RESUMEN
Over 23 months, zinc toxicosis was diagnosed in 35 baboons aged 5-12 months in one galvanized metal and concrete cage complex with conditions that led to excessive exposure to environmental zinc. Clinical signs included reduced pigmentation of hair, skin, and mucous membranes (whiteness), alopecia, dehydration, emaciation, cachexia, dermatitis, diarrhea and, in six cases, severe gangrenous dermatitis of extremities. The syndrome was characterized by pancytopenia, elevated zinc and low copper serum concentrations, low vitamin D and bone-specific alkaline phosphatase levels, and atypical myelomonocytic proliferation of bone marrow. This syndrome emphasizes the importance of proper husbandry and cage design and indicates the potential of infant baboons as a model to study the effects of excessive zinc on development. This is the first report describing the epidemiologic and clinical presentation of zinc toxicosis in infant baboons in captivity.
Asunto(s)
Exposición a Riesgos Ambientales , Vivienda para Animales , Enfermedades de los Monos/patología , Papio , Vitamina D/análogos & derivados , Zinc/envenenamiento , Alopecia/etiología , Alopecia/veterinaria , Análisis de Varianza , Anemia/etiología , Anemia/veterinaria , Animales , Huesos/diagnóstico por imagen , Cobre/sangre , Cobre/deficiencia , Proteínas de Unión al ADN/sangre , Dermatitis/etiología , Dermatitis/veterinaria , Diarrea/etiología , Diarrea/veterinaria , Citometría de Flujo/veterinaria , Cariotipificación/veterinaria , Luz , Factor de Transcripción PAX5 , Pigmentación/efectos de los fármacos , Radiografía , Radioinmunoensayo/veterinaria , Síndrome , Factores de Transcripción/sangre , Vitamina D/sangre , Zinc/sangreRESUMEN
AdUTPase gene ( du) deleted ovine lentivirus (OvLV(Deltadu)) mutant, derived from Visna/maedi virus (VMV) molecular clone KV1772, was constructed. Subsequently, a copy of the optimized green fluorescent protein ( egfp) coding region was fused into the viral pol open reading frame (ORF) at the deleted du locus to generate viral mutant, OvLV(Deltadu-egfp). OvLV(Deltadu) reverse transcriptase (RT) activity and titer of infectious virus in goat synovial membrane (GSM) cell cultures were not affected compared to that of KV1772 and OvLV-85/34 strain (p < 0.05). By contrast, OvLV(Deltadu-egfp) RT activity and virus titer were lower than for KV1772 and OvLV(Deltadu) (p < 0.05). OvLV-85/34 RT in sheep monocyte-derived macrophages (SMDM) was higher than that of KV1772, OvLV(Deltadu) and OvLV(Deltadu-egfp) (p < 0.05). The ability to prevent dUTP mis-incorporation into newly synthesized DNA was disrupted in OvLV(Deltadu) and OvLV(Deltadu-egfp) (p < 0.05). Immunoprecipitation demonstrated that GFP is expressed by OvLV(Deltadu-egfp) at a low level. OvLV(Deltadu-egfp) retained egfp after 10 passages in cell culture.OvLV(Deltadu-egfp) was re-isolated in GSM cells from peripheral blood mononuclear (PBMN) cells of three of four OvLV(Deltadu-egfp)-inoculated lambs, but by contrast to the in vitro experiments OvLV(Deltadu-egfp) lost the insert. Priming with OvLV(Deltadu-egfp) did not prevent infection with pathogenic OvLV, but cell-associated viremia in a mock-infected contact control lamb was higher than in OvLV(Deltadu-egfp)-primed lambs. OvLV serum antibody titers increased steadily in OvLV(Deltadu-egfp)-inoculated lambs, but in a lamb from which OvLV(Deltadu-egfp) was not reisolated the antibody titer surpassed the negative/positive cut-off value only after challenge with OvLV-85/34. Because OvLV(Deltadu-egfp) is attenuated for pathogenicity in vitro, replicates in vivo and stimulates an antibody response, subsequent experiments need to address the likelihood of using OvLV(Deltadu-egfp) as an attenuated, live-virus vaccine to protect sheep against OvLV-induced disease when challenged with pathogenic OvLV.
