Cultivation of Chlamydia trachomatis in cycloheximide-treated mccoy cells

Abstract

An isolation technique for Chlamydia trachomatis using McCoy cells is described. In contrast to earlier techniques employing such cells, no pretreatment of the cells was used. The glutarimide antibiotic cycloheximide was added to the culture medium used for incubating the cells after infection. Cycloheximide was used at concentrations that depressed, but did not completely inhibit, the metabolism of the eucaryotic host cells. In studies on different immunotypes of C. trachomatis cultured in the yolk sac of embryonated hen eggs, the cycloheximide technique was compared with a method using pretreatment of cells with 5-iodo-2-deoxyuridine. The cycloheximide method gave greater numbers of inclusion-forming units per cover slip for all the immunotypes of trachoma-inclusion conjunctivitis agents tested, i.e., A through I. In a study of 194 cervical and urethral specimens from women, cycloheximide treatment of McCoy cells was found to be more efficient than 5-iodo-2-deoxyuridine treatment for the isolation of C. trachomatis.