RESUMEN
Viruses can manipulate the immune response against them by various strategies to influence immune cells, i.e. by over-activation leading to functional inactivation, bypassing antigen presentation or even suppression of effector functions. Little is known, however, about how these features of immune regulation and modulation could be used for therapeutic purposes. Reasons for this include the complexity of immune regulatory mechanisms under certain disease conditions and the risks that infections with viruses pose to human beings. The orf virus (ORFV), a member of the Parapoxvirus genus of the poxvirus family, is known as a common pathogen in sheep and goats worldwide. The inactivated ORFV, however, has been used as a preventative as well as therapeutic immunomodulator in veterinary medicine in different species. Here, we review the key results obtained in pre-clinical studies or clinical studies in veterinary medicine to characterise the therapeutic potential of inactivated ORFV. Inactivated ORFV has strong effects on cytokine secretion in mice and human immune cells, leading to an auto-regulated loop of initial up-regulation of inflammatory and Th1-related cytokines, followed by Th2-related cytokines that attenuate immunopathology. The therapeutic potential of inactivated ORFV has been recognised in several difficult-to-treat disease areas, such as chronic viral diseases, liver fibrosis or various forms of cancer. Further research will be required in order to evaluate the full beneficial potential of inactivated ORFV for therapeutic immunomodulation.
Asunto(s)
Factores Inmunológicos/administración & dosificación , Inmunomodulación , Inmunoterapia/métodos , Virus del Orf/inmunología , Medicina Veterinaria/métodos , Animales , Ensayos Clínicos como Asunto , Evaluación Preclínica de Medicamentos , Cabras , Ratones , OvinosRESUMEN
The tapeworm Echinococcus granulosus is the causative agent of hydatid disease and affects sheep, cattle, dogs and humans worldwide. It has a two-stage life cycle existing as worms in the gut of infected dogs (definitive host) and as cysts in herbivores and humans (intermediate host). The disease is debilitating and can be life threatening where the cysts interfere with organ function. Interruption of the hydatid life cycle in the intermediate host by vaccination may be a way to control the disease, and a protective oncosphere antigen EG95 has been shown to protect animals against challenge with E. granulosus eggs. We explored the use of recombinant vaccinia virus as a delivery vehicle for EG95. Mice and sheep were immunized with the recombinant vector, and the result monitored at the circulating antibody level. In addition, sera from immunized mice were assayed for the ability to kill E. granulosus oncospheres in vitro. Mice immunized once intranasally developed effective oncosphere-killing antibody by day 42 post-infection. Antibody responses and oncosphere killing were correlated and were significantly enhanced by boosting mice with either EG95 protein or recombinant vector. Sheep antibody responses to the recombinant vector or to EG95 protein mirrored those in mice.
Asunto(s)
Antígenos Helmínticos/inmunología , Portadores de Fármacos , Equinococosis/prevención & control , Echinococcus granulosus/inmunología , Vectores Genéticos , Proteínas del Helminto/inmunología , Virus Vaccinia/genética , Administración Intranasal , Animales , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/genética , Modelos Animales de Enfermedad , Equinococosis/inmunología , Proteínas del Helminto/genética , Ratones , Ratones Endogámicos BALB C , Ovinos , Análisis de Supervivencia , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
The purpose of this study was to map the initiation (tsp) and termination points of transcripts arising from an open reading frame (ORF3) found in the inverted terminal repeat of the orf virus genome and also, to identify probable transcriptional control sequences. Early transcripts of approx. 0.76 kb were mapped to ORF3 and found to be transcribed toward the ends of the genome. Using the S1 nuclease and primer-extension methods, the bulk of the tsp were mapped to a position 12-13 nucleotides (nt) downstream from a sequence which resembles A + T-rich vaccinia virus early promoters. The 5' ends were 81-82 nt upstream from the first ATG in ORF3. Most of 3' ends of the transcripts mapped to a region 24-32 nt downstream from a T5NT sequence found near the ORF3 stop codon. A second transcription termination point was found 25 nt downstream from another T5NT sequence located downstream and separated by 85 nt from the first. These results infer that the A + T-rich, early transcriptional control sequences found in other poxvirus genomes have been conserved in the G + C-rich genome of orf virus.
