Inactivated Bovine Respiratory Syncytial Virus Research Articles

Investigation into the safety, and serological responses elicited by delivery of live intranasal vaccines for bovine herpes virus type 1, bovine respiratory syncytial virus, and parainfluenza type 3 in pre-weaned calves.

Despite the fact that pneumonia remains a leading cause of mortality and morbidity in pre-weaned calves, relatively little is known regarding the effects of the concurrent administration of intranasal pneumonia virus vaccines, particularly in calves with high levels of maternally derived antibodies. The objective of this study was to use a cohort of 40 dairy and dairy-beef female and male calves (27 females and 13 males) to determine serological responses to concurrent administration at 3 weeks of age (22 ± 4.85 days) of two commercially available intranasal (IN) vaccines for the viruses: bovine respiratory syncytial virus (BRSV), bovine herpes virus 1 (BoHV-1), and parainfluenza-3-virus (PI3-V). The study groups were as follows: (i) Bovilis IBR Marker Live only® (IO), (ii) Bovilis INtranasal RSP Live® only (RPO), (iii) Concurrent vaccination with Bovilis IBR Marker Live® & Bovilis Intranasal RSP Live® (CV), and (iv) a control group of non-vaccinated calves (CONT). The calves' serological response post-IN vaccination, clinical health scores, rectal temperatures, and weights were measured. Data were analyzed in SAS using mixed models and logistic regression. The CV calves had an average daily weight gain (ADG) of 0.74 (±0.02) kg, which was similar to CONT (0.77 ± 0.02 kg). Despite no significant differences in the antibody levels between study groups 3 weeks post-IN vaccination, following the administration of subsequent parenteral injections in the form of Bovilis Bovipast RSP®(antigens; inactivated BRSV, inactivated PI3-V, inactivated Mannheimia haemolytica) and Bovilis IBR Marker Live®, the antibody levels of the BRSV and PI3-V increased in both the CV and RPO study groups. Concurrent vaccination resulted in no increase in fever and no difference in health scores when compared to CONT.

The titers, duration, and residual clinical protection of passively transferred nasal and serum antibodies are similar among beef calves that nursed colostrum from vaccinated or unvaccinated dams and were challenged experimentally with bovine respiratory syncytial virus at three months of age.

To compare initial titers, duration, and residual clinical protection of passively transferred bovine respiratory syncytial virus (BRSV) nasal immunoglobulin (Ig) G-1 and IgA, and serum neutralizing (SN) antibodies. 40 three-month-old beef steers born either to unvaccinated or vaccinated cows. During the last trimester of gestation, cows were assigned randomly to either vaccinated or unvaccinated groups. Calves were grouped on the basis of whether they nursed colostrum from unvaccinated dams (NO-VACC group; n = 20) versus dams vaccinated with 2 doses of an inactivated BRSV vaccine (VACC group; n = 20). At 3 months of age, calves were challenged with BRSV. Respiratory signs were scored. Nasal BRSV IgG-1 and IgA and SN antibodies were compared before and after the challenge. The presence of BRSV in nasal secretions was evaluated by reverse transcription-PCR assays. Respiratory scores after BRSV challenge were similar between treatment groups. Nasal BRSV IgG-1 and SN antibodies were significantly greater in VACC calves at 48 hours of life; however, by 3 months of age, titers had decayed in both groups. Nasal BRSV IgA titers were minimal after colostrum intake and before the BRSV challenge, and increased in both groups after the challenge. The NO-VACC group had a significantly greater probability of shedding BRSV compared with VACC calves. At 3 months of age, titers of passively transferred BRSV antibodies in VACC and NO-VACC calves had decayed to nonprotective levels. Calves born to vaccinated dams had a decreased probability of BRSV shedding; however, this was not related to differences in SN or nasal BRSV antibody titers.

