PRELIMINARY EXAMINATION OF REGIONAL
DISTRIBUTION OF HUMAN CORONAVIRUS RNA IN POST MORTEM HUMAN BRAIN
Arlene R. Collins, Lynn Diviak,
Alicia Arnold and Mary J. Evans
State University of New York at Buffalo
In previous experiments, human
coronavirus OC43 (HcoV-OC43) was detected in the brain and cerebrospinal
fluid (CSF) of patients with multiple sclerosis, medullary atrophy, Parkinson’s
, polyneuropathy, senile dementia, and headache. HcoV strain 229E was
detected in CSF of patients with multiple sclerosis, headache, and medulary
atrophy (Cristallo, A. New Microbiologia 20:105-114, 1997; Arbour, N, J. Virol.
74:8913-8921, 2000). This research is directed at detection of
coronaviruses in postmortem brain from subjects with schizophrenia and bipolar
disorder by RT-PCR analysis. Twenty-four subjects are being studied
(Stanley Bank, Los Vegas set). Areas of the brain that are associated with
neuropathology in schizophrenia and bipolar disorder were selected for
examination, i.e. temporal lobe and cerebellum. Brain contains many
distinct cell types and is regionally heterogeneous. The outcome of these
research activities is to develop new insights into the regional biological
factors that affect manifestation of Neuropsychiatric disease.
One-step RT-PCR assay: The Perkin Elmer
EZrTth RNA PCR Assay was used. For HCoV-OC43 the primers of Arbour, N.
(JVI 74:8913-8921, 2000) were adaptable. For HCoV-229E, the primers of
Pitkaranta, A. (Pediatrics: 102:291-295, 1998) were suitable. Of the twenty-four
temporal lobe and cerebellar samples examined in duplicate, none was positive
for HCoV-OC43 or 229E strains. The integrity of all of the RNA samples was
verified by RT-PCR amplification of the cellular mRNA triosephosphate
isomerase. This result was not unexpected since Arbour, N. et al, JVI
74:8913-8921, 2000 indicated that one round of cycling would not yield
detectable PCR product. However, the same authors reported detection of
OC43 RNA in brain tissue by direct northern blotting.
Single tube “hanging droplet”
nested PCR assay: A nested PCR assay was adapted from Walsh E.E. et al (J. Med.
Virol. 63:259-263, 2001). Out of twenty-four temporal lobe tissues
examined for HCoV-OC43, ten were positive (41.6%), compared with a prevalence of
35.9% for MS patients reported by Arbour, N. et al. For HCoV-229E, two out
of twenty-four were positive (12%) compared with 51.3% for MS patients reported
by Arbour, N et al. Out of twenty-four cerebellar tissues examined for
HCoV-OC43, seven were positive (29%). For HCoV-229E, for were positive
(16.6%). In three subjects, both tissues were positive for HCoV-OC43 and
in two subjects, both tissues were positive for HCoV-229E. One subject was
positive for both viruses. The uneven tissue distribution of HCoV-OC43 may
indicate selective vulnerability to the virus.
Poster:
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Background
Early experiments to detect HCoV RNA in human brain, in which MS autopsy and biopsy specimens from plaque and from areas adjacent to plaque were examined by dot blot with antisense P32 probe to HCoV strain OC43 nucleocapsid, found no detectable RNA in 20 samples from five patients (Sorenson, O. et al. Neurology:36: 1604- 1606, 1986 ). The probe was prepared from infected mouse brain polyA+ RNA and the cellular RNA was removed by two subtractive hybridizations with mouse brain DNA. Subsequently, HCoV were detected by RT-PCR in the cerebrospinal fluid (CSF) (Table 1) (Cristallo, A. New Microbiologia 20: 105, 1997) and brain (Table 2) (Arbour, N. JVI 74: 8913, 2000). As shown in Tables 1 and 2, the OC43 strain was detected in patients with multiple sclerosis, medullary atropy, Parkinson’s, polyneuropathy, senile dementia, headache, schizophrenia. HCoV strain 229E was detected in patients with multiple sclerosis, Parkinson’s, schizophrenia, senile dementia, headache, medullary atrophy and depression. Arbour et al. (JVI 74: 8913, 2000) detected OC43 RNA in two MS autopsy samples by Northern hybridization with P32 riboprobe to OC43 nucleocapsid and in two samples, from one MS patient and one a healthy control by in situ hybridization with S35 riboprobe. No viral signal for the 229E strain was observed.
From: Cristallo, A. et al. New Microbiologia 20: 105
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