Purification, Morphology and Antigenic Characterization of Measles Virus Envelope Components

Abstract

SUMMARYMeasles virus haemagglutinin (H), fusion (F) and matrix (M) components were purified by affinity chromatography using monoclonal antibodies coupled to CNBractivated Sepharose. H and M proteins were purified to homogeneity as determined by polyacrylamide gel electrophoresis by a single cycle of adsorption-desorption. The corresponding purification of the F protein required two cycles of adsorptiondesorption. After the second adsorption an extra wash with 1 M-guanidine-HCl was employed to remove the contaminating cellular actin. Electron microscopic examination of the purified envelope proteins showed that, at pH 6.0, the H peplomers had a truncated conical shape (width 6.5 to 4 nm, length 16 nm), and the F peplomers had a club-like shape (dimensions of the oval head 6 x 9 nm, length 15 nm). Lengths of both peplomers were measured excluding their undiscernible hydrophobic part. The M component at pH 3-0 appeared as a rounded particle (diam. 8 nm, central accumulation of contrast 1.5 nm) suggested to include four to six M polypeptides. Rabbit hyperimmune sera were prepared against all three purified envelope components. These sera reacted only with the homologous antigen in radioimmunoprecipitation assays. Both antisera against the H and F components neutralized the virus and blocked virus-specific haemolysis, but only anti-H serum inhibited haemagglutination.

Measles Virus Fusion Protein Presented in an Immune-stimulating Complex (Iscom) Induces Haemolysis-inhibiting and Fusion-inhibiting Antibodies, Virus-specific T Cells and Protection in Mice

“…IAC purification of the F protein was carried out as previously described with minor modifications (Varsanyi et al, 1984;De Vries et al, 1987). Briefly, 2~ Triton X-100 was used to solubilize the membrane proteins of the concentrated virus suspension.…”

“…1), corresponding to the F~ component and the disulphide-linked F1-F2 complex of the F protein respectively (Hardwick & Bussell, 1978;Varsanyi et al, 1984). The protein observed at Mr l 10000 in the gel under non-reducing conditions is probably a dimer of the 55 000 Mr F protein, since it also reacted with monoclonal antibody F17-21 in a Western blotting experiment (not shown).…”

Haemagglutinin of Measles Virus: Purification and Storage with Preservation of Biological and Immunological Properties

“…It was also shown that the purified H glycoprotein can be stored for several days at -70 °C after lyophilization, and its biological activity (haemagglutination) and recognition by an anti-H monoclonal antibody specific for a conformational determinant can be fully conserved provided that H glycoprotein is associated with artificial membranes. The purification of measles H glycoprotein has been reported previously using lectin affinity in conjunction with gel filtration (Christie et al, 1981 ;Lund & Salmi, 1981) or immunoaffinity (Bellini et al, 1981;Varsanyi et al, 1984) chromatography. These techniques required the desorption of the material using high salt (3 M-KSCN), 8 M-urea, 1% sodium deoxycholate or low pH (0.2 M-glycine pH 3.0) solutions, i.e.…”

Immune responses in mice following immunization with chimeric synthetic peptides representing B and T cell epitopes of measles virus proteins

“…Antibodies to H inhibit haemagglutination by the virus and adsorption to host cells, and antibodies to F inhibit haemolysis and membrane fusion (Varsanyi et al, 1984;Merz et al, 1980). The importance of anti-F antibodies in vivo is highlighted by the development of atypical measles in recipients of a formalin-inactivated virus in whom no anti-F antibodies could be demonstrated (Norrby et al, 1975).…”