Hamster endogenous retrovirus (HaER) isolated from SV-40 induced tumor cells

A hamster cell line obtained from a secondary transplanted SV-40-induced tumor was found to spontaneously release type C virus particles. The particles possessed all biochemical features characteristic of retroviruses and induced syncytium formation in XC cells but were not oncogenic for hamsters. Nucleic acid hybridization assays demonstrated that the isolated virus was an endogenous retrovirus of hamster, lacking sequence homology with mouse and other animal species.     

Cloning and characterisation of a delta virus cDNA sequence derived from a human source

We report the extraction of delta virus RNA from the serum of a delta-virus-infected patient and the subsequent cloning and analysis of a 380-nucleotide-long cDNA (D380). The nucleotide sequence of D380 shows overall differences of approximately 20% when compared with previously published sequences and does not include the viroid consensus sequence previously reported (Wang et al: Nature 323:508-514, 1986). A potentially coding open reading frame extending over the whole length of the D380 has been identified. Our results demonstrate the existence of genetic heterogeneity amongst different delta virus isolates.     

Definition and variation of human endogenous retrovirus H

We defined the abundant human endogenous retrovirus group HERV-H based on pol similarity. Among 3661 pol-containing elements, 1124 integrations were similar to HERV-H RGH2 pol using translated pol sequences. A clustering procedure lessened these to 234 representatives, amenable to detailed study. Among the 1124, 926 clustered into HERV-H and 106 into adjacent HERV-H-like, the remainder being more distant to HERV-H. The HERV-H group was divided into RTVLH2-like (705 elements) and RGH2-like (77 elements) subgroups. Among 926 HERV-H, LTR differences were 1-33%, 10% had env, 78% had gag, 66% had a histidine primer binding site (PBS), and 3% (both subgroups) had a phenylalanine PBS. Allelic differences in env were studied using a convenient temperature gradient gel electrophoresis (TGGE) method and a genomic single nucleotide polymorphism (SNP) search. A pattern of abundant defective elements and less abundant less defective ones led us to formulate a "midwife" master model where more complete elements help the others in trans to transpose.     

Evolutionary implication of human endogenous retrovirus HERV-H family

The human endogenous retrovirus (HERV-H) family is most abundant and widely distributed in the human genome, with about 100–1,000 full-length or deleted elements and a similar number of solitary, long-terminal repeats. The HERV-H env ORF has been characterized in humans and in the course of primate evolution, indicating the increased possibility of biological roles in humans. Using the polymerase chain reaction approach with a human monochromosomal DNA panel, 70 env fragments belonging to the HERV-H family from chromosomes 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, X, and Y were identified and analyzed. They showed 82–99% sequence similarity to that of HERV-H (accession no. AF108843). We also identified other HERV-H env fragments in the DDBJ/EMBL/GenBank databases. The total of 120 fragments was evolutionarily analyzed. Phylogenetic analysis suggests that the HERV-H env family is divided into one major and two minor groups. The HERV-H members have been actively proliferated and evolved by intra-chromosomal spread during hominid radiation.     

Molecular characterisation of a cypovirus isolated from the western spruce budworm Choristoneura occidentalis

A novel cypovirus, assigned CoCPV, was isolated from natural populations of the western spruce budworm, Choristoneura occidentalis. The complete nucleotide sequences of genomic segments S2-S5 and S7-S10 were determined. Each segment contained a single open reading frame. Conserved motifs 5' (AGUUU......UUUGUGC) 3' were found at the ends of each segment. Analysis of S2, which encoded a putative RNA-dependent RNA polymerase protein, confirmed CoCPV belonged to the genus Cypovirus within the family Reoviridae. Further phylogenetic analysis using S10 (the polyhedrin gene) aligned this virus with species type-16, closely related to a cypovirus isolated from C. fumiferana.     