Asunto(s)
Proteínas Luminiscentes/metabolismo , Recombinación Genética , Enfermedades de las Ovejas/virología , Replicación Viral , Virus Visna-Maedi/fisiología , Animales , Anticuerpos Antivirales/sangre , Células Cultivadas , Eliminación de Gen , Cabras , Proteínas Fluorescentes Verdes , Proteínas Luminiscentes/genética , Macrófagos/virología , Monocitos/virología , Pirofosfatasas/genética , Pirofosfatasas/metabolismo , Ovinos , Membrana Sinovial/virología , Transgenes , Virus Visna-Maedi/genética , Virus Visna-Maedi/inmunología , Virus Visna-Maedi/patogenicidadRESUMEN
To investigate the adjuvant capacity of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon (IFN-gamma), we cloned these rhesus cytokines into a mammalian expression vector. Two groups of six rhesus macaques (Macaca mulatta) received intradermal immunizations of plasmid DNA coding for SIV Eng and Gag, and influenza virus nucleoprotein (Flu-NP), with or without the co-administration of plasmid DNA coding for these cytokines. Humoral immune responses to antigens of both of these viruses and SIV specific T cell proliferative responses were significantly enhanced by co-immunization with the cytokines. These twelve monkeys, and a group of six naive controls, were challenged by the oral mucosal route with the uncloned and highly pathogenic SIVmac251. All monkeys became infected. The early CD4 decline was reduced in the group co-immunized with cytokine and viral plasmids. Unexpectedly, plasma viremia set points were not different in this co-immunized group and the non-immunized control group. On the other hand, monkeys vaccinated with equivalent amounts of empty vector plasmid (i.e. no cytokine inserts) along with plasmids expressing viral antigens demonstrated a slight but significant decrease in acute viremia compared to non-immunized controls (P<0.02). However, viral loads at set points were not significantly different between both the immunized and the non-immunized control group. Thus, although the cytokine vectors demonstrated detectable enhancement of the immune response to different viral antigens, such enhanced response did not translate into better anti-viral control in our experiment. These results underscore the need for further testing of cytokines as vaccine adjuvants in relevant animal models.
Asunto(s)
Antígenos Virales/genética , Vectores Genéticos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Interferón gamma/genética , Plásmidos , Virus de la Inmunodeficiencia de los Simios/inmunología , Animales , Secuencia de Bases , Recuento de Linfocito CD4 , Cartilla de ADN , Depleción Linfocítica , Macaca mulatta , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Viremia/inmunología , VirulenciaRESUMEN
B7 is the designation of a cell clone derived from the human cell line CEMx174, which was infected with SIVsmH3 clone. B7 cells chronically produce high quantities of non-infectious virus-like particles (VLP) denominated SIVsmB7. Here we report the molecular characterization of the B7 cell line. We found that B7 cells have a single copy of the SIVsmB7 provirus integrated in a noncoding region of chromosome 20 (nt 24,957 of clone RP5-963K23 on human chromosome 20q 13.11-13.2). Similarly to HIV and SIVmac, we show that integration of SIVsm results in a characteristic five base pair sequence repeat of host DNA that flanks the proviral DNA genome. Since the SIVsmB7 genome has a deletion in the IN coding sequence, the generation of this defective proviral genome most likely occurred during a faulty process of reverse transcription. Thus, these studies reveal the molecular clonality of the SIVsmB7 VLP produced by B7 cells. These genetically homogeneous VLP are useful reagents for vaccine development. In addition, these particles have been used by others (Montelaro et al.) to study the maturation of immune system responses to SIV infection.
