Do Antiretroviral Drugs Protect From Multiple Sclerosis by Inhibiting Expression of MS-Associated Retrovirus?

Figures

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  Figure 1. MSRV/HERV-Wenv, TLR4 and TLR2 gene expression in HIV and MS patients and in healthy control (HC). MSRV/HERV-Wenv and TLR4 expression were evaluated by relative quantification RT-PCR using the 2−ΔΔCt method. Fold changes of (A) MSRV/HERV-Wenv, (B) TLR4, and (C) TLR2 expression of each individual sample in each group are illustrated in dots. HIV patients were compared to HC and MS using HC15 as a reference gene. Medians and interquartile range are indicated by bars (HC, n = 22, MS, n = 22, HIV, n = 9; Kruskal-Wallis Test with Dunn's multiple comparison test).

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  Figure 2. MSRV/HERV-Wenv, TLR4 and TLR2 gene expression in HIV patients categorized in treated (ART) and untreated patients (NO ART). MSRV/HERV-Wenv, TLR4 and TLR2 expression were evaluated by relative quantification RT-PCR using the 2−ΔΔCt method. Fold changes of (A) MSRV/HERV-Wenv, (B) TLR4, and (C) TLR2 expression of 9 HIV patients were categorized in treated (ART) and untreated (NO ART) patients are illustrated in dots. Medians and interquartile range are indicated by bars (ART, n = 6, NO ART, n = 3; Mann-Whitney Test).

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  Figure 3. Expression of HERV-WEnv protein in different immune cells in HC and MS by Flow Cytometry. (A) Representative dot plot of the expression of HERV-WEnv in gated CD20+ and CD3+ lymphocytes and CD14+ monocytes. The staining with only secondary CF 488A anti-rabbit antibody (without anti-HERV-WEnv primary Ab) was used as negative control. (B) Expression of HERV-WEnv was detected in total PBMC, CD3+ T cells, CD20+ B cells and CD14+ monocytes from 10 MS patients and 6 HC. Percentage of positive cells is illustrated in dots. Medians and interquartile range are indicated by bars (n MS = 10, n HC = 6; Mann-Whitney Test).

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  Figure 4. MSRV/HERV-Wenv expression increases in LCL and CpG activated cells. (A) MSRV/HERV-Wenv expression was evaluated by relative quantification RT-PCR using the 2-ΔΔCt method. Fold changes of MSRV/HERV-Wenv in isolated CD20+ cells, CpG activated CD20+ cells, LCL (B-lymphoblastoid cell line) and CD20– cells are illustrated in dots. Medians and interquartile range are indicated by bars (n = 5, Kruskal-Wallis Test with Dunn's multiple comparison test). (B) MSRV/HERV-Wenv expression was evaluated by relative quantification RT-PCR using the 2−ΔΔCt method. Fold changes of MSRV/HERV-Wenv in LCL from HC and MS patients are illustrated in dots. Medians and interquartile range are indicated by bars (n HC = 3, n MS = 3; Mann-Whitney Test).

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  Figure 5. Efavirenz decreases the expression of MSRV/HERV-Wenv. LCL from 3 different healthy donors were treated with 0.1, 1, and 10 μM of (A) Lamivudine, (B) Tenofovir, (C) Daranuvir, (D) Efavirenz, and (E) Raltegravir for 5 days. CTRL (Controls) represent LCL treated only with DMSO. MSRV/HERV-Wenv expression was evaluated by relative quantification RT-PCR using the 2−ΔΔCt method and is illustrated in dots (n = 3; Friedman test).

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  Figure 6. Combined ART decreases the expression of HERV-WEnv protein. (A) LCL from 6 different donors were treated with combined Lamivudine, Tenofovir, Daranuvir, Efavirenz, and Raltegravir at 10, 1, and 0.1 μM for 5 days. CTRL represents LCL treated only with DMSO. MSRV/HERV-Wenv expression was evaluated by relative quantification RT-PCR using the 2−ΔΔCt method. Each point represents an LCL (n = 6; Friedman test). (B) LCL were treated with combined drugs at 1 μM for 5 days. HERV-WEnv expression was evaluated by extracellular FC with dead cell marker included. One example is shown. (C) HERV-WEnv expression of 4 individual LCL is illustrated in dots before and after the treatment (n = 4; Wilcoxon test).