Cytotoxic T cell-resistant variants are selected in a virus-induced demyelinating disease.

C57BI/6 mice infected with mouse hepatitis virus, strain JHM (MHV-JHM) develop a chronic demyelinating encephalomyelitis. Infectious virus can be isolated only from symptomatic mice. In C57BI/6 mice, two CD8+ T cell epitopes within the MHV-JHM surface glycoprotein were previously identified. Here, we show that mutations in the RNA encoding the immunodominant of the epitopes are present in nearly all virus samples isolated from these mice. Mutations are not present in sequences flanking this epitope or in other CD8+ or CD4+ T cell epitopes. Furthermore, analysis of five peptides corresponding to variant epitopes in direct ex vivo cytotoxicity assays showed that each mutation caused a loss of epitope recognition. These results suggest that escape from CD8+ T cell recognition is necessary for enhanced virus replication and development of clinical disease in these MHV-JHM-infected mice.

The dynamics of role interaction.

Success in our respective business environments is not totally dependent on technical expertise; we must be able to also effectively interact with people. The necessity of successful leadership is assumed; however, we often fail to recognize the value of strategically subordinating ourselves to others. Both roles must be emphasized with the knowledge that preferred individual styles are valid.

Anterograde tracing of trigeminal afferent pathways from the murine tooth pulp to cortex using herpes simplex virus type 1.

Due to its predominantly nociceptive innervation, viral tracing from the tooth pulp provides a potential means for tracing central pain pathways. The neural pathways from the tooth pulp to cortex were determined using in situ hybridization to detect the anterograde transneuronal spread of herpes simplex virus type 1 strain H129 following inoculation into the murine mandibular incisor pulp. Virus first appeared in the brain at day 3 in the dorsomedial region of all three subnuclei of the spinal trigeminal nucleus and the principal sensory nucleus. By days 5-6 virus had spread to the contralateral medial nucleus of the medial geniculate complex, posterior thalamus, and ventroposteromedial thalamus. At days 7-8 virus was detected in laminae IV and Va of the primary somatosensory cortex and lamina IV of the secondary somatosensory cortex in regions previously shown to receive input from the lower jaw. Several mice also showed infection of laminae II/III of the ipsilateral dysgranular insular cortex, along with labeling for virus in the ipsilateral external lateral parabrachial nucleus, posterior thalamus, and posterior basolateral amygdala. Our results are highly consistent with previous tracing and electrophysiological studies utilizing the tooth pulp and with studies implicating the infected structures in nociception. Viral spread appeared to define two separate afferent systems with infection of structures which have been implicated in the sensory-discriminative aspects of pain, such as the ventroposteromedial thalamus and somatosensory cortex, as well as in the dysgranular insular cortex and related subcortical nuclei which may have a role in the affective-motivational aspects of pain.

Spread of MHV-JHM from nasal cavity to white matter of spinal cord. Transneuronal movement and involvement of astrocytes.

C57B1/6 mice infected intranasally with mouse hepatitis virus, strain JHM (MHV-JHM) develop hindlimb paralysis with histological evidence of demyelination several weeks after inoculation. Virus must spread from the site of inoculation, the nasal cavity, to the site of disease, the white matter of the spinal cord. It has been shown previously that after intranasal inoculation, virus enters the brain via the olfactory nerve and spreads to infect many of its neuroanatomic connections within the central nervous system (CNS). In this report, it is shown that virus infecting the spinal cord is first detected in the gray matter, with spread occurring to the white matter soon thereafter. Astrocytes are heavily infected during the process of spread from the gray to the white matter of the spinal cord. Since astrocytes are in intimate contact with neuronal synapses and are themselves connected via gap junctions, these results suggest that astrocytes may be a conduit for the spread of virus in these mice. Astrocytes provide factors for the proliferation and survival of oligodendrocytes, and widespread infection of these cells might contribute to the demyelinating process eventually observed in these mice. Additionally, since virus first appears at specific locations in the spinal cord, it should be possible to determine the source of the virus infecting the cord. While the results are not definitive, the data are most consistent with virus spreading from the ventral reticular formation to the gray matter of the cervical spinal cord.

Herpes simplex encephalitis in the temporal cortex and limbic system after trigeminal nerve inoculation.

Herpes simplex virus type 1 causes an encephalitis in humans that is primarily restricted to the temporal lobe and limbic system. The distribution of lesions suggests that virus enters the brain from a single site and then spreads transneuronally to infect connected structures. Two obvious sites of potential viral entry are the olfactory and trigeminal nerves. Trigeminal nerve entry is more likely because it innervates the oral cavity, a common site of initial infection, and the trigeminal ganglion is the most common site of viral latency. In previous reports, however, experimental trigeminal nerve infection has never led to the pattern of disease observed in humans. By directly inoculating virus into the murine tooth pulp, the mandibular division of the trigeminal nerve was selectively infected. This division, which innervates the oral cavity, is the one most commonly infected in humans. Intrapulp inoculation led to an encephalitis primarily affecting the temporal cortex and limbic system. Thus, spread via the trigeminal nerve provides an explanation for the distribution of herpes simplex virus observed in the human encephalitis.

