- Volume 74, Issue 9, 1993
Volume 74, Issue 9, 1993
- Review Article
-
-
-
Vaccinia virus glycoproteins and immune evasion
More LessIntroduction. Vaccinia virus is the vaccine that was used to eradicate smallpox. This task was accomplished in 1977 and its completion certified in 1979 by the World Health Organization (WHO) ( Fenner et al., 1988 ). Since then poxvirus infections of humans have caused little disease and have been restricted to molluscum contagiosum and rare zoonoses such as cowpox (CPV), pseudocowpox, monkeypox, orf and yaba tumour viruses. Continued interest in vaccinia virus and other poxviruses has derived in part from the development of these viruses as cloning and expression vectors ( Mackett et al., 1982 ; Panicali & Paoletti, 1982 ) that have the potential as live vaccines to combat diseases other than smallpox (Panicali et al., 1983; Smith et al., 1983 a , b , 1984 ; Paoletti et al., 1984 ).
-
-
- Bacterial
-
-
-
Phage Acm1-mediated transduction in the facultatively methanol-utilizing Acetobacter methanolicus MB 58/4
More LessPhage Acm1, generally virulent for the acidophilic facultatively methanol-utilizing strain of Acetobacter methanolicus MB 58/4, is also capable of lysogenizing its host strain at a low rate. Using amino acid-auxotrophic mutants of A. methanolicus MB 58/4 as recipient strains, transduction of His, Leu and Tyr markers could be demonstrated in this system. The ability to prepare transducing lysates by propagation of phage Acm1 on the prototrophic donor strain A. methanolicus MB 58/4, the transduction of three different markers as well as the efficiency of transduction, and the occurrence of permutations in the phage genome indicate that phage Acm1 mediates generalized transduction. Phage Acm1 might be a useful tool in genetic studies of methylotrophic A. methanolicus.
-
-
- Animal
-
-
-
Genomic human immunodeficiency virus type 1 RNA variation in mother and child following intra-uterine virus transmission
More LessIn order to study the relationship between virus populations in a human immunodeficiency virus type 1 (HIV-1)-infected mother and her infant, we analysed a 276 bp fragment, including the V3 region, of genomic HIV-1 RNA purified from serum. Samples were collected from the mother 6, 4 and 2 months prior to delivery, during delivery and 10 months after childbirth (samples MA to ME, respectively) and from the infant at birth (cord blood) and the ages of 6 weeks and 9 months. A heterogeneous sequence population was observed in the maternal samples (mean nucleotide variation of 2∙4 to 4∙2%, range 0 to 8∙3%). Until the age of 6 weeks the sequence population in the infant was highly homogeneous (mean nucleotide variation ⩽ 0∙7%, range 0 to 2∙5%). At 9 months of age, the infant’s virus population showed more heterogeneity (mean nucleotide variation of 1∙8%, range 0∙4 to 3∙6%) and a drift in the consensus sequence was observed. The evolution of the V3 region in the mother was characterized by accumulation of amino acid substitutions diverging from the virus population observed in the infant. The mean nucleotide distance between the maternal sequence populations and the sequence population of the child at birth was 2∙8, 2∙6, 3∙7, 5∙2 and 5∙3% for the samples MA, MB, MC, MD and ME, respectively. Nearly complete replacement at position 308, previously described as antigenically important, from a proline to a histidine was observed during pregnancy, whereas all clones of the child’s virus at birth and at 6 weeks contained a proline at that position. In conclusion, intra-uterine transmission is associated with a homogeneous sequence population in the child at birth, which is more closely related to the sequence population present in the mother during the first and second trimester of pregnancy than to the sequence population at delivery.
-
-
-
-
Differences in the B and T cell immune response to the envelope glycoprotein 130 (gp130) of the macaque strain of simian immunodeficiency virus (SIVmac), induced by immunization of rhesus macaques with virus-derived or vaccinia virus-expressed gp130
Rhesus macaques were immunized with purified virus-derived simian immunodeficiency virus of macaques (SIVmac) 251/32H glycoprotein 130 (gp130) or primed with recombinant vaccinia virus (VV) expressing the env gene of the SIVmac BK28 clone and boosted subsequently with virus-derived gp130. High antibody titres of at least 104 against recombinant gp140 were induced with both vaccines. Analysis of the antibody specificity with a peptide ELISA revealed that different linear epitopes were recognized after administration of virus-derived gp130 compared with those after priming with VV. Antibodies to some epitopes (peptides 10 and 49), which were also found in SIV-infected animals, were induced with both vaccines, whereas antibodies to other regions were induced by only one vaccine preparation. The analysis of the helper T cell response revealed a poor immunogenicity of the virus-derived gp130, whereas priming with VV induced a considerable helper T cell activity in all three vaccinees after the second VV infection. Using synthetic peptides, several epitopes were identified. Our observations show that immunization with a virus-derived gp130 or live recombinant VV induces a considerably different antibody and helper T cell response. These differences in immunogenicity might have important implications for further vaccine development.
-
-
-
Characteristics of a retrovirus associated with Jembrana disease in Bali cattle
A virus causing Jembrana disease in Bali cattle (Bos javanicus) was demonstrated to have characteristics of a retrovirus. Reverse transcriptase activity was detected in virus purified by sucrose gradient centrifugation. Electron microscopic examination of tissue from the affected cattle indicated that the virus matured by C-type budding through the plasma membrane and into intracytoplasmic vacuoles of cells in lymphoid tissue, with the formation of circular enveloped virus particles ranging in diameter from 96 to 124 nm with an eccentric nucleoid. Western immunoblotting using sera from recovered animals demonstrated virus proteins of M r 100K, 45K, 42K, 33K, 26K, 16K and 14K. The 26K protein of Jembrana disease virus cross-reacted in Western blots with the 26K capsid protein of bovine immunodeficiency virus (BIV). The apparent morphogenesis, protein structure and antigenic relationship with BIV suggested the virus was a lentivirus.