Asunto(s)
Cromosomas Humanos Par 20/virología , Células Clonales/virología , Linfocitos/virología , Virus de la Inmunodeficiencia de los Simios/genética , Virus de la Inmunodeficiencia de los Simios/fisiología , Integración Viral , Animales , Línea Celular , Humanos , Provirus/genética , Provirus/fisiología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virión/genética , Virión/fisiologíaRESUMEN
Interleukin-18 (IL-18), previously known as interferon-gamma (IFN-gamma)-inducing factor (IGIF), is a proinflammatory cytokine expressed by activated macrophages that acts in synergy with IL-12 as an important amplifying factor for IFN-gamma production and Th1 development. To study the effect of IL-18 on a lentiviral infection, we cloned the IL-18 gene from a rhesus macaque and constructed replication-competent simian immunodeficiency virus (SIV) that expressed either the precursor pro-IL-18 (SIV(IL-18)) or the mature form (SIV(mIL-18)) of IL-18. The predicted amino acid sequence for rhesus IL-18 had 96% homology with the human one, differing in only 8 of 193 residues. SIV(IL-18) and SIV(mIL-18) replicated more slowly than control viruses in the CEM x 174 cell line and resulted in the development of chronically infected cell lines that expressed high levels of infectious SIV. The cell line generated by SIV(IL-18) released large quantities of IL-18 into the supernatant, whereas the one obtained from SIV(mIL-18) showed the accumulation of IL-18 in the cytoplasm. Similarly, SIV(IL-18) and SIV(mIL-18) replicated more slowly than the unmodified viral vector in rhesus peripheral blood mononuclear cells (PMBC), but only SIV(IL-18) expressed biologically active IL-18. These experiments show that the precursor form of IL-18 is necessary for the efficient release of the cytokine and that IL-18 does not promote increased replication of SIV in rhesus PBMC.
Asunto(s)
Interleucina-18/metabolismo , Macaca mulatta , Virus de la Inmunodeficiencia de los Simios/fisiología , Replicación Viral , Secuencia de Aminoácidos , Animales , Línea Celular , Clonación Molecular , Vectores Genéticos/genética , Humanos , Interferón gamma/metabolismo , Interleucina-18/química , Interleucina-18/genética , Leucocitos Mononucleares/virología , Macaca mulatta/genética , Macaca mulatta/virología , Datos de Secuencia Molecular , Alineación de Secuencia , Virus de la Inmunodeficiencia de los Simios/genéticaRESUMEN
We studied the innate and adaptive immune system of rhesus macaques infected with the virulent simian immunodeficiency virus isolate SIVmac251 by evaluating natural killer (NK) cell activity, cytokine levels in plasma, humoral and virological parameters, and changes in the activation markers CD25 (interleukin 2R ¿IL-2R alpha chain), CD69 (early activation marker), and CD154 (CD40 ligand) in lymphoid cells. We found that infection with SIVmac251 induced the sequential production of interferon-alpha/beta (IFN-alpha/beta), IL-18, and IL-12. IFN-gamma, IL-4, and granulocyte-macrophage colony-stimulating factor were undetected in plasma by the assays used. NK cell activity peaked at 1 to 2 weeks postinfection and paralleled changes in viral loads. Maximum expression of CD69 on CD3(-)CD16(+) lymphocytes correlated with NK cytotoxicity during this period. CD25 expression, which is associated with proliferation, was static or slightly down-regulated in CD4(+) T cells from both peripheral blood (PB) and lymph nodes (LN). CD69, which is normally present in LN CD4(+) T cells and absent in peripheral blood leukocyte (PBL) CD4(+) T cells, was down-regulated in LN CD4(+) T cells and up-regulated in PBL CD4(+) T cells immediately after infection. CD8(+) T cells increased CD69 but not CD25 expression, indicating the activation of this cellular subset in PB and LN. Finally, CD154 was transiently up-regulated in PBL CD4(+) T cells but not in LN CD4(+) T cells. Levels of antibodies to SIV Gag and Env did not correlate with the level of activation of CD154, a critical costimulatory molecule for T-cell-dependent immunity. In summary, we present the first documented evidence that the innate immune system of rhesus macaques recognizes SIV infection by sequential production of proinflammatory cytokines and transient activation of NK cytotoxic activity. Additionally, pathogenic SIV induces drastic changes in the level of activation markers on T cells from different anatomic compartments. These changes involve activation in the absence of proliferation, indicating that activation-induced cell death may cause some of the reported increase in lymphocyte turnover during SIV infection.
