Identification of renal sympathetic preganglionic neurons in hamsters using transsynaptic transport of herpes simplex type 1 virus.

Herpes viruses have been used as retrograde transsynaptic tracers to identify pathways from the CNS to specific target tissues. We used herpes simplex virus to identify central nervous system neurons responsible for control of the kidney. Herpes simplex type 1 or herpes simplex type 2 was injected into rat kidneys and herpes simplex type 1 was microinjected into hamster and guinea pig kidneys. After three to seven days, ganglia, spinal cords and brains were examined using immunohistochemistry to visualize the virus-infected neurons. Our first experiments demonstrated that rats were not susceptible to infection with neurotropic strains of herpes simplex type 1. Injections of a wildtype strain of herpes simplex type 2 into rat kidneys led to nonspecific infection of many central nervous system neurons and glia. In contrast, herpes simplex type 1 injections in hamsters and guinea pigs caused specific infection of limited numbers of neurons in approximately one-third of the animals and the study was continued using hamsters. Sympathetic preganglionic neuron labelling was found in the ipsilateral intermediolateral cell column of the spinal cord as well as the lateral funiculus. Most infected preganglionic neurons were located in the seventh to the ninth thoracic spinal segments. Infected neurons were not found in the dorsal or ventral horn of the spinal gray matter and only one or two cells were found in the brainstem. Sympathetic preganglionic neuron morphology was usually normal, showing detailed dendritic arborizations, and lysis was infrequent. Small infected cells were sometimes observed close to sympathetic preganglionic neurons. Because herpes simplex type 1 virus was not detected immunocytochemically in ganglionic neurons in these same hamsters, the polymerase chain reaction was used in some additional hamsters to detect viral DNA in the T12 and T13 chain ganglia and splanchnic ganglia ipsilateral to the kidney injected with herpes simplex type 1. Finally, the overall distribution of renal postganglionic and splanchnic preganglionic neurons in hamsters was examined for comparison to the number and locations of virus-labelled neurons. Retrograde transport of the fluorescent dye FluoroGold demonstrated that (i) renal postganglionic neurons are distributed in the T10-L1 chain ganglia and in the prevertebral splanchnic ganglion and (ii) splanchnic preganglionic neurons are located in the T3-T12 spinal segments, predominantly in the intermediolateral and funicular spinal autonomic nuclei. In conclusion, herpes simplex type 1 virus infected an exclusive population of "renal" neurons in hamsters without lysis and with little cellular reaction to the infection after a survival period of three days, permitting these neurons to be studied in detail.

HSV-1 recovery from ocular tissues after viral inoculation into the superior cervical ganglion.

New Zealand albino rabbits were inoculated in the right superior cervical ganglion with 25 microliter of herpes simplex virus type 1 (HSV-1) (McKrae strain; 10(3) or 10(5) PFU/ml). Positive tear film swabs were detected at least once in 28/32 (88%) of ipsilateral eyes and 6/32 (19%) of contralateral eyes beginning on postinoculation (PI) day 2-6. The average HSV-1 titer in the tear film was 4.0 X 10(3) PFU in ipsilateral eyes and 2.7 X 10(3) PFU in contralateral eyes, determined from eye washes after inoculation of 25 PFU of HSV-1. In selected rabbits, the aqueous humor was positive for virus on PI days 3, 4, 5, 6, and 8. the aqueous humor in ipsilateral eyes showed positive results in 9/11 (82%) of the eyes tapped on PI 3, 13/18 (72%) on PI 4, 5/11 (45%) on PI 5, 1/6 (17%) on PI 6, and 1/2 (50%) on PI 8. No virus was detected in aqueous humor tappings in any contralateral eyes (0/65). Conjunctivitis and iritis (iris hyperemia) appeared in all ipsilateral eyes beginning as early as PI day 1. Conjunctivitis occurred in 1/21 (4.8%) of contralateral eyes. Cells and flare appeared in 18/21 (86%) of ipsilateral eyes and 2/21 (9.5%) of contralateral eyes. Hyphema was noted in 3/21 (14%) of ipsilateral eyes. Of the eyes with iritis, 12/21 (57%) developed corneal edema. Corneal dendritic ulcers were observed in 4/21 (19%) of ipsilateral eyes and 2/21 (9.5%) of contralateral eyes. No ocular fundus changes were seen in any contralateral or ipsilateral eyes.(ABSTRACT TRUNCATED AT 250 WORDS)

Variations in herpes simplex virus isolated from human ganglia and a study of clonal variation in HSV-1.

