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.





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.


Cristallo, A. et al. New Microbiologia 20: 105