Production and characterisation of antibodies against isolated placental retroviral-like particles

The mammalian genomic DNA is known to contain a variety o f endogenous proviruses but their expression is usually restricted to reproductive tissues such as the placenta and a variety of human tumour cells. More definitive characterization of retroviral gene products has been hampered by unavailability of specific biological reagents. In this study, polyclonal and a total of six monoclonal antibodies (mAbs) were raised against endogenous (intact) retroviral particles isolated from human placental villous tissue. These antibodies were characterized using immunohistochemical and biochemical methods. Five polyclonal antibodies and one mAb (RV1-17) showed strong specific immunohistochemical and immunogold staining with submembraneous structures within placental syncytiotropblast. The reactivity of these antibodies was consistent with the pattern of apical syncytiotrophoblastic budding of retroviral particles previously reported in ultrastructural analyses.     

Molecular characterization of full-length MLV-related endogenous retrovirus ChiRV1 from the chicken, Gallus gallus

We report the first full-length sequence of an endogenous retrovirus from the genome of domestic chicken, that is not related to the Avian leukemia viruses (ALV). This retrovirus, designated ChiRV1, clusters with Murine leukemia virus (MLV)-related retroviruses and hence is the first complete gammaretrovirus from the genome of a bird. Nevertheless it is not related to exogenous MLV-related retroviruses infecting chicken. The provirus is 9133 bp long and contains 90%-identical LTRs as well as reading frames for the gag, pol and env genes, interrupted by in-frame stop codons. Expression analysis showed that ChiRV1 is a transcribed provirus. Screening of the chicken genome database revealed 100 ChiRV1-related sequences that are grouped into three classes based upon LTR alignment and subsequent phylogenetic analysis.     

Efficient isolation of endogenous rhesus retrovirus from trophoblast

An examination of various rhesus monkey (Macaca mulatta) organs has shown a preference for type C viral antigen expression in the placenta. Separate cocultivations of isolated trophoblasts from 10 rhesus monkey placentas with cell lines from heterologous mammalian species led to rapid isolation of type C rhesus retrovirus in 4 of 10 cases. These four retrovirus isolates have been designated MMC-2 through MMC-5. Five of the remaining six sets of cocultivations grew simian foamy virus and were discontinued. Distinction of these viral isolates from the initial rhesus isolate (MMC-1) and the previous isolate from the stumptail monkey, Macaca arctoides (MAC-1), could be made by liquid DNA hybridization, although not by limited restriction endonuclease digestion. Both MAC-1 and MMC-1 were obtained in single long-term cocultivation experiments (over 7 months). The present isolates MMC-2 through MMC-5 were detected in 2 to 5 weeks. Consequently, primary trophoblast cells represent a useful differentiated cell type for isolation of infectious retrovirus from this primate species.     

Molecular mimicry and immunomodulation by the HRES-1 endogenous retrovirus in SLE

Genetic and environmental factors are believed to influence development of systemic lupus erythematosus (SLE). Endogenous retroviruses (ERV) correspond to the integrated proviral form of infectious retroviruses, which are trapped within the genome due to mutations. ERV represent a key molecular link between the host genome and infectious viral particles. ERV-encoded proteins are recognized by antiviral immune responses and become targets of autoreactivity. Alternatively, ERV protein may influence cellular processes and the life cycle of infectious viruses. As examples, the HRES-1 human ERV encodes a 28-kDa nuclear autoantigen and a 24-kDa small GTP-ase, termed HRES-1/Rab4. HRES-1/p28 is a nuclear autoantigen recognized by cross-reactive antiviral antibodies, while HRES-1/Rab4 regulates surface expression of CD4 and the transferrin receptor (TFR) through endosome recycling. Expression of HRES-1/Rab4 is induced by the tat gene of HIV-1, which in turn down-regulates expression of CD4 and susceptibility to re-infection by HIV-1. CD4 and the TFR play essential roles in formation of the immunological synapse (IS) during normal T-cell activation by a cognate MHC class II peptide complex. The key intracellular transducer of T-cell activation, Lck, is brought to the IS via binding to CD4. T-cell receptorzeta (TCRzeta) chain binds to the TFR. Abnormal T-cell responses in SLE have been associated with reduced lck and TCRzeta chain levels. HRES-1 is centrally located on chromosome 1 at q42 relative to lupus-linked microsatellite markers and polymorphic HRES-1 alleles have been linked to the development of SLE. 1q42 is one of the three most common fragile sites in the human genome, and is inducible by DNA demethylation, a known mechanism of retroviral gene activation. Molecular mimicry and immunomodulation by a ERV, such as HRES-1, may contribute to self-reactivity and abnormal T and B-cell functions in SLE.     