Asunto(s)
Genes Virales , Virus del Orf/genética , Regiones Promotoras Genéticas , Regiones Terminadoras Genéticas , Transcripción Genética , Virus Vaccinia/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Regulación Viral de la Expresión Génica , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Mapeo Restrictivo , Moldes GenéticosRESUMEN
Three parapoxviruses which cause orf or related diseases in humans and animals and the orthopoxvirus, vaccinia virus, were tested for their in vitro sensitivity to cidofovir. The 50% inhibitory concentration for the three parapoxviruses was between 0.21 and 0.27 microg/ml and for vaccinia was 1.32 microg/ml. The selectivity index varied from 198 to 264 for the parapoxviruses and was 42 for vaccinia virus. Virus yield assays confirmed the ability of cidofovir to reduce ortho- and parapoxvirus replication. The efficacy of cidofovir against parapoxviruses justifies its evaluation as a candidate drug for the treatment of parapoxvirus infections in humans and animals.
Asunto(s)
Antivirales/farmacología , Citosina/farmacología , Organofosfonatos , Compuestos Organofosforados/farmacología , Parapoxvirus/efectos de los fármacos , Replicación Viral/efectos de los fármacos , Animales , Línea Celular , Cidofovir , Citosina/análogos & derivados , Parapoxvirus/fisiología , Infecciones por Poxviridae/virología , Ovinos , Vaccinia/virología , Virus Vaccinia/efectos de los fármacosRESUMEN
Hysterectomy-procured, barrier maintained lambs were immunised with either of virus or vaccinia virus and subsequently challenged with both viruses. Under these conditions lambs were protected from challenge with the homologous virus but no cross-protection was observed. The feeding of colostrum that contained antibodies to orf virus had no effect on the duration of viral lesions. Immunoblotting analysis and ELISA of serum samples taken during the course of the experiment indicated that the animals mounted antibody responses to both viruses. The cross recognition of 3 vaccinia virus antigens by the hyperimmune anti-orf virus serum was revealed by immunoblotting.
Asunto(s)
Calostro/inmunología , Virus del Orf/inmunología , Ovinos/inmunología , Vacunación/veterinaria , Virus Vaccinia/inmunología , Animales , Anticuerpos Antivirales/biosíntesis , Western Blotting/veterinaria , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Vida Libre de Gérmenes/inmunología , Embarazo , Ovinos/virologíaRESUMEN
Attitude surveys of registered nurses were conducted in 1984 (just prior to implementation of prospective payment) and in 1989 (after implementation of changes responsive to prospective payment and increased competition) in an academic medical center. Results indicate more negative attitudes toward hospital administration, pay and promotional opportunities in 1989. However, overall job satisfaction, job variety, job market alternatives, participation in decision making, and intention to leave were unchanged while job variety and perceptions of job market alternatives were more positive in 1989. Implications for health-care management and future research are discussed.
Asunto(s)
Actitud del Personal de Salud , Satisfacción en el Trabajo , Personal de Enfermería en Hospital/psicología , Hospitales Públicos , Hospitales de Enseñanza , Estudios Longitudinales , Administración de Personal en Hospitales/normas , Administración de Personal en Hospitales/estadística & datos numéricos , Sistema de Pago Prospectivo/organización & administración , Análisis de Regresión , Encuestas y Cuestionarios , Estados Unidos , Recursos HumanosAsunto(s)
Seguro de Responsabilidad Civil/estadística & datos numéricos , Mala Praxis/estadística & datos numéricos , Pautas de la Práctica en Medicina/estadística & datos numéricos , Recolección de Datos , Estudios de Evaluación como Asunto , Modelos Estadísticos , Análisis Multivariante , Proyectos Piloto , Análisis de Regresión , TennesseeRESUMEN
The GIF protein of orf virus (ORFV) binds and inhibits the ovine cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-2 (IL-2). An equivalent protein has so far not been found in any of the other poxvirus genera and we therefore investigated whether it was conserved in the parapoxviruses. The corresponding genes from both the bovine-specific pseudocowpox virus (PCPV) and bovine papular stomatitis virus (BPSV) were cloned and sequenced. The predicted amino acid sequences of the PCPV and BPSV proteins shared 88 and 37 % identity, respectively, with the ORFV protein. Both retained the six cysteine residues and the WSXWS-like motif that are required for biological activity of the ORFV protein. However, an analysis of the biological activity of the two recombinant proteins revealed that, whilst the PCPV GIF protein bound to both ovine and bovine GM-CSF and IL-2 with very similar binding affinities to the ORFV GIF protein, no GM-CSF- or IL-2-binding activity was found for the BPSV protein.
