TNP-enzyme conjugates for the detection of anti-TNP antibody producing cells in vivo.

After injection of TNP-KLH in mice and TNP-BGG-PEN in rabbits, anti-TNP antibody-forming cells were observed in the spleen. When cryostat sections of the stimulated spleen were incubated with TNP-HRP conjugate, and then treated for HRP cytochemistry, the cytoplasm of anti-TNP-forming cells was stained red. When similar sections were incubated with TNP-AP conjugate, and then treated for AP cytochemistry, the cytoplasm of anti-TNP-forming cells was stained blue. After simultaneous incubation with TNP-AP conjugate and PEN-HSA-HRP conjugate, followed by treatment for HRP cytochemistry and AP cytochemistry, anti-TNP-forming cells with a blue cytoplasm and anti-PEN-forming cells with a red cytoplasm could be distinguished in the same spleen section.