 0.001), whereas delaying T cell depletion until day 30 when recipients had severe TEC H/P had no effect (p > 0.1). N= 9 (Con, Day 30); 10 (Con, Day 60); 5 (anti-CD4 day 6); 9 (anti-CD4 and 8, day 6,) C, 5 (anti-CD4 and 8 day 30). D, Splenocytes from IFN-γ−/− CD28−/− females were cultured 72 hr. Cells were separated into CD4+ or CD8+ subsets as described in Methods, and transferred to SCID females. Recipients were given NaI water and thyroids were removed 28 days later. Splenocytes and CD4+T cells, but not CD8+, T cells induced severe TEC H/P in recipient mice. N= 8 (spl); 12 (CD4+); 11 (CD8+) and 8 (CD4 and CD8). Results are representative of two (C) or three (A, B) separate experiments. Thyroids and spleens of recipients of CD4+ T cells had no detectable CD8+ T cells (not shown).