A recombinant subunit vaccine for bovine RSV and Histophilus somni protects calves against dual pathogen challenge

Bovine respiratory syncytial virus (BRSV) and Histophilus somni synergize to cause respiratory disease in cattle. These pathogens cause enhanced disease during dual-infection and an IgE response to antigens of H. somni in dual-infected but not singly infected calves. Vaccines containing whole inactivated BRSV or H. somni have been associated with IgE responses A vaccine strategy that avoids stimulation of IgE antibodies would provide superior protection from dual infection. We hypothesized that a subunit vaccine consisting of the nucleoprotein (NP) from BRSV and the recombinant antigen IbpA DR2 (a surface antigen of H. somni with two toxic fic motifs) in Quil A adjuvant would elicit protection without disease enhancement. Three groups of calves were vaccinated twice with either: Formalin inactivated BRSV (FI) plus Somnivac®, NP & IbpA DR2 plus Quil A or Quil A alone, followed by BRSV and H. somni challenge. Clinical scores and antibody levels (to whole pathogens and to the subunits) were evaluated. Lungs were examined at necropsy on day 23 after infection. Clinical scores were significantly greatest for the FI & Somnivac® group and both clinical scores and lung pathology were lowest for the subunit group. All calves shed BRSV in nasal secretions. FI & Somnivac® induced IgE antibodies to H. somni and BRSV, but not to NP or DR2. The subunit vaccine did not induce an IgE antibody response to IbpA DR2 antigen and induced little IgE to H. somni. It did not induce an IgG antibody response to BRSV and H. somni, but stimulated production of IgG antibodies against the subunits. In summary, the subunit vaccine, consisting of the BRSV NP and H. somni IbpA DR2 in Quil A, protected against severe clinical signs and decreased lung pathology but did not prevent viral shedding. Importantly it prevented synergistic disease expression in response to dual infection.

Cell-mediated immune responses of lambs to challenge with bovine respiratory syncytial virus.

The lamb is a good model to study the pathogenesis and immune responses to infections with respiratory syncytial virus (RSV) as lambs experimentally infected with bovine or human RSV may develop overt clinical disease. In the present study the development of cellular cytotoxic responses was studied in splenic, pulmonary and peripheral blood mononuclear cells obtained from lambs after primary and secondary infection with bovine RSV. Infection with bovine RSV was followed by the appearance of cytotoxic cells in the peripheral blood, the spleen and lung lavage fluids. These effector cells lysed virus-infected targets in a self-restricted manner. Depletion techniques revealed that cytotoxic activity was largely due to OvCD8+ cells. When effector cells obtained from primed lambs were stimulated with inactivated bovine RSV or with virus-infected cells in vitro, virus-specific cytotoxicity was significantly increased.

Antibody responses in mice after immunization with inactivated bovine respiratory syncytial virus using different adjuvants

Bovine respiratory syncytial virus (BRSV) causes pneumonia in young cattle. Modified-live-virus (MLV) and inactivated vaccines are currently used for the control of clinical effects of BRSV infections in cattle. On the present research, the stimulation of specific anti-BRSV immunoglobulin isotypes was investigated, through the use of different commercially available adjuvants (Water-in-oil emulsion, Quil A, Aluminum-hydroxide) in inbred mice (Balb/C and C57BL/6). BRSV antibodies were measured using an enzyme-linked immunosorbent assay (ELISA) and the results were compared to the antibody levels induced by immunization of animals using live-BRSV-virus. Water-in-oil emulsion and alum- adjuvant preparations induced higher levels of IgG1 immunoglobulins, whereas Quil A favored the production of IgG2 antibodies, this last being a more appropriate response profile for the specific case of BRSV. Not using adjuvants resulted in poor levels of specific antibodies. The isotype profile of specific antibodies obtained varied greatly depending on the adjuvants used. This information may be useful for the formulation of more effective BRSV inactivated vaccines; however, these findings have to be confirmed in cattle.

Antibody responses in mice after immunization with inactivated bovine respiratory syncytial virus using different adjuvants

Bovine respiratory syncytial virus (BRSV) causes pneumonia in young cattle. Modified-live-virus (MLV) and inactivated vaccines are currently used for the control of clinical effects of BRSV infections in cattle. On the present research, the stimulation of specific anti-BRSV immunoglobulin isotypes was investigated, through the use of different commercially available adjuvants (Water-in-oil emulsion, Quil A, Aluminum-hydroxide) in inbred mice (Balb/C and C57BL/6). BRSV antibodies were measured using an enzyme-linked immunosorbent assay (ELISA) and the results were compared to the antibody levels induced by immunization of animals using live-BRSV-virus. Water-in-oil emulsion and alum- adjuvant preparations induced higher levels of IgG1 immunoglobulins, whereas Quil A favored the production of IgG2 antibodies, this last being a more appropriate response profile for the specific case of BRSV. Not using adjuvants resulted in poor levels of specific antibodies. The isotype profile of specific antibodies obtained varied greatly depending on the adjuvants used. This information may be useful for the formulation of more effective BRSV inactivated vaccines; however, these findings have to be confirmed in cattle.

