Peyers patches of rats

In rats, cells from immune lymph nodes are an excellent source of specific B cells and we use routinely cells taken from the mesenteric nodes of animals immunized via the Peyer s patches. As with spleen cells, the animals are re-challenged via the Peyer s patches three days before removing the mesenteric nodes for fusion. Using this route for immunization we have successfully prepared rat x rat IgA-secreting hybridomas following a short immunization schedule whereas hyperimmunization yielded high affinity IgG antibodies (14). 

Generation of Immune spleen or lymph node cells  

Soluble proteins or conjugated peptides emulsified in FCA or FIA (see Protocols 1 and 2) 

Anaesthetize the animals and take a blood sample from the tail vein (mice) or jugular (rats) into a capped 0.5 ml or 1.5 ml microcentrifuge tube to act as a pre-immune sample. Allow to clot, centrifuge (1500 g), remove serum, and store at -20 C. 

For fusions that will use the mesenteric nodes of rats, the antigens are injected into the Peyer s patches that lie along the small intestine. The surgical procedure is described in Protocol 4. 

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