Alcelaphine herpesvirus 1 genes A7 and A8 regulate viral spread and are essential for malignant catarrhal fever.

Author summary Gammaherpesvirus entry into immune cells can result in latent infection which is associated with viral persistence and severe lymphoproliferative diseases. Gammaherpesviruses enter target cells during primary infection via a complex machinery of envelope glycoproteins. Alcelaphine herpesvirus 1 (AlHV-1) is a gammaherpesvirus carried by wildebeests without causing any clinical sign but induces malignant catarrhal fever (MCF) upon transmission to several species of ruminants including cattle. MCF is a deadly lymphoproliferative disease developing after a prolonged incubation period. In the present study, we demonstrated that the genes A7 and A8 of AlHV-1 encode envelope glycoproteins that are orthologs of Epstein-Barr virus gp42 and gp350, which regulate cell tropism switch. Impairment of A7 or A8 expression in a pathogenic strain of AlHV-1 strongly altered viral propagation in vitro. We further showed using bovine respiratory cell lines in vitro that AlHV-1 uses A7 to mediate cell-to-cell spread whereas A8 is necessary for cell-free viral propagation. Then, infection of rabbits as an experimental model to induce MCF with recombinant viral strains demonstrated that both A7 and A8 are essential for the induction of MCF. Thus, this study highlights an essential role for gp42 and gp350 orthologs in the pathogenesis of a gammaherpesvirus-induced lymphoproliferative disease. Alcelaphine herpesvirus 1 (AlHV-1) is a gammaherpesvirus that is carried asymptomatically by wildebeest. Upon cross-species transmission to other ruminants, including domestic cattle, AlHV-1 induces malignant catarrhal fever (MCF), which is a fatal lymphoproliferative disease resulting from proliferation and uncontrolled activation of latently infected CD8(+) T cells. Two laboratory strains of AlHV-1 are used commonly in research: C500, which is pathogenic, and WC11, which has been attenuated by long-term maintenance in cell culture. The published genome sequence of a WC11 seed stock from a German laboratory revealed the deletion of two major regions. The sequence of a WC11 seed stock used in our laboratory also bears these deletions and, in addition, the duplication of an internal sequence in the terminal region. The larger of the two deletions has resulted in the absence of gene A7 and a large portion of gene A8. These genes are positional orthologs of the Epstein-Barr virus genes encoding envelope glycoproteins gp42 and gp350, respectively, which are involved in viral propagation and switching of cell tropism. To investigate the degree to which the absence of A7 and A8 participates in WC11 attenuation, recombinant viruses lacking these individual functions were generated in C500. Using bovine nasal turbinate and embryonic lung cell lines, increased cell-free viral propagation and impaired syncytia formation were observed in the absence of A7, whereas cell-free viral spread was inhibited in the absence of A8. Therefore, A7 appears to be involved in cell-to-cell viral spread, and A8 in viral cell-free propagation. Finally, infection of rabbits with either mutant did not induce the signs of MCF or the expansion of infected CD8(+) T cells. These results demonstrate that A7 and A8 are both essential for regulating viral spread and suggest that AlHV-1 requires both genes to efficiently spread in vivo and reach CD8(+) T lymphocytes and induce MCF.