Anti-idiotypic antibodies to bovine herpesvirus-1 inhibit virus infection in cell cultures

Abstract

A panel of murine monoclonal antibodies (MAbs) to bovine herpesvirus-1 (BHV-1) was prepared. Three of them were neutralizing MAbs and reacted against 130/75/50 kDa, 77 kDa, or 97 kDa glycoproteins (gp). A fourth non-neutralizing MAb recognized the 97 kDa gp. Competition radioimmunoassay demonstrated that each of the four MAbs reacted against a different virus epitope. Anti-idiotypic antibodies (anti-id) to the four MAbs were produced in rabbits and purified by sequential immunoaffinity chromatography. Each anti-id inhibited the binding of its respective MAb to BHV-1 in competitive ELISA and blocked BHV-1 neutralizing activity of the MAb. This inhibition suggested that the anti-ids were specific for the antigen binding site of the MAbs. Treatment of MDBK cells with anti-ids inhibited BHV-1 infection, which suggested that the anti-ids block a cellular component essential for virus infection. Absence of significant cross-reactivity among the anti-ids for heterologous MAbs indicated that they recognized unique determinants on the antigen binding site of the homologous MAb.

Comparison of Bovine Immune Responses to Affinity-Purified Bovine Herpesvirus-1 Antiidiotypes and Glycoproteins

“…In the present experiments, we have shown that anti-Id immunizations did not protect calves from BHV-1 challenge. Several characteristics of the anti-Id suggested that they could elicit a protective immune response, i.e., competition with BHV-1 in MAb binding or neutralization assays (1,30), inhibition of BHV-1 infection in tissue culture cells (1), and ability of one of the anti-Id (anti-R54) to elicit BHV-1-neutralizing antibodies in BALB/c mice (31). A polyclonal anti-Id preparation is expected to consist of a mixture of all types of Ab2 (7,19,24), including internal image anti-Id (Ab2ß).…”

“…Production (in BALB/c mice), purification, and characterization of gl and gIV-specific MAb (R54, 11 IB, and 83) used for anti-Id antibody production have been described (1,30). Recently, using immunoprecipitation and immunoblotting, we determined that R54 is specific for gIV rather than gill.…”

“…Sera were collected before immunization and at 3-7 day intervals during immunization and after challenge, heat inactivated at 56CC for 30 min, and stored at -20°C. Antibodies against BHV-1 and anti-Id were detected by ELISA and serum neutralization tests (1,30). Total glycoproteinspecific antibody production was assessed by ELISA using purified BHV-1 gl, gill, or gIV on the ELISA plates as previously described (37).…”

“…We have prepared and characterized rabbit polyclonal anti-Id reagents to BHV-1-neutralizing monoclonal antibodies (MAbs) to the three major BHV-1 envelope glycoproteins (1,30). Anti-Id antibodies to a BHV-1 -neutralizing MAb (R54) induced production of BHV-1 -neutralizing antibodies in BALB/c mice (31).…”

Enfeksiyöz Bovine Rhinotracheitis Virusun Tespitinde IBRV Suş SD Rekombinant Glikoprotein D Kullanılarak İndirekt ELISA Yönteminin Geliştirilmesi ve Uygulanması

“…The whole virus ELISA method experiences a risk of virus contamination although with a better antigenicity. The neutralization antibody against gD largely produced in the host has the best performance to antagonize IBRV, hence is favored as the top choice for manufacturing diagnostic reagents and subunit vaccines [11] . This study utilizes a prokaryotic system to express recombinant gD.…”