Studies on vaccination against papillomaviruses: a comparison of purified virus, tumour extract and transformed cells in prophylactic vaccination

Calves were vaccinated with two preparations made from one cutaneous fibropapilloma induced by bovine papillomavirus type 2 (BPV-2). One vaccine consisted of homogenised tumour; the other contained purified virus only. Both produced resistance to a heavy challenge infection of BPV-2. One calf in the vaccinated group developed a small tumour and rejected it earlier than the control calves. It would appear likely that the prophylactic immune response was induced by viral structural proteins only and that tumour-specific antigens are unnecessary. Bovine fibroblasts were transformed in vitro by BPV-2 and administered as a vaccine; immunity was not induced.     

Effects of DNA dose,route of vaccination,and coadministration of porcine interleukin-6 DNA on results of DNA vaccination against influenza virus infection in pigs

OBJECTIVE: To examine the effects of DNA dose, site of vaccination, and coadministration of a cytokine DNA adjuvant on efficacy of H1-subtype swine influenza virus hemagglutinin (HA) DNA vaccination of pigs. ANIMALS: 24 eight-week-old mixed-breed pigs. PROCEDURE: 2 doses of DNA were administered 27 days apart by use of a particle-mediated delivery system (gene gun). Different doses of HA DNA and different sites of DNA administration (skin, tongue) were studied, as was coadministration of porcine interleukin-6 (pIL-6) DNA as an adjuvant. Concentrations of virus-specific serum and nasal mucosal antibodies were measured throughout the experiment, and protective immunity was assessed after intranasal challenge with homologous H1N1 swine influenza virus. RESULTS: Increasing the dose of HA DNA, but not coadministration of pIL6 DNA, significantly enhanced virus-specific serum antibody responses. Pigs that received DNA on the ventral surface of the tongue stopped shedding virus 1 day sooner than pigs vaccinated in the skin of the ventral portion of the abdomen, but none of the vaccinated pigs developed detectable virus-specific antibodies in nasal secretions prior to challenge, nor were they protected from challenge exposure. Vaccinated pigs developed high virus-specific antibody concentrations after exposure to the challenge virus. CONCLUSIONS AND CLINICAL RELEVANCE: Co-administration of pIL-6 DNA did not significantly enhance immune responses to HA DNA vaccination or protection from challenge exposure. However, HA DNA vaccination of pigs, with or without coadministration of pIL-6 DNA, induced strong priming of the humoral immune system.     

Clinical signs, histology, and CD3-positive cells before and after treatment of dogs with chronic enteropathies

Background: Histopathology is widely used for the diagnosis of inflammatory bowel disease in dogs. Variations in lesions and unavailability of uniform grading systems limit the usefulness of histologic examination.
Hypothesis: CD3 cell numbers in chronic enteropathies of dogs correlate with clinical activity of the disease and with severity of histopathologic changes.
Animals: Nineteen client-owned dogs examined because of chronic diarrhea, vomiting, or both.
Methods: Samples of duodenal and colonic mucosa were collected endoscopically before and after treatment. Dogs that responded to a hypoallergenic diet were grouped as food-responsive diarrhea dogs (FRD, n = 10). Dogs with no clinical improvement after 10 days of treatment then received prednisolone (immunosuppressive doses) and were grouped as steroid-responsive diarrhea dogs (SRD, n = 9). Histopathologic assessment with a standardized grading system was performed retrospectively on the intestinal samples. Histologic score, total number of infiltrating cells, and CD3-positive cells were counted and compared with the clinical scoring.
Results: No statistically significant difference was detected among histologic grading, total number of cells in the lamina propria, and T-cell numbers in biopsies before and after treatment in either group (FRD and SRD).
Conclusions and Clinical Importance: Currently used histopathologic grading scores, total numbers of cells, and numbers of CD3-positive cells did not allow differentiation between FRD and SRD and did not correlate with clinical response to therapy. Based on these results, new grading scores assessing other criteria than total cell numbers and CD3-positive cells should be evaluated in the future.     

