Archana Thakur

Shows Phenotype, Viability, and Cytotoxicity of the Product.

A and B. Evaluation of Vβ repertoire. The three distinct patterns were observed for Vβ repertoire in CD3+/IFN-γ+ T cells post IT. Pattern 1) the proportions of Vβ repertoire noticeably higher after IT relative to preIT PBMC (solid black arrows); Pattern 2) the proportions of Vβ repertoire that were marginally higher at mid- and/or post-IT relative to pre-IT PBMC (dashed black arrows); Pattern 3) large proportions of Vβ repertoire were either lower or similar at mid-IT or post-IT relative to preIT PBMC in all four patients tested. PreIT (pretherapy), midIT (preinfusion 5) postIT (1 week post infusion 8 of BATs).

A-C. FFPE sections of PC were stained by multiplex staining panel for CD4, CD8, FoxP3 (T regulatory cells), CD19 (B cells), CD68 (macrophages). Stained sections were imaged on the Hamamatsu slide scanner NanoZoomer S360 and images were viewed on the NDP viewer software. Analysis of images were performed on online Visiopharm software. Using the low power image, high-powered Fields (HPFs) were selected from specific tumor regions. Data shows the distribution of CD4+ (teal), CD8+ (purple), FoxP3 (brown), CD19+ (green) and CD68+ (yellow) cells on two representative whole scanned slide images (KCI 04 and KCI 11) are shown. Top panel shows whole scanned tumor sections, inset 1 and 2 are shown below at high magnification (40x). B and C) Show the second set of serial sections of tumor specimens stained for IFN-γ and IL-10 from two patients shown in the upper panel, both IFN-γ (brown) and IL-10 (brown) were stained individually.

Representativeness of Study Participants.

Shows the persistence of anti-CD3 x anti-HER2 bispecific antibody armed activated T cells (HER2 BATs) in the peripheral blood of two out of four patients.

Tumor clearance by T cells is impaired by insufficient tumor antigen recognition, insufficient tumor infiltration, and the immunosuppressive tumor microenvironment. Although targeted T cell therapy circumvents failures in tumor antigen recognition, suppression by the tumor microenvironment and failure to infiltrate the tumor can hinder tumor clearance. Checkpoint inhibitors (CPIs) promise to reverse T cell suppression and can be combined with bispecific antibody armed T cell (BAT) therapy to improve clinical outcomes. We hypothesize that adoptively transferred T cell function may be improved by the addition of CPIs if the inhibitory pathway is functionally active. This study develops a kinetic-dynamic model of killing of hormone receptor–positive breast cancer cells mediated by BATs using single-cell transcriptomic and temporal protein data to identify T cell phenotypes and quantify inhibitory receptor expression. LAG3, PD-1, and TIGIT were identified as inhibitory receptors expressed by cytotoxic effector CD8 BATs upon exposure to hormone receptor–positive breast cancer cell lines. These data were combined with real-time tumor cytotoxicity data in a multivariate statistical analysis framework to predict the relevant contributions of T cells expressing each receptor to tumor reduction. A mechanistic kinetic-dynamic mathematical model was developed and parametrized using protein expression and cytotoxicity data for in silico validation of the findings of the multivariate statistical analysis. The model corroborated the predictions of the multivariate statistical analysis which identified LAG3+ BATs as the primary effectors, while TIGIT expression dampened cytotoxic function. These results inform CPI selection for BATs combination therapy and provide a framework to maximize BATs antitumor function.

Figure S7 from Peripheral Blood IFN Responses to Toll-Like Receptor 1/2 Signaling Associate with Longer Survival in Men with Metastatic Prostate Cancer Treated with Sipuleucel-T

Figure S5 from Peripheral Blood IFN Responses to Toll-Like Receptor 1/2 Signaling Associate with Longer Survival in Men with Metastatic Prostate Cancer Treated with Sipuleucel-T

Figure S3 from Peripheral Blood IFN Responses to Toll-Like Receptor 1/2 Signaling Associate with Longer Survival in Men with Metastatic Prostate Cancer Treated with Sipuleucel-T

Figure S4 from Peripheral Blood IFN Responses to Toll-Like Receptor 1/2 Signaling Associate with Longer Survival in Men with Metastatic Prostate Cancer Treated with Sipuleucel-T