Characterization of trimethoprim- and sulphisoxazole-resistant Chlamydia trachomatis

Trimethoprim and sulphisoxazole were used as selective agents in culture to isolate, by a stepwise procedure, a series of Chlamydia trachomatis L2 populations resistant to the cytotoxic effects of the drugs. Two trimethoprim-resistant populations, L2TriR-60 and L2TriR-100, and one sulphonamide-resistant population, L2SulfR-100, were characterized in more detail. In addition to being resistant to trimethoprim, L2TriR-60 was cross-resistant to methotrexate, sensitive to sulphisoxazole and displayed a ribonucleotide auxotrophy similar to that of its parental wild type, C. trachomatis L2. Surprisingly, L2TriR-100 and L2SulfR-100 appeared phenotypically identical. Both mutants were highly resistant to trimethoprim, sulphisoxazole, and methotrexate. In contrast to wild-type C. trachomatis L2, these populations were sensitive to 5-fluorouracil. L2TriR-100 and L2SulfR-100 were incapable of taking pyrimidine ribonucleotides from the host cell and no longer synthesized thymidine nucleotides de novo. The pyrimidine requirement of these mutants was met by salvaging host-cell uracil and thymidine, a property which can account for their drug-resistant characteristics. L2TriR-100 and L2SulfR-100 could also salvage adenine and guanine. Using L2TriR-100 as a starting stock, a mutant population resistant to the cytotoxic effects of trimethoprim and 5-fluorouracil (L2Tri/5-FU) was selected. L2Tri/5-FU was resistant to 5-fluorouracil because it had regained the capacity to take pyrimidine ribonucleotides from the host cell.