Peroxidase Immunocytochemistry for the Detection of Specific Anti-Hapten (Penicilloyl) Antibody Producing Cells in the Spleen, After Injection of a Hapten-Carrier Conjugate

Cells producing specific antibodies against the hapten penicilloyl (Pen) are demonstrated in tissue sections of the spleen, using peroxidase immunocytochemistry. Horseradish peroxidase-human serum albumin-penicilloyl (HRP-HSA-Pen) conjugate, prepared by coupling Pen to HRP-HSA conjugate, was used for detection of anti-Pen producing cells in the spleen after intravenous injection of bovine gamma globulin-penicilloyl (BGG-Pen) conjugate. HRP-HSA conjugate was used as a control to exclude the possibility that positive cells contained antibody against a common determinant in both HSA and BGG. The method may be applied for the detection of cells producing antibodies against other haptens as well.