Epstein-Barr virus DNA modulates regulatory T-cell programming in addition to enhancing interleukin-17A production via Toll-like receptor 9 - Fig 1

<p><b>Relative gene expression of (A) IL-17A and (B) CD4 in BALB/c mouse splenic tissues.</b> BALB/c mice were intraperitoneally injected with sterile distilled water, EBV DNA (144x10³ copies) or <i>S</i>. <i>epidermidis</i> DNA (28.3pg). Spleens were collected on days 3, 6 and 9 post-injection and relative gene expression was assessed by reverse transcriptase real-time PCR. Levels were normalized to those on day 3 post-injection with sterile distilled water. * indicates p<0.05 compared to water-injected group on respective day of injection.</p

Average serum levels of IL-17A in RA patients and non-RA controls.

<p>Serum was collected from 24 RA patients and 24 non-RA controls. IL-17A levels were then assessed by ELISA. * indicates p<0.05 compared to controls.</p

Primers used for real-time PCR.

<p>Primers used for real-time PCR.</p

IL-17A levels from BALB/c mouse PBMC.

<p>PBMCs were cultured with EBV DNA (9000 copies), TLR9 inhibitor alone (1.4μM), EBV DNA and TLR9 inhibitor, or with <i>Staphyloccocus epidermidis</i> DNA. Untreated cells were examined as controls. After a 24-hour culture period IL-17A levels were assessed in the culture medium by ELISA. * indicates p<0.05 when compared to untreated cells; ** indicates p<0.05 when compared to cells cultured with EBV DNA.</p

Relative gene expression of RORγT in BALB/c mouse splenic tissues.

<p>BALB/c mice were intraperitoneally injected with sterile distilled water, EBV DNA (144x10³ copies) or <i>S</i>. <i>epidermidis</i> DNA (28.3pg). Spleens were collected on days 3, 6 and 9 post-injection and RORγT relative gene expression was assessed by reverse transcriptase real-time PCR. Levels were normalized to those on day 3 post-injection with sterile distilled water. * indicates p<0.05 compared to water-injected group on respective day of injection.</p

Average levels of EBV DNA in RA patients and non-RA controls.

<p>PBMCs were separated from blood collected from 24 RA patients and 24 non-RA controls. DNA was extracted from subject cells and then EBV DNA copy numbers were determined by qPCR. * indicates p<0.05 compared to controls.</p

Relative gene expression of CTLA4 in BALB/c mouse splenic tissues.

<p>BALB/c mice were intraperitoneally injected with sterile distilled water, EBV DNA (144x10³ copies) or <i>S</i>. <i>epidermidis</i> DNA (28.3pg). Spleens were collected on days 3, 6 and 9 post-injection and CTLA4 relative gene expression was assessed by reverse transcriptase real-time PCR. Levels were normalized to those on day 3 post-injection with sterile distilled water. * indicates p<0.05 compared to water-injected group on respective day of injection.</p

Epstein-Barr virus DNA modulates regulatory T-cell programming in addition to enhancing interleukin-17A production via Toll-like receptor 9 - Fig 8

<p><b>Correlation between EBV DNA copy numbers and serum IL-17A levels in (A) RA patients and (B) non-RA controls.</b> EBV DNA copy numbers and IL-17A levels were assessed in RA and non-RA control subjects by qPCR and ELISA respectively.</p