Asunto(s)
Citocinas/biosíntesis , Células Asesinas Naturales/inmunología , Activación de Linfocitos/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunología , Animales , Anticuerpos Antivirales/inmunología , Biomarcadores , Línea Celular , Productos del Gen env/inmunología , Productos del Gen gag/inmunología , Inmunofenotipificación , Linfocitos/inmunología , Macaca mulatta , Masculino , Linfocitos T/inmunologíaRESUMEN
Simian immunodeficiency virus (SIV) infection of macaques is a model for human immunodeficiency virus (HIV) infection. We have previously reported the construction and characterization of an SIV vector with a deletion in the nef gene (SIV(delta nef)) and expressing gamma interferon (SIV(HyIFN)) (L. Giavedoni and T. Yilma, J. Virol. 70:2247-2251, 1996). We now show that rhesus macaques vaccinated with SIV(HyIFN) have a lower viral load than a group similarly immunized with SIV(delta nef). Viral loads remained low in the SIV(HyIFN)-vaccinated group even though SIV expressing gamma interferon could not be isolated after 6 weeks postimmunization in these animals. All immunized and two naive control macaques became infected when challenged with virulent SIV(mac251), at 25 weeks postvaccination. In contrast to the two naive controls that died by 12 and 18 weeks postchallenge, all vaccinated animals remained healthy for more than 32 weeks. In addition, postchallenge cell-associated virus load was significantly lower in SIV(HyIFN)-immunized animals than in the group vaccinated with SIV(delta nef). These findings indicate that cytokine-expressing viruses can provide a novel approach for development of safe and efficacious live attenuated vaccines for AIDS.
Asunto(s)
Interferón gamma/genética , Vacunas contra el SIDAS , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/genética , Carga Viral , Animales , Expresión Génica , Humanos , Interferón gamma/inmunología , Macaca mulatta , Virus de la Inmunodeficiencia de los Simios/inmunología , VacunaciónRESUMEN
To better define the molecular epidemiology of bluetongue virus (BTV) infection, the genetic characteristics and phylogenetic relationships of the S3 genes of the five U.S. prototype strains of BTV, the commercially available serotype 10 modified live virus vaccine, and 18 field isolates of BTV serotypes 10, 11, 13, and 17 obtained in California during 1980, 1981, 1989, and 1990 were determined. With the exception of the S3 gene of the U.S. prototype strain of BTV serotype 2 (BTV 2), these viruses had an overall sequence homology of between 95 and 100%. Phylogenetic analyses segregated the prototype U.S. BTV 2 strain to a unique branch (100% bootstrap value), whereas the rest of the viruses clustered in two main monophyletic groups that were not correlated with their serotype, year of isolation, or geographical origin. The lack of consistent association between S3 gene sequence and virus serotype likely is a consequence of reassortment of BTV gene segments during natural mixed infections of vertebrate and invertebrate hosts. The prototype strain of BTV 13, which is considered an introduction to the U.S. like BTV 2, presents an S3 gene which is highly homologous to those of some isolates of BTV 10 and especially to that of the vaccine strain. This finding strongly suggests that the U.S. prototype strain of BTV 13 is a natural reassortant. The different topologies of the phylogenetic trees of the L2 and S3 genes of the various viruses indicate that these two genome segments evolve independently. We conclude that the S3 gene segment of populations of BTV in California is formed by different consensus sequences which cocirculate and which cannot be grouped by serotype.
