Measurement of systemic and local respiratory cell-mediated immunity after influenza infection in chickens.

The detection of ChIFN production after ex vivo antigenic-stimulation of T-lymphocytes has been evaluated for the first time, as a tool to assess cell-mediated immunity (CMI) after avian influenza (AI) infection in 10-day-old SPF chickens. Preliminarily, recall antigens have been produced either by concentrating and inactivating the whole virus or by dissociating the viral proteins. Biologically and structurally intact forms of the viral proteins were isolated by non-ionic detergents while heating, chemical agents and ionic detergent used for virus inactivation altered the antigenic viral components. The n-octyl-B-D-gluco-pyranoside treatment at low temperature was very efficient to produce AI antigenic proteins used for evaluation of ChIFN production after ex vivo antigenic-stimulation of splenic and peripheral lymphocytes. In addition, protocols to isolate lymphocytes from the respiratory tract - the trachea and the lung - have been adapted for local CMI evaluation after similar ex vivo recall assay. Specific AI CMI in the spleen, the blood and the lung was detected for 5 weeks after low pathogenic AI (LPAI) infection in chickens, while further development is needed for tracheal CMI measurement.

The positive adjuvant effect of chitosan on antigen-specific cell-mediated immunity after chickens vaccination with live Newcastle disease vaccine.

The development of safe, novel strong adjuvants is necessary to maximize the efficacy of and the immune response induced by new and/or available vaccines administered through the mucosal route to chickens. Chitosan is a non-toxic, biocompatible, biodegradable and natural polysaccharide derived from the exoskeleton of crustaceans and insects. It has been demonstrated to be an effective absorption enhancer to improve mucosal delivery of peptide and protein drugs in human and mice. In poultry, mucosal administration of live vaccine has been already explored with success. However, the effects of the use of the chitosan as adjuvant for mucosal vaccination in birds have not been investigated yet. To this aim, we explored its potential as adjuvant given by oculo-nasal route to one-day-old chickens with live Newcastle disease (ND) vaccine. The immune response has been evaluated during three independent vaccination experiments on specific pathogen free (SPF) chickens. It was shown that chitosan enhanced the antigen-specific cell-mediated immune response in the spleen. New protocols were developed to measure the chicken IFNgamma production after ex vivo antigen-stimulation of peripheral blood and duodenal lamina propria lymphocytes. It was then observed than the peripheral cellular immune response was earlier and stronger, while the local cellular immune response in digestive tract as shorter when chitosan was used as adjuvant. On the other hand, the chitosan had no effect on the systemic, lachrymal and digestive antibody-mediated immunity. This study indicates thus that the chitosan is a cell-promising adjuvant for the mucosal delivery of live vaccine in poultry, by enhancing the Th1 pathway of immunity. However, further investigations are required to explore its mechanism of action and to evaluate the inferred protection.

Improved vaccination against Newcastle disease by an in ovo recombinant HVT-ND combined with an adjuvanted live vaccine at day-old.

The continuous outbreaks of fatal Newcastle disease (ND) in commercial poultry flocks demonstrate that current vaccination strategies are not fully efficacious and should be improved by new generation of vaccines. In this context, maternally immune conventional layer chickens were vaccinated in ovo with a turkey herpesvirus recombinant expressing the fusion (F) gene of NDV (rHVT-ND) and/or at day-old with an apathogenic enterotropic live ND vaccine co-administrated or not with chitosan by oculo-nasal route. The induced vaccinal immune responses and conferred protection against a challenge with a circulating NDV velogenic viscerotropic strain were evaluated. The innovative rHVT-ND/live ND-chitosan vaccination regimen provided the best protection against mortality and morbidity as well as the strongest reduction of virus shedding that could be related to the higher measured cellular immune response and digestive antibody-mediated immunity.