-
-
-
The involvement of a spliceosome component in internal initiation of human rhinovirus RNA translation
More LessHuman rhinoviruses (HRVs) and encephalomyocarditis virus (EMCV) belong to different genera of the picornavirus family, but the translation of the RNAs of both viruses is by the same mechanism, that is, internal ribosome entry. In rabbit reticulocyte lysates this translation initiation is efficient for mRNAs bearing the EMCV 5′ untranslated region (5′ UTR), but very inefficient for mRNAs bearing the HRV 5′ UTR, unless factors from HeLa cells are added. The copurification of the HeLa cell translation stimulatory activity with proteins which can be specifically cross-linked to the HRV 5′ UTR by u.v. irradiation has been examined. Both the EMCV and HRV 5′ UTRs can be cross-linked to a 58/60K protein doublet present in HeLa cell extracts in higher amounts than in reticulocyte lysates, which is shown to be very similar, if not identical to the polypyrimidine tract binding protein (PTB) previously identified as a component of a multi-subunit complex necessary for pre-mRNA splicing. However, the activity in HeLa cell extracts that specifically stimulates translation initiation on mRNAs with the HRV 5′ UTR does not copurify with the majority of the 58/60K protein present in these extracts, but copurifies with a minor fraction of these proteins and with a 97K protein which can be cross-linked to the HRV 5′ UTR but not to the EMCV 5′ UTR, and which is absent from reticulocyte lysates. It is proposed that the specific translation initiation stimulatory activity found in HeLa cells is due to a high M r complex containing the 97K polypeptide and PTB.
-
-
-
A temperature-sensitive mutation in the acidic polymerase gene of an influenza A virus alters the regulation of viral protein synthesis
More LessThe temperature-sensitive defect of mutant ts 263 of fowl plague virus (FPV) is located in the acidic polymerase (PA) gene and is due to a single base substitution (C2036T), which leads to an amino acid replacement (Ala671 to Val) in a highly conserved region of the protein. During passage at 33 °C ts 263 stably carries over a ninth RNA segment, which consists of a truncated PA gene. Although the deletion is in-frame and it is transcribed into mRNA, no corresponding protein is detected in vivo. After reversion to wild-type this extra RNA segment is immediately lost. At the nonpermissive temperature of 40 °C no significant viral products of ts 263 are synthesized. Under semi-permissive conditions there is a relative, but very significant over-production of the M1 protein, which is not accompanied by a corresponding elevated M1 mRNA synthesis. These results are in agreement with the idea that the PA protein is involved in the regulation of viral protein synthesis at the level of expression of mRNA. Preinfection of chicken embryo cells with ts 263 at a semi-permissive temperature interferes with the replication of FPV wild-type indicating that premature availability of M1 might be detrimental for influenza virus replication.
-
-
-
Sequence determination of the nucleocapsid protein gene of the porcine epidemic diarrhoea virus confirms that this virus is a coronavirus related to human coronavirus 229E and porcine transmissible gastroenteritis virus
More LessThe nucleotide sequence of 1.7 kbp cDNA, comprising the region nearest the 3′ end of the genome of the porcine epidemic diarrhoea virus (PEDV), has been independently determined for two European isolates of PEDV. Almost identical results were obtained for the two isolates, which were derived from cases of PEDV infection in Belgium and Britain in 1977 and 1987, respectively. The sequences contained a 1323 nucleotide (nt) open reading frame (ORF), which showed moderate identity to the nucleocapsid (N) gene of other coronaviruses. The greatest similarity at both the nucleic acid and protein levels was to the human coronavirus 229E. The PEDV N gene was, however, notably larger than that of the human 229E and porcine transmissible gastroenteritis viruses. This reflects the presence of a putative insertion of approximately 135 nt located towards the middle of the N gene. A second 336 nt ORF, which might encode a leucine-rich protein similar to, but shorter than, the bovine coronavirus internal protein was found within the PEDV N gene. Several RNA motifs typical of coronaviruses were also observed. These results confirm the earlier provisional classification of PEDV as a coronavirus.
-
-
-
A novel small RNA virus isolated from the cotton bollworm, Helicoverpa armigera
More LessA small RNA virus with novel characteristics has been isolated from laboratory-bred larvae of Helicoverpa armigera. Infection by the H. armigera stunt virus causes severe retardation of larval development and subsequent death. Its particles are isometric, 38 nm in diameter, and have a buoyant density of 1.296 g/ml in caesium chloride. The viral capsid has two major non-glycosylated protein components with M rs of 65000 and 6000, and contains a genome composed of two non-polyadenylated single-stranded RNA molecules with lengths of 2.4 kb and 5.5 kb. The 5′ termini of these RNAs are capped; their 3′ termini are unblocked. In vitro translations of the viral RNAs showed synthesis of large proteins of sizes near the maximum coding capacity of each strand along with synthesis of numerous smaller proteins; no evidence for processing of precursors was seen. The physicochemical properties of the virus are most similar to those of the Nudaurelia ω virus, a provisional member of the Tetraviridae, although no antigenic relationship was observed between the two viruses. The bipartite genome and distinct capsid structure of these two viruses indicate the existence of a previously unrecognized virus group.
-
-
-
A gene encoding a highly expressed spindle body protein of Heliothis armigera entomopoxvirus
More LessThe gene encoding the most abundant protein of purified preparations of Heliothis armigera entomopoxvirus (HaEPV) has been cloned and sequenced. The gene sequence encodes a 40.1K polypeptide with a putative N-terminal 20 amino acid leader peptide, and a single potential N-glycosylation site. Analysis of the protein, which has an apparent M r of 50K on polyacrylamide gels, confirmed post-translational loss of the leader peptide, but showed no evidence of glycosylation. The protein is related to others previously described from Choristoneura biennis EPV (63% identity) and Autographa californica nuclear polyhedrosis virus (42% identity). Polyclonal antiserum raised against a bacterial fusion protein containing the majority of the HaEPV protein specifically labelled HaEPV spindle bodies; confocal laser scanning microscopy suggests that the protein is distributed throughout those viral structures.
-
-
-
Analysis of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus trans-activators IE-1 and IE-2 using monoclonal antibodies
More LessWe have produced monoclonal antibodies against two Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) transcriptional trans-activators, IE-1 and IE-2. Temporal analysis of IE-1 and IE-2 proteins have shown that IE-1 continues to increase in steady-state levels from 0 to 120 h post-infection, whereas IE-2 declines by 36 h post-infection. At least five different electrophoretic forms of IE-1 are present in OpMNPV-infected LD652Y cells of which three appear to be from the non-spliced IE-1 gene. Cotransfection experiments also showed that IE-1 causes a reduction in the levels of IE-2 protein but concurrently IE-2 increases IE-1 expression. Western blot analysis indicated that a form of IE-1 copurified with budded virions but did not copurify with the polyhedron-derived virus.