Asunto(s)
Virus de la Lengua Azul/genética , Genes Virales , California , Filogenia , Estados UnidosRESUMEN
We report the construction and characterization of several replication-competent simian immunodeficiency virus (SIV) vectors with a deletion in the viral nef gene (SIV(delta nef)) that express gamma interferon (IFN-gamma). The expression of the cytokine gene was controlled either by the simian virus 40 early promoter or by the SIV 5' long terminal repeat regulatory sequences, utilizing the nef gene splice signals. To enhance the expression of IFN-gamma, the two in-frame nef start codons were mutated without altering the Env amino acid sequence (SIV(HyIFN)). Plasmids containing full-length proviral genomes were used to obtain high-titer stocks of each recombinant virus in cell cultures. Expression of IFN-gamma by SIV(HyIFN) reached levels as high as 10(6) U/ml after 11 days in culture. The IFN-gamma gene was unstable and sustained deletions after serial passage of SIV(delta nef) vectors in CEM-X-174 cells. The degree of instability appears to depend on size and orientation of the insert and the expression of IFN-gamma. Only one virus, SIV(HyIFN), expressed detectable levels of IFN-gamma up to the sixth passage. Prospects for the use of IFN-gamma and other lymphokines to enhance the safety and efficacy of live attenuated vaccines are discussed.
Asunto(s)
Vectores Genéticos , Interferón gamma/genética , Virus de la Inmunodeficiencia de los Simios/genética , Secuencia de Bases , Línea Celular , Cartilla de ADN , Vectores Genéticos/genética , Humanos , Interferón gamma/biosíntesis , Datos de Secuencia Molecular , Virus de la Inmunodeficiencia de los Simios/fisiología , Replicación ViralAsunto(s)
Anestésicos Locales/administración & dosificación , Bupivacaína/administración & dosificación , Náusea/prevención & control , Dolor Postoperatorio/prevención & control , Desprendimiento de Retina/cirugía , Vitrectomía , Anestesia General , Humanos , Inyecciones , Cuidados Intraoperatorios , Persona de Mediana Edad , Órbita , Dimensión del Dolor , Curvatura de la Esclerótica , Resultado del TratamientoRESUMEN
As a safe alternative to inactivated and live-attenuated whole-virus SIV vaccines, we have evaluated the potential of SIVmac239 gp160 expressed by recombinant vaccinia virus (vSIVgp160) and baculovirus (bSIVgp160) to protectively immunize rhesus macaques against intravenous (i.v.) infection with pathogenic SIVmac isolates. Macaques were immunized with live vSIVgp160 and/or bSIVgp160 protein partially purified from insect cells. The challenge viruses, propagated in rhesus peripheral blood mononuclear cells, consisted of the molecular clone SIVmac239 and another genetically similar, uncloned isolate, SIVmac251. Although antibodies that bind gp130 were induced in all animals following immunization with SIVgp160, neutralizing antibodies were undetectable 1 week prior to virus challenge. These results differ from those for macaques vaccinated with inactivated, whole SIV. All animals became infected after i.v. inoculation with 1-10 AID50 of either challenge virus. For animals challenged with SIVmac251, but not those challenged with SIVmac239, the cell-free infectious virus load in plasma of vSIVgp160-primed, bSIVgp160-boosted macaques was significantly lower than in unimmunized controls at 2 weeks postchallenge. Virus virulence, immunization regimen, and challenge with homologous or heterologous virus are factors critical to the outcome of the study. Immunization with surface glycoprotein may not necessarily provide protective immunity against infection but may reduce virus load. The relationship between reduction in virus load by vaccination and delay in onset of disease remains to be determined.