A single vaccination with an inactivated bovine respiratory syncytial virus vaccine primes the cellular immune response in calves with maternal antibody

BackgroundThe efficacy of a single dose of an inactivated bovine respiratory syncytial virus (BRSV) - Parainfluenaza type 3 (PI3) - Mannheimia haemolytica (Mh) combination vaccine, in calves positive for maternal antibodies, was established in a BRSV infection study.ResultsAs expected the single vaccination did not have any effect on the decline of BRSV-specific neutralising or ELISA antibody. The cellular immune system was however primed by the vaccination. In the vaccinated group virus excretion with nasal discharge was reduced, less virus could be re-isolated from lung tissues and the lungs were less affected.ConclusionsThese results indicate that a single vaccination with an inactivated BRSV vaccine was able to break through the maternal immunity and induce partial protection in very young calves. It can be speculated that the level and duration of protection will improve after the second dose of vaccine is administered. A two-dose basic vaccination schedule is recommended under field conditions.

Field efficacy of combination vaccines against bovine respiratory pathogens in calves

The efficacy of an inactivated bovine respiratory syncytial virus (BRSV)--bovine parainfluenza type 3 (PI3)--Mannheimia haemolytica (Mh) combination vaccine was examined in two field studies. Calves were vaccinated (i) with the inactivated vaccine, (ii) a modified live/killed viral combination vaccine, or (iii) left unvaccinated. The efficacy of the vaccines was judged by the (i) number of treated animals, (ii) number of individual antibiotic treatments per calf and (iii) mortality rates. After vaccination with the inactivated vaccine, the number of calves requiring antibiotic treatment was significantly lower than in the unvaccinated group (odds ratios: 0.26 first study and 0.53 second study), but differences between vaccination with live/killed combination vaccines and controls were not significant (odds ratios: 0.56 and 0.90, respectively). In both studies, a number of unvaccinated controls died due to respiratory disease (4.6% first and 6.7% second study). By contrast, none of the animals vaccinated with the inactivated vaccine died in the first study and only 3.3% in the second study. The mortality rates for the groups vaccinated with the live vaccine (1.3% and 7.8%) were similar to the unvaccinated controls. In summary, these data demonstrate the efficacy of the inactivated vaccine under field conditions.

Response of calves to challenge exposure with virulent bovine respiratory syncytial virus following intranasal administration of vaccines formulated for parenteral administration

To determine whether single-fraction and combination modified-live bovine respiratory syncytial virus (BRSV) vaccines commercially licensed for parenteral administration could stimulate protective immunity in calves after intranasal administration. Randomized controlled trial. 39 calves. Calves were separated from dams at birth, fed colostrum with a minimal concentration of antibodies against BRSV, and maintained in isolation. In 2 preliminary experiments, 9-week-old calves received 1 (n = 3) or 2 (3) doses of a single-component, modified-live BRSV vaccine or no vaccine (8 control calves in each experiment), and were challenged with BRSV 21 days after vaccination. In a third experiment, 2-week-old calves received combination modified-live virus (MLV) vaccines with or without BRSV and calves were challenged with BRSV 8 days later. Calves were euthanized, and lung lesions were measured. Immune responses, including serum and nasal antibody and nasal interferon-alpha concentrations, were assessed. BRSV challenge induced signs of severe clinical respiratory tract disease, including death and pulmonary lesions in unvaccinated calves and in calves that received a combination viral vaccine without BRSV. Pulmonary lesions were significantly less severe in BRSV-challenged calves that received single or combination BRSV vaccines. The proportion of calves that shed virus and the peak virus titer was decreased, compared with control calves. Protection was associated with mucosal IgA antibody responses after challenge. Single and combination BRSV vaccines administered intranasally provided clinical protection and sparing of pulmonary tissue similar to that detected in response to parenteral delivery of combination MLV and inactivated BRSV vaccines previously assessed in the same challenge model.

