Figure 4.

MV infection of the NY-ESO-1 donor cell lines sensitizes presenting cell lines to recognition by the NY-ESO-1-specific CD4+ T cell clone NY67. (A-E) NY-ESO-1 donor cell lines, M6 or M18, were cultured alone, infected for 3 days with MV at an MOI of 2, or underwent 3 freeze/thaw cycles (FT). They were then cultured for 24 h alone or with presenting cell lines in the presence or absence of IFN-γ. The NY-ESO-1-specific CD4+ T cell clone NY67 was added to these cultures in presence of brefeldin A. 5 h later, IFN-γ response of the clone was measured by flow cytometry after CD4/IFN-γ staining. (A) Example of CD4/IFN-γ staining of the clone NY67 obtained in one experiment where M18 is the NY-ESO-1 donor cell line and M88 and M199 are the presenting cells lines. (B and C) Results are expressed as mean ± SEM of the percentage of IFN-γ-producing CD4+ T cells obtained from three independent experiments with M88 (B) or M199 (C) as presenting cell lines, respectively. (D) Example of CD4/IFN-γ staining of the clone NY67 obtained in one experiment where M6 is the NY-ESO-1 donor and also the presenting cell line. (E) Results are expressed as mean ± SEM of the percentage of IFN-γ-producing CD4+ T cells obtained from three independent experiments with M6.