Supplementary Data from NL-201 upregulates MHC-I expression and intratumoral TCR diversity, and demonstrates robust antitumor activity as monotherapy and in combination with PD-1 blockade

Supplementary Table S1. Neo-2/15 maximum pSTAT5 signaling (maximal Mean fluorescent intensity, MFI) in human PBMCs and mouse splenocytes. Supplementary Table S2. IL-2 Receptor Signaling in Primary Human and Cynomolgus Monkey CD8+ T Cells induced by IL-2, NL-201, and Neo2/15. Supplementary Figure S1. A-B, Flow cytometric analysis of human PBMCs (A) and mouse splenocytes (B) for cell surface expression of IL-2Rα (CD25), IL-2Rβ (CD122), and IL-2Rγc (CD132). Supplementary Figure S2. Flow cytometric analysis of CT26-hHER2 and CT26 cell death assessed by Annexin V labeling after a 6-hour co-culture with isolated Balb/c NK cells and indicated concentrations of IL-2R agonists. Supplementary Figure S3. NL-201 demonstrates improved in vivo efficacy as compared to Neo-2/15. Supplementary Figure S4. A-B, Flow cytometric analysis of MHC-I (A) and PD-L1 (B) surface expression on in vitro B16F10 cells cultured with the indicated concentrations of cytokines for 1 day. Supplementary Figure S5. A-C, Tumor volume measurements of individual mice bearing subcutaneous B16F10 melanoma (A), EMT6 mammary carcinoma (B), and Renca renal adenocarcinoma (C) tumors. Supplementary Figure S6. A-B, B16F10 tumor-bearing mice were given a single dose of NL-201 by intravenous administration and then harvested 5 days later for ex vivo flow cytometric analysis to measure the abundance of CD45+ cells (A) and CD3+ T cells of CD45+ gate (B).