Efficient Anti-Idiotypic Immunization with Homologous, Virus Neutralizing Monoclonal Antibodies Conjugated with KLH and Combined with Quil A
Recently, we described an enzyme immunoassay (EIA) on cell cultures for the rapid demonstration of Semliki Forest virus (SFV) neutralizing antibodies in serum (Tiel et al, 1986). In that assay, the spike proteins of SFV, either E1 or E2, can be detected in infected L cell monolayers with a horse radish peroxidase (HRPO-) labeled anti-E1 or anti-E2 specific monoclonal antibody (MA). Preincubation of the virus inoculum with SFV neutralizing serum reduces virus multiplication and thereby the appearance of the spike proteins on the surface of the infected L cells which are detected by inhibition of absorbance in the EIA. In Fig. 1, the strong neutralizing capacity of MA UM 5.1 (IgG2a) is demonstrated by this assay. The EIA is very useful for the determination of homologous anti-idiotypic activity in sera of mice. Preincubation of a critically neutralizing dose of MA, e.g. -10log dilution 5.5 of MA UM 5.1 (Fig. 1), with anti-idiotypic immune serum leads to a diminished capacity of this MA to neutralize and thereby to a rise in absorbance value in the EIA. The anti-idiotypic response was provoked in (female) BALB/c mice. The animals were injected intra/subcutaneously with a mixture of the saponin Quil A and protein A Sepharose-purified MA conjugated with keyhole limpet haemocyanin (KLH).
KeywordsAscitic Fluid Semliki Forest Virus Booster Injection Spike Protein Normal Mouse Serum
Unable to display preview. Download preview PDF.
- Bona, C., Hooghe, R., Cazenave, D.A., Leguern, C. and Paxil, W.E., 1979, Cellular basis of regulation of expression of idiotype. II. Immunity to anti-MOPC-460 idiotype antibodies increases the level of anti-Trini trophenyl antibodies bearing 460 idiotype, J.exp.Med., 149:815.PubMedCrossRefGoogle Scholar
- Choi, K.L. and Sauder, D.N., 1986, The role of Langerhans cells and keratinocytes in epidermal immunity, J.of Leukocyte Biol., 39:343.Google Scholar
- Jerne, N.K., 1974, Towards a network theory of the immune system, Ann.Inst.Pasteur Immunol., 125C:373.Google Scholar
- Tiel, F.H. van, Harmsen, T., Wagenaar, M., Boere, W.A.M., Kraaijeveld, C.A. and Snippe, H., 1986, Rapid determination of neutralizing antibodies in serum by enzyme immunoassay in cell culture using virus specific peroxidase-labeled monoclonal antibodies, J.of Clin. Microbiol., 24:665.Google Scholar
- Urbain, J., Brait, M., Bruyns, C., Demeur, C., Dubois, P., Francotte, M., Franssen, J.D., Hiernaux, J., Leo, O., Marvel, J., Meyers, P., Moser, M., Slaoui, M., Tassignon, J., Urbain-Vansanten, G., Van-Acker, A., Wikler, M., Willems, F. and Wuilmart, C., 1985, The idiotypic network: Order from the beginning or order out of chaos? Current Topics in Microbiol, and Immunobiol., 119:127.CrossRefGoogle Scholar