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. Author manuscript; available in PMC: 2016 Jan 15.
Published in final edited form as: Clin Cancer Res. 2015 Mar 31;21(14):3241–3251. doi: 10.1158/1078-0432.CCR-14-3197

Figure 5. HERV-K env-specific CAR+ T cells recognize shed TAA.

Figure 5

(A) Immunoblot (n = 3) revealing presence of HERV-K env in conditioned culture supernatant harvested from tumor cells. (B) Immunoblot assay (n = 3) shows specificity of 6H5 mAb by using recombinant protein (K10) to block binding to shed TAA. Conditioned supernatant from tumor cells containing shed HERV-K env protein was incubated with 6H5 and K10. (C) Immunoblot showing the absence of HERV-K env particles in the flow through after conditioned culture media was concentrated through a 100 kDa membrane. (D) A 4 hour CRA (n = 3; effector: target is 1: 10) using HERV-K env-specific CAR+ T cells as effectors against A375-SM tumor targets incubated with concentrated A375-SM tumor supernatant. Concentrated RPMI media treated were used as control. (E) IFN-γ production by HERV-K env-specific and CD19-specific CAR+ T cells upon incubation with EL4 targets in the presence of concentrated supernatant harvested from EL4 cells or AaPC(clone 4). Data represent mean ± SD from three healthy donors (average of triplicate measurements for each donor). One-way ANOVA for CRA and IFN-γ production assay with Tukey’s multiple comparison test was used *p < 0.05.