-
-
-
Discovery of a novel point mutation changing the HDAg expression of a hepatitis delta virus isolate from Central African Republic
More LessNone of the mutations so far discovered in several hepatitis delta virus (HDV) isolates appears to determine important changes in HDV specific protein (HDAg) expression, except for a putative mutation at nucleotide 1012 converting an amber stop codon (TAG) to a codon for tryptophan (TGG). Here we present the characterization of an HDV obtained from the liver of a woodchuck inoculated with sera from fulminant HDV patients in Central African Republic (CAR). By restriction enzyme analysis and sequencing of HDAg-coding region cDNA clones, we found that this HDV isolate bears a novel mutation (T to A) at nucleotide 1013 which converts the amber stop codon (TAG) to a codon for lysine (AAG). Comparison of these nucleotide sequences with those available from American, Japanese, Taiwanese, French, Italian and Nauru isolates showed a variability of 1.7 to 21.5% and 1.9 to 28.7% at the nucleic acid and amino acid levels, respectively. The HDAg-encoding sequence of the CAR isolate is closely related to that of the Italian HDV isolate. The in vitro expression of this HDV isolate resulted in a unique HDAg species (28K) which was identical with that characterized in vivo.
-
-
-
Nucleotide and predicted amino acid sequences of Marek′s disease virus homologues of herpes simplex virus major tegument proteins
More LessThe DNA sequence of an 8.4 kbp BamHI-EcoRI fragment of Marek′s disease virus (MDV) strain GA was determined. Three of the predicted polypeptides are homologous to UL47, UL48 and UL49 encoding the major tegument proteins of herpes simplex virus type 1 (HSV-1), and four are homologous to HSV-1 UL45, UL46, UL49.5 and UL50. These seven genes are found in the long unique region of the MDV genome and are collinear with homologues in HSV-1 and varicellα-zoster virus (VZV). Northern blot analysis revealed different transcriptional patterns from those of HSV-1 and VZV. MDV homologues of UL49.5, UL49 and UL47 lack a poly(A) signal immediately downstream of their coding regions. Amino acid conservation between MDV and HSV-1, and between MDV and VZV is as high as that between HSV-1 and VZV. The MDV homologue of UL48 shows 60% similarity to its HSV-1 counterpart. Amino acid sequence comparison reveals that the MDV homologue of UL48 lacks an acidic carboxyl terminus. This homologue, like the VZV homologue of UL48, may be involved in the trans-activation of immediate early genes and may function as an important component of the structural proteins.
-
-
-
Identification and expression of the human herpesvirus 6 glycoprotein H and interaction with an accessory 40K glycoprotein
More LessIn herpes simplex virus (HSV) the small secreted glycoprotein gL forms a heterodimer with the transmembrane envelope glycoprotein gH. Here we identify the human herpesvirus 6 (HHV-6) gL gene, express HHV-6 gL and gH homologues, and examine interactions between HHV-6 gH and gL. The HHV-6 gL gene encoded a glycoprotein with an amino acid sequence which showed closest similarity to the human cytomegalovirus (HCMV) gL homologue (18% identity). Products of HHV-6 gH and gL genes were characterized in an in vitro transcription-translation system and in a transient in vivo expression system. Both gH and gL were transcribed and translated in vitro to give products of apparent Mr of 65K and 28K in SDS-PAGE, and these could be processed by addition of microsomes to 110K and 40K, respectively. To study gH/gL interactions, gH was tagged with the nine amino acid epitope for monoclonal antibody LP14 (anti-HSV-1 gD). LP14 and a human serum sample specifically immunoprecipitated gH and a stable complex of gH and gL co-expressed in an in vivo vaccinia virus-T7 system. The gH and gL produced in this in vivo expression system corresponded to the Mr s of the fully processed glycoproteins identified in the in vitro system. The gH expressed together with gL was recognized by human sera more easily than when examined on its own in immunofluorescence assays. Dual expression of gH and gL in transfected T lymphocytes (JJhan) caused reactions with 75% of human sera tested (12 HHV-6-positive, HCMV-negative serum samples), but gL expressed alone was not recognized by these sera. The immunofluorescence studies also showed that the glycoproteins were localized in Golgi-like bodies in fibroblasts, but occurred throughout the endoplasmic reticulum in T lymphocytes, the normal cellular target for HHV-6. These results show the identification of the HHV-6 homologue to the HCMV and HSV gL genes, identification and production of HHV-6 gH and gL expressed both in vitro and in vivo, complex formation between these glycoproteins, and evidence that this complex may be localized differently in fibroblasts as compared to T lymphocytes and that it is immunogenic.
-
-
-
In vivo characterization of site-directed mutations in the promoter of the herpes simplex virus type 1 latency-associated transcripts
Transient expression assays in PC12 cells showed that the cAMP response element (CRE) and the TATA box of the herpes simplex virus type 1 latency-associated transcripts (LATs) promoter are essential for basal expression. Recombinant viruses were generated containing site-specific mutations in these motifs. The abilities of these recombinants to replicate, express LATs and reactivate from latency were compared with wild-type and marker-rescued viruses in a murine ocular model. The acute replication of these TATA and CRE mutant viruses was at a level equivalent to their respective marker-rescued viruses. The reactivation of virus was unaffected by mutation in the TATA box as compared with wild-type or marker-rescued viruses. In situ hybridization of TATA box mutant virus-infected ganglia, however, showed threefold fewer LAT-positive neurons than wild-type virus-infected ganglia, with consistently weaker hybridization signals. Thus, this TATA box is required for normal expression of the LATs but not for efficient reactivation. The LATs CRE mutant reactivated with slightly but reproducibly reduced frequency and delayed kinetics relative to marker-rescued virus. By in situ hybridization, however, the percentage and intensity of LATs-positive neurons were found to be comparable for the CRE mutant- and wild-type virus-infected ganglia, suggesting that the CRE is dispensable for abundant LATs expression but that a reactivation function of the LATs may depend upon the presence of the CRE. Finally, using a modified assay for examining the timing of reactivation, we showed that the induction of viral reactivation by addition of exogenous cAMP can occur independently of the LATs.
-
-
-
Epitope mapping identifies an exposed loop between the unique amino- and conserved carboxy-domains of the large subunit of herpes simplex virus type 1 ribonucleotide reductase
More LessThe large subunits of herpes simplex virus types 1 and 2 ribonucleotide reductases contain unique aminoterminal regions comparising 311 and 318 residues respectively, which are not found in ribonucleotide reductases from other sources. We report the mapping of the epitope recognized by monoclonal antibody 1026, which is specific for the large subunit (R1) of HSV-1, and then deduce the structural relationship of the amino-terminal region of R1 with the rest of the protein. A panel of 10 fusion proteins containing sequences spanning the entire R1 subunit were constructed. They were used together with proteolytic fragments of R1 and several synthetic peptides to show that the epitope is discontinuous and appears to be a loop structure centred on a previously located trypsin-sensitive site at residue 305. The existence of the loop was suggested by the observation that reactivity of the antibody with R1 could be blocked by peptides corresponding to residues 289 to 303 and 308 to 313 which flank the trypsin-sensitive site. Our results suggest that the unique amino-terminal region of R1 consists of a structurally distinct domain which is linked to the conserved carboxy region by an exposed loop.