Growth of human sensory ganglia in culture has led to the reactivation of herpes simplex virus from over 50% of cases studied. Infected cell polypeptide and restriction enzyme analysis has led to the conclusion that each individual has one unique latent strain of HSV-1 that can be present in more than one ganglion in the body. Analysis of 91 isolates has shown that the long region of the genome is variable in terms of DNA restriction sites, DNA sequences, and in coding for the majority of variable polypeptides. The short region is stable with only polypeptides Vmw 21 and 22, restriction sites HindIII-(M-N) and BglII-(G-H) and the DNA sequence BamHI-1' having been found to vary. The insertion and deletion of small DNA sequences at specific locations allows individual reactivation events to be distinguished. Detection of information able to complement and produce ts+ virus on superinfection of otherwise negative ganglia with ts mutants, has led to the conclusion that ganglion cells may harbor herpes virus-related information that is only detectable by use of such genetic probes.

Herpes zoster ophthalmicus.

Herpes zoster ophthalmicus occurs worldwide, usually in healthy adults, but, increasingly in patients who are immunocompromised. After primary varicella infection (chickenpox), the virus lies dormant in the sensory ganglion until it becomes reactivated as zoster. Involvement of the ophthalmic branch of the trigeminal nerve is characterized early by corneal dysesthesia and dendritiform keratopathy, and these are self-limited. However, smoldering disease may cause pathological changes in the ocular structures through direct invasion of virus, secondary inflammation, and alterations of autoimmune mechanisms. Antiviral agents have demonstrated some success in resolving early signs and symptoms, but their role in preventing and treating late complications remains to be fully studied. Until a definitive antiviral agent is established, the benefits of steroid use in certain acute inflammatory processes outweight its risk of reducing host immunity. Corneal complications of herpes zoster ophthalmicus sometimes require surgical intervention.

Binding, uptake and retrograde axonal transport of herpes virus suis in sympathetic neurons.

Newborn rat dissociated sympathetic neurons were grown in a chamber culture system, where a Teflon ring sealed with silicon grease separated the axonal plexus from the corresponding nerve cell bodies. The binding of 35S-labeled herpes virus suis (HVS) to the neurites was partially inhibited by an excess of unlabeled HVS as well as by concanavalin A, indicating the presence of specific binding sites for the virus. Specific binding was a prerequisite for the subsequent uptake and retrograde transport of HVS to the nerve cell bodies. Predominantly free nucleocapsids were detected by electron microscopy in the axons at the time of retrograde transport, both in culture and in vivo, suggesting the possibility that nucleocapsids without lipid membrane and not contained in cellular membrane compartments can be transported by retrograde axonal transport.

Intraocular herpes simplex virus injection in neonatal rats induces sympathetic nerve cell destruction: effect of nerve growth factor.

The effect of herpes simplex virus (HSV) injection on the sympathetic nerve system of newborn rats was studied at structural, ultrastructural, and immunohistochemical levels. It was found that HSV injected into the anterior eye chamber is retrogradely transported and reaches the nerve cell bodies of the ipsilateral superior cervical ganglion (SCG) after 18-24 hr, causing complete cell destruction within 3-4 days. In subsequent days, nerve cells of the contralateral SCG, spinal sensory ganglia, chromaffin cells and brain cells also become infected and are eventually killed by the virus. Pretreatment with nerve growth factor (NGF) produces an initial protection from viral cell destruction, but does not block the final, lethal effect of the virus. These investigations demonstrate that sympathetic nerve cell destruction can be induced in newborn rodents by HSV, and that NGF treatment renders the cells, for a time-limited period, more resistant to the virus.

Interferon protects neurons in culture infected with vesicular stomatitis and herpes simplex viruses.

The protective effect of crude rat interferon against infection by vesicular stomatitis virus (VSV) and herpes simplex virus (HSV) was assessed in two culture systems: the PC12 cell line and dissociated rat neurons derived from the superior cervical ganglion (SCG). Interferon was induced in rat embryo cells by inactivated Newcastle disease virus, and its effect was assessed by reduction of viral yields and prevention of viral cytopathology. Interferon protected PC12 cells, both in the presence and absence of nerve growth factor (NGF), against infection by both viruses, although at differing concentrations: protection against VSV was noted at approximately a 10-fold lower interferon concentration than that required to inhibit HSV infection. Dissociated SCG neurons were also protected, but higher interferon concentrations were required. These results demonstrate that the antiviral state can be established in neurons in response to interferon.

