Immunological Relationships Between Homologous Structural Polypeptides of Measles and Canine Distemper Virus

Abstract

SUMMARYThe major structural polypeptides of measles and canine distemper virus (CDV) were isolated by SDS-polyacrylamide slab gel electrophoresis. Rabbit hyperimmune sera directed against the isolated HA, P, NP, F and M polypeptides were prepared. In addition, rabbit hyperimmune sera directed against purified native internal components of measles virus and against purified virions of measles and CDV, untreated or treated with trypsin, were prepared. These sera were used to study the immunological relationships between measles and CDV in immune precipitation tests with aSS-methionine-labelled extracellular virions and intracellular virus polypeptides and by use of complement fixation and mixed haemadsorption tests.An immunological relationship between the M polypeptides was demonstrated with sera against both native and partially denatured antigens. A reciprocal immunological cross-reactivity between the NP polypeptides was demonstrated by the use of sera directed against native antigens. However, antisera directed against the NP polypeptides isolated from gels did not show any cross-reaction, indicating that they lacked antibodies against the shared antigenic determinants. The immunological cross-reactivity between envelope components of the two viruses was analysed. A close reciprocal immunological relationship between the F polypeptides was demonstrated by immune precipitation of labelled polypeptides. Immunological cross2reactivity between the HA polypeptides of the two viruses could not be demonstrated by use of the same technique neither with sera directed against denatured antigen nor with sera against native antigen when extracellular denatured 3~S-methionine-labelled purified virions were used in immune precipitation. A slight cross-reactivity could, however, be demonstrated with sera against native antigens in immune precipitation tests with aSS-methioninelabelled intracellular antigens.It is concluded that all the major structural polypeptides of measles and CDV show varying degrees of reciprocal immunological cross-reactions. The M polypeptides of the two viruses appear most closely related, whereas the HA polypeptides show the most pronounced differences.

Immunological relationships between phocid and canine distemper virus studied with monoclonal antibodies

“…Vero cell-grown purified virions, from mink passage 4a (Blixenkrone-M611er et al, 1989) of PDV, at a protein concentration of 0.5 mg to 1.0 mg per ml, were treated with 1 of the non-ionic detergent octyl-fl-D-glucopyranoside (octyl glycoside, Calbiochem-Behring) for 15 min at room temperature. After the treatment, two consecutive ultracentrifugations were performed, and the superuatant, which contained purified H and F proteins, was used for immunization (Orvell & Norrby, 1980). The proceciure for immunization of BALB/c mice and for establishment of mouse hybridoma cell lines have been previously described (0rvell & Grandien, 1982), with the exception that the fusions were performed with the myeloma cell line SP2/0.…”

“…The samples were processed as described previously (Orvell & Norrby, 1980). The details of the procedures for polyacrylamide gel electrophoresis and scintillation autofluorography have been described previously (Orvell, 1978).…”

“…[35S]Methionine-labelled purified virions, prepared as previously described, were used as the antigen in RIPA (Orvell & Norrby, 1980). The extracellular [35S]meth.…”

Characterization of Four Parainfluenza Virus Type 3 Proteins by Use of Monoclonal Antibodies

“…Extracellular virions (ECV) were pelleted from the clarified supernatant in the same centrifuge at 27000 g for 90 min. Subsequently, they were purified on a discontinuous sucrose gradient (30/65~) and the interphase was collected, resuspended and pelleted as previously described (Orvell & Norrby, 1980).…”

Immunoassays for Measles Virus Nucleocapsid Antigen: Effect of Antigen-Antibody Complexes

“…2 shows, the less-related mumps and parainfluenza 2 nucleocapsid antigens did not react at all in concentrations at least 10 000 times higher than measles virus nucleocapsid. Although distemper virus is relatively closely related to measles virus and there is clear cross-reactivity between their nucleocapsids (0rvell & Norrby, 1980), a more than 1000-fold greater reactivity was seen here in the homologous system. Viral antigens can be released from infected cells by different treatments including detergents.…”