Marek's disease herpesvirus is a vaccine vector of great promise for chickens; however, complete protection against foreign infectious diseases has not been achieved. In this study, two herpesvirus of turkey recombinants (rHVTs) expressing large amounts of infectious bursal disease virus (IBDV) VP2 antigen under the control of a human cytomegalovirus (CMV) promoter or CMV/␤-actin chimera promoter (Pec promoter) (rHVT-cmvVP2 and rHVT-pecVP2) were constructed. rHVT-pecVP2, which expressed the VP2 antigen approximately four times more than did rHVT-cmvVP2 in vitro, induced complete protection against a lethal IBDV challenge in chickens, whereas rHVT-cmvVP2 induced 58% protection. All of the chickens vaccinated with rHVT-pecVP2 had a protective level of antibodies to the VP2 antigen at the time of challenge, whereas only 42 and 67% of chickens vaccinated with rHVT-cmvVP2 or the conventional live IBDV vaccine, respectively, had the antibodies. The antibody level of chickens vaccinated with rHVT-pecVP2 increased for 16 weeks, and the peak antibody level persisted throughout the experiment. The serum antibody titer at 30 weeks of age was about 20 or 65 times higher than that of chickens vaccinated with rHVT-cmvVP2 or the conventional live vaccine, respectively. rHVT-pecVP2, isolated consistently for 30 weeks from the vaccinated chickens, expressed the VP2 antigen after cultivation, and neither nucleotide mutations nor deletion in the VP2 gene was found. These results demonstrate that the amount of VP2 antigen expressed in the HVT vector was correlated with the vaccine efficacy against lethal IBDV challenge, and complete protective immunity that is likely to persist for the life of the chickens was induced. Marek's disease (MD) virus (MDV) is a cell-associated, lymphotropic alphaherpesvirus of chickens that causes the most-common, highly contagious T-cell lymphoma (6), and all three serotypes of MDV have been completely sequenced (1,19,22,41). The MDV vaccine strains, which are serotypes 1 (MDV1), MDV2, and MDV3 (herpesvirus of turkey [HVT]) (6), have merits as a distinguished vector (7,15,24,30). MDV vaccines can overcome the inhibition of maternal antibodies (28, 35) and might induce long-term protective immunity in chickens. Down-regulation of major histocompatibility complex class I expression is a common mechanism of herpesviruses, including MDV, used to evade cellular immunity and persist in their hosts (17,20). MDV1 has high vaccine efficacy against MD but grows slowly in cell culture, whereas HVT has a relatively low vaccine efficacy but is highly safe for chickens and grows remarkably well in cell culture. Despite the high potential of the MDV vectors, attempts to elicit complete protection against infections in chickens have not been successful (8, 13, 15, 27, 28, 31-33, 35, 39). The lack of effective MDV1 recombinants is likely due to a variety of factors such as the difficulty in making recombinants without attenuating the virus.Infectious bursal disease (IBD) virus (IBDV), a member of the Birnaviridae famil...

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).

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Antibody response of recombinant fowlpox virus (FPV) was studied in chickens inoculated with the virus in the presence or absence of antibodies against Newcastle disease virus (NDV) or FPV. In the case of NDV, high hemagglutination-inhibition titers to NDV were obtained when the antibody was present. No immune response to NDV was observed in the chickens previously vaccinated with FPV.

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