Priming of multiple T cell subsets by modified-live and inactivated bovine respiratory syncytial virus vaccines

T cell activity is a critical component of immunity to bovine respiratory syncytial virus (BRSV). We tested the effects of immunization by modified-live and inactivated BRSV vaccines on cell-mediated and humoral immunity in young calves. The two forms of vaccine stimulated similar serum neutralizing antibody production, although the early kinetics of those responses differed. CD4 +, CD8 +, and γδ T cells were analyzed before and after immunization for BRSV-specific in vitro recall responses, as evaluated by CD25 upregulation measured by flow cytometry. Modified-live virus (MLV) primed each of the three subsets for statistically significant in vitro responses to antigen. Inactivated vaccine also primed each T cell population for significant antigen-driven CD25 upregulation, including responses by CD4 + and γδ T cells that were stronger and longer-lasting than those primed by MLV. Monoclonal antibody was used in additional assays to block MHC class I during incubation of BRSV antigen with peripheral blood mononuclear cells from an animal in the inactivated vaccine group. The recall response by CD8 + T cells was more inhibited by this treatment than the other subsets, further suggesting that the inactivated vaccine had primed antigen-specific CD8 + T cells. In summary, the data indicate that balanced BRSV-specific T cell responses can be induced by inactivated, as well as modified-live, conventional vaccines, which may implicate an alternative pathway of MHC class I antigen presentation.

Cytokine responses of bovine dendritic cells and T cells following exposure to live or inactivated bovine respiratory syncytial virus

We compared the effects of live or inactivated bovine respiratory syncytial virus (BRSV) on cytokine production by bovine monocyte-derived dendritic cells (MoDC). We also investigated the response of resting memory CD4(+) T cells to MoDC exposed to both viral preparations. Although BRSV did not appear to replicate in MoDC or to affect expression of major histocompatibility complex (MHC) class I, MHC class II, or CD80/86, a higher percentage of cells exposed to live virus appeared to undergo apoptosis/necrosis. To investigate how the interaction of BRSV with MoDC affects the immune response, a multiplex, real-time, polymerase chain reaction was established to analyze transcription of bovine cytokines. Exposure of MoDC to live BRSV induced more interleukin (IL)-10 mRNA and markedly less IL-12p40 and IL-15 mRNA than did heat-inactivated virus. To determine whether these differences might influence the T cell response, CD4(+) memory T cells primed in vivo were restimulated in vitro by MoDC pulsed with heat-inactivated or live BRSV. Stimulation of CD4(+) T cells induced similar levels of IL-2-and IL-4-like activity and interferon-gamma. These observations suggest that while IL-10, produced by MoDC as a result of exposure to live BRSV, may affect IL-12 and IL-15 synthesis by MoDC, it does not appear to affect the cytokine response of BRSV-specific memory CD4(+) T cells. It is possible, however, that differences in the pattern of cytokines produced by MoDC exposed to live or inactivated virus may influence the development of the primary CD4(+) T cell response in vivo.

Bovine Respiratory Syncytial Virus (BRSV) Pneumonia in Beef Calf Herds Despite Vaccination

The present report describes the clinical, pathological, serological and virological findings in calves from 2 larger Danish beef herds experiencing outbreaks of pneumonia. The calves had been vaccinated with an inactivated bovine respiratory syncytial virus (BRSV) vaccine 2 months prior to the outbreak. The clinical signs comprised nasal discharge, pyrexia, cough and increased respiratory rates. A total of 28 calves died in the 2 herds. The laboratory investigations revealed that BRSV was involved and probably initiated both outbreaks. Furthermore, the serological results suggested that the vaccine induced only sparse levels of antibodies probably due to the presence of maternally derived antibodies at the time of vaccination. Necropsy findings in 5 calves revealed changes typical for infectious pneumonia with involvment of BRSV. In conclusion, vaccination of calves against BRSV in 2 Danish beef herds failed to protect the calves against severe or even fatal BRSV mediated respiratory disease 2 months later.

Efficacy of an Inactivated BRSV Vaccine Against a Virulent BRSV Experimental Challenge in Young Calves

The objective of this study was to determine the efficacy of a vaccine containing inactivated Bovine Respiratory Syncytial Virus (BRSV) against a highly virulent BRSV challenge in calves.

High mortality rate associated with bovine respiratory syncytial virus (BRSV) infection in Belgian white blue calves previously vaccinated with an inactivated BRSV vaccine.