-
-
-
Effect of indomethacin on ultraviolet radiation-induced recurrent herpes simplex virus disease in guinea-pigs
More LessExposure to u.v. radiation increases the local level of prostaglandins which may play a role in u.v. radiation-induced herpes simplex virus (HSV) recurrences. We used the guinea-pig model of u.v. radiation-induced recurrent genital HSV-2 disease for examining the effects of indomethacin, a prostaglandin inhibitor, on u.v.-induced recurrences. In the first experiment, performed 100 days after HSV-2 inoculation, treatment with indomethacin for 5 days begun 24 h before u.v.-irradiation decreased the proportion of animals developing HSV disease recurrences from 11/13 (84∙6%) to 2/13 (15∙4%) (P < 0∙001). In the second experiment, performed 135 days after HSV-2 inoculation, treatment with indomethacin for 5 days begun 24 h before u.v.-irradiation decreased the number of animals developing recurrences from 12/21 (57∙1%) to 5/21 (23∙8%) (P < 0∙05). Five days of indomethacin treatment begun 4 h after u.v.-irradiation, however, did not reduce the percentage of animals developing disease recurrences but did decrease the mean number of days with recurrent lesions in animals that developed recurrences. Our data suggest that indomethacin may modify u.v. radiation-induced recurrent lesions by decreasing viral reactivation when given before u.v. radiation exposure or by reducing prostaglandin-induced immunosuppression when given before or after exposure. Future studies are needed for evaluating indomethacin prophylaxis for recurrent HSV disease when prolonged u.v. radiation exposure is anticipated.
-
-
-
An epitope within the DNA-binding domain of the herpes simplex virus immediate early protein Vmw175 is conserved in the varicella-zoster virus gene 62 protein
More LessWe have isolated a panel of monoclonal antibodies that recognize the DNA-binding domain of the herpes simplex virus type 1 (HSV-1) immediate early polypeptide Vmw175. The mice used for the fusions had been immunized with the isolated Vmw175 DNA-binding domain. This had been purified from bacteria that carried a phage T7 expression plasmid with the DNA-binding domain coding region. The epitopes recognized by the monoclonal antibodies were mapped by using a family of truncated versions of the DNA-binding domain, which had also been expressed in the bacterial expression system. The monoclonal antibodies divided into at least four different groups according to this mapping. Several of the monoclonal antibodies recognized Vmw175 expressed in infected BHK cells by HSV-1 strain 17 in Western blots. One of them also recognized the corresponding protein of varicella-zoster virus gene 62. This is further illustration of the relatedness of the two polypeptides.
-
-
-
The DNA sequence of the equine herpesvirus 4 gene encoding glycoprotein gp17/18, the homologue of herpes simplex virus glycoprotein gD
More LessThe nucleotide sequence of the gene to the left of the gI gene of equine herpesvirus 4 (EHV-4) was determined. The gene encodes a peptide of 402 amino acids with an unprocessed Mr of 45323. The predicted polypeptide has several features of a glycoprotein including a hydrophobic signal sequence, a membrane spanning domain and four potential N-linked glycosylation sites within the proposed external domain. The predicted amino acid sequence of EHV-4 gD shows 83% identity with that of equine herpesvirus 1 gD. Conservation of the tertiary structure is suggested by the alignment of six cysteine residues with those of the gD of six other alphaherpesviruses. Screening a λgt11/EHV-4 expression library with monoclonal antibodies against several of the most abundant EHV-4 glycoproteins unequivocally identified the protein encoded by the EHV-4 gD gene as gp17/18.
-
-
-
Inhibition of human cytomegalovirus major immediate early gene expression by antisense RNA expression vectors
More LessWe have used antisense oligonucleotides and expression vectors to inhibit human cytomegalovirus (HCMV) major immediate early (IE) gene expression. We find that oligonucleotides complementary to the HCMV 72K IE protein (IE1) coding region do inhibit HCMV infection, but this is non-specific. However, the use of certain antisense expression vectors, which express short oligonucleotides complementary to IE1, specifically inhibits IE1 expression at the protein level after introduction of IE expression vectors into cells or after HCMV infection.
-
-
-
Three African swine fever virus genes encoding proteins with homology to putative helicases of vaccinia virus
More LessSequence analysis of the SalI g, h, i and j restriction fragments of the African swine fever virus (ASFV) genome from the virulent isolate Malawi (LIL20/1) identified three open reading frames (ORFs) encoding predicted proteins of 125.0K (g10L), 80.4K (j10L) and 58.0K (j11L) which showed homology to members of the DNA and RNA helicase superfamily. ORF j10L was related to protein 4 of the Kluyveromyces lactis killer plasmid pKG12 and to two putative helicases, D6R and D11L, of vaccinia virus. ORF g10L was most closely related to ASFV j10L and to vaccinia virus D11L. ORF j11L was homologous to A18R, a third putative helicase of vaccinia virus. The possible functions of these genes in the replication of ASFV are discussed and the evolutionary implications are considered.
-
-
-
Localization of the RNA-binding domain of mouse hepatitis virus nucleocapsid protein
More LessThe 454-amino acid nucleocapsid (N) protein of mouse hepatitis virus (MHV) binds the leader RNA sequence located at the 5′ ends of all plus-sense genomic and subgenomic viral mRNAs. Purified N protein was cleaved with formic acid to determine which domain interacts with the leader RNA sequence. Incubation at 42 °C resulted in partial cleavage into two fragments of M rs of approximately 32K and 37K and three fragments of 17K, 16K and 14K. Incubation at 56 °C resulted in complete cleavage yielding only the three lower molecular mass products. Both the 32K and 37K partial cleavage products and one of the complete cleavage products bind MHV leader RNA, suggesting that the central region of the N protein contains the RNA-binding domain. Monoclonal antibody mapping of the cleavage products confirmed that the MHV leader RNA binding domain is contained within the central 140-amino acid fragment, comprising amino acids 169 to 308. Analysis of the amino acids within this domain indicates no similarity to any previously described RNA-binding protein, suggesting that N protein may possess a unique RNA-binding motif.
