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R Eberle

Publications and source records attributed to R Eberle.

At least 37 records · Page 2Linked to original sources
PubMed1998

Molecular evidence for distinct genotypes of monkey B virus (herpesvirus simiae) which are related to the macaque host species.

Although monkey B virus (herpesvirus simiae; BV) is common in all macaque species, fatal human infections appear to be associated with exposure to rhesus macaques (Macaca mulatta), suggesting that BV isolates from rhesus monkeys may be more lethal to nonmacaques than are BV strains indigenous to other macaque species. To determine if significant differences that would support this supposition exist among BV isolates, we compared multiple BV strains isolated from rhesus, cynomolgus, pigtail, and Japanese macaques. Antigenic analyses indicated that while the isolates were very closely related to one another, there are some antigenic determinants that are specific to BV isolates from different macaque species. Restriction enzyme digest patterns of viral DNA revealed marked similarities between rhesus and Japanese macaque isolates, while pigtail and cynomolgus macaque isolates had distinctive cleavage patterns. To further compare genetic diversity among BV isolates, DNA sequences from two regions of the viral genome containing genes that are conserved (UL27 and US6) and variable (US4 and US5) among primate alphaherpesviruses, as well as from two noncoding intergenic regions, were determined. From these sequence data and a phylogenetic analysis of them it was evident that while all isolates were closely related strains of BV, there were three distinct genotypes. The three BV genotypes were directly related to the macaque species of origin and were composed of (i) isolates from rhesus and Japanese macaques, (ii) cynomolgus monkey isolates, and (iii) isolates from pigtail macaques. This study demonstrates the existence of different BV genotypes which are related to the macaque host species and thus provides a molecular basis for the possible existence of BV isolates which vary in their levels of pathogenicity for nonmacaque species.

Animals↗
PubMed1998

Shedding and transmission of baboon Herpesvirus papio 2 (HVP2) in a breeding colony.

Baboons in a captive breeding colony were monitored twice a year, and new additions were screened on arrival for shedding of Herpesvirus papio 2 (HVP2) and serologic reactivity to the agent. For 128 individual animals tested over a period of 1.5 years, shedding of infective virus was detected in 13 of 342 swab specimens (3.8%), each of these incidents representing shedding by a different animal. Among long-term colony animals, infective virus was recovered on only two occasions (5 of 236 swab specimens from five individuals). In all but one instance, animals shedding virus were infants, not adults, and all animals were shedding virus in the oral cavity. One of these five instances was an isolated case, but four (three infants and one adult) were clustered within a single breeding group. Molecular analyses of the HVP2 isolates from this cluster indicated that they likely arose from a single common source, probably the mother of one of the infants. None of 31 wild-caught baboons added to the colony during this period were found to be shedding infective virus, despite 93.5% of them being seropositive for HVP2. In contrast, 6 of 18 adult baboons (all seropositive) transferred into the colony from another breeding colony were found to be shedding HVP2 either orally (3 of 6) or genitally (3 of 6). In addition, 2 of 8 juvenile baboons in this shipment were found to be shedding virus in the oropharynx. Overall, 10 of 13 instances of HVP2 isolation were from the oropharynx rather than the genital tract, and 6 of 13 baboons shedding virus were infants or juveniles rather than adults. These results suggest that, although venereal transmission of HVP2 occurs among adult animals, oral infection of young, sexually immature baboons is not uncommon.

Aging↗
PubMed1997

Genetic and functional complementation of the HSV1 UL27 gene and gB glycoprotein by simian alpha-herpesvirus homologs.

Utilizing co-transfection of DNA from glycoprotein gB- strain of HSV1 and cloned fragments of several simian alpha-herpesviruses containing the UL26, UL27 (gB glycoprotein), and UL28 gene homologs, replication-competent recombinant viruses were produced. Genetic analysis of one HSV1/SA8 recombinant (HSV1/SgB) demonstrated the presence of SA8 DNA comprising the entire UL27 (gB) gene and parts of the UL28 and UL26 ORFs in an otherwise HSV1 genome. The recombinant was shown to express the SA8 gB and p40 proteins (UL27 & UL26.5 gene products, respectively); all other proteins were indistinguishable from those of HSV1. The recombinant behaved like SA8 in gB-specific virus neutralization and cell surface antibody binding assays, while plaque morphology and replication kinetics were very similar to HSV1. Despite its overwhelming HSV1 genetic constitution, the recombinant displayed a pathogenic phenotype in mice very different from the parental HSV1. While HSV1 produced corneal disease in ocularly infected mice and readily spread to the nervous system. HSV1/SgB was markedly impaired in both respects. These results demonstrate the functional equivalency of the cercopithecine monkey virus gB glycoproteins and genes (including transcriptional regulatory elements) in HSV1, the functional nature of HSV1/SA8 chimeric UL28 and UL26 genes/proteins, and that UL28, gB and/or p40 proteins may effect the pathogenicity of HSV1.

