RESUMEN
Histophilosis, a mucosal and septicemic infection of cattle is caused by the Gram negative pathogen Histophilus somni (H. somni). As existing vaccines against H. somni infection have shown to be of limited efficacy, we used a reverse vaccinology approach to identify new vaccine candidates. Three groups (B, C, D) of cattle were immunized with subunit vaccines and a control group (group A) was vaccinated with adjuvant alone. All four groups were challenged with H. somni. The results demonstrate that there was no significant difference in clinical signs, joint lesions, weight change or rectal temperature between any of the vaccinated groups (B,C,D) vs the control group A. However, the trend to protection was greatest for group C vaccinates. The group C vaccine was a pool of six recombinant proteins. Serum antibody responses determined using ELISA showed significantly higher titers for group C, with P values ranging from < 0.0148 to < 0.0002, than group A. Even though serum antibody titers in group B (5 out of 6 antigens) and group D were significantly higher compared to group A, they exerted less of a trend towards protection. In conclusion, the vaccine used in group C exhibits a trend towards protective immunity in cattle and would be a good candidate for further analysis to determine which proteins were responsible for the trend towards protection.
Asunto(s)
Vacunas Bacterianas/inmunología , Enfermedades de los Bovinos/prevención & control , Infecciones por Haemophilus/veterinaria , Haemophilus somnus/inmunología , Vacunas de Subunidad/inmunología , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/microbiología , ADN Bacteriano/genética , Ensayo de Inmunoadsorción Enzimática , Genoma Bacteriano , Infecciones por Haemophilus/microbiología , Infecciones por Haemophilus/prevención & control , Haemophilus somnus/aislamiento & purificación , Inmunización , Proteínas Recombinantes/inmunología , Vacunación , VirulenciaRESUMEN
The selection of an optimal adjuvant to enhance the potency and longevity of antigen specific immune responses has always been imperative for the development of more effective and safer vaccines. A balanced type of immune enhancing ability of a new adjuvant known as polyphosphazene (PCEP) has been demonstrated in mice. In the present study we have compared the humoral and cellular immune responses to vaccine formulations containing Actinobacillus pleuropneumoniae outer membrane antigen (OmlA) with PCEP or Emulsigen as adjuvants. Our data showed a significant improvement and a shift toward more balanced immune responses when OmlA antigen was formulated with PCEP and CpG ODN. Moreover, in contrast to Emulsigen, immunization with PCEP did not result in local injection site reactions highlighting its potential as a safe and effective adjuvant for pigs. Further, the ease of formulation, administration and relatively low per dose cost of PCEP make it a promising adjuvant for pigs.
Asunto(s)
Infecciones por Actinobacillus/veterinaria , Actinobacillus pleuropneumoniae/inmunología , Adyuvantes Inmunológicos/farmacología , Proteínas de la Membrana Bacteriana Externa/inmunología , Vacunas Bacterianas/inmunología , Lipoproteínas/inmunología , Fenilpropionatos/farmacología , Polímeros/farmacología , Enfermedades de los Porcinos/inmunología , Infecciones por Actinobacillus/inmunología , Infecciones por Actinobacillus/microbiología , Infecciones por Actinobacillus/prevención & control , Animales , Anticuerpos Antibacterianos/sangre , Proteínas de la Membrana Bacteriana Externa/genética , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/genética , Ensayo de Inmunoadsorción Enzimática/veterinaria , Lipoproteínas/genética , Porcinos , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/prevención & control , Células TH1/inmunología , Células Th2/inmunología , Vacunación/métodos , Vacunación/veterinariaRESUMEN
Immunostimulatory CpG oligodeoxynucleotide (ODN) can protect mice against infection by many pathogens but the mechanisms mediating disease protection are not well defined. Furthermore, the mechanisms of CpG ODN induced disease protection in vivo have not been investigated in other species. We investigated the induction of antiviral effector molecules in sheep treated with a class B CpG ODN (2007). Subcutaneous injection of ODN 2007 induced a dose-dependent increase in serum levels of the antiviral effector molecule, 2'5'-A synthetase. Peak levels of enzyme were observed 4 days following ODN injection and enzyme levels remained elevated for the following 3-5 days. Repeated ODN injections induced a more sustained elevation of serum 2'5'-A synthetase activity. Finally, formulation of ODN 2007 in emulsigen increased the level of serum 2'5'-A synthetase activity and this response was CpG-specific. Elevated serum 2'5'-A synthetase activity suggested that CpG ODN acted through the induction of either interferon (IFN)-alpha or IFN-gamma. ODN 2007 did not induce detectable levels of IFN-alpha or IFN-gamma when incubated with peripheral blood mononuclear cells, but both IFN-alpha and IFN-gamma were detected following stimulation of lymph node cells with ODN 2007. CpG ODN induction of 2'5'-A synthetase in vitro correlated with the secretion of both IFN-alpha and IFN-gamma. Furthermore, immunohistochemical staining of skin revealed a marked cellular infiltration at the site of ODN 2007 injection. This cellular infiltration was CpG-specific and consisted of primarily CD172(+) myeloid cells. Many of the cells recruited to the site of ODN 2007 injection expressed IFN-alpha and some IFN-gamma. These observations support the conclusion that localized cell recruitment and activation contribute to CpG ODN induction of antiviral effector molecules, such as interferon and 2'5'-A synthetase.
Asunto(s)
2',5'-Oligoadenilato Sintetasa/sangre , Antivirales/sangre , Oligodesoxirribonucleótidos/inmunología , Oligodesoxirribonucleótidos/metabolismo , 2',5'-Oligoadenilato Sintetasa/metabolismo , Adyuvantes Inmunológicos/metabolismo , Animales , Bovinos , Células Cultivadas , Femenino , Inmunoglobulinas/inmunología , Inmunoglobulinas/metabolismo , Interferón-alfa/metabolismo , Interferón gamma/metabolismo , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/metabolismo , Ganglios Linfáticos/citología , Masculino , Ovinos , Piel/citología , Piel/metabolismoRESUMEN
Examples exist in the literature that demonstrate that treatment with immunostimulatory cytosine-phosphate-guanosine (CpG)-DNA can protect mice against infection by intracellular pathogens. There are, however, few studies reporting that CpG-DNA offers similar disease protection in other species. In this study, we assessed the potential of a class A and class B CpG oligodeoxynucleotide (ODN) to induce innate immune responses in sheep, an outbred species. Using peripheral blood mononuclear cells, we have for the first time demonstrated CpG-ODN-induced innate immune responses, including natural-killer-like activity [non-major histocompatibility complex (MHC)-restricted cytotoxicity], interferon-alpha secretion and 2'-5'A oligoadenylate synthetase activity, that could contribute to immune protection in sheep. The type and magnitude of these responses were dependent on ODN class and non-MHC-restricted killing was not associated with interferon-gamma production. The latter observation is in contrast with observations reported for mice and humans. These observations support the conclusion that differences in CpG-ODN-induced responses exist among species and that specific ODN sequences can significantly influence innate immune responses.