Herpes B Research Articles

Competitive radioimmunoassay to detect antibodies to herpes B virus and SA8 virus

A monoclonal competitive radioimmunoassay (CompRIAm) which detects antibody to herpesvirus simiae (B virus) in monkey and human sera and antibody to SA8 virus in monkey sera but not antibody to herpes simplex virus in human sera is described. Of 232 serum samples from wild-caught cynomolgus monkeys, 117 serum samples were positive when tested by CompRIAm. The results were in close agreement (97.5%) with B virus neutralizing antibody results on the same sera. Sera from 97 wild-caught rhesus monkeys and 92 wild-caught baboons were also tested. The CompRIAm was able to differentiate between sera that had neutralizing antibody to B virus and SA8 virus and those that did not, although the discrimination was not as clear as that in the tests on cynomolgus monkey sera. Sequential sera from two humans with confirmed cases of B virus infection were tested by CompRIAm. B virus antibody was detected in sera from both humans. None of 237 other serum samples from blood donors and patients attending sexually transmitted disease clinics reacted in the CompRIAm.

Polymerase chain reaction for detection of herpesvirus simiae (B virus) in clinical specimens.

A polymerase chain reaction (PCR) was designed which is specific to Macaca fascicularis (cynomolgus monkey) isolates of B virus. The PCR primers produced the expected 188 basepair product from the Cyno 2 strain and seven other cynomolgus monkey isolates of B virus. Oligomer hybridization with a 31-mer oligonucleotide was used to confirm the origin of this product. The PCR failed to amplify DNA of Epstein-Barr virus, cytomegalovirus, varicella-zoster virus, and other alphaherpesviruses (herpes simplex virus types 1 and 2, four SA 8 isolates and three rhesus isolates of B virus). PCR testing of swabs obtained from four orally-infected cynomolgus monkeys confirmed the presence of B virus DNA in samples previously shown to be positive by culture. In addition, PCR detected B virus in several swabs from infected monkeys that were culture negative. Total DNA extracts from the trigeminal and sacral ganglia of these animals were tested by nested PCR and B virus DNA was detected in the trigeminal ganglia of 3 of the 4 orally-infected cynomolgus monkeys. Nested PCR did not detect B virus DNA in total DNA extracts obtained from the brains of the four monkeys.

Monoclonal antibodies for the identification of herpesvirus simiae (B virus).

To differentiate between B virus and HSV isolates from monkeys and man monoclonal antibodies (mabs) were produced to herpesvirus simiae (B virus) and herpes simplex type 1 and 2 (HSV-1 and HSV-2). Mabs were tested by indirect immunofluorescence (IFAT) for reactivity against herpesviruses from Asiatic monkeys (B virus), African monkeys (SA 8 virus), and man (HSV-1, HSV-2, varicella-zoster virus, cytomegalovirus, and Epstein-Barr virus). Mabs could be divided into groups A-E displaying specific reactivity for B virus (A); reactivity with both B virus and SA 8 but not HSV (B); reactivity with B virus, SA 8 virus and HSV strains (C); specific reactivity with HSV-1 (D); and specific reactivity with HSV-2 (E). Two of the B virus specific mabs were able to differentiate between cynomolgus and rhesus strains of B virus. None of the mabs reacted with human varicella-zoster virus, cytomegalovirus, or Epstein-Barr virus. A panel of mabs for the unequivocal identification of B virus isolates from monkey or man is proposed.

Primate Viral Diseases in Perspective

The recent occurrence of fatal Herpesvirus simiae (B virus) infection in human subjects has again focused the attention of primatologists on this virus. B virus, however, is only one of a number of viral diseases that plays a role in primate colony management. This report is to emphasize to the primatologist a number of viruses other than H. simiae, with high morbidity and mortality rates, of importance for health management of nonhuman primate animal colonies. This concept is supported by the recent occurrence in colonies of nonhuman primates of simian hemorrhagic fever virus, SA8, herpesvirus, respiratory syncytial virus, encephalomyocarditis virus, Ebola virus, and simian immunodeficiency viruses.