3T3 Cells↗
PubMed1997

Effect of ethanol on postprandial duodenojejunal motility in humans.

The effect of ethanol on postprandial small bowel motility was investigated in eight healthy volunteers using perfusion of nutrient solutions (17% proteins, 59% carbohydrates, 24% lipids) into the descending duodenum (5 ml/min for 120 min). An ethanol-containing solution (4% w/v, 4.06 kcal/min, 1190 mosmol/kg) was compared with the corresponding ethanol-free solution (2.64 kcal/min, 160 mosmol/kg) and another ethanol-free hyperosmolar solution adapted in caloric load and osmolality (4.06 kcal/min, 1160 mosmol/kg). Motility was recorded with a data logger and six pressure transducers at 3-cm intervals around the duodenojejunal flexure. Clustered contractions (27 +/- 4/hr) migrating aborally through the whole recording segment were the predominant motor pattern with ethanol compared with the ethanol-free (10 +/- 2/hr; P < 0.01) and the ethanol-free hyperosmolar solution (6 +/- 3/hr; P < 0.001). Other motility parameters with ethanol were not different from the ethanol-free solution, whereas the ethanol-free hyperosmolar solution showed a much less intense motor response. We conclude that ethanol does modify human postprandial duodenojejunal motility by inducing propagative motor patterns.

Adult↗
PubMed1997

Detection and differentiation of primate alpha-herpesviruses by PCR.

A rapid method for detection and differentiation of 5 primate alpha-herpesviruses (human herpes simplex virus types 1 and 2 [HSV1, HSV2], green monkey simian agent 8, baboon herpesvirus 2 [HVP2], and macaque B virus [BV]) was developed utilizing the polymerase chain reaction (PCR). PCR primers were located in conserved regions of the gene encoding the glycoprotein B, which flanks an intervening region that is highly divergent among the 5 viruses. Amplified PCR products from the 5 viruses were readily differentiated by their unique restriction enzyme digestion patterns. No variation in digestion patterns was noted among strains of HSV1, HSV2, or HVP2. One clinical isolate of BV exhibited variation in a single restriction site, but its overall restriction pattern remained typical of BV. This method (PCR/RFLP) allowed the presence of herpesvirus DNA in clinical swabs from primates to be readily detected and the virus unambiguously identified.

Animals↗
PubMed1997

Prevalence of Herpesvirus papio 2 in baboons and identification of immunogenic viral polypeptides.

The prevalence of Herpesvirus papio 2 (HVP2) in several groups of captive and wild-caught baboons was determined by detection of anti-HVP2 antibodies in 133 sera of adult baboons. Over 90% of newly imported (wild-caught) adult olive baboons (Papio anubis) from Kenya and chacma baboons (P. ursinus) from South Africa were found to have anti-HVP2 titers. Similarly, approximately 85% of captive breeding colony baboons (P. anubis and P. cynocephalus) were seropositive for HVP2. Infected animals were generally easily identifiable by enzyme-linked immunosorbent assay because anti-HVP2 IgG titers in immune animals were usually high (16,000 to 64,000). There was little variation in the relative reactivity patterns of individual HVP2-immune sera when tested against herpes simplex viruses 1 and 2, monkey B virus, H. cercopithecus 2, and HVP2, or against different HVP2 strains. Also, differences were not detected between reactivity of olive and chacma baboon immune sera. Analysis of the polypeptide specificity of immune sera by western blot identified four viral antigens that were consistent targets of immune sera. These antigens were the gB glycoprotein, a pair of unidentified glycoproteins of 80 to 100 kDa, the gD glycoprotein, and a series of smaller capsid proteins. Additional viral proteins were variably recognized by individual immune sera. The results of this study indicate that HVP2 is a common infection of baboons; there is little antigenic variation among HVP2 strains; and there are several HVP2 antigens that represent consistent targets of the anti-HVP2 immune response of baboons.

Animals↗
PubMed1996

Characterization of virus-specific and cross-reactive monoclonal antibodies to Herpesvirus simiae (B virus).

