The incidence of cutaneous melanoma (CM) rises despite advances in immunotherapy and targeted therapy. Studies have shown that interferon-gamma (IFN-γ), a pro-tumorigenic cytokine in CM, induces the expression of programmed death-ligand 1 (PD-L1), leading to the escape of melanoma cells from immunosurveillance. Our previous study suggests that neuronal nitric oxide synthase (nNOS)-mediated nitric oxide (NO) signaling is crucial for melanoma progression and forms a mechanistic rationale behind nNOS inhibition as a novel treatment strategy for CM. An additional study demonstrated that IFN-γ interacts with nNOS signaling in melanoma and novel nNOS inhibitors (nNOSi) effectively diminished the induction of PD-L1 post IFN-γ exposure. In this study, we determined the effects of nNOSi on melanoma-induced immunosuppression of T cells. Fresh whole blood samples were collected from 11 disease-free human donors (7 male, 10 Caucasian) between the ages of 20 – 40 years old. CD3+ T cells derived from peripheral blood mononuclear cells (PBMCs) were co-incubated with human metastatic melanoma cells in the presence or absence of IFN-γ (250 U/ml). A selective nNOSi MAC-3-190 was added to suppress nNOS-NO signaling in melanoma cells. IL-2+ T cells detected by flow cytometry represent the activation of CD4+ T cells and CD8+ T cells. Our ex vivo study demonstrated that co-incubation with melanoma significantly reduced the average relative percent of IL-2+ in CD4+ and CD8+ T cell populations to below 0.5-fold compared to T cell alone (p+ and CD8+ T cell populations remained inactivated when co-incubated with IFN-γ-treated melanoma cells as indicated by the decrease in IL-2+ T cells to 0.5-fold of control (p+ T cells as shown by the increase in IL-2+ T cells to 0.8-fold of control compared to IFN-γ-treated melanoma (p
","abstract_html":"<p>The incidence of cutaneous melanoma (CM) rises despite advances in immunotherapy and targeted therapy. Studies have shown that interferon-gamma (IFN-γ), a pro-tumorigenic cytokine in CM, induces the expression of programmed death-ligand 1 (PD-L1), leading to the escape of melanoma cells from immunosurveillance. Our previous study suggests that neuronal nitric oxide synthase (nNOS)-mediated nitric oxide (NO) signaling is crucial for melanoma progression and forms a mechanistic rationale behind nNOS inhibition as a novel treatment strategy for CM. An additional study demonstrated that IFN-γ interacts with nNOS signaling in melanoma and novel nNOS inhibitors (nNOSi) effectively diminished the induction of PD-L1 post IFN-γ exposure. In this study, we determined the effects of nNOSi on melanoma-induced immunosuppression of T cells. Fresh whole blood samples were collected from 11 disease-free human donors (7 male, 10 Caucasian) between the ages of 20 – 40 years old. CD3<sup>+</sup> T cells derived from peripheral blood mononuclear cells (PBMCs) were co-incubated with human metastatic melanoma cells in the presence or absence of IFN-γ (250 U/ml). A selective nNOSi MAC-3-190 was added to suppress nNOS-NO signaling in melanoma cells. IL-2<sup>+</sup> T cells detected by flow cytometry represent the activation of CD4<sup>+</sup> T cells and CD8<sup>+</sup> T cells. Our <em>ex vivo</em> study demonstrated that co-incubation with melanoma significantly reduced the average relative percent of IL-2<sup>+</sup> in CD4<sup>+</sup> and CD8<sup>+</sup> T cell populations to below 0.5-fold compared to T cell alone (<em>p</em>+ and CD8<sup>+</sup> T cell populations remained inactivated when co-incubated with IFN-γ-treated melanoma cells as indicated by the decrease in IL-2<sup>+</sup> T cells to 0.5-fold of control (<em>p</em>+ T cells as shown by the increase in IL-2<sup>+</sup> T cells to 0.8-fold of control compared to IFN-γ-treated melanoma (<em>p</em></p>","abstract_has_math":false,"creators":["Lozada, Kate Alison"],"institution":null,"degree_name":"Master of Science (MS)","degree_level":"Thesis","degree_discipline":"Pharmaceutical Sciences","degree_department":null,"school":null,"contributors":["Sun Yang","Rennolds Ostrom","Jennifer Totonchy","Miao Zhang"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2022,"date_issued":"2022-08-01T07:00:00Z","date_published":"2022-08-01T07:00:00Z","updated_at":"2026-07-24T01:38:24Z","subjects":["melanoma","immunotherapy","nNOS inhibitors","IL-2","T cell","PD-L1","Other Pharmacy and Pharmaceutical Sciences","Pharmaceutics and Drug Design"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/25","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Sun Yang","Rennolds Ostrom","Jennifer Totonchy","Miao Zhang"]},{"key":"dc:creator","label":"Author","values":["Lozada, Kate Alison"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2024-09-01T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Pharmaceutical Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["melanoma","immunotherapy","nNOS inhibitors","IL-2","T cell","PD-L1","Other Pharmacy and Pharmaceutical Sciences","Pharmaceutics and