Two neurotropic viruses, herpes simplex virus type 1 and mouse hepatitis virus, spread along different neural pathways from the main olfactory bulb.

Several neurotropic viruses enter the brain after peripheral inoculation and spread transneuronally along pathways known to be connected to the initial site of entry. In this study, the pathways utilized by two such viruses, herpes simplex virus type 1 and mouse hepatitis virus strain JHM, were compared using in situ hybridization following inoculation into either the nasal cavity or the main olfactory bulb of the mouse. The results indicate that both viruses spread to infect a unique and only partially overlapping set of connections of the main olfactory bulb. Both quantitative and qualitative differences were observed in the patterns of infection of known primary and secondary main olfactory bulb connections. Using immunohistochemistry for tyrosine hydroxylase combined with in situ hybridization, it was shown that only herpes simplex virus infected noradrenergic neurons in the locus coeruleus. In contrast, both viruses infected dopaminergic neurons in the ventral tegmental area, although mouse hepatitis virus produced a more widespread infection in the A10 group, as well as infecting A8 and A9. The results suggest that differential virus uptake in specific neurotransmitter systems contributes to the pattern of viral spread, although other factors, such as differences in access to particular synapses on infected cells and differences in the distribution of the cellular receptor for the two viruses, are also likely to be important. The data show that neural tracing with different viruses may define unique neural pathways from a site of inoculation. The data also demonstrate that two viruses can enter the brain via the olfactory system and localize to different structures, suggesting that neurological diseases involving disparate regions of the brain could be caused by different viruses, even if entry occurred at a common site.

The olfactory nerve and not the trigeminal nerve is the major site of CNS entry for mouse hepatitis virus, strain JHM.

Several viruses, including mouse hepatitis virus strain JHM (MHV-JHM), enter the brain after intranasal inoculation and spread transneuronally to other parts of the central nervous system (CNS). Both the olfactory and trigeminal nerves innervate the nasal cavity and are potential portals of virus entry into the CNS. To evaluate the relative importance of each nerve for MHV infection, mice were infected under conditions that discriminated between trigeminal and olfactory nerve entry. When olfactory nerve entry was selectively eliminated by surgical removal of both olfactory bulbs or by chemical destruction of the olfactory epithelium, MHV-JHM spread into the CNS was completely prevented. On the other hand, direct inoculation into the olfactory bulb, which eliminates all entry via the trigeminal nerve, had no effect on the pattern of virus infection. Thus MHV-JHM enters the CNS via the olfactory nerve after intranasal inoculation while entry via the trigeminal nerve is an insignificant part of this process.

Correlates of postural hypotension in a community sample of elderly blacks and whites.

Postural hypotension is thought to be prevalent among the elderly, but few community-based studies of this condition have been conducted. In addition, little is known about postural hypotension in blacks despite well documented racial differences in hypertension and stroke. Data on 659 elderly (greater than or equal to 60 years of age) participants in a survey of two rural, biracial townships were analyzed to describe the frequency and correlates of postural hypotension. Twelve percent of the 659 adults experienced a drop of 10 mmHg or greater in systolic blood pressure on going from sitting to standing (supine measures were not available). This degree of postural hypotension was twice as common for whites as for blacks (14.5% vs 7.5%, P = 0.01). Postural hypotension was associated with elevated sitting blood pressure and showed positive but statistically non-significant relationships with anti-hypertensive medications and leanness. The association between race and postural hypotension persisted after adjusting for these and other risk factors (OR = 2.2, 95% CI:1.2,4.0).

Schizophrenic deficit in span of apprehension.

We investigated two hypotheses about the span of apprehension task in schizophrenia: (a) schizophrenics show performance deficit on the forced-choice (FC) version but not on the full-report (FR) version, and (b) schizophrenic impairment on the FC version is greater when the display subtends a wide visual angle than when it subtends a narrow one. Schizophrenic (n = 21) and normal (n = 22) groups were tested on 3 versions of the task. A narrow-angle FC version was matched psychometrically with a wide-angle one by use of a greater number of noise letters in the narrow version. Schizophrenics reported fewer correct letters than normals on the FR version but did not differ on either the wide or the narrow FC versions. The results imply that schizophrenic deficit is not specific to the FC version and that on the FC version, visual angle is not more important than number of noise letters for demonstrating schizophrenic deficit.