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. 2019 Aug 14;10:1939. doi: 10.3389/fimmu.2019.01939

Figure 3.

Figure 3

Vaccination with HER2-DC1 enhances anti-HER2 Th1 immune response and increases T cell infiltration in TUBO bearing mice. (A) Splenocytes from class I HER2-DC1 vaccinated mice were re-stimulated with p66 rHER2 peptide. Culture supernatants were collected after 3 days and IFN-γ secretion was measured by standard IFN-γ ELISA. (B) Splenocytes from class II HER2-DC1 vaccinated mice were re-stimulated with (class II) rat HER2 peptides (rHER2); p5 (ELAAWCRWGFLLALLPPGIAG), p435 (IRGRILHDGAYSLTLQGLGIH), and p1209 (SPPHPSPAFSPAFDNLYYWDQ) individually and IFN-γ secretion was measured by standard IFN-γ ELISA. (C) Flow gating strategy and analysis of T cell infiltration in tumors by flow cytometry. Tumors were harvested on day 30 and processed as described in Materials and Methods section. Single cell suspension was stained for live/dead near IR, CD3, CD4, and CD8. Data were acquired on an LSRII flow cytometer and analyzed by FlowJo software. (D) Bar graphs represent T cell infiltration gated on live cells per mg of tumor and (E) percent T cells of all live cells within the tumor single cell suspension;. ***p < 0.001, **p < 0.01, *p < 0.05 using Student t-test.