Asunto(s)
Secuencia Conservada , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Interleucina-2/metabolismo , Parapoxvirus , Proteínas Virales , Secuencia de Aminoácidos , Animales , Bovinos , Clonación Molecular , Variación Genética , Datos de Secuencia Molecular , Virus del Orf/genética , Virus del Orf/metabolismo , Parapoxvirus/clasificación , Parapoxvirus/genética , Parapoxvirus/metabolismo , Virus de la Seudoviruela de las Vacas/genética , Virus de la Seudoviruela de las Vacas/metabolismo , Análisis de Secuencia de ADN , Ovinos , Proteínas Virales/química , Proteínas Virales/genética , Proteínas Virales/metabolismoAsunto(s)
Enfermedades de los Gatos/virología , Enfermedades del Pie/veterinaria , Infecciones por Poxviridae/veterinaria , Virus de la Seudoviruela de las Vacas/aislamiento & purificación , Enfermedades Cutáneas Virales/veterinaria , Animales , Enfermedades de los Gatos/patología , Gatos , Enfermedades del Pie/patología , Enfermedades del Pie/virología , Masculino , Infecciones por Poxviridae/patología , Infecciones por Poxviridae/virología , Enfermedades Cutáneas Virales/patología , Enfermedades Cutáneas Virales/virología , Factores de TiempoRESUMEN
CASE HISTORY: Three cats were presented with single proliferative lesions affecting one foot, which failed to heal after medical treatment, and recurred despite surgical resection. PATHOLOGICAL FINDINGS: Histologically, the lesions were proliferative and papillary. There was marked acanthosis, rete peg formation, and compact orthokeratosis, with large numbers of bacteria in the orthokeratotic scale. Some biopsies had multifocal keratinocyte swelling of the stratum granulosum, and amphophilic intracytoplasmic inclusions were present in some of the swollen cells. The dermis consisted of a light fibrous stroma with marked capillary proliferation. Parapoxviruses were detected in the lesions of all cats by electron microscopic examination. PCR analysis detected orf virus (contagious ecthyma virus) in two cats, and orf virus was cultured from one cat. DIAGNOSIS: Parapoxvirus infection in cats. CLINICAL RELEVANCE: Parapoxvirus infection should be considered as a differential diagnosis when dealing with proliferative, non-healing lesions on the feet of cats, especially cats in rural areas. The recovery of orf virus from a cat with typical poxvirus lesions extends the range of species affected by this virus.
Asunto(s)
Enfermedades de los Gatos/virología , Infecciones por Poxviridae/veterinaria , Enfermedades Cutáneas Virales/veterinaria , Secuencia de Aminoácidos , Animales , Enfermedades de los Gatos/patología , Gatos , Femenino , Masculino , Datos de Secuencia Molecular , Infecciones por Poxviridae/patología , Infecciones por Poxviridae/virología , Enfermedades Cutáneas Virales/patología , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genéticaRESUMEN
Inoculation of lambs with a New Zealand strain of orf virus (NZ 2) failed to protect them against subsequent infection with the Lister strain of vaccinia virus. Similarly, in the reciprocal test, vaccinia virus failed to protect against subsequent orf virus infection. Inoculation with either orf virus or vaccinia virus alone afforded protection against reinfection with the same virus. These results have relevance to the use of vaccinia virus gene vectors in sheep.
Asunto(s)
Virus del Orf/inmunología , Poxviridae/inmunología , Virus Vaccinia/inmunología , Animales , Inmunización/métodos , OvinosRESUMEN
There are three accepted members of the parapoxvirus genus, orf virus (OV), papular stomatitis virus (PSV), and pseudocowpox virus (PCV). OV is maintained in sheep and goats and PSV and PCV in cattle. Restriction endonuclease profiles of the DNA derived from representatives of these established members of the genus were compared with profiles from a parapoxvirus recently isolated from red deer. In no case did the profile of this latter virus (DPV) resemble those generated from the other parapoxviruses. Southern blot hybridization using total DPV DNA as a probe revealed homology between DPV and the central regions of the genomes of the other parapoxviruses but not to their terminal regions. These results indicate that the genome of DPV is as different from the genomes of the three accepted members of the genus as the latter are from each other and argue for the inclusion of DPV as a new member of the parapoxvirus genus.