Asunto(s)
Productos del Gen env/administración & dosificación , Productos del Gen env/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Virus de la Inmunodeficiencia de los Simios/inmunología , Vacunas Virales/administración & dosificación , Animales , Células Cultivadas , Células HeLa , Humanos , Inmunización , Macaca mulatta , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/microbiología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Virus Vaccinia/inmunología , Vacunas Virales/inmunologíaRESUMEN
Rinderpest is a highly contagious viral disease of ruminants and has greater than 95% morbidity and mortality. The etiological agent, rinderpest virus (RPV), is a member of the family Paramyxoviridae and the genus Morbillivirus. Immune responses to both the hemagglutinin (H) and the fusion (F) antigens of morbilliviruses play an important role in the prevention of infection, and only attenuated live vaccines have been shown to provide protective immunity against the group. The lack of protection with inactivated vaccines has been attributed to the denaturation of the F glycoprotein of the virus. Our previous study, however, demonstrated complete protection of cattle vaccinated with infectious vaccinia virus recombinants expressing the H (vRVH) or F (vRVF) protein alone, even in the presence of only 4 U of serum-neutralizing (SN) antibody to RPV (T. Yilma, D. Hsu, L. Jones, S. Owens, M. Grubman, C. Mebus, M. Yamanaka, and B. Dale, Science 242:1058-1061, 1988). We have constructed recombinant baculoviruses that express the F (Fb) and H (Hb) glycoproteins of RPV. Furthermore, we have analyzed the immune responses of mice and cattle to these antigens. Cattle vaccinated with Fb or Hb or a mixture of both antigens were not protected from challenge inoculation with RPV, even when the SN titer was greater than in cattle vaccinated with vRVF alone. This lack of protection, in the presence of SN antibody, would indicate that live attenuated and recombinant vaccines induce immune responses necessary for protection (e.g., cell-mediated immunity) that are not generated by subunit or inactivated whole-virus vaccines.
Asunto(s)
Anticuerpos Antivirales/inmunología , Bovinos/inmunología , Glicoproteínas/inmunología , Inmunoterapia Activa , Peste Bovina/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Antivirales/sangre , Formación de Anticuerpos , Baculoviridae/genética , Glicoproteínas/genética , Hemaglutininas Virales/inmunología , Proteínas de la Membrana , Ratones , Pruebas de Neutralización , Proteínas Recombinantes/inmunología , Transfección , Vacunas de Productos Inactivados/inmunología , Vacunas Sintéticas/inmunología , Proteínas Virales de Fusión/genética , Proteínas Virales de Fusión/inmunología , Proteínas Virales/genéticaRESUMEN
Simian immunodeficiency virus (SIV) infection of rhesus macaques is a model for human immunodeficiency virus (HIV) infection in humans. Inactivated and modified live whole-virus vaccines have provided limited protective immunity against SIV in rhesus macaques. Because of safety concerns in the use of inactivated and live whole-virus vaccines, we evaluated the protective immunity of vaccinia virus recombinants expressing the surface glycoprotein (gp130) of SIVmac and subunit preparations of gp130 expressed in mammalian cells (CHO). Three groups of animals were immunized with recombinant SIV gp130. The first group received SIV gp130 purified from genetically engineered CHO cells (cSIVgp130), the second group was vaccinated with recombinant vaccinia virus expressing SIVmac gp130 (vSIVgp130), and the third group was first primed with vSIVgp130 and then given a booster immunization with cSIVgp130. Although anti-gp130 binding antibodies were elicited in all three groups, neutralizing antibodies were transient or undetectable. None of the immunized animals resisted intravenous challenge with a low dose of cell-free virus. However, the group primed with vSIVgp130 and then boosted with cSIVgp130 had the lowest antigen load (p27) compared with the other groups. The results of these studies suggest that immunization of humans with HIV type 1 surface glycoprotein may not provide protective immunity against virus infection.
