PMC9835940_01
TIL treatment and correlation with immunological and clinical responses. (A) Treatment schedule. Time point of each TIL infusion was defined as day 0. TIL-1 and TIL-2 infusions were generated from the same initial biopsy, TIL-3 infusion from an independent second tumor biopsy. The patient received conditioning preparative non-myeloablative lymphodepleting chemotherapy (CTX) with cyclophosphamide (60 mg/kg) on day -1 (TIL-1) and (30 mg/kg) on day -4 (TIL-2 and TIL-3) before intravenous infusion of the TIL supported by administration of IL-2 (Aldesleukin) at 600 000 IU/kg. IL-2 was infused four and five times every 12 hours starting 16 hours after each TIL infusion. Four doses of pembrolizumab (1 mg/kg) were administered within the third TIL treatment (TIL-3) on days -7, +15, + 59, and + 80. (B) PSA values: After TIL-1 treatment PSA decreased from 33 microg/L to 18 microg/L within a period of 5 weeks and recurred. After TIL-2 treatment PSA decreased from 34 microg/L to 5 microg/L within a period of 7 weeks and recurred. After TIL-3 treatment PSA levels continuously decreased from 38 microg/L to <0.01 microg/L within 8 weeks and remained there. (C) NY-ESO-1 serum antibody. The patient showed a high antibody titer before TIL treatment. During and after the course of TIL treatments the antibody continuously decreased and is currently detectable only at a residual level. ELISA OD values are shown at 1:400 serum dilution. ( ): cut off OD in healthy donor. (D) MR images over the course of TIL treatment. The tumor has significantly regressed 8 weeks after TIL-3 infusion and has completely resolved 18 months after TIL-3 infusion. The patient continued to demonstrate complete response (CR) 3.5 years after TIL-3 treatment. CR, complete remission; MR, magnetic resonance; NY-ESO-1, New York esophageal squamous cell carcinoma 1; OD, optical density; PSA, prostate-specific antigen; TIL, tumor-infiltrating lymphocyte.
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F1
An overview of the treatment schedule is provided in figure 1A. Concomitantly, the high pre-TIL NY-ESO-1 serum antibody titer continuously decreased over the course of TIL treatment (figure 1B, C). TIL-3. The patient's most recent imaging studies show complete and sustained tumor remission lasting for now 3.5 years (figure 1D).
Specificity of TIL. (A) NY-ESO-1 reactivity measured by IFN-gamma secretion was detectable in all three TIL infusions, in particular for the HLA-A2 restricted epitope NY-ESO-1 p157-165, for NY-ESO-1 20mer peptides p81-100 and p91-110, and for NY-ESO-1 24mer peptide p119-143. (B)Specific cytotoxicity of TIL-2 and TIL-3 against NY-ESO-1/HLA-A2-expressing tumor cells in standard chromium-51 (Cr51) release assay. (C)TIL reactivity specific for several mutational peptides measured by IFN-gamma secretion: KKRCLFRSF derived from gene PRPF8 (TIL 1,2,3), YLGEITYPF from gene TRPS1 (TIL 1,3), FVSGKYKCL derived from gene ZNF512 (TIL 1,3), NQVVQASMEK from gene GFM1 (TIL 2,3), CPSKPSPLV from gene SGIP1 (TIL 3) and peptide VKWAKALPDCERAAS derived from a part of the unique C-terminus of the V12 splice variant AR-V12 (TIL 1,2,3). (D)IFN-gamma secretion of TIL-3 was significantly higher for the mutant versus the wild type peptides derived from PRPF8 and TRPS1 genes. Values represent IFN-gamma secretion by 10 000 TIL and of three independent tests. AR, androgen receptor; HLA, human leukocyte antigen; IFN-gamma, interferon gamma; NY-ESO-1, New York esophageal squamous cell carcinoma 1; TIL, tumor-infiltrating lymphocyte; IFN-gamma, interferon gamma
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F2
Specific reactivity for NY-ESO-1 was found in all three TIL infusions, in particular for the HLA-A2 restricted peptide NY-ESO-1 p157-165, for NY-ESO-1 20mer peptides p81-100 and p91-110 that include 9mer peptides p92-100 and p96-104, known to be presented on HLA-Cw3 and for NY-ESO-1 24mer peptide p119-143 (figure 2A). In addition, TIL showed specific cytotoxicity against NY-ESO-1/HLA-A2 expressing tumor cells in standard chromium-51 (Cr51) release assay (figure 2B). GFM1 (TIL 2,3), CPSKPSPLV from gene SGIP1 (TIL3) and peptide VKWAKALPDCERAAS derived from a part of the unique C-terminus of the V12 splice variant AR-V12 (TIL 1,2,3) (figure 2C). Given that IFN-gamma production was significantly higher for the mutant versus the wild type peptide (figure 2D).