A panel of 13 monoclonal antibodies (MAbs) was produced that detect B virus (BV) proteins. Several of these MAbs were highly specific for BV, while the remainder cross-reacted in varying degrees with the other primate alphaherpesviruses. Utilizing western blot and radioimmunoprecipitation analysis, the MAbs were found to detect at least four distinct BV-infected cell antigens, several of which were composed of multiple polypeptides. One target antigen has been identified as the BV glycoprotein B (gB) homologue and was recognized by both virus-specific and cross-reactive MAbs. Although gB is an essential protein, none of the anti-gB MAbs neutralized infectious virus.

Animals↗
PubMed1995

Herpesvirus papio 2, an SA8-like alpha-herpesvirus of baboons.

Several SA8 isolates obtained from baboons were compared to the prototype SA8 herpesvirus of African green monkeys. SDS-PAGE and restriction enzyme analyses revealed definite differences between green monkey and baboon isolates. DNA and amino acid sequences of the gB, gD and gJ glycoprotein genes exhibited substantial differences in variable regions. For the gB and gD, the amount of amino acid substitutions between SA8 and the baboon viruses was comparable to levels observed between analogous genes of SA8 & B virus or HSV1 & HSV2. Although a high degree of antigenic cross-reactivity was apparent, virus-specific antigenic determinants were also readily detected. Phylogenetic analyses supported separation of the baboon isolates and SA8 as distinct viruses. Taken together these results suggest that although closely related to SA8, the baboon viruses represent a distinct simian alpha-herpesvirus which we propose be designated Herpesvirus papio 2.

Amino Acid Sequence↗
PubMed1995

The simian herpesviruses.

Increased use of monkeys in biomedical research has led to an intensified awareness of potential dangers posed by zoonotic infections. Zoonoses have an impact not only upon human health and safety but also upon continued availability of nonhuman primate resources for the biomedical community. Neurotropic herpesviruses indigenous to primates are significant owing to their potential for causing severe or fatal infections when transmitted between human and nonhuman primates or between different species of monkeys. Although the macaque herpesvirus (B virus) is known to many investigators, other simian herpesviruses have remained relatively obscure in spite of reports of disease-causing potential. In this review we summarize what is known about the natural history and pathogenic potential of simian alpha-herpesviruses. Recent research into the molecular biology of this group of viruses is also reviewed, and recent advances toward development of diagnostic tests based on these data are discussed.

Animals↗
PubMed1994

Use of chromosome microdissection, the polymerase chain reaction, and dot blot hybridization to analyze double minute chromosomes.

The potential usefulness of chromosome microdissection, the polymerase chain reaction (PCR), and dot blot hybridization as a quick screening method for determining the genetic composition of double minute chromosomes (DMs) was evaluated. DMs or abnormally banding regions (ABRs) were microdissected from multidrug-resistant hamster cell lines and amplified with PCR using primers specific for the hamster multidrug-resistance (MDR) gene, pgp 1. The microdissected-PCR-amplified products were shown to (a) hybridize to a 32P-labeled pCHP1 probe for the hamster MDR gene by using dot blot or Southern blot analysis and also (b) hybridize back to the chromosome region from which they were originally dissected by using fluorescent in situ hybridization. Microdissected/PCR-amplified DMs were also shown to hybridize to ABRs. When microdissected DMs and ABRs were amplified using hamster specific Alu primers, the resulting material was shown to hybridize with probes for hamster MDR and Alu. These results suggest that the DMs contained in these MDR hamster cell lines contain Alu-like sequences and the chromosome microdissection-PCR-hybridization approach might be used as a quick screening method for identifying genes amplified in DMs and ABRs in cell lines and human tumor samples.

Animals↗
PubMed1993

Gene mapping and sequence analysis of the unique short region of the simian herpesvirus SA 8 genome.

A 10.5 kbp BamHI restriction fragment representing most of the unique short (Us) region of the genome of the simian alpha-herpesvirus SA8 was identified and cloned. Partial sequencing of this DNA fragment identified regions of sequence homology with eight open reading frames (ORFs) of HSV1 and/or HSV2. Sequence and size analysis of subcloned fragments of the SA8 Us region and comparison with homologous HSV Us sequences determined that the number, order, size, and orientation of SA8 Us ORFs are comparable to those of HSV. Based on the location of transcriptional control elements, transcription of SA8 Us genes appears to be organized into 3' co-terminal mRNA sets as in HSV, although the grouping of the gene sets is different. The SA8 US4 (gG) ORF is more similar to that of HSV2 than HSV1, both in size and predicted amino acid sequence. Complete sequences were determined for five SA8 genes which represent homologs of the HSV gD, gE, gI, US5, and US9 genes. The predicted polypeptides encoded by SA8 are similar to the corresponding HSV polypeptides. All SA8 Us genes were more closely related to those of HSV than to related gene homologs of other mammalian alpha-herpesviruses.