-
-
-
Molecular characterization of the S protein gene of human coronavirus OC43
More LessThe gene encoding the spike protein of the OC43 strain of human coronavirus (HCV-OC43) was cloned and sequenced. The complete nucleotide sequence revealed an open reading frame of 4062 nucleotides encoding a protein of 1353 amino acids with a predicted M r of 150078. Structural features include 22 N-glycosylation sites, an N-terminal hydrophobic signal sequence of 17 amino acids, an hydrophilic cysteine-rich sequence of 35 amino acids near the C terminus, and a potential proteolytic cleavage site (RRSR) between amino acid residues 758 and 759, yielding S1 and S2 segments of 84730 and 65366 M r, respectively. The predicted amino acid sequence of the spike protein of HCV-OC43 has 91% identity with that of the Mebus strain of bovine coronavirus, revealing more sequence divergence in the putative bulbous part (S1) than in the predicted stem region (S2).
-
-
-
Fusion protein gene nucleotide sequence similarities, shared antigenic sites and phylogenetic analysis suggest that phocid distemper virus type 2 and canine distemper virus belong to the same virus entity
Nucleotide sequencing of the fusion protein (F) gene of phocid distemper virus-2 (PDV-2), recently isolated from Baikal seals (Phoca sibirica), revealed an open reading frame (nucleotides 84 to 2075) with two potential in-frame ATG translation initiation codons. We suggest that the second in-frame ATG triplet at positions 264 to 266 initiates the translation, resulting in a protein of 537 amino acid residues with a calculated M r of 63035. The putative F1/F2 cleavage site, located approximately 100 amino acid residues from the N terminus, is identical to those of the F proteins of phocid distemper virus-1 (PDV-1) isolated from European harbour seals (Phoca vitulina) and of canine distemper virus (CDV). A full scale comparison of morbillivirus F genes reveals that the conserved F0 extracellular protein-encoding region contains a large number of non-expressed mutations, suggesting that this part of the protein is under strong functional constraints. Phylogenetic analysis of morbillivirus F gene nucleotide sequences revealed a closer evolutionary relationship between PDV-2 and CDV than between PDV-1 and PDV-2. These data were supported by cross-reactivity patterns of PDV-2 and CDV obtained with monoclonal antibodies to structural proteins of PDV-1 and CDV, and suggest that PDV-2 is a strain of CDV, resulting from a trans-species infection.
-
-
-
Development of a novel subunit vaccine that protects cotton rats against both human respiratory syncytial virus and human parainfluenza virus type 3
A cotton rat model of experimental human respiratory syncytial virus (RSV) and human parainfluenza virus type 3 (PIV-3) infection was used to examine the efficacy of FRHNP, a novel chimeric glycoprotein which contains the extracellular regions of the fusion glycoprotein of RSV and the attachment glycoprotein of PIV-3, as a single subunit vaccine against these two viruses. This work was prompted by previous cotton rat studies that demonstrated that the major protective antigens of the two viruses were these glycoproteins. FRHNP was expressed in insect cells using a recombinant baculovirus. Vaccination with FRHNP resulted in induction of both RSV and PIV-3 neutralizing antibody and doses of 200 ng completely protected rats from either RSV or PIV-3 challenge. These results demonstrate that in the cotton rat animal model a single chimeric glycoprotein can be an effective vaccine against both RSV and PIV-3.
-
-
-
Sequence variability of the glycoprotein gene of bovine respiratory syncytial virus
More LessSequence variation in the attachment glycoprotein G of bovine respiratory syncytial virus (BRSV) was determined. The nucleotide sequences of the G mRNAs of the A51908, VC464 and FS-1 strains of BRSV were compared with the published sequence of the BRSV strain 391-2. Nucleotide sequence alignment showed that overall they are highly conserved, with 90 to 97% identity. In addition, the coding region of strain A51908 was longer by 18 nucleotides at the 3ʹ end. An 84 to 95% level of identity was observed among the deduced amino acid sequences of the G proteins of BRSV strains. A maximum divergence of 19% was found when the extracellular domains of the G proteins were compared. The level of diversity would be consistent, by analogy to the human respiratory syncytial viruses, with these BRSV strains forming a single subgroup.
-
-
-
Topographical analysis of canine parvovirus virions and recombinant VP2 capsids
The distribution of epitopes defined by monoclonal antibodies (MAbs) on the surface of canine parvovirus (CPV) virions and recombinant VP2-capsids was established using immunoelectron microscopy. A correlation appeared to exist between the linear position, neutralizing activity and immunogold staining. Both viral capsids and recombinant capsids gave similar patterns of immunostaining. The neutralizing MAbs that recognized epitopes not previously identified by Pepscan or immunoblotting gave a clear staining. However, MAbs 3C9 and 3C10, identified by Pepscan and immunoblotting as recognizing linear epitopes, did not show any labelling (3C9) or only scattered labelling (3C10). MAb 3C9 recognizes an N-terminal domain of VP2. MAb 4AG6, which recognizes the same linear epitope as 3C10, did not bind to the capsids, indicating a different orientation. An immunofluorescence assay was performed to supplement the B cell epitope characterization. In contrast to other MAbs that gave nuclear and cytoplasmic staining, MAb 3C9 gave a preferential nuclear staining. Based on these results, it is hypothesized that the N terminus of VP2 is barely, or not at all, exposed on the surface of the native virions, but becomes accessible after some virion steric change (e.g. after attachment to the cell receptor).
-
-
-
Trans-activation of the long terminal repeat of human immunodeficiency virus type 1 by the parvovirus B19 NS1 gene product
More LessPersistent parvovirus B19 infections in human immuno-deficiency virus type 1 (HIV-1)-infected patients have been reported. The two viruses could share common target cells. The NS1 protein of B19 regulates B19 expression and we have investigated its possible effect on the long terminal repeat (LTR) of HIV-1. In transient transfection experiments, NS1 trans-activated the expression of reporter genes under the control of the HIV-1 LTR. The effect of NS1 was apparent only in the presence of the HIV-1 Tat protein, and required intact TAR and TATA box sequences.
-
-
-
Electron microscopic evidence for budding process-independent assembly of double-shelled rotavirus particles during passage through endoplasmic reticulum membranes
More LessSlowing down of the maturation process of human rotavirus particles on ice allowed the clear demonstration of two different assembly pathways through the endoplasmic reticulum (ER) membrane. One was the ‘enveloped’ and single-shelled (ss) particle assembly pathway, in which a transient envelope is acquired through the budding of subviral particles from the cytoplasm to the ER lumen, and later these ‘enveloped’ particles are released as ss particles in the ER lumen. The other was a double-shelled particle assembly pathway by which subviral particles acquire the outer capsid proteins during their transport across the ER membrane.