Fatal Herpesvirus Simiae (B Virus) Infection in a Patas Monkey (Erythrocebus Patas)

Herpesvirus simiae (B virus) occurs naturally in Asian macaques, particularly rhesus, bonnet, and cynomolgus monkeys 7,8 The term B virus was coined after a physician (W.B.) died from encephalitis 3 days after being bitten by a clinically normal rhesus monkey. Herpesvirus simiae was isolated from the patient’s brain, medulla, spinal cord, and spleen. The B virus is well adapted to its natural simian host, frequently causing only transient, mild oral lesions. Like many herpesviruses, a latent form occurs in recovered animals, and the virus can be recovered from kidney, oral secretions, and the trigeminal ganglia in clinically normal monkeys. Infected animals may not show clinical signs or have detectable antibody to the B virus. Surveys of wild and captive monkeys reveal a high variability in B virus infection rates. Approximately 10% of young recently captured animals were seropositive for B virus, whereas 70-80% of adults had positive titers. In addition to age, population density also influenced the number of positive animals. In Indonesia, 77% of those monkeys tested had positive titers, in Malaysia, 64%, and in the Philippines, 28%. Sera obtained from various laboratories utilizing primates revealed approximately 19% of the animals tested had antibody titers to B virus. The major interest in B virus has been the disease it causes in humans. Infected individuals develop an almost invariably fatal ascending myelitis and encephalitis. Fatal disease suspected to be caused by B virus has been reported in patas and colobus monkeys. This report describes a case of fatal B virus infection in a pet patas monkey. An adult male patas monkey was submitted for necropsy following a brief illness. The monkey had been normal the previous morning, yet developed anorexia and lethargy the evening prior to its demise. The monkey had been purchased from an unknown dealer in Missouri 3 weeks previously. He had been housed in an outside enclosure until 1 week prior to his death, when he was moved into an 8x 12x 6-foot indoor cage located in the basement of a house. There were 2 pairs of rhesus monkeys also in the basement. Their cages were separated from the patas monkey’s cage by a solid partition. All cages were cleaned at 2-3-day intervals. The patas monkey’s cage had never been used for holding other primates. All monkeys were fed a commercial diet formulated for primates and supplemented with various fruits. The patas monkey was fed an all fruit diet for 1 week prior to his death.

Parvovirus Infection in Pigs with Exudative Skin Disease

rier status of the rhesus monkeys and the high mortality rate 2. Centers for Disease Control: 1989, B virus infection in humansin humans clinically infected with B virus. He was advised Michigan. MMWR 38:453-454. to limit human contact with the remaining monkeys and 3. Hillard JK, Munoz RM, Lipper SL, Eberle R: 1986, Rapid idenconsult with his physician. Family members were seronegatification of Herpesvirus simiae (B virus) DNA from clinical tive for B virus antibodies. The owner, who was the primary isolates in nonhuman primate colonies. J Virol Methods 13:55caretaker of the monkeys, refused serologic testing. Upon the 62.

Ocular Histopathologic Findings in a Case of Human Herpes B Virus Infection

A 37-year-old male laboratory technician who sustained a cutaneous penetrating wound from a rhesus monkey developed a progressive ascending encephalomyelitis due to culture-proven herpes B virus (Herpesvirus simiae) infection. He died 6 weeks after his injury despite acyclovir and ganciclovir treatment that was initiated after central nervous system symptoms developed. Histopathological examination of the patient's left eye revealed a multifocal necrotizing retinitis associated with a vitritis, optic neuritis, and prominent panuveitis. Herpes-type virus was identified in the involved retina by electron microscopy. Postmortem vitreous cultures taken from both eyes and retinal cultures taken from the right eye were positive for herpes B virus. Herpes B virus produces infection and destruction of retinal tissues similar to other herpesviruses. To our knowledge, this case represents the first histopathologic demonstration of herpes B virus infection in a human eye.

In vitro sensitivity of herpesvirus simiae (B-virus) to twelve antivirals effective against herpesvirus hominis (H. simplex) : K.F. Soike and J.K. Hillard, Delta Regional Primate Research Center, Covington, LA and Southwest Foundation for Biomedical Research, San Antonio, TX

In vitro sensitivity of herpesvirus simiae (B-virus) to twelve antivirals effective against herpesvirus hominis (H. simplex) : K.F. Soike and J.K. Hillard, Delta Regional Primate Research Center, Covington, LA and Southwest Foundation for Biomedical Research, San Antonio, TX

Seroprevalence of B Virus (Herpesvirus simiae) Antibodies in a Naturally Formed Group of Rhesus Macaques

Eighty-two percent of a group of rhesus monkeys removed from Cayo Santiago were seropositive for B virus (Herpesvirus simiae) antibodies. Similar results were obtained from the Cayo Santiago macaque population two decades ago and from feral Indian rhesus monkeys. Thus it is likely that B virus has been enzootic in the Cayo Santiago population since 1938, when the colony was established with stock imported from India.