In a group of 60 Belgian White Blue calves less than 8 months old still housed in barns, a bovine respiratory syncytial virus (BRSV) outbreak was revealed on the basis of a direct diagnosis (immunofluorescence and virus isolation) performed on the lungs of dead animals, and the kinetics of BRSV neutralizing antibodies. Clinical signs, macroscopical and microscopical pulmonary lesions were also compatible with a BRSV infection. This outbreak is peculiar because the 35 oldest calves (204 +/- 29 days old) had been vaccinated 3-4 months before with an inactivated BRSV vaccine and 30% of these animals had died of respiratory distress. While they experienced a mild respiratory symptomatology, no death was recorded among the 25 youngest calves (69 +/- 29 days old) which had been left unvaccinated. Another peculiarity was found at the histological level where a massive infiltration of eosinophils was demonstrated in the pulmonary tissues of the dead animals. Together these data parallel the dramatic story described 30 years ago in children previously vaccinated with a formalin-inactivated human RSV (HRSV) vaccine upon a natural HRSV challenge. This illustrates that an immunopathological phenomenon also takes place after BRSV vaccination in cattle.

Cell-mediated immune responses of lambs to experimental infection with bovine respiratory syncytial virus

Primary infection of lambs with bovine respiratory syncytial virus (RSV) was characterised by the presence of virus-specific cytotoxic cells in the peripheral blood as early as 5 days post-infection. These effector cells lysed virus-infected autologous targets in a self-restricted manner. Depletion techniques revealed that cytotoxic activity was largely due to OvCD8 + cells. Neutrophils obtained from experimentally infected lambs 5 to 10 days post-inoculation exhibited significant cytotoxic activity in the presence of bovine RSV-specific antiserum. During the same period there was a significant lymphoproliferative response to live or inactivated bovine RSV. Lymphoproliferative and cytotoxic activity coincided with the clearance of the virus from nasal secretions.

Bovine respiratory syncytial virus-specific immune responses in cattle following immunization with modified-live and inactivated vaccines. Analysis of the specificity and activity of serum antibodies

Cattle were immunized with vaccines containing modified-live or inactivated bovine respiratory syncytial virus (BRSV) and serum antibody responses were analyzed. Compared with preinculation values, at Day 14 after two biweekly immunizations with modified-live or inactivated vaccines there were significant increases in BRSV-specific titers in the sera of cattle that received both types of vaccines, as determined by a whole cell ELISA. Using a blocking ELISA and radioimmune precipitation it was determined that there was recognition of the fusion (F) protein by antibodies from cattle that received both types of BRSV antigens: however, virus neutralization assays revealed that only cattle that received modified live virus, either in monovalent or polyvalent vaccines, developed neutralizing antibodies to BRSV after two immunizations. These results indicate that inactivation of BRSV can lead to a dissociation between serological recognition of the F protein and virus neutralization in vaccinated cattle.

Bovine respiratory syncytial virus-specific immune responses in cattle following immunization with modified-live and inactivated vaccines. Analysis of proliferation and secretion of lymphokines by leukocytes in vitro

Cattle were immunized with vaccines containing modified-live or inactivated bovine respiratory syncytial virus (BRSV) and lymphocyte proliferative responses and cytokine secretion were monitored sequentially. Compared to pre-inoculated values, significant increases in proliferative responses to modified-live BRSV were detectable by Day 7 after the primary immunization with the vaccine containing inactivated BRSV, and by 7 days after the second immunization with modified-live virus. After a third immunization with the respective vaccines, proliferative responses to live BRSV were significantly higher in the group that received modified-live vaccine compared to the group that received inactivated vaccine. Proliferative responses to live BRSV corresponded with the presence of interleukin-2 (IL-2) in the supernatants from BRSV-stimulated leukocyte cultures and there were significantly higher levels of IL-2 in cultures from the group that received modified-live BRSV. An interferon species with the characteristics of interferon-α was also present in the supernatants from leukocyte cultures and there were no significant differences between the groups of vaccines. The predominant phenotype of proliferating cells in BRSV-stimulated leukocyte cultures derived from both groups of bovine vaccines was a BoCD4+ T-lymphocyte. These in vitro data suggest that both types of vaccines are capable of stimulating cell-mediated immune responses to BRSV in cattle.