Asunto(s)
Anticuerpos Antivirales/inmunología , Productos del Gen env , Inmunoterapia Activa , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Vacunas Sintéticas/uso terapéutico , Proteínas del Envoltorio Viral/inmunología , Animales , Formación de Anticuerpos , Células CHO , Cricetinae , Modelos Animales de Enfermedad , Infecciones por VIH , Macaca mulatta , Proteínas Recombinantes/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunología , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Virus Vaccinia/genética , Proteínas del Envoltorio Viral/biosíntesis , Proteínas del Envoltorio Viral/genética , VirulenciaRESUMEN
Peste des petits ruminants (PPR) is a viral disease of goats and sheep characterized by necrotizing and erosive stomatitis, enteritis and pneumonia. The causative agent, PPRV, is a member of the family Paramyxoviridae and the genus Morbillivirus. Other members of the genus include rinderpest (RPV), measles, canine distemper and phocid distemper viruses. PPR has a very high rate of morbidity and mortality, and effective control of this disease is of economic importance in Africa, Asia and the Middle East. Currently, there is no safe and effective vaccine available against the disease. The tissue culture rinderpest vaccine (TCRV) protects small ruminants against severe disease; there are, however, clinical problems associated with vaccination. This laboratory has recently developed several effective vaccinia virus recombinant vaccines for rinderpest. These vaccines are easy to administer, inexpensive to produce and heat-stable. Goats were vaccinated with a vaccinia virus double recombinant expressing the haemagglutinin and fusion genes of RPV. Although vaccinated animals developed antibodies (neutralizing and ELISA) to RPV, and not to PPRV, they were completely protected against challenge inoculation with virulent PPRV. This would indicate that protection is most probably due to cell-mediated immunity. Use of the rinderpest double recombinant vaccinia virus in areas of the world where PPRV is endemic would aid in the control and eradication of PPR.
Asunto(s)
Anticuerpos Antivirales/biosíntesis , Glicoproteínas/inmunología , Enfermedades de las Cabras/prevención & control , Cabras/inmunología , Paramyxoviridae/inmunología , Infecciones por Respirovirus/veterinaria , Virus de la Peste Bovina/inmunología , Vacunas Sintéticas/inmunología , Virus Vaccinia , Proteínas Virales de Fusión/inmunología , Proteínas Virales/inmunología , Vacunas Virales/inmunología , Animales , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Células Cultivadas , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Glicoproteínas/genética , Enfermedades de las Cabras/inmunología , Enfermedades de las Cabras/microbiología , Hemaglutininas Virales , Inmunidad Activa , Proteínas de la Membrana , Pruebas de Neutralización , Infecciones por Respirovirus/inmunología , Infecciones por Respirovirus/microbiología , Infecciones por Respirovirus/prevención & control , Virus de la Peste Bovina/genética , Proteínas Virales de Fusión/genética , Proteínas Virales/genéticaAsunto(s)
Síndrome de Inmunodeficiencia Adquirida del Felino/prevención & control , Virus de la Inmunodeficiencia Felina/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Virus de la Inmunodeficiencia de los Simios/inmunología , Vacunación , Vacunas Virales/inmunología , Animales , Gatos , Modelos Animales de Enfermedad , Femenino , Productos del Gen env/inmunología , Inyecciones Intravenosas , Macaca mulatta/inmunología , Masculino , Vacunas Atenuadas/inmunología , Vacunas de Productos Inactivados/inmunología , Vacunas Sintéticas/inmunología , VaginaRESUMEN
We have developed a method for attenuating vaccinia virus recombinants by expressing a fusion protein of a lymphokine and an immunogen. Chimeric genes were constructed that coded for gamma interferon (IFN-gamma) and structural proteins of the human immunodeficiency virus type 1 (HIV-1). In this study, we describe the biological and immunological properties of vaccinia virus recombinants expressing chimeric genes of murine or human IFN-gamma with glycoprotein gp120, gag, and a fragment of gp41. All fusion proteins retained the antigenic characteristics of both IFN-gamma and HIV as shown by immunoblot analysis. However, the antiviral activity of IFN-gamma could be demonstrated only for the IFN-gamma-gag fusion protein. In contrast, the attenuating activity of IFN-gamma for nude mice was retained by all of the recombinants, albeit at various rates. Unlike the antiviral activity, the attenuating activity of IFN-gamma was not species specific. Implications for the development of attenuated live recombinant vaccines for AIDS are discussed.