Nonhuman primate bites

Nonhuman primate (monkey) bites to researchers and attending animal care staff may present problems in patient management. Such inoculations can transmit serious bacterial and viral infections to the human handlers. Significant local and systemic manifestations can subsequently develop following such an injury. Since Herpesvirus simiae (B virus) is enzootic in Asiatic monkeys of the genus Macaca, and since B virus infection in humans is usually fatal, additional prophylactic and therapeutic measures must be taken when persons are bitten by macaque monkeys. Primate bites require early aggressive intervention.

Relatedness of glycoproteins expressed on the surface of simian herpesvirus virions and infected cells to specific HSV glycoproteins

The antigenic relatedness of the surface glycoprotein antigens of six herpesviruses indigenous to human and nonhuman primates was examined. Binding of anti-viral sera to viral antigens expressed on the surface of infected cells demonstrated that the surface antigens of herpes simplex virus type 1 (HSV 1), HSV 2, simian agent 8 (SA8), and Herpesvirus simiae (B virus) exhibit extensive cross-reactivity. Surface antigens of two viruses isolated from South American primates, H. saimiri 1 (HVS 1) and H. ateles 1 (HVA 1), were comparatively more virus-specific in their antigenic reactivity. Endpoint neutralization tests performed in the presence and absence of complement confirmed these results. Immunoprecipitation of viral proteins was used to identify those representing cross-reactive surface antigens. A glycoprotein of approximately 110,000-125,000 Daltons (110-125 k) was immunoprecipitated from cells infected with each of the six primate herpesvirus by antisera to each of the viruses. Using monospecific antisera, these glycoproteins were shown to be antigenically related to the gB glycoproteins of HSV. Although these glycoproteins were antigenically conserved among all six viruses, antibodies to the gB glycoproteins did not cross-neutralize heterologous viruses. A glycoprotein of approximately 60-70 k was precipitated from HSV 1, HSV 2, SA8, and B virus infected cells by antisera to each of these four viruses. These SA8 and B virus glycoproteins were shown to be antigenically related to the gD glycoproteins of HSV 1 and HSV 2 and to be involved in cross-neutralization among these viruses. Antisera to HVS 1 and HVA 1 did not recognize these gD glycoproteins nor was a glycoprotein of similar molecular weight precipitable from HVS 1 or HVA 1 infected cells by antisera to the other four viruses. Southern blot hybridizations using probes for HSV glycoprotein genes confirmed the conservation of the gB glycoproteins among all the simian viruses and of the gD gene in SA8 and B virus. A glycoprotein of approximately 75-80 k was, however, precipitated from HVS 1 and HVA 1 infected cells by antisera to either of these two viruses. In addition, at least one glycoprotein which appeared to be predominantly virus-specific in its reactivity was identified for five of the viruses.

Herpesvirus simiae Contamination of primary rhesus monkey kidney cell cultures

Herpesvirus simiae Contamination of primary rhesus monkey kidney cell cultures

Discrimination between twenty isolates of herpesvirus simiae (B virus) by restriction enzyme analysis of the viral genome

Twenty isolates, obtained from adult breeding monkeys, were all identified as herpesvirus simiae (B virus) by neutralisation with polyclonal B virus antiserum. Subsequent analysis of restriction enzyme profiles produced by digestion of DNA from the isolates enabled discrimination to be made between them. In particular Cynomolgus monkey isolates could be distinguished from those of Rhesus animals. One isolate (isolate 9) could not be typed either as B virus or as the antigenically related herpesvirus SA8, despite neutralisation by B virus antiserum. Unlike herpes simplex virus, B virus isolates could not be divided into oral and genital types on the basis of restriction enzyme profiles.

Simian alphaherpesviruses and their relation to the human herpes simplex viruses

Biochemical and immunological properties of structural and non-structural polypeptides of the human simplex viruses (HSV1 and HSV2) and four related herpesviruses of non-human primates [Herpesvirus simiae (B virus), H. cercopithicus (SA8), H. saimiri 1 (HVS 1), and H. ateles 1 (HVA 1)] were compared. Using a radioimmunoassay (RIA), the presence of antigenic determinants shared among all six viruses was demonstrated. The relative degree of antigenic cross-reactivity among these viruses was further assessed by competition RIA. Antigenically, HSV 1 and HSV 2 were most closely related to each other although both SA 8 and B virus were also very closely related to HSV 1. Considerably less cross-reactivity existed between either HVS 1 or HVA 1 and the other four primate herpesviruses. Cross-hybridization between simian and human herpesvirus genomes demonstrated that extensive homology exists between each of the simian viruses and both HSV1 and HSV 2. Viral polypeptides bearing common antigenic determinants were identified by immune precipitation of infected cell polypeptides and by immunoblotting. Among the polypeptides of HSV which were recognized by antisera to simian viruses were the VP 5 and p40 proteins, both of which are structural components of the virion nucleocapsid. Using recombinant plasmids containing sequences of the HSV 1 VP5, p40, DNA polymerase, major DNA binding protein, and TK enzyme genes, homologous sequences were detected in all four simian viruses. Together, these results demonstrate that HSV 1, HSV 2, SA 8, and B virus form a closely related sub-group of the primate herpesviruses; HVS 1 and HVA 1 are also related to the other four primate herpesviruses, albeit more distantly.