Drug Design"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/25"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The incidence of cutaneous melanoma (CM) rises despite advances in immunotherapy and targeted therapy. Studies have shown that interferon-gamma (IFN-γ), a pro-tumorigenic cytokine in CM, induces the expression of programmed death-ligand 1 (PD-L1), leading to the escape of melanoma cells from immunosurveillance. Our previous study suggests that neuronal nitric oxide synthase (nNOS)-mediated nitric oxide (NO) signaling is crucial for melanoma progression and forms a mechanistic rationale behind nNOS inhibition as a novel treatment strategy for CM. An additional study demonstrated that IFN-γ interacts with nNOS signaling in melanoma and novel nNOS inhibitors (nNOSi) effectively diminished the induction of PD-L1 post IFN-γ exposure. In this study, we determined the effects of nNOSi on melanoma-induced immunosuppression of T cells. Fresh whole blood samples were collected from 11 disease-free human donors (7 male, 10 Caucasian) between the ages of 20 – 40 years old. CD3+ T cells derived from peripheral blood mononuclear cells (PBMCs) were co-incubated with human metastatic melanoma cells in the presence or absence of IFN-γ (250 U/ml). A selective nNOSi MAC-3-190 was added to suppress nNOS-NO signaling in melanoma cells. IL-2+ T cells detected by flow cytometry represent the activation of CD4+ T cells and CD8+ T cells. Our ex vivo study demonstrated that co-incubation with melanoma significantly reduced the average relative percent of IL-2+ in CD4+ and CD8+ T cell populations to below 0.5-fold compared to T cell alone (p+ and CD8+ T cell populations remained inactivated when co-incubated with IFN-γ-treated melanoma cells as indicated by the decrease in IL-2+ T cells to 0.5-fold of control (p+ T cells as shown by the increase in IL-2+ T cells to 0.8-fold of control compared to IFN-γ-treated melanoma (p
"]},{"key":"dc:source","label":"Dc Source","values":["Lozada, KAJ. Effects of nNOS Inhibitors on Melanoma-Induced Immunosuppression. [master’s thesis]. Irvine, CA: Chapman University; 2022. https://doi.org/10.36837/chapman.000401"]},{"key":"dc:title","label":"Title","values":["Effects of nNOS Inhibitors on Melanoma-Induced Immunosuppression"]}]}],"canonical_facts":{"dc:contributor":["Sun Yang","Rennolds Ostrom","Jennifer Totonchy","Miao Zhang"],"dc:creator":["Lozada, Kate Alison"],"dc:date.available":["2024-09-01T07:00:00Z"],"dc:description.abstract":["The incidence of cutaneous melanoma (CM) rises despite advances in immunotherapy and targeted therapy. Studies have shown that interferon-gamma (IFN-γ), a pro-tumorigenic cytokine in CM, induces the expression of programmed death-ligand 1 (PD-L1), leading to the escape of melanoma cells from immunosurveillance. Our previous study suggests that neuronal nitric oxide synthase (nNOS)-mediated nitric oxide (NO) signaling is crucial for melanoma progression and forms a mechanistic rationale behind nNOS inhibition as a novel treatment strategy for CM. An additional study demonstrated that IFN-γ interacts with nNOS signaling in melanoma and novel nNOS inhibitors (nNOSi) effectively diminished the induction of PD-L1 post IFN-γ exposure. In this study, we determined the effects of nNOSi on melanoma-induced immunosuppression of T cells. Fresh whole blood samples were collected from 11 disease-free human donors (7 male, 10 Caucasian) between the ages of 20 – 40 years old. CD3+ T cells derived from peripheral blood mononuclear cells (PBMCs) were co-incubated with human metastatic melanoma cells in the presence or absence of IFN-γ (250 U/ml). A selective nNOSi MAC-3-190 was added to suppress nNOS-NO signaling in melanoma cells. IL-2+ T cells detected by flow cytometry represent the activation of CD4+ T cells and CD8+ T cells. Our ex vivo study demonstrated that co-incubation with melanoma significantly reduced the average relative percent of IL-2+ in CD4+ and CD8+ T cell populations to below 0.5-fold compared to T cell alone (p+ and CD8+ T cell populations remained inactivated when co-incubated with IFN-γ-treated melanoma cells as indicated by the decrease in IL-2+ T cells to 0.5-fold of control (p+ T cells as shown by the increase in IL-2+ T cells to 0.8-fold of control compared to IFN-γ-treated melanoma (p
"],"dc:identifier":["https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/25"],"dc:source":["Lozada, KAJ. Effects of nNOS Inhibitors on Melanoma-Induced Immunosuppression. [master’s thesis]. Irvine, CA: Chapman University; 2022. https://doi.org/10.36837/chapman.000401"],"dc:subject":["melanoma","immunotherapy","nNOS inhibitors","IL-2","T cell","PD-L1","Other Pharmacy and Pharmaceutical Sciences","Pharmaceutics and Drug Design"],"dc:title":["Effects of nNOS Inhibitors on Melanoma-Induced Immunosuppression"],"thesis:degree_discipline":["Pharmaceutical Sciences"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["Master of Science (MS)"]},"updated_at":"2026-07-24T01:38:24Z"}