A Method for the Analysis of a Large Number of Specific and Multispecific B Cell Hybridomas Derived from Primary Immunized Lymph Nodes

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Abstract
A previously described efficient short term immunization protocol enables analysis of the specificities of large numbers of monoclonal antibodies in murine lymph nodes. In this paper we measure the B cell responses against two highly immunogenic antigens, rat type II collagen (NRC) and chick ovalbumin (OVA), as well as against the poorly immunogenic denatured rat type II collagen (DRC). We found that relatively large numbers of specific B cell hybridomas could be produced after immunization with NRC (28 %) and lower numbers with OVA (6 %) and DRC (3%). However, in all immunizations there were also generated large numbers of hybridomas producing multispecific antibodies (5-15%). The multispecificity was sustained when the hybridomas were subcloned. These results shew that large numbers of immunogen-reactive hybridomas can easily be obtained with the present method. They also show that different immunogens differ in their immunogenicity. The very high efficiency of native type II collagen to induce an antibody response, which is widely crossreactive with autologous type II collagen, could possibly be explained if it is assumed that the natural occurring repertoire is based on stimulation of autoantigens.