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. 2020 Apr 21;9:e53244. doi: 10.7554/eLife.53244

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© 2020, Murata et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 1. — (A, B) Staining of HLA-A*02:01⁺ (A) and C*05:01⁺ (B) M87 TILs with pHLA multimers whose peptides were previously known or predicted by publicly available algorithms. The peptides employed for the A*02:01 and C*05:01 multimers are shown in Supplementary files 1b and f, respectively. (A) The TILs showed positivity to the A*02:01/ABCB5_700-708 (No. 14), A*02:01/gp100_154-162 (No. 51), A*02:01/tyrosinase_369-377 (No. 66), A*02:01/wild-type MART1_27-35 (No. 83), and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. (B) The TILs showed positivity to the C*05:01/tyrosinase_460-468 (No. 36) multimer. The A*02:01/HIV pol_476-484 (No. 87 in A), A*02:01/HTLV-1 tax_11-19 (No. 88 in A), A*02:01/unexchanged (No. 89 in A), C*05:01/HIV rev_67-75 (No. 50 in B), and C*05:01/unexchanged (No. 51 in B) multimers were used as negative controls. (C–E) Positive staining of TILs with a panel of pHLA multimers for HLA-A*02:01⁺ M25, M31, M37, M40, M66, and M96 TILs (C), HLA-A*24:02⁺ M37 TILs (D), and HLA-B*07:02⁺ M68 TILs (E). All the high-throughput staining data are shown in Figure 1—figure supplement 1. (C) M25 and M96 TILs showed positivity to the A*02:01/wild-type MART1_27-35 (No. 83) and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. M31 TILs showed positivity to the A*02:01/wild-type NY-ESO-1_157-165 (No. 85) and A*02:01/heteroclitic NY-ESO-1_157-165 (No. 86) multimers. M37 TILs showed positivity to the A*02:01/SSX2_41-49 (No. 48) multimer. M40 TILs showed positivity to the A*02:01/SSX2_41-49 (No. 48), A*02:01/wild-type NY-ESO-1_157-165 (No. 85), and A*02:01/heteroclitic NY-ESO-1_157-165 (No. 86) multimers. M66 TILs showed positivity to the A*02:01/gp100_154-162 (No. 51), A*02:01/gp100_209-217 (No. 54), A*02:01/gp100_280-288 (No. 55), A*02:01/wild-type MART1_27-35 (No. 83), and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. (D) M37 TILs showed positivity to the A*24:02/gp100-intron4 (No. 13) multimer. (E) M68 TILs showed positivity to the B*07:02/NY-ESO-1_60-72 (No. 4) and B*07:02/MAGE-A1_289-297 (No. 51) multimers. The A*02:01/HTLV-1 tax_11-19 (No. 88 in C), A*24:02/HTLV-1 tax_301-309 (No. 64 in D), and B*07:02/HIV nef_128-137 (No. 59 in E) multimers were used as negative controls. The red squares highlight positive TIL staining with pHLA multimers. The percentage of multimer⁺ cells in CD8⁺ T cells is shown. The data shown are representative of two independent experiments.

Figure 1—figure supplement 1. — (A, B) Staining of HLA-A*02:01⁺ (A) and C*05:01⁺ (B) M87 TILs with pHLA multimers whose peptides were previously known or predicted by publicly available algorithms. The peptides employed for the A*02:01 and C*05:01 multimers are shown in Supplementary files 1b and f, respectively. (A) The TILs showed positivity to the A*02:01/ABCB5_700-708 (No. 14), A*02:01/gp100_154-162 (No. 51), A*02:01/tyrosinase_369-377 (No. 66), A*02:01/wild-type MART1_27-35 (No. 83), and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. (B) The TILs showed positivity to the C*05:01/tyrosinase_460-468 (No. 36) multimer. The A*02:01/HIV pol_476-484 (No. 87 in A), A*02:01/HTLV-1 tax_11-19 (No. 88 in A), A*02:01/unexchanged (No. 89 in A), C*05:01/HIV rev_67-75 (No. 50 in B), and C*05:01/unexchanged (No. 51 in B) multimers were used as negative controls. (C–E) Positive staining of TILs with a panel of pHLA multimers for HLA-A*02:01⁺ M25, M31, M37, M40, M66, and M96 TILs (C), HLA-A*24:02⁺ M37 TILs (D), and HLA-B*07:02⁺ M68 TILs (E). All the high-throughput staining data are shown in Figure 1—figure supplement 1. (C) M25 and M96 TILs showed positivity to the A*02:01/wild-type MART1_27-35 (No. 83) and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. M31 TILs showed positivity to the A*02:01/wild-type NY-ESO-1_157-165 (No. 85) and A*02:01/heteroclitic NY-ESO-1_157-165 (No. 86) multimers. M37 TILs showed positivity to the A*02:01/SSX2_41-49 (No. 48) multimer. M40 TILs showed positivity to the A*02:01/SSX2_41-49 (No. 48), A*02:01/wild-type NY-ESO-1_157-165 (No. 85), and A*02:01/heteroclitic NY-ESO-1_157-165 (No. 86) multimers. M66 TILs showed positivity to the A*02:01/gp100_154-162 (No. 51), A*02:01/gp100_209-217 (No. 54), A*02:01/gp100_280-288 (No. 55), A*02:01/wild-type MART1_27-35 (No. 83), and A*02:01/heteroclitic MART1_26-35 (No. 84) multimers. (D) M37 TILs showed positivity to the A*24:02/gp100-intron4 (No. 13) multimer. (E) M68 TILs showed positivity to the B*07:02/NY-ESO-1_60-72 (No. 4) and B*07:02/MAGE-A1_289-297 (No. 51) multimers. The A*02:01/HTLV-1 tax_11-19 (No. 88 in C), A*24:02/HTLV-1 tax_301-309 (No. 64 in D), and B*07:02/HIV nef_128-137 (No. 59 in E) multimers were used as negative controls. The red squares highlight positive TIL staining with pHLA multimers. The percentage of multimer⁺ cells in CD8⁺ T cells is shown. The data shown are representative of two independent experiments.