Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Evaluation of the cytotoxic activity of NK cells treated with anti-PSMA Ab in vitro . (A, B) The killing rates of NK cells against 22RV1 cells (PSMA strongly positive) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the Cell Counting Kit-8 (CCK-8) assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (A) , 1:1 (B) , respectively; (C, D) The killing rates of NK cells against PC3 cells (PSMA negative) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the CCK-8 assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (C) , 1:1 (D) , respectively; (E, F) The killing rates of NK cells against RWPE-1 cells (PSMA moderately positive) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the CCK-8 assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (E) , 1:1 (F) , respectively; (G) PSA levels in the culture supernatant of NK cells co-cultured with 22RV1 cells in the control and treatment groups, including NK, NK + IgG, and NK + anti-PSMA Ab (10 μg/mL), measured via ELISA at 2, 6, 12, and 24 h after co-culturing (n = 3); (H) Representative flow cytometry plots and summary data (n = 3) of the MFI for CD107a expression in NK cells co-cultured with 22RV1 cells in the presence of anti-PSMA Ab (10 μg/mL) or IgG control (10 μg/mL). CD107a expression in NK cells was set as a negative control. Degranulation of NK cells was induced upon interaction with 22RV1 cells at a 1:1 ratio with anti-PSMA Ab or IgG for 6 h at 37 °C, the GolgiStop protein transport inhibitor was added during the final 2 h of the culture and NK cells were collected for cytometry measurement; (I, J) Comparison of supernatant perforin (I) and granzyme B (J) levels among NK cells co-cultured with 22RV1 cells and their counterparts co-cultured with 22RV1 cells in the presence of IgG control (10 μg/mL) or anti-PSMA Ab (10 μg/mL) at E:T of 1:1 after 6h coculture (n = 3). NK cells alone were set as negative control; (K, L) Comparison of IFN-γ (K) and TNF-α (L) levels among NK cells co-cultured with 22RV1 cells and their counterparts co-cultured with 22RV1 cells in the presence of IgG control (10 μg/mL) or anti-PSMA Ab (10 μg/mL) at E:T of 1:1 after 6h co-culture (n=3). NK cells alone were set as negative control; Data expressed as means ± SD were plotted, and ANOVA followed by a Tukey’s post hoc test was used to compare three or more groups (A–L) . *p < 0.05; ns, not significant. anti-PSMA Ab, anti-prostate-specific membrane antigen antibody; PSA, prostate-specific antigen; ELISA, enzyme-linked immunosorbent assay; IFN-γ, interferon-γ; TNF-α, tumor necrosis factor-α; E:T, effector-to-target ratio.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: Activity Assay, In Vitro, Cell Counting, CCK-8 Assay, Cell Culture, Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Negative Control, Cytometry, Comparison, Co-Culture Assay, Membrane

Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Development of PDO PCa models and cytotoxicity of combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells against the PDO. (A) Representative hematoxylin-and-eosin images of PCa tissue-derived organoid derived from PCa specimens (scale bar 200 μm). The left image was the panorama, and the right four images were local magnifications part by part (a→a’, b→b’, c→c’, d→d’); (B) Representative PSMA immunohistochemistry images of PCa tissue-derived organoid derived from PCa specimens (scale bar 200 μm). The left image was the panorama, and the right four images were local magnifications part by part (a→a’, b→b’, c→c’, d→d’); (C) Representative bright-field image of coculture of NK cells with PCa tissue-derived organoid in the presence of IgG or the constructed anti-PSMA antibody after 2 h and 6h coculture, PDOs alone were set as controls (n = 3); (D) Cytotoxicity of NK cells with IgG or anti-PSMA antibody (10 μg/mL) against PCa tissue-derived organoid at E/T ratio of 5:1 after 2 h and 6h coculture measured using LDH assay (n = 3); (E) IFN-γ levels of the supernatant after the NK cells were co-cultured with PCa tissue-derived organoid at E/T ratio of 5:1 after 2 h and 6h coculture measured using ELISA (n = 3). Data are shown as mean ± SD. Statistical significance was determined using an unpaired t-test (D, E) . *p < 0.05; ns, not significant. PDO, patient-derived organoid; PCa, prostate cancer; PSMA, prostate-specific membrane antigen; IgG, immunoglobulin G; E/T, effector-to-target ratio; LDH, lactate dehydrogenase; IFN-γ, interferon-gamma; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: Derivative Assay, Immunohistochemistry, Construct, Lactate Dehydrogenase Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Membrane

Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Anti-tumor effect of NK cells against CRPC in combination with anti-PSMA Ab in a subcutaneous tumor model in vivo . (A) Experimental protocol for the CRPC model used in (B–I) : mice were injected with PBS, anti-PSMA Ab or isotype-matched control mAb (10 mg/mg) intraperitoneally (i.p.) on days 10, 18 and injected with NK cells (1 × 10 7 ) intravenously (i.v.) on days 11, 15, 19, and 23 after injections of 2 × 10 6 22RV1 cancer cells subcutaneously (s.c.) on day 0 (n = 6 per group); (B) Tumor volumes at various times (horizontal axis) after tumor inoculation in the control, NK, NK + IgG, and NK + anti-PSMA Ab groups. Tumor volumes were calculated according to the formula L × W 2 /2, where L and W represent the longest and shortest diameters measured using a caliper, respectively (n = 6 per group); (C) Body weights in the control and treatment groups over the whole treatment course (n = 6 per group); (D) Serum PSA levels of mice in the control, NK, NK + IgG, and anti-PSMA Ab groups (n = 6 per group) on days 10, 16, 22, and 28; (E) Images of tumors in mice 28 days after tumor inoculation (n = 5–6 in each group); (F) Tumor weights corresponding to each group when harvested on day 28 (n = 5–6 in each group); (G) HE examination of tumor specimen in the control and treatment groups on day 28 (n = 5–6 in each group); (H) Serum IL-6 levels in the control and treatment groups on day 28; (I) Cumulative Kaplan–Meier survival curves for mice (n = 6 per group) after tumor implantation. Data expressed as the means ± SD were plotted, and ANOVA followed by a Tukey’s post hoc test was used for multiple group comparisons (B-D, F, H) . The Kaplan–Meier method was used to estimate survival functions, and the log-rank test was used for group comparisons (I) . *p < 0.05; ns, not significant. CRPC, castration-resistant prostate cancer; PSMA, prostate-specific membrane antigen; Ab, antibody; PSA, prostate-specific antigen; IL-6, interleukin-6.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: In Vivo, Injection, Control, Tumor Implantation, Membrane