Asunto(s)
Ciervos/virología , Parapoxvirus/clasificación , Polimorfismo de Longitud del Fragmento de Restricción , Animales , ADN Viral/análisis , Nueva Zelanda , Parapoxvirus/genética , Parapoxvirus/aislamiento & purificaciónRESUMEN
Orf virus is an epitheliotropic DNA parapoxvirus with a worldwide distribution that induces acute pustular lesions in the skin of sheep, goats and man. Genetic mapping and sequencing of the orf virus genome have revealed that orf virus has a typical poxvirus distribution of genes, with those essential for viral DNA synthesis, replication and packaging located in the central region, and those involved in virulence concentrated in the terminal regions. The immune and inflammatory response to orf virus infection in the skin and local lymph is vigorous and typical of an anti-viral response, involving CD4+ helper and CD8+ cytotoxic T cells, interferons and antibodies. In spite of this, the virus can repeatedly infect sheep. Host acquired immunity involving CD4+ T cells and interferons is effective in controlling the extent of viral replication, but does not prevent reinfection. Several virus putative virulence genes have been identified. These are: viral homologues of ovine vascular endothelial growth factor (VEGF); ovine IL-10; vaccinia virus E3L interferon resistance gene; and in addition a viral activity that inhibits the cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). These may be responsible for rescuing orf virus, at least temporarily, from host immunity and aiding viral replication in epidermal cells.
Asunto(s)
Ectima Contagioso/inmunología , Enfermedades de las Cabras/inmunología , Virus del Orf/inmunología , Animales , Anticuerpos Antivirales/biosíntesis , Ectima Contagioso/prevención & control , Ectima Contagioso/virología , Genoma Viral , Enfermedades de las Cabras/prevención & control , Enfermedades de las Cabras/virología , Cabras , Humanos , Inmunidad Celular , Sistema Linfático/inmunología , Virus del Orf/genética , Virus del Orf/patogenicidad , Ovinos , Piel/inmunología , Piel/patología , Piel/virología , Linfocitos T Citotóxicos/inmunología , Vacunación/veterinaria , Proteínas Virales/inmunología , VirulenciaRESUMEN
The ability of different metal ions to promote transformation of Pseudomonas aeruginosa by deoxyribonucleic acid of the plasmid RP1 was examined. CaCl2, MgCl2, and MnCl2 were found to promote such transformation, although at different frequencies and with the optimum response at different concentrations. Only MgCl2 promoted transfection of P. aeruginosa by the linear deoxyribonucleic acid of phage F116. CaCl2 was demonstrated to allow adsorption and entry into the cell of F116 deoxyribonucleic acid such that it became resistant to exogenous deoxyribonuclease, but phage production occurred only when MgCl2 was provided. Inactivation of linear phage deoxyribonucleic acid taken up in the absence of MgCl2 was observed. The transfection frequencies at various concentrations of MgCl2 were compared, and the optimum response occurred at the concentration which promoted the highest frequency of transformation by RP1 deoxyribonucleic acid.