Mucosal vaccination against influenza: protection of pigs immunized with inactivated virus and ether-split vaccine

Effective vaccinations against swine influenza reduce the economic loss of pig industries, and also may minimize the possibility of emergence of new pandemic viruses, since pigs are intermediate hosts to generate reassortant viruses among avian and mammalian influenza viruses. In this study, we showed that intranasal immunization of pigs with formalin-inactivated or ether-split influenza vaccine (A/Aichi/2/68) induced virus-specific IgG, IgM, and IgA antibodies in their nasal secretions and sera, resulting in complete protection from virus challenge. Antibody response to the challenge virus was not observed in the immunized pigs, suggesting that the replication of the virus in the primary targets, respiratory epithelial cells, was inhibited. The present results indicate that intranasal immunization of pigs with inactivated vaccines is effective to control swine influenza, and also provide a good model, as well as a mouse model, to evaluate an intranasal application of influenza vaccine for humans.     

Malignant catarrhal fever virus in rabbits—Reproduction of clinical disease by cell-free virus and partial protection against such disease by vaccination with inactivated virus

The C500 strain of the herpesvirus of African malignant catarrhal fever (MCFV-C500) was able to kill rabbits. Some rabbits ($\text{4}\text{6}$) were protected against 320 intravenous LD50 of cell-free virus by immunisation with inactivated virus.     

Intranasal vaccination of pigs against pseudorabies: absence of vaccinal virus latency and failure to prevent latency of virulent virus

The avirulent Bartha's K strain of pseudorabies virus (PRV) was used to vaccinate 8 pigs at 10 weeks of age by the intransal route (experiment 1). On postvaccination days (PVD) 63 and 91, pigs were treated with corticosteroids. Viral shedding could not be detected. Explant cultures of trigeminal ganglia and tonsils did not produce virus. Four pigs with maternal antibody were vaccinated intranasally with Bartha's (attenuated) K strain of PRV at 10 weeks of age and were challenge exposed with a virulent strain of PRV on PVD 63 (experiment 2). Corticosteroid treatment, starting on postchallenge exposure day 70 (PVD 133) resulted in viral shedding in 1 of 4 pigs. In another pig of these 4, a 2nd corticosteroid treatment was required to trigger reactivation. In both pigs, sufficient reactivated virus was excreted to infect susceptible sentinel pigs. Restriction endonuclease analysis indicated that viruses isolated from the 2 pigs after challenge exposure and corticosteroid treatment were indistinguishable from the virulent virus. Evidence was not obtained for simultaneous excretion of vaccinal and virulent virus. Of 4 pigs without maternal antibody vaccinated twice with 1 of 2 inactivated PRV vaccines, challenge exposed on PVD 84, and treated with corticosteroids on postchallenge exposure day 63 (PVD 147), 1 was latently infected, as evidenced by the shedding of PRV (experiment 3). However, its sentinel pig remained noninfected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Enhanced protective response and immuno-adjuvant effects of porcine GM-CSF on DNA vaccination of pigs against Aujeszky’s disease virus

This study was conducted to investigate whether the co-delivery of DNA encoding porcine cytokines would enhance a protective immune response in pigs to a Pseudorabies virus (PRV; or Aujeszky’s disease virus) DNA vaccine. Aujeszky’s disease in pigs results in respiratory and nervous symptoms with important economic losses. To evaluate cytokine effects, eukaryotic expression vectors were constructed for porcine GM-CSF, IL-2 and IFN-γ. cDNA for each of these cytokines was inserted under the control of a CMV promoter in the pcDNA3 plasmid and cytokine expression was confirmed after DNA transfection in various mammalian cell cultures by bioassays (GM-CSF and IL2) and ELISA (IFN-γ). Pigs were vaccinated by single intramuscular injection with plasmid DNA encoding PRV gB and gD along with various combinations of cytokine plasmid constructs. Pig serum was tested for the production of antibody by isotype specific anti-PRV ELISA. Pigs were then challenged with the highly virulent PRV strain NIA3 on day 21 after vaccination. The survival and growth rate of pigs were monitored for seven days after the viral challenge. The co-administration of GM-CSF plasmid increased the immune response induced by gB and gD PRV DNA vaccine. This immune response was characterized by an earlier appearance of anti-PRV IgG2, a significantly enhanced anti-PRV IgG1 and IgG2 antibody response, a significantly decreased and shortened viral excretion in nasal swabs and an improved protection to the viral challenge. In contrast, the co-administration of porcine IL-2 or IFN-γ had no adjuvant effects. Our results thus demonstrate for the first time that the application of porcine GM-CSF gene in a DNA vaccine formulation can exert immuno-adjuvant and protective effects with single vaccination in the natural host pig against Aujeszky’s disease.     

