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. 2020 Sep 17;75(11):1004–1008. doi: 10.1136/thoraxjnl-2020-215027

Figure 3.

Figure 3

Coculture of the biphasic malignant pleural mesothelioma (MPM) cell cultures: MESO-174 and MESO-392 with HLA-A2-restricted, SSX-2 specific, CD8+ T cells-induced T cell degranulation, killing capacity and production of the cytotoxic cytokines interferon γ (IFNγ) and tumour necrosis factor α (TNFα) without the peptide pulse. (A) Barplot presenting the percentage of CD107a+ T cells on coculture with the epithelioid: MESO-044, MESO-278 and the biphasic: MESO-174, MESO-392 MPM cell cultures, without the SSX2-peptide loading. The biphasic cell cultures induced production of CD107a. Data are summarised as mean±SEM. The table below shows the comparisons by one-way analysis of variance (ANOVA) with Tukey’s correction for multiple testing. Quadruplicate samples were used per cell culture/condition. (B) Graph presenting the killing capacity of the T cells on coculture with the epithelioid: MESO-044, MESO-278 and the biphasic: MESO-174, MESO-392 MPM cell cultures, without the peptide loading. The vertical axis shows the percentage of MPM cell death and the horizontal the MPM cell culture. Notably, the biphasic MPM cells triggered T cell cytotoxicity. Data are summarised as mean±SEM. The table below shows the comparisons by one-way ANOVA with Tukey’s correction for multiple testing. Quadruplicate samples were used per cell culture/condition. (C) Barplots of IFNγ (left) and TNFα (right) expression by the T cells on coculture with the epithelioid: MESO-044, MESO-278 and the biphasic: MESO-174, MESO-392 MPM cell cultures, without the peptide pulse. The biphasic MPM cell cultures induced the expression of the cytotoxic cytokines IFNγ and TNFα. Data are summarised as mean±SEM. The tables below show the comparisons by one-way ANOVA with Tukey’s correction for multiple testing. Quadruplicate samples were used per cell culture/condition. (D) SSX-2 mRNA expression by qPCR of the epithelioid cell cultures: MESO-044 and MESO-278 and the biphasic: MESO-174 and MESO-392 corrected to GAPDH and compared with MESO-044 expression. The two biphasic cell cultures displayed increased SSX2 expression levels compared with the epithelioid ones. These data combined suggest the existence of an SSX2+ subpopulation within the biphasic MPM cell cultures MESO-174 and MESO-392. Data are summarised as mean±SEM. The table on the right shows the comparisons by one-way ANOVA with Tukey’s correction for multiple testing. Triplicate samples were used per cell culture/condition.