Amino Acid Sequence↗
PubMed1993

Sequence analysis of herpes simplex virus gB gene homologs of two platyrrhine monkey alpha-herpesviruses.

Homologs of the herpes simplex virus gB gene were identified in two alpha-herpesviruses of platyrrhine monkeys, Herpesvirus saimiri 1 (HVS 1) and H. ateles 1 (HVA 1). These genes were cloned and sequenced in their entirety. Analysis of the predicted amino acid sequences indicated that the gB glycoproteins of these two viruses are of similar size, have 10 Cys residues and 5 potential N-linked glycosylation sites which align exactly with those in other primate alpha-herpesvirus gB polypeptides, and have a similar distribution of predicted secondary structural features, all of which indicate a conserved structure of the gB polypeptide. Alignment of these two gB sequences with those of four other primate alpha-herpesviruses (SA 8, B virus, HSV 1 and HSV 2) revealed localized regions of extensive sequence divergence as well as highly conserved regions. On comparison of the six primate virus gB sequences, the gBs of the two platyrrhine monkey viruses form a subgroup separate from that of the four catarrhine virus gBs. The degree of relatedness of the HVA 1 and HVS 1 gB sequences to each other was equivalent to the degree of relatedness between the human and the cercopithecine monkey virus gB sequences.

Amino Acid Sequence↗
PubMed1993

Rapid detection of B virus (herpesvirus simiae) DNA by polymerase chain reaction.

Rapid diagnosis of B virus (herpesvirus simiae) infection in humans followed by early antiviral treatment is essential for the patient's survival. To improve laboratory diagnosis of B virus infections, a polymerase chain reaction (PCR)-based test using synthetic oligonucleotide primers and probe was developed to detect B virus DNA in clinical samples. After the specificity of the PCR was assessed for detection of several B virus isolates, the method was used to investigate human and monkey specimens, and results were compared with those obtained by viral culture. PCR appeared to be more sensitive than conventional virus isolation and thus of practical use for a rapid identification of B virus infection when conventional viral cultures are negative.

Animals↗
PubMed1992

Evidence for an alpha-herpesvirus indigenous to mountain gorillas.

Sera from wild mountain gorillas were screened for antibodies reactive with primate alpha-herpesviruses. Four of seven individuals tested (58%) were positive. In all four sera the highest titers were to HSV-2 followed by HSV-1 and SA8. Immunoblot analysis confirmed a preferential reactivity with HSV-2 antigen. Further analysis by competition ELISA indicated that these gorillas had experienced infection with a virus antigenically similar but not identical to HSV-2. These results represent the first evidence for an alpha-herpesvirus indigenous in a free ranging, nonhuman anthropoid species.

Animals↗
PubMed1991

The simian herpesvirus SA8 homologue of the herpes simplex virus gB gene: mapping, sequencing, and comparison to the HSV gB.

The genomic location and DNA sequence of the simian herpesvirus SA8 gene encoding a homologue of the HSV1 gB glycoprotein was determined. Using a cloned gB gene of herpes simplex virus type 1 (HSV1) as probe in Southern blot hybridizations, the SA8 gB gene was localized to a 10-kbp KpnI fragment mapping in the unique long part of the genome. A 2.8 kbp, 68.4% GC segment of this fragment was sequenced. It contained a 2649 nucleotide ORF possibly encoding a 98.4 kDa polypeptide. The predicted amino acid sequence of the SA8 gB polypeptide is 78.4% and 78.9% identical to the sequence of the HSV1 and HSV2 gBs, respectively, and was 88.4% similar or identical to both HSV gB sequences. Structural characteristics predicted for the SA8 gB polypeptide were very similar to those of HSV1 gB. These included a hydrophobic signal sequence of 29 amino acids, conservation of all 10 cysteine residues and 5 of 6 potential N-linked glycosylation sites present in the HSV1 gB, a triple hydrophobic transmembrane domain, and a highly charged cytoplasmic tail region. Both hierarchical cluster analysis and phylogenetic analysis of sequences for gB polypeptides of 12 different herpesviruses demonstrated that the gB glycoprotein of SA8 is most closely related to the HSV gB glycoproteins. Comparison of these closely related gB sequences identified four regions in which non-conservative amino acid substitutions were clustered. Localized regions of the gB polypeptide were identified which are likely to be associated with the conserved structure/function of the polypeptide.

Amino Acid Sequence↗