PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST AVIAN REOVIRUS STRAIN S1133

Monoclonal antibodies (MAbs) were produced against the avian reovirus strain S1133. Mabs were isotyped and used to develop diagnostic tests. Splenocytes from immunized mice were screened by enzyme-linked immuno sorbent assay (ELISA). Two hybridomas secreted MAbs against avian reov irus S1133. One MAb secreted IgG1 and the other secreted IgG2a. All MA b light chains were kappa. Specificity of MAbs was rested against four avian reovirus strains: S1133, 1733, CO8, and 2408. Strains S1133, 17 33, and 2408 viruses were in the same subtype; the CO8 virus belonged to a different subtype. The MAbs reacted with all reovirus strains by ELISA, dot blot, immunofluorescence assay, and immunoblotting. No MAb had neutralizing activity against the rested reoviruses. Immunoblot an alysis showed the one MAb bound to protein sigma A with molecular weig ht of 39,000 Daltons for all reovirus strains. Another MAb bound to th e protein sigma C with an approximate molecular mass of 32,000 Daltons . An indirect immunoperoxidase (IF) procedure was developed using a MA b to detect reovirus in formalin-fixed, paraffin-embedded tissues from infected chickens and chicken embryo fibroblast cell cultures. The IP test was simple, fast, and economical and enabled simultaneous evalua tion of viral antigen-producing cells with tissue pathologic changes c onfirming that the reovirus caused the lesions.