-
-
-
Productive and non-productive phases during long-term persistence of influenza C virus
More LessPersistent infection with a variant of influenza C/Ann Arbor/1/50 virus in MDCK cells has been previously reported. However, the precise molecular mechanism of persistence is still unknown. We show that the release of active progeny virus, as tested for by haemagglutination and acetylesterase profiles, does not take place in freshly seeded MDCK cells. Productive virus replication occurs simultaneously with massive production of structural proteins as shown by immunoprecipitation and immunofluorescence. PCR for the HEF structural protein-encoding segment 4 revealed that positive-sense RNA is present only during virus multiplication whereas negative-sense RNA appears to be constantly detectable. In this study we give initial evidence that influenza C virus can persist in the form of its genomic minus strand RNA, and plus strand transcription, protein synthesis and virus replication remain restricted to productive phases.
-
-
-
Antigenic characterization of serogroup ‘A’ of infectious pancreatic necrosis virus with three panels of monoclonal antibodies
More LessMonoclonal antibodies (MAbs) were produced against three serotypes of infectious pancreatic necrosis virus (IPNV): A1 (LWVRT 60-1, U.S.A.), A2 (d'Honnincthun, France) and A9 (Jasper, Canada). Each panel of MAbs (identified as LW, HF and JA) was analysed by ELISA with the 10 proposed serotypes of IPNV and their specificity defined by immunoprecipitation and Western immunoblotting analysis. A first group of MAbs, directed against the outer capsid protein VP2, reacted with linear or conformational epitopes. A second group of MAbs, directed against the internal protein VP3, reacted with linear epitopes. There was no relationship between the neutralizing property of anti-VP2 MAb and the configuration of the epitope that it recognized. The MAbs were used for antigenic characterization of serogroup A. Each panel of MAbs showed a characteristic pattern of reactivity. The European HF series was predominantly cross-reactive and detected conserved epitopes among the 10 serotypes for both VP2 and VP3. The North American LW and JA series identified a group of conserved epitopes on VP3 and new specific epitopes on VP2 and VP3. The higher variability observed for VP2 in comparison with VP3 is one example of how external pressures may promote natural selection of those epitopes required for virus survival. Our results are consistent with an ancestral relationship of the European to the North American strains, the latter having developed new antigenic determinants upon evolution in their new geographical location.
-
-
-
Assessment of the antigenic structure of tick-borne encephalitis virus by the use of synthetic peptides
The feasibility of using synthetic peptides for the identification of individual monoclonal antibody (MAb)-defined epitopes was assessed on the basis of a structural model of the tick-borne encephalitis (TBE) virus envelope glycoprotein E. For this purpose a series of 19 synthetic peptides was prepared, covering most of the E protein sequence. Each of the peptides was tested by ELISA for reactivity with 19 protein E-specific MAbs raised against TBE virus strain Neudoerfl. Specific reactivity was observed with three MAbs and two peptides (representing amino acids 1 to 22 and 221 to 240, respectively), thus providing new information on the location of the corresponding epitopes. Specificity was confirmed in a competition ELISA by the ability of the peptides to block MAb binding to TBE virus antigen. However, in contrast to the other MAbs, these peptide-reactive MAbs were not blocked by native virus particles in the competition ELISa, indicating that they do not recognize the native conformation of the E protein. These three MAbs also showed increased reactivity with denatured forms of the virus in a dot blot assay. Additionally, they reacted only in ELISA systems in which the virus was directly coated to the solid phase and thereby presumably partially denatured, but not when a capture antibody was used, which preserves the native antigen conformation. We have thus identified two classes of MAbs, those which recognize the native form and those which recognize the denatured form of protein E. The latter may be useful for the analysis of sites probably involved in protein folding and oligomerization.
-
- Plant
-
-
-
Novel rod-shaped viruses isolated from garlic, Allium sativum, possessing a unique genome organization
More LessRod-shaped flexuous viruses were partially purified from garlic plants (Allium sativum) showing typical mosaic symptoms. The genome was shown to be composed of RNA with a poly(A) tail of an estimated size of 10 kb as shown by denaturing agarose gel electrophoresis. We constructed cDNA libraries and screened four independent clones, which were designated GV-A, GV-B, GV-C and GV-D, using Northern and Southern blot hybridization. Nucleotide sequence determination of the cDNAs, two of which correspond to nearly one-third of the virus genomic RNA, shows that all of these viruses possess an identical genomic structure and that also at least four proteins are encoded in the viral cDNA, their Mr s being estimated to be 15K, 27K, 40K and 11K. The 15K open reading frame (ORF) encodes the core-like sequence of a zinc finger protein preceded by a cluster of basic amino acid residues. The 27K ORF probably encodes the viral coat protein (CP), based on both the existence of some conserved sequences observed in many other rod-shaped or flexuous virus CPs and an overall amino acid sequence similarity to potexvirus and carlavirus CPs. The 11K ORF shows significant amino acid sequence similarities to the corresponding 12K proteins of the potexviruses and carlaviruses. On the other hand, the 40K ORF product does not resemble any other plant virus gene products reported so far. The genomic organization in the 3ʹ region of the garlic viruses resembles, but clearly differs from, that of carlaviruses. Phylogenetic analysis based upon the amino acid sequence of the viral capsid protein also indicates that the garlic viruses have a unique and distinct domain different from those of the potexvirus and carlavirus groups. The results suggest that the garlic viruses described here belong to an unclassified and new virus group closely related to the carlaviruses.
-
-
-
-
Changes in populations of cauliflower mosaic virus DNA and RNA forms during turnip callus proliferation
More LessCauliflower mosaic virus (CaMV) nucleic acids accumulate in the cell in different structural conformations related to their roles in gene expression, replication and virion assembly. We have characterized changes in the population CaMV DNA and RNA replication products which occur following culture of infected turnip leaves under conditions where callus proliferates. After only 5 days in culture, a significant increase in the level of genome-length and subgenomic supercoiled (SC) DNA forms was observed by two-dimensional (2D) gel electrophoresis. Open circular (OC) molecules, corresponding to these SC DNAs, with mobilities consistent with the presence of a single break in each strand, were also detected after 5 days culture. By 10 days culture, the proportion of OC molecules with only one break per double-stranded molecule had increased. After 34 days culture, SC DNA with a range of sizes predominated in the unencapsidated DNA fraction. The change in pattern of OC and SC DNA forms during callus proliferation suggests a possible precursor/product relationship involving generation of deleted molecules from gap-containing virion DNA-like molecules followed by sequential repair of the gaps to produce SC DNA. Moreover, heterogeneity in the mobility of OC DNAs in the neutral dimension of 2D electrophoresis, a feature exhibited by twisted CaMV virion DNA, changed during the time-course suggesting that untwisting occurs during gap repair. Although the relative abundance of SC DNA increased during callus proliferation, CaMV polyadenylated 35S and 19S transcripts declined together with immediate reverse transcription products. We suggest that cellular changes during callus growth lead to a decline in authentic CaMV transcripts in the cytoplasm resulting in cessation of synthesis of viral products and progeny DNA genomes. In consequence, pre-existing virion DNAs return to the nucleus, possibly as a result of a relaxation in a cytoplasmic control mechanism, where they are assembled into various forms of SC DNA. The presence of CaMV SC DNAs in replicating cells might also enhance illegitimate integration into host chromosomes, as hybridization of CaMV DNA to high M r DNA was observed.