Proteomic alteration of Marc-145 cells and PAMs after infection by porcine reproductive and respiratory syndrome virus

Viral infections usually result in alterations in the host cell proteome, which determine the fate of infected cells and the progress of pathogenesis. To uncover cellular protein responses in porcine reproductive and respiratory syndrome virus (PRRSV), infected pulmonary alveolar macrophages (PAMs) and Marc-145 cells were subjected to proteomic analysis involving two-dimensional electrophoresis (2-DE) followed by MALDI-TOF-MS/MS identification. Altered expression of 44 protein spots in infected cells was identified in 2D gels, of which the 29 characterised by MALDI-TOF-MS/MS included 17 up-regulated and 12 down-regulated proteins. Some of these proteins were further confirmed at the mRNA level using real-time RT-PCR. Moreover, Western blot analysis confirmed the up-regulation of HSP27, vimentin and the down-regulation of galectin-1. Our study is the first attempt to analyze the cellular protein profile of PRRSV-infected Marc-145 cells using proteomics to provide valuable information about the effects of PRRSV-induced alterations on Marc-145 cell function. Further study of the affected proteins may facilitate our understanding of the mechanisms of PRRSV infection and pathogenesis.     

Phenotypic and functional modulation of bone marrow-derived dendritic cells by porcine reproductive and respiratory syndrome virus

It is well documented that there is a delay in the development of effective immunity to porcine reproductive and respiratory syndrome virus (PRRSV) in infected and vaccinated pigs. This suggests that PRRSV might possess some inherent properties to evade host defense mechanisms during the early stage of infection. Dendritic cells (DCs) play a crucial role in the activation and control of T-cells in response to viral antigens. In this study, we investigated the phenotypic and functional property changes of bone marrow-derived immature DCs (BM-imDCs) that take place after infection by PRRSV. Results showed that BM-imDCs were permissive to PRRSV infection, as productive replication took place in these cells. A down-regulated expression of MHC I molecules along with an up-regulated expression of CD80/86 is observed at 48 h following infection. Also at 48 h following PRRSV infection, a significant increase of IL-10 secretion by BM-imDCs was noticed. Results suggest that the inhibited expression of MHC I and the enhanced secretion of IL-10 by BM-imDCs after PRRSV infection might be among the strategies used by the virus to evade the host immune defenses.     

Protection conferred by vaccination with Blacksburg and Komarov strains of Newcastle disease virus against Newcastle disease in Bangladesh

Summary An evaluation was undertaken of the efficacy of vaccination of day-old chicks with the Blacksburg (B1) strain of Newcastle disease virus (NDV) followed at various times by vaccination with the Komarov (K) strain. Antibody was detected by the haemagglutination inhibition (HAI) test one week after vaccination with B1 and titres peaked at three weeks and had declined to undetectable levels by nine weeks.After subsequent vaccination with K strain at five, seven or eight weeks of age levels of HAI antibody (titre 80 to 640) were detected after three weeks. Birds vaccinated at seven weeks were tested for antibody and resistance to challenge beyond 19 weeks of age. In this group the HAI titres remained constant (80 to 640) up to 32 weeks of age and then steadily declined to 10 to 20 at 44 weeks of age.A linear relationship between HAI titre and virus neutralising index (VNI) was demonstrated with a range of selected sera. Only birds with an HAI titre of 80 or greater resisted artificial challenge. It is recommended that, following B1 vaccincation at day-old and K vaccination at seven weeks old, revaccination with K strain should be performed at intervals of not more than seven months.

