Abstract
We have analysed the suppressive effect of cyclophosphamide, when given in vivo, on the antibody response of BALB/c mice against Shigella dysenteriae antigens using ELISA and immunoblot. Out of various protocols tried, it was found that co-injection of cyclophosphamide at 150 mg/kg bodyweight, i.p., at the time of antigen administration and then after a lapse of 24 h during both primary and secondary immunizations, was the most effective in suppressing antibody response of mice. Analysis of sera by ELISA demonstrated the presence of some antibodies to S. dysenteriae antigens after secondary immunization, but immunoblot analysis using the same sera revealed complete suppression of antibody response. Animals whose antibody response was almost completely suppressed after two immunizations with co-injection of cyclophosphamide, when immunized again after the lapse of 14 days from the date of secondary immunization with Shigella antigens but without administration of cyclophosphamide, partially recovered their ability to respond to the same antigens. This protocol can now be used in mice to analyse the hierarchy of immunogenic epitopes present in a complex mixture of antigens.
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