Detection of infectious laryngotracheitis virus antibodies by glycoprotein-specific ELISAs in chickens vaccinated with viral vector vaccines.

Two glycoproteins of infectious laryngotracheitis virus (ILTV), gI and gB, were expressed in baculovirus and purified for the development of ILTV recombinant protein-based ELISAs. The ability of gB and gI ELISAs to detect ILTV antibodies in chickens vaccinated with viral vector vaccines carrying the ILTV gB gene, Vectormune FP-LT (the commercial fowlpox vector laryngotracheitis vaccine) and Vectormune HVT-LT (commercial turkey herpesvirus vector laryngotracheitis vaccine), was evaluated using serum samples from experimentally vaccinated and challenge chickens. The detection of gB antibodies in the absence of gI antibodies in serum from chickens vaccinated with FP-LT indicated that the gB ELISA was specific for the detection of antibodies elicited by vaccination with this viral vector vaccine. The gB ELISA was more sensitive than the commercial ILTV ELISA to detect seroconversion after vaccination with the FP-LT vaccine. Both gI and gB antibodies were detected in the serum samples collected from chickens at different times postchallenge, indicating that the combination of these ELISAs was suitable to screen serum samples from chickens vaccinated with either recombinant viral vector FP-LT or HVT-LT vaccines. The agreement between the gI ELISA and the commercial ELISA to detect antibodies in serum samples collected after challenge was robust. However, further validation of these ELISAs needs to be performed with field samples.

Protection and antibody response caused by turkey herpesvirus vector Newcastle disease vaccine.

Newcastle disease (ND) is prevalent worldwide and causes significant clinical and economic losses to the poultry industry. Current vaccine programs using live attenuated vaccines and inactivated vaccines have limitations, and new vaccines with distinct features are needed. To offer an alternative solution to control ND, a turkey herpesvirus vector Newcastle disease vaccine (HVT/ND) expressing the fusion gene of Newcastle disease virus (NDV) has been developed. First, immunogenicity of the HVT/ND was evaluated in specific-pathogen-free layer chickens after vaccination by the in ovo route to 18-day-old embryos or by the subcutaneous route to 1-day-old chicks. Antibodies against NDV were detected at 24 days of age using a commercial NDV enzyme-linked immunosorbent assay (ELISA) kit and the hemagglutination inhibition test. At least 90% of chickens were protected against challenge with velogenic neurotropic NDV Texas GB strain (genotype II; pathotype velogenic) at 4 wk of age, while none of the nonvaccinated, challenged controls were protected from challenge. Second, the age at which a vaccinated chicken elicits an immunologic response to the HVT/ND prepared for this study, and thus is protected from ND virus, was assessed in commercial broiler chickens after in ovo vaccination of 18-day-old embryos. Challenge was conducted using a low-virulence NDV strain (genotype II; pathotype lentogenic) via the respiratory tract each week between 1 and 5 wk of age, in order to mimic the situation in areas where virulent NDV strains do not normally exist and low-virulence strains cause mild respiratory symptoms leading to economic losses. Protection was evaluated by the presence or absence of isolated virus from tracheal swabs at 5 days postchallenge. Partial protection was observed at 3 wk of age, when 6 out of 10 (60%) chickens were protected. Full protection was obtained at 4 and 5 wk of age, when 9 out of 10 (90%) and 10 out of 10 (100%) chickens were protected, respectively. Finally, protection against challenge with virulent Texas GB strain at 19 wk of age was evaluated in commercial female layer chickens vaccinated at 1 day of age with HVT/ND. All of the vaccinated chickens were protected, while all of the challenge controls succumbed to the challenge. Furthermore, anti-NDV antibodies measured by ELISA were maintained through 50 wk of age.

Safety and efficacy of a turkey herpesvirus vector laryngotracheitis vaccine for chickens.

Turkey herpesvirus vector laryngotracheitis vaccine (HVT/LT) expressing the glycoprotein B gene of laryngotracheitis virus (LTV) has been developed. In vitro growth kinetics of HVT/LT were similar to those of parental turkey herpesvirus (HVT), FC-126 strain. Genetic and phenotypic stabilities of HVT/LT after in vitro (in cell culture) or in vivo (in chickens) passage were confirmed by various assays, including Southern blot analysis, western blot analysis, and an indirect immunofluorescence assay. Safety of HVT/LT was assessed by an overdose study as well as by a backpassage study in specific-pathogen-free (SPF) chickens. The overdose study indicated that HVT/LT did not cause any adverse effects in chickens. The backpassage study confirmed that HVT/LT does not revert to virulence after five passages in chickens. The vaccine did not transmit laterally from vaccinated chickens to commingled nonvaccinated chickens. Efficacy of HVT/LT was evaluated in SPF layer chickens after vaccination by the subcutaneous route at 1 day of age. The majority of the vaccinated chickens (92%-100%) were protected against challenge with virulent LTV at 7 wk of age. Efficacy of HVT/LT was further evaluated in broiler chickens from a commercial source after in ovo vaccination to embryos at 18 days of incubation. After challenge with virulent LTV at 21 and 35 days of age, 67% and 87% of HVT/LT-vaccinated chickens did not develop LT clinical signs, respectively, while 100% (21 days of age) and 73% (35 days of age) of the challenge control chickens showed clinical signs of LT. These results suggest that HVT/LT is a safe and efficacious vaccine for control of laryngotracheitis (LT).