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

Abstract

SUMMARYMonoclonal antibodies directed against four structural components of the ATCC strain C243 of parainftuenza virus type 3 were produced. The specific reaction of the antibodies with individual structural components was determined by radioimmune precipitation assay. In the collection of monoclonal antibodies, 21 reacted with the haemagglutinin-neuraminidase (HN) glycoprotein (mol. wt. 72000), eight with the fusion (F) glycoprotein (mol. wt. 64000), 27 with the nucleocapsid (NP) protein (mol. wt. 69000) and 24 with the matrix (M) protein (tool. wt. 40000). The F-specific monoclonal antibodies precipitated two proteins which were interpreted to represent intact F protein and the large cleavage product F1 (mol. wt. 52000). The numbers of epitopes were determined in a competition ELISA with the monoclonal antibodies. The epitopes found were six for the HN, two for the F, six for the NP and six for the M protein. The six groups of antibodies reacting with different epitopes on the HN molecule showed varying capacities to inhibit biological activities. Two exhibited high neutralization (NT), haemagglutination inhibition (HI) and haemolysis inhibition (HLI) activity. Three groups had somewhat lower NT, lower HI and no detectable HLI activity. One group showed no activity in these tests. Of the eight monoclonal antibodies directed to the F protein two had demonstrable HLI activity.

The structural proteins of a porcine paramyxovirus (LPMV)

“…This was carried out essentially as described by Rydbeck et al (1986). Mock-infected cells, LPMVinfected cells and purified virus, labelled as above, were disrupted on ice in L buffer (0-02M-potassium phosphate, 0-1 M-NaC1, 0-5% deoxycholate 1 ~ Triton X-100, 0-1% SDS and 1 mM-phenylmethylsulphonyl fluoride in 0-01 M-Tris-HC1 pH 7-8) for 2 h, followed by centrifugation at 50000g for 10 rain.…”

“…methionine was done essentially as described by Rydbeck et al (1986). Confluent IBRS cells grown in Eagle's MEM were infected with LPMV at an m.o.i, of 5 to 10 p.f.u./cell.…”

Antigenic Analysis of Human and Bovine Parainfluenza Virus Type 3 Strains with Monoclonal Antibodies

“…band in (a), lane 7, is interpreted to be a dimer of the HN protein. t Epitope group determined by competition ELISA (Rydbeck et al, 1986b).…”

Antigenic variation of human and bovine parainfluenza virus type 3 strains

“…Two different strains have been identified (Abinanti et al, 1961), one human (Chanock et al, 1958) and one bovine (Reisinger et al, 1959) both causing respiratory tract disease (Wenzel et al, 1972;Frank & Marshall, 1973). PIV3 consists of at least six structural proteins, the large (L), haemagglutinin-neuraminidase (HN), fusion (F), phospho (P), nucleocapsid (N), and matrix (M) proteins (Galinski et al, 1986;Rydbeck et al, 1986;Sanchez & Banerjee, 1985;Shibuta et al, 1979). Human and bovine PIV3 share neutralizing epitopes but show distinct antigenic properties (Coelingh et al, 1986;Glezen et al, 1984;Muchmore et al, 1981).…”