Fig. 2 Adoptive transfer of adult Ox40−/− splenocytes into adult HBVEnvRag−/− mice alters hepatic inflammation, HBsAg clearance, HBsAb seroconversion,

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Fig. 2 Adoptive transfer of adult Ox40−/− splenocytes into adult HBVEnvRag−/− mice alters hepatic inflammation, HBsAg clearance, HBsAb seroconversion, T cell responses, and cytokine production. Adoptive transfer of adult Ox40−/− splenocytes into adult HBVEnvRag−/− mice alters hepatic inflammation, HBsAg clearance, HBsAb seroconversion, T cell responses, and cytokine production. (A) Plasma alanine aminotransferase (ALT) values, reflecting liver injury, were determined for HBVEnvRag−/− mice receiving wild-type (WT) (Ox40+/+) splenocytes (open squares, solid line) or Ox40−/− splenocytes (closed triangles, dotted line) at given time points after adoptive transfer (n ≥ 5). (B) Hematoxylin and eosin staining of liver sections from adult HBVEnvRag−/− mice 8 days after adoptive transfer. Arrows indicate necrotic hepatocytes. Scale bar, 25 μm. (C) Composite score of hepatic necrosis, portal inflammation, and intraparenchymal inflammation in the liver sections determined by a pathologist blinded to sample identity. Statistical significance was determined using Mann-Whitney two-tailed (***P = 0.0008; n = 8 mice). (D) Plasma HBsAg loss and (E) HBsAb titer in HBVEnvRag−/− mice at given time points after adoptive transfer. Statistical significance was determined using the chi-square test (***P < 0.001; n ≥ 5 mice). (F and G) Presence of hepatitis B virus (HBV)–specific T cells was determined using IFN-γ enzyme-linked immunospot (ELISpot) 8 days (F) and 84 days (G) after adoptive transfer (data pooled from n = 5 mice). Threshold defining a positive response is ≥2× the baseline (dashed line). (H and I) Percentages of total lymphocyte subpopulations were determined by flow cytometry in the liver 8 days after adoptive transfer: T cells are CD19− and TCRβ+; CD4+ T cells are CD4+, CD8−, TCRβ+, and CD19−; TFH cells are CD4+, CD8−, TCRβ+, CD19−, CXCR5+, and ICOS+; CD8+ T cells are CD4−, CD8+, TCRβ+, and CD19−; B cells are TCRβ− and CD19+; NK cells are TCRβ−, CD19-, and NK1.1+. Statistical significance was determined using the unpaired two-tailed t test (**P < 0.01 and ***P < 0.001; n = 5 mice). Bars represent means ± SEM. (J) Expression of various cytokine mRNAs relative to Gapdh was detected by RT-PCR on mRNA from liver lymphocytes 8 days after adoptive transfer (n ≥ 5). Bars represent means ± SEM. Statistical significance was determined using the unpaired two-tailed t test (*P < 0.05). Jean Publicover et al., Sci Transl Med 2018;10:eaah5766 Published by AAAS