-
-
-
Defective cell-to-cell movement of cowpea mosaic virus mutant N123 is efficiently complemented by sunn-hemp mosaic virus
More LessDuring an infection with cowpea mosaic virus (CPMV) both virion assembly and formation of tubules associated with plasmodesmata are required for cell-to-cell movement. These functions are encoded by the M-RNA of CPMV. To study the mechanism of CPMV movement, mutant N123 was used in complementation studies with sunn-hemp mosaic virus (SHMV), a legume-infecting tobamovirus. Previous studies have shown that N123 fails to spread in cowpea plants because of mutation(s) in its M-RNA. However, the mutant was efficiently replicated in cowpea protoplasts, in which virions were formed and tubular transport structures were induced. After high-dose inoculation of cowpeas with N123, only a few infected protoplasts could be isolated, indicating that cell-to-cell transport of N123 was greatly impaired, if not completely abolished. Upon coinoculation with SHMV, mutant N123 infected cowpea plants systemically and accumulated to levels which were comparable to those of wild-type CPMV. In contrast, separate B-RNA of CPMV and a CPMV deletion mutant lacking the tubule-inducing function, were complemented by SHMV to only low levels. It is concluded that SHMV-facilitated spread of CPMV in the non-virion tobamovirus mode is inefficient and that spread of mutant N123 is probably in the CPMV mode, SHMV providing an as yet unidentified helper function.
-
-
-
Failure of long-distance movement of southern bean mosaic virus in a resistant host is correlated with lack of normal virion formation
More LessSunn-hemp mosaic tobamovirus (SHMV) facilitated the spread of the cowpea strain of southern bean sobemovirus (SBMV-C) only in inoculated leaves of common bean (Phaseolus vulgaris L. cv. Bountiful), a resistant host for SBMV-C. Tissue prints of bean primary leaves doubly inoculated with SHMV and SBMV-C, developed by Western blotting, showed the presence of the SBMV-C capsid antigen in the mesophyll and epidermis, but no antigen was detected in the conducting bundles. Typical SBMV-C virions were not seen in electron micrographs of immunogold-labelled mesophyll cells; instead, specifically labelled, amorphous protein clumps were found in the vacuole. Particles of smaller diameter than that of typical SBMV-C virions were specifically trapped by SBMV antibodies following immunosorbent electron microscopy of extracts from doubly infected leaves. SBMV-C coat protein from infected Vigna unguiculata L. (cowpea) and bean plants showed no difference in its mobility following electrophoresis in denaturing SDS–polyacrylamide gels. Lack of efficient assembly of SBMV-C virions does not impede cell-to-cell movement of the virus in doubly infected leaves of bean, yet it is probably an important factor in determining the inability of SBMV-C to move into and/or through the vascular system of this host.
-
-
-
Electrotransfection of turnip yellow mosaic virus RNA into Brassica leaf protoplasts and detection of viral RNA products with a non-radioactive probe
More LessWe describe here a convenient and efficient system for studying turnip yellow mosaic virus (TYMV) replication in leaf protoplasts. Inoculation of rapeseed (Brassica napus ) or Chinese cabbage (B. sinensis ) protoplasts was achieved via electroporation, and sensitive detection of viral RNA products was performed by Northern blot analyses using a non-radioactive digoxigenin-labelled cDNA probe. Virus replication was detected when 1.5×106 rapeseed protoplasts were inoculated with 20 ng of TYMV RNA. Electrotransfection of TYMV RNA was more efficient in rapeseed than in Chinese cabbage protoplasts, and gave somewhat higher signals than those of TYMV virions. TYMV RNA appeared to replicate equally well whether the protoplasts were incubated in the dark or under constant light.
-
-
-
Genome organization of grapevine fanleaf nepovirus RNA2 deduced from the 122K polyprotein P2 in vitro cleavage products
More LessThe full-length transcript of grapevine fanleaf virus (GFLV) RNA2 produces a primary product of 122K when translated in the rabbit reticulocyte system. This 122K polyprotein is completely processed in vitro by the RNA1-encoded 24K proteinase. The positions of the cleavage sites within the polyprotein have been mapped and the genome organization of GFLV-F13 RNA2 has been established. The order of mature proteins in the 122K polyprotein is the amino-terminal 28K protein, the 38K protein followed by the 56K coat protein at the carboxy terminus. These proteins represent the final cleavage products of the 122K polyprotein. A 66K protein which yields 28K and 38K proteins constitutes the major maturation intermediate. Microsequencing of the amino extremity of radioactively labelled 38K protein allowed identification of the Cys257/Ala258 site as the cleavage site recognized by the GFLV proteinase between the 28K and the 38K proteins in the 66K protein in addition to the Arg605/Gly606 site between the 38K protein and the coat protein.
-
-
-
Complete nucleotide sequence of the genome of an apple isolate of apple chlorotic leaf spot virus
More LessThe complete nucleotide sequence of the genome of an apple isolate of apple chlorotic leaf spot virus (ACLSV-A) was determined. The genome is 7552 nucleotides excluding the poly(A) tail and contains three open reading frames (ORFs 1, 2 and 3), encoding proteins with M r values of 216503 (216∙5K), 50453 (50∙4K) and 21394 (21∙4K), respectively. Nucleotide sequence comparisons between ACLSV-A and the previously sequenced ACLSV from plum (ACLSV-P) showed that the sequence identity at the nucleotide level was 79∙8%. Amino acid sequence identities of ORFs 1 and 2 between both isolates were 88∙4% and 79∙9%, respectively. The 21.4K protein encoded by ORF 3 of ACLSV-A had an amino acid sequence identity of 88∙6% with the 28∙3K protein encoded by ORF 3 of ACLSV-P. Immunoblot analysis of the 21∙4K protein expressed in Escherichia coli showed that this protein is the coat protein of ACLSV-A.