Sensory and sympathetic ganglia in HIV-1 infection: immunocytochemical demonstration of HIV-1 viral antigens, increased MHC class II antigen expression and mild reactive inflammation.

Sensory and sympathetic ganglia from 12 cases of human immunodeficiency virus type 1 (HIV-1) infection, all but one without clinical evidence of peripheral nerve disease, were studied immunocytochemically for their content of lymphocytes, macrophages, MHC Class II antigens and HIV-1 and cytomegalovirus antigens. They were compared with ganglia from 7 normal and peripheral nerve disease control cases. Compared with normal controls, many of the ganglia from the majority of HIV-1-infected subjects contained more T lymphocytes and macrophages and enhanced MHC class II expression. A few also showed occasional neuronal degeneration which was not present in the normal controls. In 7 cases HIV-1 gp41 envelope protein and/or p24 core protein antigens were detected in intraganglionic macrophages. Sensory ganglia contained more gp41 HIV-1 antigen than sympathetic ganglia. There was no clear correlation between detection of HIV-1 antigens in ganglia and in the CNS. Detection of HIV-1 antigens in ganglia was more common in cases of HIV-1 infection that had progressed to clinical AIDS by the time of death (71%) than in those that had not done so (40%). It is concluded that there is commonly a mild ganglionitis which is asymptomatic in the absence of detailed clinical testing and frequently associated with local presence of HIV-1 antigens in sensory and sympathetic ganglia in AIDS.

Coxsackievirus B4 infection of sympathetic ganglia in squirrel monkeys.

Lumbar sympathetic ganglionitis was found by light microscopy in 2 of 17 (12%) squirrel monkeys (Saimiri sciureus) experimentally infected with Coxsackievirus B4. This finding shows that viruses can cause ganglionitis which, in turn, must cause autonomic nervous system dysfunction. Such viral ganglionitis may explain some diseases, including cardiovascular ones, of poorly understood or unknown etiology which present with manifestations of dysfunction of the sympathetic nervous system.

[Herpes simplex type I virus observed in the superior cervical ganglion from human cadavers and autopsy materials]. / Insan kadavra ve otopsi materyallerinden elde edilen ganglion cervicale superius'larda herpes simplex tip I virusunun arastirilmasi.

Herpes Simplex Type I (HSV I) causes some infections such as herpes labialis, encephalitis, keratoconjunctivitis and also some cranial nerve syndromes such as acute vestibular neuritis, migraine and Meniere's disease in human. We used 4 fixated and 16 fresh cadavers to isolate HSV I virus from the Superior Cervical Ganglia. The ganglia materials are inoculated to PRK (primary rabbit kidney), VERO (African Green Monkey Kidney) and BHK 21 (Baby Hamster Kidney) cell lines in order to isolate the virus. We isolated HSV I virus from 12 fresh cadavers' cervical ganglia (75%) and neutralisation test is performed in order to characterize HSV I. But we could not isolate the virus from any of the fixated cadavers.

Acute and latent herpes simplex virus neurological disease in mice immunized with purified virus-specific glycoproteins gB or gD.

Groups of 5-week-old BALB/c mice were immunized intraperitoneally with approximately 10 micrograms of purified alum-precipitated glycoprotein gB or gD of either herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) origin. Control mice received injections of alum-precipitated 1% bovine serum albumin (BSA). Following a second immunization 4 weeks later, seroconversion was confirmed by demonstrating the presence of glycoprotein-specific antibody by immune precipitation. All animals were challenged with lethal doses of either HSV-1 or HSV-2 by footpad inoculation and assessed for acute virus-induced neurological disease and the development of ganglionic latency. Whereas 70% of control (BSA-immunized) HSV-1-infected animals developed ascending myelitis and died, 100% of mice immunized with either gB-1, gB-2, gD-1, or gD-2 antigens remained free of clinical illness and survived HSV-1 challenge. In contrast, gB-1-or gB-2-immunized mice were not protected against acute HSV-2-induced neurological disease and showed a mortality rate of 60-90% (equivalent to that seen in controls), although mean survival times were prolonged. However, significant protection against HSV-2 challenge was observed with gD-1 or gD-2 immunization. When sacral ganglia were removed from surviving mice 9-12 months after virus challenge, latent virus was detected in all gB- or gD-immunized animals, although the extent of latent infection was restricted. These results provide evidence that glycoprotein gD might be superior to glycoprotein gB as an immunogen for the control of acute HSV-1 and HSV-2 neurological disease in mice. However, neither glycoprotein prevents ganglionic latency, the source of virus for recurrent herpesvirus infections.

