Mapping of T-Helper Epitopes of Rinderpest Virus Hemagglutinin Protein

Rinderpest virus (RPV) is a highly contagious and often fatal disease of domestic and wild ruminants, caused by rinderpest virus of the genus Morbillivirus under the family Paramyxoviridae. Hemagglutinin (H) and fusion (F) proteins of this enveloped virus confer protective immunity against experimental challenge with virulent rinderpest virus. We have earlier demonstrated that immunization with a single dose of recombinant extracellular baculovirus expressing H protein elicits H-specific humoral and lymphoproliferative responses in cattle. The lymphoproliferative responses are predominantly BoLA class II restricted. In this work, we have analyzed lymphoproliferative responses of peripheral lymphocytes from immunized cattle to truncated H protein fragments expressed in E. coli for locating domains harboring T epitopes. One region (aa 113-182) recognized by immune $T_h$ cells is conserved in the H protein of measles virus, which was earlier shown to contain a dominant $T_h$ epitope in mouse. Synthetic peptides within this region of measles virus H protein were used to identify a $T_h$ epitope conserved in the H protein of RPV virus (aa 123-137) in cattle. A second $T_h$ epitope located at the C-terminus of RPV-H was mapped to the region corresponding to aa 512-609 using truncated protein fragments expressed in E. coli. The C-terminal epitope (aa 575-583) was mapped using synthetic peptides corresponding to measles virus H as well as RPV-H protein.