Journal:

Article Title: Tc17 cells are capable of mediating immunity to vaccinia virus by acquisition of a cytotoxic phenotype

doi: 10.4049/jimmunol.1000818

Figure Lengend Snippet: Tc17 cells are induced in response to infection with vaccinia virus. (A) CD8+ T cells were isolated from the spleens of Balb/c, C57BL/6, or C3H/HeJ mice and stimulated for 48 h before cell free supernatants were used to measure IL-17 and IFN-γ protein levels using ELISA. Results are presented as the mean ± SEM of CD8+ T cells from 3 mice. (B, C) C57BL/6 mice were infected i.p. with 5x106 pfu VV per mouse. Spleens and ovaries were respectively isolated for cytokine analysis following re-stimulation (B) or viral titers (C) at 5, 7, 10, 14, and 21 days postinfection. The data are presented as an average of values from 4 mice per time point ± SEM. (D) C57BL/6 mice were infected on days 0 and 21 with 1x106 pfu VV, i.p., per mouse at each time point. Five days after the second infection, CD8+ and CD4+ cells were isolated from the spleens and re-stimulated before analyzing IL-17 and IFN-γ production using ICS. The data shown are an average of values from 2–4 mice ± SEM. (E–F) BoyJ homozygous (CD45.1+) or heterozygous (CD45.1+CD45.2+) mice were injected i.v. with 1x106 OT-I / Rag1−/− CD8+ (CD45.2+) T cells. One day later, mice were infected with either 2x106 pfu VV-SIINFEKL i.n., or 2x107 pfu VV-SIINFEKL, i.p. (E) Five days after infection, splenocytes were stimulated with SIINFEKL peptide or left unstimulated. CD45.1-CD45.2+ IL-17+ and IFN-γ+ cells were detected using ICS. The data are the average ± SEM of 5 mice. (F) Splenocytes were stimulated with SIINFEKL peptide for 48 h and IL-17 and IFN-γ protein levels in cell free supernatants were measured using ELISA. Statistics in B and D were performed using a Student’s T test. *, p<0.05 compared to unstimulated condition.

Article Snippet: C57BL/6 and Balb/c mice were purchased from Harlan Bioscience (Indianapolis, IN), OT-I / Rag1 −/− mice were purchased from Taconic Farms (Hudson, NY), and Ifng −/− and C3H/HeJ mice were purchased from Jackson Laboratories (Bar Harbor, ME).

Techniques: Infection, Isolation, Enzyme-linked Immunosorbent Assay, Injection