Asunto(s)
Cloruro de Calcio/farmacología , Magnesio/farmacología , Pseudomonas aeruginosa/genética , Transfección/efectos de los fármacos , Transformación Bacteriana/efectos de los fármacos , Bacteriófagos/genética , Manganeso/farmacología , Plásmidos , Pseudomonas aeruginosa/efectos de los fármacosRESUMEN
About 47% identity was observed between the deduced amino acid sequences of a protein encoded by a gene of the parapoxvirus orf virus (OV) strain NZ2 and a 6 kDa protein of unknown function reported to be produced by an open reading frame expressed early after infection by the orthopoxvirus Western Reserve vaccinia virus (VAC); the open reading frame is absent from VAC strain Copenhagen. Examination of sequences reported for variola virus (VAR) strains Bangladesh, India, Congo- 1970, Somalia- 1977 and Garcia- 1966 revealed each encoded a correlate 58 amino acid protein. The open reading frame was not reported in the original analyses of these sequences because a lower limit of 60 amino acids was used to identify potential encoded proteins. Inspection of partial reading frames reported for cowpox virus (CWV) and ectromelia virus (EMV) suggested that these viruses might also code for a correlate of the VAC WR protein. DNA sequencing of cloned fragments of CWV and EMV confirmed that both these orthopoxviruses encode closely related, full length variants of the VAC and VAR open reading frames. The OV homologue is coded in the OV strain NZ2 BamHI-E fragment E2L open reading frame, which we reported is transcribed early postinfection; moreover, analysis of an NZ2 variant showed E2L was absent, indicating that E2L, like the VAC cognate, is nonessential for virus replication in cell culture. The parapoxvirus and orthopoxvirus correlates have about 20% amino acid sequence resemblance to African swine fever virus DNA binding protein p10, suggesting an ancestral relation of genes.
Asunto(s)
Genes Virales , Virus del Orf/genética , Virus de la Viruela/genética , Secuencia de Aminoácidos , Secuencia de Bases , ADN Viral , Datos de Secuencia Molecular , Orthopoxvirus/genética , Homología de Secuencia de Aminoácido , Virus Vaccinia/genéticaRESUMEN
A homolog of the vaccinia virus (VAC) interferon resistance gene E3L has been discovered in orf virus strain NZ-2, a parapoxvirus that infects sheep, goats and humans. The gene is located 20 kb from the left terminus of the orf virus genome and is transcribed towards this terminus. RNase protection studies have been used to define the limits of the gene and Northern analysis revealed that it is expressed early in infection. The predicted amino acid sequence of the orf virus protein shares 31% identity (57% similarity) with the VAC E3L protein. Four of the six residues identified as being essential to dsRNA binding in the vaccinia virus protein are conserved in the orf virus protein whilst the other two amino acid changes are conservative substitutions. The orf virus gene has been sequenced in two other orf virus strains which vary markedly in their ability to produce experimental lesions in vivo. Their predicted protein sequences vary by less than 3% from the NZ-2 protein. The recombinant orf virus protein, expressed as a fusion protein in E. coli, bound double-stranded (ds)RNA but not dsDNA, single-stranded (ss)DNA or ssRNA . This is the first demonstration of a VAC E3L-like gene encoded by a parapoxvirus.
Asunto(s)
Antivirales/farmacología , Interferón-alfa/farmacología , Virus del Orf/genética , Proteínas de Unión al ARN/genética , Proteínas Virales/genética , Secuencia de Aminoácidos , Secuencia de Bases , Farmacorresistencia Microbiana , Datos de Secuencia Molecular , Virus del Orf/metabolismo , ARN Bicatenario/metabolismo , Proteínas de Unión al ARN/química , Proteínas de Unión al ARN/metabolismo , Proteínas Recombinantes , Análisis de Secuencia de ADN , Transcripción Genética , Proteínas Virales/química , Proteínas Virales/metabolismoRESUMEN
Two BamHI fragments from the right-hand terminal region of the orf virus genome have been sequenced. The bulk of the inverted terminal repetition (ITR) sequence is contained within these fragments and makes up 3388 bp of the 4425-bp sequence reported. The overall base composition of the larger sequence is 59.4% G + C and of the ITR, 60.2% G + C. An extremely G/C-rich (83.2%) block of sequence was found spanning the ITR/unique sequence junction. The bulk of the ITR could be divided into three blocks of directly repeated sequences. One block begins about 250 nucleotides from the terminus and is a direct repeat 15 bp long, repeated 14 times. The other blocks contain seven sequence sets ranging from 16 to 36 nucleotides which are repeated 2 to 4 times, interspersed with one another, interrelated in sequence, and sometimes separated by unique sequence. Eight open reading frames (ORFs), each with the potential to code for polypeptides of 50 residues or more, were identified. Three were found within the ITR, four spanned the ITR/unique sequence junction and one was found outside the ITR. A search for putative poxvirus transcriptional control signals indicated that three of the eight ORFs are likely to be transcribed early, all in the same direction toward the right end of the genome. Sequences of the type T(A)3-5T were found only twice in the sequence and only one preceded an ORF.