-
-
-
The genomic sequence of cardamine chlorotic fleck carmovirus
More LessThe complete genomic sequence of cardamine chlorotic fleck carmovirus (CCFV) has been determined. The genome is a positive-sense ssRNA molecule 4041 nucleotides in length, and has 47 to 64% sequence identity with turnip crinkle, carnation mottle and melon necrotic spot carmoviruses. CCFV and these other carmoviruses have four similar open reading frames (ORFs), and CCFV has large regions of amino acid identity in all of these ORFs with a European isolate of turnip crinkle virus. CCFV, which replicates well in Arabidopsis thaliana, has only been found so far in Australia in the wild perennial brassica Cardamine lilacina.
-
-
-
Nucleotide sequence of tobamovirus Ob which can spread systemically in N gene tobacco
More LessThe genomic RNA sequence of tobamovirus Ob (Ob), which can spread systemically in tobacco carrying the N gene, was determined. It consists of 6507 nucleotides and contains four open reading frames, exactly corresponding to the genomic organization of tobamoviruses known so far, i.e. encoding the 130K, 180K, 30K and coat proteins. There were no nucleotide overlaps between any open reading frames. The Ob nucleic acid sequence, predicted protein sequences and gene organization were compared with those of other tobamoviruses reported previously. This virus was originally reported as a tomato mosaic virus; however, the nucleotide sequence data given here refute this classification. The determinants that allow tobamovirus Ob to overcome the N gene, a feature peculiar to this virus, were not identified apart from sequence data. This virus should be regarded as a new tobamovirus. The determinants interacting with the tentative N gene product have not yet been analysed.
-
-
-
Association of the non-structural P3 viral protein with cylindrical inclusions in potyvirus-infected cells
More LessAntibodies raised against the 42K non-structural P3 protein encoded by the RNA genome of a potyvirus (tobacco vein mottling virus, TVMV) have been used with immunogold labelling procedures to determine the subcellular location of P3 in infected Nicotiana tabacum cells. P3-specific gold label was found almost exclusively associated with the cylindrical inclusions typically formed in the cytoplasm of potyvirus-infected cells by another non-structural viral protein, the cylindrical inclusion protein. The P3 antibodies reacted with inclusions in both the transverse (pinwheel-like) and longitudinal (bundle-like) orientations of these structures. Immunocytological examination of TVMV-infected protoplasts showed the association of P3 with cylindrical inclusions during the early stages of formation of these structures and suggests that the P3 may be involved in the replication of potyviral RNA.
-
- Corrigenda
-
-
-
A model for reverse transcription by a dimeric enzyme
Peter R. Cook. Journal of General Virology (1993), 74, 691–697.
Errors occurred during the preparation of Fig. 1. The corrected versions of parts (i) and (j) are given below.
-
-
-
-
Prophylactic and therapeutic vaccination against a mucosal papillomavirus
M. Saveria Campo, G. Joan Grindlay, Brian W. O’Neil, Lata M. Chandrachud, Gail M. McGarvie and William F. H. Jarrett. Journal of General Virology (1993), 74, 945–953.
Reference to L2b in the footnotes to Table 1 should be deleted. The first sentence of the first footnote should read ‘The animals were inoculated twice with 1 mg of protein, except in group 2, expt 5 where they received two inoculations of 100 µg each.’
-
Volumes and issues
-
Volume 105 (2024)
-
Volume 104 (2023)
-
Volume 103 (2022)
-
Volume 102 (2021)
-
Volume 101 (2020)
-
Volume 100 (2019)
-
Volume 99 (2018)
-
Volume 98 (2017)
-
Volume 97 (2016)
-
Volume 96 (2015)
-
Volume 95 (2014)
-
Volume 94 (2013)
-
Volume 93 (2012)
-
Volume 92 (2011)
-
Volume 91 (2010)
-
Volume 90 (2009)
-
Volume 89 (2008)
-
Volume 88 (2007)
-
Volume 87 (2006)
-
Volume 86 (2005)
-
Volume 85 (2004)
-
Volume 84 (2003)
-
Volume 83 (2002)
-
Volume 82 (2001)
-
Volume 81 (2000)
-
Volume 80 (1999)
-
Volume 79 (1998)
-
Volume 78 (1997)
-
Volume 77 (1996)
-
Volume 76 (1995)
-
Volume 75 (1994)
-
Volume 74 (1993)
-
Volume 73 (1992)
-
Volume 72 (1991)
-
Volume 71 (1990)
-
Volume 70 (1989)
-
Volume 69 (1988)
-
Volume 68 (1987)
-
Volume 67 (1986)
-
Volume 66 (1985)
-
Volume 65 (1984)
-
Volume 64 (1983)
-
Volume 63 (1982)
-
Volume 62 (1982)
-
Volume 61 (1982)
-
Volume 60 (1982)
-
Volume 59 (1982)
-
Volume 58 (1982)
-
Volume 57 (1981)
-
Volume 56 (1981)
-
Volume 55 (1981)
-
Volume 54 (1981)
-
Volume 53 (1981)
-
Volume 52 (1981)
-
Volume 51 (1980)
-
Volume 50 (1980)
-
Volume 49 (1980)
-
Volume 48 (1980)
-
Volume 47 (1980)
-
Volume 46 (1980)
-
Volume 45 (1979)
-
Volume 44 (1979)
-
Volume 43 (1979)
-
Volume 42 (1979)
-
Volume 41 (1978)
-
Volume 40 (1978)
-
Volume 39 (1978)
-
Volume 38 (1978)
-
Volume 37 (1977)
-
Volume 36 (1977)
-
Volume 35 (1977)
-
Volume 34 (1977)
-
Volume 33 (1976)
-
Volume 32 (1976)
-
Volume 31 (1976)
-
Volume 30 (1976)
-
Volume 29 (1975)
-
Volume 28 (1975)
-
Volume 27 (1975)
-
Volume 26 (1975)
-
Volume 25 (1974)
-
Volume 24 (1974)
-
Volume 23 (1974)
-
Volume 22 (1974)
-
Volume 21 (1973)
-
Volume 20 (1973)
-
Volume 19 (1973)
-
Volume 18 (1973)
-
Volume 17 (1972)
-
Volume 16 (1972)
-
Volume 15 (1972)
-
Volume 14 (1972)
-
Volume 13 (1971)
-
Volume 12 (1971)
-
Volume 11 (1971)
-
Volume 10 (1971)
-
Volume 9 (1970)
-
Volume 8 (1970)
-
Volume 7 (1970)
-
Volume 6 (1970)
-
Volume 5 (1969)
-
Volume 4 (1969)
-
Volume 3 (1968)
-
Volume 2 (1968)
-
Volume 1 (1967)