Protective immunity against murine hepatitis virus (MHV) induced by intranasal or subcutaneous administration of hybrids of tobacco mosaic virus thatcarries an MHV epitope

Hybrids of tobacco mosaic virus (TMV) were constructed with the use of fusi on to the coat protein peptides of 10 or 15 amino acids, containing the 5B1 9 epitope from the spike protein of murine hepatitis virus (MHV) and giving rise to TMV-5B19 and TMV-5B19L, respectively, The TMV hybrids were propaga ted in tobacco plants, and the virus particles were purified. Immunogold la beling, with the use of the monoclonal MAb5B19 antibody, showed specific de coration of hybrid TMV particles, confirming the expression and display of the MHV epitope on the surface of the TMV. Mice were immunized with purifie d hybrid viruses after several regimens of immunization. Mice that received TMV-5B19L intranasally developed serum IgG and IgA specific for the 5B19 e pitope add for the TMV coat protein. Hybrid TMV-5B19, administered by subcu taneous injections, elicited high titers of serum IgG that was specific for the 5B19 epitope and for coat protein, but IgA that was specific against 5 B19 was not observed. Mice that were immunized with hybrid virus by subcuta neous or intranasal routes of administration survived challenge with a leth al dose (10 x LD50) of MHV strain JHM, whereas mice administered wild-type TMV died 10 d post challenge. Furthermore, there was a positive correlation between the dose of administered immunogen and protection against MHV infe ction. These studies show that TMV can be an effective vaccine delivery veh icle for parenteral and mucosal immunization and for protection from challe nge with viral infection.