Recurrence of herpes simplex in the mouse requires an intact nerve supply to the skin.

The nerves supplying the pinna of the ear of mice latently infected in the 2nd and 3rd cervical ganglia were sectioned. Immediately after neurotomy, or some days later, the denervated ears were stripped with cellophane tape to induce recurrent disease. The cervical ganglia and skin of the ears were tested for the presence of infectious virus at different times after neurotomy. Nerve section induced a low incidence of reactivation of virus in ganglia. After neurotomy, infectious virus was isolated from the skin very rarely and recurrent disease was not seen.

Acute and recurrent herpes simplex in several strains of mice.

Acute and recurrent herpes simplex was studied after infection in the ear of two outbred and five inbred strains of mice. In all strains tested there was clinical evidence of infection, and a proportion of the mice became latently infected in the cervical ganglia. Six weeks after infection, when attempts were made to induce recurrent desease by stripping the ears of the mice with cellophange tape, a proportion of animals of each strain developed recurrent disease, characterized by erythema in the skin. At monthly intervals thereafter, the ears were stripped again and, on each occasion, a proportion of the animals developed recurrent disease, with the exception of Balb/c mice. The different reaction of Balb/c and other inbred strains might prove useful in studies on the mechanism of control of recurrent herpes simplex.

Recovery of herpes simplex type 1 from the celiac ganglion after renal transplantation.

Herpes simplex type 1 was recovered from the celiac ganglion of a renal transplant patient who had esophageal lesions consistent with herpes simplex esophagitis as well as peptic ulcers requiring operation. We speculate that the virus got to the celiac ganglion via retrograde spread from the stomach and/or duodenum. Further studies are needed to determine the frequency of HSV involvement in visceral autonomic ganglia and whether by reactivation it could lead to subsequent gastrointestinal ulceration.

Effect of acyclovir on recurrence of herpes simplex skin lesions in mice.

Acyclovir (ACV) was effective in preventing recurrence of herpes simplex in mice whose skin was stripped with cellophane tape. Treatment with ACV did not eliminate latent herpes simplex virus from the cervical ganglia.

Immunological memory to herpes simplex virus type 1 glycoproteins B and D in mice.

Mouse L cell lines constitutively expressing glycoproteins B or D of herpes simplex virus type 1 (LTKgB and LTKgD respectively) were used to study the longevity of the immune response to these viral glycoproteins in mice. Two immunizations with the cell lines were necessary to induce a persisting antibody response (present for over 200 days). Only LTKgD induced a neutralizing at antibody response in mice and this also remained at high titres over 200 days after two inoculations. The presence in mice of precursor cytotoxic T lymphocytes specific for gB expressed in the L cells was also shown up to 270 days after immunization. Mice immunized with the cell lines showed an increased rate of virus clearance from the ear pinna, inoculation with LTKgD resulting in more clearance than LTKgB at 7 days post-immunization. This type of protection was reduced with time after inoculation, until by day 161 there was no significant difference in virus titres between immunized and control groups. However, LTKgD immunization protected against the establishment of latent infections in the ganglia of mice even up to 186 days post-inoculation.

Detection of herpes simplex virus-specific DNA sequences in latently infected mice and in humans.

Herpes simplex virus-specific DNA sequences have been detected by Southern hybridization analysis in both central and peripheral nervous system tissues of latently infected mice. We have detected virus-specific sequences corresponding to the junction fragment but not the genomic termini, an observation first made by Rock and Fraser (Nature [London] 302:523-525, 1983). This "endless" herpes simplex virus DNA is both qualitatively and quantitatively stable in mouse neural tissue analyzed over a 4-month period. In addition, examination of DNA extracted from human trigeminal ganglia has shown herpes simplex virus DNA to be present in an "endless" form similar to that found in the mouse model system. Further restriction enzyme analysis of latently infected mouse brainstem and human trigeminal DNA has shown that this "endless" herpes simplex virus DNA is present in all four isomeric configurations.