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. 2021 Jul 13;12:687806. doi: 10.3389/fimmu.2021.687806

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Copyright © 2021 Cabanas, Muraki, Eaton-Fitch, Staines and Marshall-Gradisnik

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Modulation of PregS- evoked currents with Ononetin in NK cells from ME/CFS patients taking LDN compared to HC. Data were obtained under whole-cell patch clamp conditions. (A) A representative time-series of current amplitude at +100 mV and −100 mV showing the effect of 10 µM ononetin (grey) on ionic currents in the presence of 100 µM PregS in NK cells from HC. (B) I–V before and after application of ononetin in the presence of PregS in a cell as shown in (A). (C) Scatter plots representing change of each current amplitude before and after application of ononetin in presence of PregS in all NK cells from HC. Each cell represented as red lines had reduction in currents by ononetin. (D) A representative time-series of current amplitude at +100 mV and −100 mV showing the effect of 10 µM ononetin (grey) on ionic currents in the presence of 100 µM PregS in NK cells from ME/CFS patients taking LDN. (E) I–V before and after application of ononetin in the presence of PregS in a cell as shown in (D). (F) Scatter plots representing change of each current amplitude before and after application of ononetin in presence of PregS in all NK cells from ME/CFS patients taking LDN. Each cell represented as red lines had reduction in currents by ononetin. (G) Table summarizing data for sensitive and insensitive cells to the application of 10 µM ononetin in presence of PregS in ME/CFS patients taking LDN (N = 9; n = 49) compared to HC (N = 9; n = 56). Data are analysed with Fisher’s exact test. (H). Bar graphs representing TRPM3 current amplitude at + 100 mV after stimulation with 100 mu;Mu; PregS and application of 10 µM ononetin in presence of PregS in isolated NK cells from ME/CFS patients taking LDN (N = 9; n = 25) compared with HC (N = 9; n = 35). Only the records sensitive to ononetin have been included from (G). The ROUT outlier test has been applied and any outliers identified have been removed. Data are represented as mean ± SEM. HC, healthy controls; LDN, low dose naltrexone; ME/CFS, myalgic encephalomyelitis/chronic fatigue syndrome; NK, natural killer; PregS, Pregnenolone sulfate.