Monoclonal antibodies to the glycoprotein of vesicular stomatitis virus: comparative neutralizing activity

Abstract

Nineteen independently isolated hybridomas producing monoclonal antibodies to the glycoprotein of vesicular stomatitis virus were isolated and studied for their capacity to neutralize viral infectivity. By measuring competitive binding of 125ilabeled monoclonal antibodies in a radioimmunoassay, 11 different, non-crossreacting antigenic determinants were identified on the vesicular stomatitis virus G protein. All monoclonal antibodies reacting with determinants 1, 2, 3, and 4 resulted in viral neutralization, whereas those binding to the other seven determinants did not neutralize infectivity. A mixture of two monoclonal antibodies binding to different determinants resulted in a more rapid neutralization than either antibody alone, suggesting that different antibodies can exert a synergistic effect on viral neutralization. Kinetic experiments revealed biphasic neutralization curves similar to those expected for heterologous antibody. No evidence could be obtained to relate biphasic kinetics of viral neutralization to heterogeneous populations either of antibody molecules or of virus. The possible significance of the kinetic data with monoclonal antibodies is discussed.

Identification of epitopes associated with different biological activities on the glycoprotein of vesicular stomatitis virus by use of monoclonal antibodies

“…The M A b s assigned to four epitopes (Ep 2, Ep 4, Ep 5, and Ep 7) had VSV-neutralizing activity, although of varying efficiency. Previous reports also demonstrated the same number of neutralizing epitopes on G protein of VSV-Indiana [14,36].…”

“…a 1A7 (Ep 1); b P2F9 (Epl); e VllA2 (Ep2); d V17E8 (Ep2); e 2B9 (Ep 3 a); f V20B12 (Ep 3 b); g 3F4 (Ep 4); h 5El 1 (Ep 4); i 5C6 (Ep 5); j P2E11 (Ep 6); k V12B3 (Ep 6); l P2F3 (Ep 7); m Ig G1 (control); n Ig G2a (control); o MAb free; p mock. Inhibition of fusion was observed in g, h, i, and 1 marized in Table 3 b + and -HI titers against 4 H A U of >t 32 and ~< 16, respectively, given in the 4th column of Table 3 c _ No inhibition or slight inhibition of adsorption (65-125%); e enhancement of adsorption (~> 135%) in the virus binding assay shown in [36] and Le-Francois and Lyles [14,15], w h o d e m o n s t r a t e d 11 and 10 epitopes on G protein of VSV-Indiana, respectively. The enhancement of binding was f o u n d also by LeFrancois and Lyles [14,15].…”

“…One approach to the structure-function relationship of surface glycoproteins of viruses is to analyze for the sites and effects of the monoclonal antibody (MAb) binding [4,13,31,33]. Production of MAbs against G proteins of two major serotypes of VSV (Indiana and New Jersey) has been reported by two research groups [5,14,15,36]. These MAbs have mainly been used to map neutralization and non-neutralization epitopes on G protein and to analyze the mutation leading to antigenic variations of G protein [8,11,[16][17][18]35].…”

Topographical mapping of epitopes on the glycoproteins of murine hepatitis virus-4 (strain JHM): Correlation with biological activities

“…Binding curves for these nonneutralizing antibodies were similar to those of the neutralizing anti-bodies suggesting that failure to neutralize was not due to gross differences in titer or binding avidity. Nonneutralizing antibodies to viral attachment proteins have now been demonstrated for the retroviruses (Massey and Schochetman, 1981;Bruck et ak, 1982), togaviruses (Roehrig et a& 1980;Kimura-Kuroda and Yasui, 1983), orthomyxoviruses (Breschkin et al, 1981;Carter et d, 1982), rhabdoviruses (Volk et aL, 1982), and reoviruses (Burstin et &, 1982). We found that antibodies to El or E2 which failed to neutralize in vitro also did not alter the disease process in tivo (Buchmeier et al, 1984), in contrast to nonneutralizing MAb to Sindbis virus which prevented lethal encephalitis in viva (Schmaljohn et uk, 1982).…”

Mapping of a region of dengue virus type-2 glycoprotein required for binding by a neutralizing monoclonal antibody

“…One approach involves the use ofmAbs as specific reagents for defining single epitopes on the complex antigenic structure of protein molecules. Using methods involving screening of purified proteolytic fragments of a protein molecule and/or screening a collection of overlapping synthetic peptides, the antigenic determinants of a number of viral glycoproteins have been studied (Emini et al, 1982;Lubeck and Gerhard, 1982;Massey and Schochetman, 1981;Mehra et al, 1986;Roehrig et al, 1982;Volk et al, 1982). Using RNA recombination as a genetic tool, the antigenic determinants involved in neutralization, as well as virus neuropathogenicity of the peplomer protein encoded by gene C of murine coronavirus were localized at the C-terminal one third of the peplomer (Makino et al, 1987).…”