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Proteccion Conferida Mediante Vacunacion Con Las Cepas Blacksburg Komarov Del Virus De Newcastle En Bangladesh

Resumen Se llevó a cabo una evaluación de la eficacia de la vacunación de pollitos de un día, con la cepa Blacksburg (B1) del virus de la enfermedad de Newcastle, seguida de varias aplicaciones vacunales periódicas con la cepa Komarov (K). Se detectaron anticuerpos mediante la prueba de inhibición de la hemaglutinación, una semana después de la vacunación con B1, los títulos alcanzando el pico a las tres semanas y declinaron a niveles no detectables a las nueve semanas.Después de aplicaciones vacunales periódicas con la cepa K, a las cinco, siete u ocho semanas de edad, se detectaron anticuerpos mediante la prueba de inhibición de la hemaglutinación (títulos 80 a 640), después de tres semanas. Las aves vacunadas a las siete semanas, recibieron una descarga y se les hizo prueba de anticuerpos, después de las 19 semanas de edad. En este grupo los títulos de anticuerpos permanecieron constantes (80 to 640) hasta las 32 semanas de edad y después declinaron lentamente a 10 ó 20 a las 44 semanas de edad.Se demostró una relación lineal entre los títulos hallados y el índice viral neutralizante, en un rango de sueros seleccionados. Solamente las aves con títulos de 80 o más altos resistieron la descarga artificial. Se recomienda que, seguidamente a la vacunación con B1 a un día de edad y con K a las siete semanas, las revacunaciones con la cepa K debería hacerse con intervalos de no más de siete meses.

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Protection Conferee Contre La Maladie De Newcastle Au Bangladesh Par La Vaccination A L'Aide Des Souches Blacksburg Et Komarov De Virus De La Maladie De Newcastle

Résumé Une evaluation de l'efficacité de la vaccination de poussins d'un jour à l'aide des souches Blackburg (B1) du virus de la maladie de Newcastle, suivie à différents moments par la vaccination à l'aide de la souche Komarov (K), a été entreprise. Les anticorps furent détéctés par inhibition de l'hemagglutination (HAI), une semaine après la vaccination avec la souche B1 et le titre atteignit un pic à 3 semaines pour décliner jusqu'à des niveaux indétectables à neuf semaines. A la suite de vaccinations consécutives avec la souche K à cinq, sept ou huit semaines, les niveaux d'anticorps HAI furent détectés après trois semaines. Les oiseaux vaccinés à sept semaines furent examinés pour les anticorps et la résistance à une épreuve au delà de 19 semaines d'âge. Dans ce groupe les titres HAI sont restés constants (80 à 640) jusqu'à l'àge 32 semaines, puis ont diminué régulièrement jusqu'à 10 à 20 à l'âge de 44 semaines.Une relation linéaire entre le titre HAI et l'index de neutralisation du virus (VNI) a été démontrée sur une gamme de sérums choisis. Seuls les oiseaux ayant un titre HAI de 80 ou plus ont résisté à l'épreuve artificielle. On recommande que, après une vaccination B1 à un jour, et une vaccination K à l'âge de sept semaines, une revaccination K soit réalisée à des intervalles n'excédant pas sept mois.

     

The effect of vaccination of lambs with live parainfluenza virus type 3 on pneumonia produced by parainfluenza virus type 3 and Pasteurella haemolytica

Groups of 20 lambs were vaccinated with parainfluenza virus type 3 PI₃ by the intranasal (I.N.) or intramuscular route (I.M.).Approximately 6 weeks later, vaccinated and non-vaccinated lambs were challenged sequentially with PL₃, and Pasteurella haemolytica. At slaughter, 10 to 12 days after challenge, 65% of non-vaccinates and 45% of I.M. vaccinates had pneumonic lesions whereas lesions were not observed in any of the I.N. vaccinates.     

