Increase of IFN-γ and TNF-γ production in CD107a + NK-92 cells co-cultured with cervical cancer cell lines pre-treated with the HO-1 inhibitor

Abstract

BackgroundNatural killer (NK) cells eliminate virus-infected and tumor cells through the release of perforins and granzymes; they also produce Interferon gamma (IFN-γ) and Tumor necrosis factor alpha (TNF-α), which induce apoptosis in target cells. Many tumors express Heme oxygenase 1 (HO-1), and this expression has been associated with avoiding immunosuppression and apoptosis. In this work, HO-1+ Cervical cancer cell (CCC) lines were pre-treated with HO-1 inhibitor and we assessed whether this inhibition enhanced the sensitivity of CCC to NK cell activity.MethodsWe assessed the expression of HO-1 in HeLa, SiHa, and C-33A CCC by Flow cytometry (FC). CCC were pre-treated with SnPP or ZnPP HO-1 inhibitors. After that, NK-92 cells were co-cultured with HeLa, SiHa, and C-33A CCC pre-treated or not with HO-1 inhibitors, and the expression of IFN-γ, TNF-α, CD107a, Granzyme B, NKp44, NKp46, NKp30, and NKG2D was evaluated by FC.ResultsCCC lines HeLa, SiHa, and C-33A expressed HO-1. Inhibition of HO-1 in these cells increased the expression of IFN-γ and TNF-α in CD107a + NK-92 cells. We observed a reduction in the expression of NKG2D, NKp46, and NKp30 in NK cells co-cultured with HeLa and SiHa cells, and when HeLa and SiHa cells were pre-treated with the HO-1 inhibitors, the expression of NKG2D and NKp30 in NK cells was restored. We observed a similar effect in NK cells co-cultured with C-33A cells in NKp30 expression.ConclusionInhibition of HO-1 in CCC induces an increase in IFN-γ and TNF-α production in CD107a + NK-92 cells and restores NKG2D, NKp46 and NKp30 downmodulation in NK cells.

Combination Cancer Therapy Using Chimeric Antigen Receptor-Engineered Natural Killer Cells as Drug Carriers

“…In addition to sensitizing tu-mor cells to NK cytotoxicity, 65 IFN-g release by both primary NK cells and NK cell lines 66 signals to surrounding immune cells, including T cells, dendritic cells, monocytes, and macrophages, initiating broader adaptive and innate immune responses. 67,68 Our in vivo biodistribution study further supports that CAR.NK cells enhance nanoparticle accumulation within the tumor site. Mice treated with cMLV(DiD) without a cell chaperone had significantly greater cMLV accumulation in the liver, likely indicating hepatic clearance as commonly observed with larger liposomes.…”

Regulación de hemooxigenasa e indoleamina por citocinas en células de cáncer cervicouterino y actividad citotóxica de células natural killer

“…ej., IL-10 y factor de crecimiento transformante β [TGF-β], quimiocinas receptor de quimiocina tipo 2 (CCR2) o enzimas tales como IDO y HO-1 23,24 . Se ha reportado la sobreexpresión de HO-1 e IDO en diferentes tipos de cáncer, incluido el cáncer cervicouterino 20,25,26 . Los mecanismos de regulación de estas enzimas van desde factores de transcripción, activados por estímulos externos, hasta los productos de la actividad de las enzimas 27 .…”

“…Asimismo, la inhibición de esta enzima conduce a la reducción del crecimiento tumoral y a un incremento en la sensibilidad a la quimioterapia 19 . En nuestro laboratorio observamos que en células NK en cocultivo con células de cáncer cervicouterino tratadas previamente con un inhibidor específico de HO-1, la producción de IFN-γ y TNF-α por las células NK se incrementó en comparación con aquéllas que estuvieron en contacto con líneas celulares sin tratamiento 20 . Sin embargo, se desconoce el efecto que puedan tener citocinas como IL-4, IL-10, TNF-α e IFN-γ en líneas celulares de cáncer de cervicouterino sobre la expresión de IDO y HO-1, así como la actividad citotóxica de células NK en células de cáncer cervicouterino pretratadas con estos inhibidores.…”

NK cell‑produced IFN‑γ regulates cell growth and apoptosis of colorectal cancer by regulating IL‑15

“…The immune system is responsible for eliminating cancerous cells and foreign infections (4). In particular, natural killer (NK) cells are primarily responsible for eliminating tumor cells through contact-dependent cytotoxicity and cytokine production (5).…”

“…For instance, NK-92 cells attack cancer cells and the tumors grown within the control of the organism (6). One of those cytokines, interferon gamma (IFN-γ), is secreted by NK cells and has been previously reported to promote the apoptosis and cytolysis of target tumor cells (4,7). IFN-γ has immunoregulatory, antiviral and anti-tumor properties (8).…”