Bilateral Posterior Parietal Atrophy in an Infant Rhesus Monkey (Macaca mulatto)

A case of severe neurological deficiency syndrome marked by visual impairment, motor disturbances, and bilateral extensive glial scarring of the posterior parietal cortex is described in an infant rhesus monkey. Although morphologically resembling perinatal asphyxia, the possibility of a perinatal or postnatal Herpesvirus simiae infection is discussed.

Leads from the MMWR. Guidelines for prevention of Herpesvirus simiae (B virus) infection in monkey handlers

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Herpesvirus simiae (B virus): replication of the virus and identification of viral polypeptides in infected cells.

The events and products of replication of Herpesvirus simiae (B virus) in Vero cells were studied. The time course of the synthetic events of DNA replication and protein synthesis were found to be similar to the processes of the herpes simplex viruses and SA 8. Infectious progeny virus were detected by 4 hours post infection and were first found extracellularly between 6 and 8 hours post infection (PI). As in the case of SA 8, all cell lines tested were permissive for lytic infection by B virus. Analyses of B virus-infected cells by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) revealed approximately 50 infected cell polypeptides (ICP) ranging in molecular weight from about 26,000 to 239,000 daltons. The kinetics of synthesis of the ICPs were also identified. At least nine glucosamine-containing glycopeptides were noted ranging from 133,000 to 29,000 daltons.

ELISA for detection of group-common and virus-specific antibodies in human and simian sera induced by herpes simplex and related simian viruses

A rapid (3.5 h) enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibodies to herpes simplex virus type 1 (HSV-1) and to two antigenically related monkey viruses, simian agent 8 (SA8) and Herpesvirus simiae (B virus). Crude preparations of detergent solubilized infected cells and similarly treated control mock-infected cells served as antigens for coating wells in microplates. Biotinylated protein A and avidin-conjugated alkaline phosphatase were used to detect antibodies in sera from different species (humans, monkeys and rabbits). Three prototype assays are described with three degrees of specificity. Common or specific determinants on the viral antigens could be assayed in simple competition tests using similar antigen preparations to those coating the wells. The specific assays permitted rapid differential serodiagnosis of antibodies to human and simian herpesviruses.

Rapid identification of herpesvirus simiae (B virus) DNA from clinical isolates in nonhuman primate colonies

A rapid, simple technique, based on restriction endonuclease analysis ofradioactively labeled infected cell DNA, is described for identification of Herpesvirus simiae (B virus) infection in clinical isolates from nonhuman primates. Isolates can be screened within 2–3 days from the time of collection of a specimen from a suspect lesion to final viral identification. Isolates were obtained from eight animals with suspected B virus infections. The results indicated the presence of B virus in each of the eight animals, each isolate unique from the others, but with the dominant prototypic pattern of the laboratory strain of B virus (E2490) and not HSV-1 (KOS), HSV-2 (186), or SA8 (3264).

Herpes simplex virus complement fixing antibody and herpes B virus serum neutralizing antibody in sera of wild and laboratory-bred cynomolgus monkeys.

The result of the complement fixation (CF) test for the antibody to herpes simplex virus (HSV) in sera of the cynomolgus monkeys was compared with that of the neutralization test (NS) for the antibody to herpes B virus (HBV) in the same sera. Fifty-seven (74%) of 77 wild-originated monkeys were positive for HSV-CF, while 65 (84%) of the 77 animals were positive for HBV-SN. All of the 57 CF positive cases were also positive for HBV-SN. On the other hand, 30 (75%) of 40 laboratory-bred monkeys had neither HSV-CF antibody nor HBV-SN antibody. Remaining 10 of the 40 laboratory-bred animals were positive for HSV-CF. However, no HBV-SN antibody was detected in nine of the 10 HSV-CF positive animals. These results suggest that the HSV-CF test may be as satisfactory as the HSV-SN test as a practical measure for rough screening of HBV infection in the cynomolgus monkey in laboratories having no containment unit for handling HBV.