Embryo vaccination of specific-pathogen-free chickens with infectious bursal disease virus: tissue distribution of the vaccine virus and protection of hatched chickens against disease

Vaccination of specific-pathogen-free chickens as 18-day embryos with the BVM isolate of infectious bursal disease virus (IBDV) resulted in extensive replication of the vaccine virus in the embryonic tissues. The virus was recovered from lung, thymus, proventriculus, liver, kidney, and spleen of embryos 1 day postvaccination, and recoverable virus persisted for at least 7 days. Replication and spread of the vaccine virus in chickens vaccinated as 18-day embryos was compared with that in chickens vaccinated at hatch. Distribution of the virus in tissues was more extensive, virus levels in tissues were generally higher, and detectable virus persisted longer in chickens vaccinated as 18-day embryos than in those vaccinated at hatch. Effective vaccine response could be initiated with 6.2 median embryo lethal doses, the lowest dose tested. Chickens immunized as embryos developed neutralizing antibody against IBDV and resisted challenge with pathogenic IBDV at 4, 6, 8, and 10 weeks of age.     

Co-infection of porcine dendritic cells with porcine circovirus type 2a (PCV2a) and genotype II porcine reproductive and respiratory syndrome virus (PRRSV) induces CD4(+)CD25(+)FoxP3(+) T cells in vitro

Porcine circovirus associated disease (PCVAD) is currently one of the most economically important diseases in the global swine industry. Porcine circovirus type 2 (PCV2) is the primary causative agent, however co-infection with other swine pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV) is often required to induce the full spectrum of clinical PCVAD. While the specific mechanisms of viral co-infection that lead to clinical disease are not fully understood, immune modulation by the co-infecting viruses likely plays a critical role. We evaluated the ability of dendritic cells (DC) infected with PRRSV, PCV2, or both to induce regulatory T cells (T(regs)) in vitro. DCs infected with PCV2 significantly increased CD4(+)CD25(+)FoxP3(+) T(regs) (p<0.05) and DCs co-infected with PRRSV and PCV2 induced significantly higher numbers of T(regs) than with PCV2 alone (p<0.05). Cytokine analysis indicated that the induction of T(regs) by co-infected DCs may be dependent on TGF-β and not IL-10. Our data support the immunomodulatory role of PCV2/PRRSV co-infection in the pathogenesis of PCVAD, specifically via T(reg)-mediated immunosuppression.     

Interplay of foot-and-mouth disease virus, antibodies and plasmacytoid dendritic cells: virus opsonization under non-neutralizing conditions results in enhanced interferon-alpha responses

ABSTRACT: Foot-and-mouth disease virus (FMDV) is a highly infectious member of the Picornaviridae inducing an acute disease of cloven-hoofed species. Vaccine-induced immune protection correlates with the presence of high levels of neutralizing antibodies but also opsonising antibodies have been proposed as an important mechanism of the immune response contributing to virus clearance by macrophages and leading to the production of type-I interferon (IFN) by plasmacytoid dendritic cells (pDC). The present study demonstrates that the opsonising antibody titres mediating enhanced IFN-α responses in pDC were similar to neutralizing titres, when antigenically related viruses from the same serotype were employed. However, sera cross-reacted also with non-neutralized isolates of multiple serotypes, when tested in this assay. Both uncomplexed virus and immune complexed virus stimulated pDC via Toll-like receptor 7. An additional finding of potential importance for strain-specific differences in virulence and/or immunogenicity was that pDC activation by FMDV strongly differed between viral isolates. Altogether, our results indicate that opsonising antibodies can have a broader reactivity than neutralizing antibodies and may contribute to antiviral responses induced against antigenically distant viruses.