The effect of mycoplasma and mycoplasma removal agent on the hydrolase activity in fibroblasts of patients with lysosomal diseases

Abstract

Este estudio fue disenado para evaluar el efecto de la contaminacion por micoplasmas sobre la actividad de hidrolasas acidas y la accion del agente de eliminacion de micoplasmas (MRA) en cultivos de fibroblastos humanos de pacientes con enfermedades lisosomales. Se midio la actividad de la b-galactosidasa, arilsulfatasa B (ASB), hexosaminidasa A y a-glucosidasa en estos cultivos. La actividad de estas enzimas en los fibroblastos contaminados por micoplasmas se midio antes y despues de la adicion de MRA. Los resultados se compararon con los obtenidos en cultivos libres de contaminacion. Solo la enzima ASB demostro alteracion significativa en la actividad, tanto en presencia de micoplasmas como con la adicion de MRA. Las enzimas restantes no sufrieron alteraciones significativas en presencia de micoplasmas, ni tras la adicion de MRA. La actividad medida para la enzima ASB aumento significativamente en presencia de micoplasmas y MRA, lo que podria conducir a un diagnostico dudoso. Por lo tanto, sugerimos evitar la contaminacion con micoplasmas mediante el uso de tecnicas asepticas y la utilizacion de MRA en los cultivos de fibroblastos que no se puedan descartar.

This study was designed to evaluate the effect of mycoplasma contamination on acid hydrolase activity and the action of the mycoplasma removal agent (MRA), in cultures of human fibroblasts from individuals with lysosomal diseases. For this purpose, we measured the activity of the b-galactosidase, arylsulphatase B (ASB), hexosaminidase A and a-glucosidase enzymes. The activity of the above mentioned enzymes in fibroblasts contaminated by mycoplasma was measured before and after the addition of the MRA. The results were then compared to the enzymatic activity in contamination-free cultures. Only the ASB enzyme showed significant alteration in activity both in the presence of mycoplasma and MRA. The remaining enzymes did not suffer significant interference by the presence of the two agents. Of the four enzymes tested, three did not suffer significant alterations by the presence of the mycoplasma nor from the MRA. However, the activity measured in the ASB enzyme increased significantly in the presence of mycoplasma and MRA and could lead to a doubtful diagnosis. Therefore, we suggest that contamination should be prevented by using aseptic techniques as well as the MRA in those fibroblast cultures that cannot be discarded.