The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP 2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

• Published in: Journal of translational medicine Vol. 19; no. 1; p. 306
• Main Authors: Eaton-Fitch, Natalie, Cabanas, Hélène, du Preez, Stanley, Staines, Donald, Marshall-Gradisnik, Sonya
• Format: Journal Article
• Language: English
• Published: England 15.07.2021
• Subjects: Fatigue Syndrome, Chronic - drug therapy; Flow Cytometry; Humans; Interleukin-2; Killer Cells, Natural; TRPM Cation Channels; Myalgic Encephalomyelitis; Transient receptor potential melastatin 3; IL-2; Natural killer cells; Chronic Fatigue Syndrome; PIP2
• ISSN: 1479-5876

Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a serious multifactorial disorder. The origin remains ambiguous, however reduced natural killer (NK) cell cytotoxicity is a consistent immunological feature of ME/CFS. Impaired transient receptor potential melastatin 3 (TRPM3), a phosphatidylinositol dependent channel, and impaired calcium mobilisation have been implicated in ME/CFS pathology. This investigation aimed to examine the localisation of TRPM3 at the NK cell plasma membrane and co-localisation with phosphatidylinositol 4,5-bisphosphate (PIP ). The effect of IL-2 priming and treatment using pregnenolone sulfate (PregS) and ononetin on TRPM3 co-localisation and NK cell cytotoxicity in ME/CFS patients and healthy controls (HC) was also investigated. NK cells were isolated from 15 ME/CFS patients and 15 age- and sex-matched HC. Immunofluorescent technique was used to determine co-localisation of TRPM3 with the NK cell membrane and with PIP of ME/CFS patients and HC. Flow cytometry was used to determine NK cell cytotoxicity. Following IL-2 stimulation and treatment with PregS and ononetin changes in co-localisation and NK cell cytotoxicity were measured. Overnight treatment of NK cells with PregS and ononetin resulted in reduced co-localisation of TRPM3 with PIP and actin in HC. Co-localisation of TRPM3 with PIP in NK cells was significantly reduced in ME/CFS patients compared with HC following priming with IL-2. A significant increase in co-localisation of TRPM3 with PIP was reported following overnight treatment with ononetin within ME/CFS patients and between groups. Baseline NK cell cytotoxicity was significantly reduced in ME/CFS patients; however, no changes were observed following overnight incubation with IL-2, PregS and ononetin between HC and ME/CFS patients. IL-2 stimulation significantly enhanced NK cell cytotoxicity in HC and ME/CFS patients. Significant changes in co-localisation suggest PIP -dependent TRPM3 function may be impaired in ME/CFS patients. Stimulation of NK cells with IL-2 significantly enhanced cytotoxic function in ME/CFS patients demonstrating normal function compared with HC. A crosstalk exists between IL-2 and TRPM3 intracellular signalling pathways which are dependent on Ca influx and PIP . While IL-2R responds to IL-2 binding in vitro, Ca dysregulation and impaired intracellular signalling pathways impede NK cell function in ME/CFS patients.