Asunto(s)
ADN Viral/genética , Virus del Orf/genética , Poxviridae/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Southern Blotting , Clonación Molecular , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Regiones Promotoras Genéticas , Biosíntesis de Proteínas , Secuencias Repetitivas de Ácidos Nucleicos , Mapeo RestrictivoRESUMEN
A 3.3-kb BamHI fragment from the center of the orf virus (OV) NZ2 genome has been sequenced, revealing three major open reading frames (ORFs) with homology to vaccinia virus (VAC) genes. These ORFs have been designated F2L, F3R, and F4R and the proteins they encode were found to be homologous to VAC genes H4L (RNA polymerase-associated protein RAP94), H5R (35-kDa virion envelope antigen) and H6R (topoisomerase), respectively. The OV ORFs are arranged on the genome in an almost identical manner to their VAC counterparts revealing the common evolutionary origin of the two viruses despite the extreme difference in their G+C content. Like its VAC counterpart, F3R was shown to be transcribed early and late during infection. S1 and primer extension analysis located the 5' ends of F3R early transcripts to a position 15-16 nt and 5-10 nt, respectively, downstream from an AT-rich sequence resembling a VAC early promoter. The 5' ends of F3R late transcripts were located to an A within the sequence 5'-TAAAG, 41 nt downstream from the early promoter and 17 nt upstream from the initiation codon. S1 analysis of F2L, which is transcribed only late in infection, revealed transcripts initiating from within the sequence 5'-TAAATG. No transcriptional start point could be detected for F4R but the VAC late transcriptional initiation sequence TAAAT was found close to the predicted translational start point. Another late promoter-like sequence, 5'-TAAATG, was found at the 3' end of F2L. This preceded a short ORF tentatively designated as F1L and predicted to be the beginning of a homologue of VAC H3L.
Asunto(s)
Genes Virales , Virus del Orf/genética , Virus Vaccinia/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Línea Celular , Secuencia Conservada , ADN Viral , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Mapeo Restrictivo , Homología de Secuencia de Aminoácido , Transcripción Genética , Proteínas Virales/genéticaRESUMEN
The nucleotide sequence of a near-terminal region of orf virus DNA was determined. Examination of the sequence revealed an open reading frame encoding a peptide with significant amino acid homology to the pseudoprotease domains recently identified in a number of retroviruses including mouse mammary tumor virus, simian Mason-Pfizer virus, maedi-visna virus, and equine infectious anaemia virus. The orf virus pseudoprotease shares up to 28% amino acid homology with retroviral pseudoproteases and appears to be a discrete transcriptional unit rather than a subunit of a larger polypeptide as is the case in retroviruses. The sharing of amino acid composition across such wide taxonomic boundaries suggests that this polypeptide has a functional significance in both retroviruses and poxviruses.
Asunto(s)
ADN Viral/genética , Virus del Orf/genética , Péptido Hidrolasas/genética , Poxviridae/genética , Retroviridae/enzimología , Animales , Secuencia de Bases , Clonación Molecular , Datos de Secuencia Molecular , Virus del Orf/enzimología , Retroviridae/genética , Homología de Secuencia de Ácido NucleicoRESUMEN
DNA sequence analysis of a 1.55-kb region located 10 kb from the left end of the orf virus NZ-2 strain (OV NZ2) genome revealed an open reading frame, B2L, encoding a protein with a predicted molecular weight of 41.67 kDa. This protein (p42K) shows 42% amino acid sequence identity to the vaccinia virus (VAC) major envelope antigen p37K. In addition, p42K shows homology to a protein encoded by molluscum contagiosum virus (42.8% identity) and another encoded by fowlpox virus (38.3% identity). These proteins are themselves homologues of the VAC p37K. B2L is actively transcribed after the onset of DNA replication and S1 nuclease analysis mapped the 5' end of the transcript to within the sequence TAAATG. A VAC recombinant capable of expressing the p42K gene was constructed and used as an antigen in radioimmune precipitations and lymphocyte transformation assays. These assays demonstrated that OV p42K is one of a limited number of OV proteins to which sheep mount a strong antibody response and which stimulate lymphocytes derived from draining lymph nodes following a natural infection with OV NZ2.