A targeted mutation within the feline leukemia virus (FeLV) envelope protein immunosuppressive domain to improve a canarypox virus-vectored FeLV vaccine.

We previously delineated a highly conserved immunosuppressive (IS) domain within murine and primate retroviral envelope proteins that is critical for virus propagation in vivo. The envelope-mediated immunosuppression was assessed by the ability of the proteins, when expressed by allogeneic tumor cells normally rejected by engrafted mice, to allow these cells to escape, at least transiently, immune rejection. Using this approach, we identified key residues whose mutation (i) specifically abolishes immunosuppressive activity without affecting the "mechanical" function of the envelope protein and (ii) significantly enhances humoral and cellular immune responses elicited against the virus. The objective of this work was to study the immunosuppressive activity of the envelope protein (p15E) of feline leukemia virus (FeLV) and evaluate the effect of its abolition on the efficacy of a vaccine against FeLV. Here we demonstrate that the FeLV envelope protein is immunosuppressive in vivo and that this immunosuppressive activity can be "switched off" by targeted mutation of a specific amino acid. As a result of the introduction of the mutated envelope sequence into a previously well characterized canarypox virus-vectored vaccine (ALVAC-FeLV), the frequency of vaccine-induced FeLV-specific gamma interferon (IFN-γ)-producing cells was increased, whereas conversely, the frequency of vaccine-induced FeLV-specific interleukin-10 (IL-10)-producing cells was reduced. This shift in the IFN-γ/IL-10 response was associated with a higher efficacy of ALVAC-FeLV against FeLV infection. This study demonstrates that FeLV p15E is immunosuppressive in vivo, that the immunosuppressive domain of p15E can modulate the FeLV-specific immune response, and that the efficacy of FeLV vaccines can be enhanced by inhibiting the immunosuppressive activity of the IS domain through an appropriate mutation.

Cross-clade immunity in cats vaccinated with a canarypox-vectored avian influenza vaccine.

Several felid species have been shown to be susceptible to infection with highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype. Infection of felids by H5N1 HPAI virus is often fatal, and cat-to-cat transmission has been documented. Domestic cats may then be involved in the transmission of infection to other animals but also to humans. A particular concern is the hypothetical role of the cat in the adaptation of the virus to mammalian species, thus increasing the pandemic risk. Therefore, the development of a HPAI vaccine for domestic cats should be considered a veterinary and also a public health priority. Here we show that vaccination of cats with a recombinant canarypox (ALVAC)(1)) virus, expressing the hemagglutinin (HA) of influenza virus A/chicken/Indonesia/03 (H5N1) confers protection against challenge infection with two antigenically distinct H5N1 virus isolates from humans. Despite low hemagglutination inhibiting (HI) antibody titers at the time of challenge, all vaccinated cats were protected against mortality and had reduced histopathological changes in the lungs. Importantly, viral shedding was reduced in vaccinated cats as compared to controls, suggesting that vaccination of cats could reduce the risk of viral transmission. In conclusion this study showed that the recombinant canarypox virus protected cats against homologous and heterologous H5N1 HPAI virus challenges.