Asunto(s)
Genes Sintéticos , VIH-1/genética , Interferón gamma/inmunología , Proteínas Recombinantes de Fusión/inmunología , Vacunas Atenuadas , Virus Vaccinia/inmunología , Proteínas Estructurales Virales/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , Quimera , Genes Virales , Humanos , Interferón gamma/genética , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Oligodesoxirribonucleótidos , Homología de Secuencia de Ácido Nucleico , Virus Vaccinia/genética , Proteínas Estructurales Virales/genéticaRESUMEN
Rinderpest is a highly contagious viral disease of ruminants with greater than 95% morbidity and mortality. We have constructed an infectious vaccinia virus recombinant that expresses both the fusion (F) gene and the hemagglutinin (H) gene of rinderpest virus. The Wyeth strain of vaccinia virus was used for the construction of the recombinant. Cattle vaccinated with the recombinant virus were 100% protected from challenge inoculation with greater than 1000 times the lethal dose of rinderpest virus. No transmission of recombinant vaccinia virus from vaccinated animals to contact animals was observed. The lyophilized form of vaccinia virus is thermostable and allows circumvention of the logistical problems associated with the distribution and administration of vaccines in the arid and hot regions of Asia and Africa. The insertional inactivation of both the thymidine kinase and the hemagglutinin genes of vaccinia virus led to increased attenuation of the virus; this was manifested by the lack of detectable pock lesions in vaccinated animals. This approach may have wide application in the development of safe and efficacious recombinant vaccines for humans and animals. This becomes quite relevant with the concern of the use of vaccinia virus in a population with high incidence of the human immunodeficiency virus.
Asunto(s)
Enfermedades de los Bovinos/prevención & control , Hemaglutininas Virales/inmunología , Virus de la Peste Bovina/inmunología , Peste Bovina/prevención & control , Proteínas Virales de Fusión/inmunología , Vacunas Virales/genética , Animales , Anticuerpos Antivirales/biosíntesis , Antígenos Virales/genética , Bovinos , Clonación Molecular , Genes Virales , Hemaglutininas Virales/genética , Mapeo Restrictivo , Virus de la Peste Bovina/genética , Vacunas Sintéticas , Virus Vaccinia/inmunología , Proteínas Virales de Fusión/genética , Proteínas Estructurales Virales/genéticaRESUMEN
Major immunogenic sites of foot-and-mouth disease virus (FMDV) have been mapped to the C-terminal third of capsid protein VP1; we studied the immunogenicity of a series of TrpE-FMDV fusion proteins containing this region of FMDV strain O1 Campos. Fusion protein TrpE-dCN, which contains a dimer of VP1 amino acid sequences consisting of amino acids 200 to 213 linked by a diproline spacer to amino acids 141 to 158 (200-213 approximately P-P-G approximately 141-158), induced the best response. A single inoculation of guinea-pigs with 100 micrograms TrpE-dCN elicited high levels of neutralizing antibodies and protected all the animals against challenge infection with homologous virus. Although the closely related FMDV strains O1 Campos and O1 Caseros induced high levels of cross-protection, TrpE-dCN-vaccinated guinea-pigs were poorly protected against challenge infection with heterologous FMDV strain O1 Caseros. Nucleotide sequence analysis revealed that amino acid differences at residues 149 and 152 were critical for the induction of cross-protection and that neutralizing epitopes not present in TrpE-dCN are likely to be responsible for conferring a high level of cross-protection between FMDV strains O1 Campos and O1 Caseros.