Pathogenesis of Chronic Enterovirus Infection in Myalgic Encephalomyelitis (ME/CFS) – in vitro and in vivo Studies of Infected Stomach TissuesJohn K. Chia, M.D.
Andrew Chia, Rabiha El-Habbal. EV Med Research. Lomita, CA
Objectives:
Chronic enterovirus infection has been implicated in the pathogenesis of ME/CFS. Previously, we demonstrated enteroviral protein (VP1), RNA and non-cytopathic viruses from the stomach biopsies of ME/CFS patients. The basis of viral persistence has not been clearly defined. Enterovirus can form double-stranded RNA (dsRNA) in tissue cultures and in muscles of infected mice and human. We evaluated the presence of dsRNA in the stomach biopsies and possible infectivity of stomach tissues in SCID mice.
Method:
Archived, paraffin-embedded stomach biopsies from CFS patients and controls were stained for dsRNA using antidsRNA monoclonal antibody and immunoperoxidase technique. 9 cryopreserved ,VP1+ and dsRNA+ stomach biopsy samples were injected ip into SCID mice; and 2 boiled RNA+ samples, 4 VP1-neg and dsRNA-neg samples and one sample of culture medium were injected ip into other 7 SCID mice as controls. Mice were sacrificed 3-4 weeks after infection, and organs processed for viral cultures, EV RNA and viral protein staining.
Results:
108/132 (82%) and 84/132 (64%) of the stomach biopsies from ME/CFS patients stained positive for VP1 and dsRNA respectively, whereas 4/40 (10%) of the control specimens were positive for dsRNA (p<0.01, ?2 test). Pretreatment with RNase III of selective samples diminished or abolished the dsRNA staining; higher concentrations of enzyme and incubation period were required for specimens from sicker patients. 21/23 (91%) of stomach biopsies previously tested positive for EV RNA by RT-PCR or had grown non-cytopathic virus were positive for dsRNA. Of organs taken from SCID mice injected with stomach biopsies, 7/9 (78%) spleen specimens were positive whereas 0/7 controls were positive for VP1 protein by immunoperoxidase staining (p<0.01, ?2 test). 4/9 lung specimens 0/8 of heart, liver and kidney sections demonstrated VP1 staining. All tissue homogenates were negative for EV RNA or growth of virus in BGMK-DAF and WI-38 cells.
Conclusion:
DsRNA was frequently demonstrated in the VP 1+ stomach biopsies taken from ME/CFS patients, and most EV RNA+ samples had detectable dsRNA. In pilot experiments, dsRNA+ samples were infectious in SCID mice, as compared to control samples. Enteroviral dsRNA may play a central role in the pathogenesis of chronic EV infection and ME/CFS, and the mechanism should be further investigated.
Källa: Abstract fron IACFS/ME konferensen hösten 2011:
http://www.iacfsme.org/LinkClick.aspx?fileticket=%2bG6GTkbP33I%3d&tabid=499