Distribution of human endogenous retrovirus type W receptor in normal human villous placenta

BACKGROUND: The fusion of trophoblast cells into the villous syncytiotrophoblast is crucial for appropriate placental function and fetal development. Fusion occurs following the interaction of syncytin-1, an envelope protein of the endogenous retrovirus HERV-W, and the RD114/mammalian type D retrovirus receptor (RDR/ASCT2) on adjacent cell membranes. This process must be tightly regulated in order to maintain the proliferative pool of cytotrophoblast cells as well as the function of the syncytia. AIM: We sought to investigate whether syncytial fusion of placental cytotrophoblast cells may be regulated via modulation of RDR/ASCT2 expression. METHODS: Expression of RDR/ASCT2 in term and first trimester villous placenta was assessed along with a number of molecular markers using immunofluorescent staining. In a complementary approach, Western blotting was used to investigate RDR/ASCT2 expression in a panel of choriocarcinoma cell lines before and after stimulation of fusion. RESULTS: Villous placental RDR/ASCT2 expression was found to be restricted to the cytotrophoblast compartment, being largely absent in the syncytiotrophoblast. Local variations in RDR/ASCT2 expression were not associated with the proliferative status of cytotrophoblast cells. RDR/ASCT2 expression was also shown to be down-regulated in BeWo choriocarcinoma cells after stimulation of syncytial fusion. CONCLUSION: This first report of the localisation and distribution of RDR/ASCT2 in human placental villi suggests that the fusion of placental trophoblast cells is not regulated by local or temporal variations of RDR/ASCT2 expression in villous cytotrophoblast cells.     

Molecular characterisation of an avihepadnavirus isolated from Psittacula krameri (ring-necked parrot)

Avihepadnaviruses have been documented previously in ducks, herons, geese, storks and cranes. Here, we describe the full genome of a new avihepadnavirus isolated from Psittacula krameri (ring-necked parrot) in Poland. The parrot hepatitis B virus (PHBV) genome (3042 bp) shares <76% sequence identity with other avihepadnavirus isolates and is phylogenetically most closely related to heron and stork hepatitis B viruses isolates. PHBV has a genome organization similar to that of other hepadnaviruses and contains ORFs for a preC/C, preS/S and polyprotein. Additionally, we identified an X-like ORF in the genome of PHBV. The full-genome data will be useful in developing screening tools for avihepadnaviruses in parrots.     

Genetic characterisation of a Hantavirus isolated from a laboratory-acquired infection

Seoul (SEO) viruses belong to the Hantavirus genus (family Bunyaviridae), cause the moderate form of haemorrhagic fever with renal syndrome, and have been associated with laboratory-acquired infections in many countries. To investigate the pedigree of an isolate, designated IR461, which was obtained from a laboratory-acquired infection in a UK research institute, we determined the nucleotide sequences of the small (S) and medium (M) genome segments. In addition, we determined the sequences of the S segments of two Chinese isolates (R22 and L99) and an American isolate (Tchoupitoulas [TCH]). The S segments range within 1769-1785 nucleotides in length and showed identities of >88% in nucleotide sequence and 97% in amino acid sequence to those of published S segment sequences. The M RNA segment of IR461 is 3651 nucleotides long and shows >84% identity at the nucleotide level and >98% at the amino acid level to the M segments of other SEO viruses. These data confirm that SEO viruses show the least diversity within the Hantavirus genus. Phylogenetic analyses of the sequences showed geographic clustering of the Chinese SEO viruses, and that IR461 was more closely related to SEO viruses isolated in the New World than to those from the Far East.