A recombinant fowl adenovirus (FAdV) fiber protein, derived from a FAdV-8a strain, was tested for its efficacy to protect chickens against inclusion body hepatitis (IBH). FAdV-E field isolates belonging to both a homotypic (FAdV-8a) and heterotypic (-8b) serotype were used as challenge. Mechanisms underlying fiber-induced protective immunity were investigated by fiber-based ELISA, virus neutralization assays and flow cytometry of peripheral blood mononuclear cells, monitoring the temporal developments of humoral and cellular responses after vaccination and challenge exposure. Birds were clinically protected from the homologous challenge and showed a significant reduction of viral load in investigated target organs, whereas fiber-based immunity failed to counteract the heterologous serotype infection. These findings were supported in vitro by the strictly type-specific neutralizing activity of fiber immune sera. In protected birds, fiber vaccination prevented a post-challenge drop of peripheral B cells in blood. Furthermore, fiber immunization stimulated CD4⁺ T lymphocyte proliferation while moderating the CD8α⁺ T cell response and prevented challenge-induced changes in systemic monocytes/macrophages and γδ⁺ T cell subpopulations. Both vaccinated and adjuvant-only injected birds experienced a priming of systemic B cells and TCRγδ⁺ T lymphocytes, which masked possible pre-challenge effects due to the antigen. In conclusion, within FAdV-E, recombinant fiber represents a vaccine candidate to control the adverse effects of homotypic infection by eliciting an effective humoral immunity and regulating B and T cell response, whereas the failure of heterotypic protection suggests a primordial role of humoral immunity for this vaccine.

In the past decades, fowl adenovirus (FAdV)-related diseases became an increasing concern for the poultry industry worldwide. Various immunization strategies against FAdVs have been experimentally investigated, with a particular focus on subunit vaccines against hepatitis-hydropericardium syndrome (HHS), caused by FAdV serotype 4, and inclusion body hepatitis (IBH), caused by serotypes 2, 8a, 8b and 11. In this study, we extended our innovative concept of recombinant chimeric fiber proteins to design a novel chimera combining epitopes from two distinct serotypes, FAdV-4 and -11, and we investigated its efficacy to simultaneously protect chickens against HHS and IBH. Specific pathogen-free chickens were vaccinated with the novel recombinant chimeric fiber and subsequently challenged with either a HHS- or IBH-causing strain. Vaccinated/challenged birds exhibited a reduction of clinical signs, limited hepatomegaly and lower levels of AST compared to the respective challenge controls. Furthermore, the vaccine prevented atrophy of HHS-affected lymphoid organs, such as thymus and bursa of Fabricius, and viral load in the target organs was significantly reduced. Clinical protection was associated with high levels of pre-challenge antibodies measured on ELISA plates coated with the vaccination antigen. Interestingly, the development of neutralizing antibodies was limited against FAdV-11 and absent against FAdV-4, indicating that protection granted by such an antigen may be linked to different immunization pathways. In conclusion, we proved that the concept of chimeric fiber vaccines can be extended across viral species boundaries and represents the first single-component FAdV subunit vaccine providing comprehensive protection against different FAdV-associated diseases.

In recent years, fowl adenovirus (FAdV)-induced diseases became a global problem with considerable impact on chicken health and welfare. This has prompted numerous studies to focus on experimental immunization strategies using whole virus formulations (live or killed vaccines), some of them modified as recombinantly constructed vector vaccines. In addition, FAdV capsid proteins were frequently reported as immunizing antigens (subunit vaccines), with fiber proteins being amongst the most successful candidates. To date, there is no standardized protocol to assess vaccine efficacy in experimental FAdV protection studies, with the consequence that the experimental settings present several degrees of variations even when sharing similar premises. Differences in formulation preparations, route of inoculation, antigen dose, vaccination scheme, choice of challenge strain, or type and age of the birds are capable to greatly influence the magnitude of the immune response and the consequent protective efficacy, altogether addressing remaining challenges. Beyond the antigen composition of a vaccine, the epidemiology of FAdVs with the potential of vertical transmission of virus and/or antibodies from breeders to progenies has a substantial impact on protection strategies. The goal of this review is to outline a broad overview of the findings made thus far regarding immunization strategies against diseases associated to FAdV infections, considering the literature published since the appearance of hepatitis-hydropericardium syndrome (HHS) in the late Eighties, in order to emphasize the current knowledge on FAdV vaccines and highlight fields of future research and intervention.