Antibiotic Effect of Tylosin on a Mycoplasma Contaminant in A Tissue Culture Leukemia Cell Line.

Abstract

In view of the frequent contamination of tissue cultures with organisms of the pleuropneumonia group (PPLO), we are reporting the effectiveness of tylosin tartrate in elminating such a contaminant from our culture.During the attempt to plate in agar one of our lines of murine leukemia cells(l), we detected a filterable microorganism which may be related to Mycoplasma granu1arum.f Several chemotherapeutic and antibiotic agents, namely tetracyclin, kanamycin, erythromycin, chloromyce tin, polymixin, s tovarsol and iodine had been previously tested with uniformly negative results. Although the cultures frequently seemed ('cured" when examined immediately after treatment, PPLO reappeared in the subsequent passages.Materials and methods. Purina tylan soluble (tylosin tartrate), a product of Eli Lilly and Co., is distributed by Ralston Purina Co. Prior to the use of tylan in tissue cultures, it was tested with bacteriologic media for activity against M . pulmonis of murine infectious catarrh and M . granularurn, recovered from tissue cultures. These two species are readily cultivable in 20% horse serum-nutrient bouillon and 48 hours-old cultures were employed. Transfers (.OS ml) were made to serum-bouillon and serum-agar containing remectively 100, 50. and 2 5 pg/ml of tylan. The two higher concentrations completely suppressed growth of both mycoplasmas in both media, as did also the lowest one with M . pulmonis. Growth of M . granularum was not completely inhibited, however, in the presence of 25 pg/ml and a few atypical colonies aDpeared on serum-agar. A concentration of 50 pg/ml was used, therefore, as a triaI *

Spontaneous human lymphocyte-mediated cytotoxicity against tumor target cells

“…However, it is well established that the presence of EBV-determined antigens is not necessary for the target cell to be lysed. On the chance that the isolation technique was inadequate, the cell line was treated with the antimycoplasma agent Tylocine accord-H. F. Pross and M. G. Baines: A Review of SLMC ing to Friend et al (1966). It is also possible, however, that in the process of in vitro transformation, the cells have become contaminated with mycoplasma, or a virus against which normal donors are reactive.…”

Quantitative aspects ofMycoplasma pneumoniae-cell relationships in cultures of lung diploid fibroblasts

“…The feasibility of extending this model to the quantitative assay of antibiotics is discussed. According to recent observations the inhibitory effect of antibiotics may be strongly affected by the milieu in which mycoplasmas are perhaps protected by their close association with cells (Gori & Lee, 1964;Friend, Patuleia & Nelson, 1966). That cell attachment involves an active biological process on behalf of either cell or mycoplasma has recently been reported by Fogh & Fogh (1967), who have also convincingly demonstrated that quantitative models of mycoplasma-cell interactions may lend themselves to more meaningful studies of effects of antibiotics.…”

Characteristics of a New Sterol-nonrequiringMycoplasma

“…MATERIALS AND METHODS Mycoplasma. The S-410 and S-743 strains were recovered from two murine leukemia tissue culture cell lines (5,6) by Charlotte Friend. The cell cultures were grown in basal media (Eagle) with Earle's balanced salt solution, supplemented with 15% fetal calf serum.…”