Infectivity of wild-type rubella virus in fibrochondrocyte cells

Abstract

This study describes the rapid growth of the rubella virus in samples of a primary fibrochondrocyte cell culture with the development of a cytopathic effect (CPE), in response to infection by the rubella virus. The cells were isolated from the meniscus joint of a rabbit after enzymatic extraction and incubated at 37°C with a Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% fetal calf serum. A total of six clinical samples from urine, blood and cerebrospinal fluid were inoculated in the fibrochondrocyte and the cell lines of the African green monkey kidney -ATCC CCL-81 (Vero). The fibrochondrocyte cell showed CPE after 24 hours and virus growth was confirmed by immunohistochemistry and Nested PCR. The cells inoculated with samples were examined by phase contrast microscopy and showed characteristic rounding, along with bipolar and multipolar cells. The infection curve increased during the five days of observation, showing that the titers in fibrochondrocyte cells were then higher than those observed in Vero cell lines. Fibrochondrocytes Rubella virus Isolation resumoEste trabalho descreve o rápido crescimento do vírus da rubéola em amostras clínicas em cultura primária de fibrocondrócitos com desenvolvimento de efeito citopático em resposta a infecção pelo vírus da rubéola. As células foram isoladas do menisco do joelho do coelho após extração enzimática e incubadas a 37°C em meio DMEM suplementado com 10% de soro fetal bovino. Seis amostras clínicas de urina, sangue e liquor foram inoculadas na cultura primária de fibrocondrócitos e na linhagem Vero. As células de fibrocondrócitos apresentaram efeito citopático após 24 horas de incubação e o vírus foi detectado por imunoistoquímica e Nested PCR. As células infectadas apresentaram aspecto arredondado, com formação de alguns prolongamentos citoplasmáticos e sincícios, produzindo células multinucleadas. A curva de crescimento da infecciosidade do vírus foi mais alta quando comparada com a linhagem celular Vero. Fibrocondrócitos Vírus da rubéola Isolamento unitermos y key words ARTIGO ORIGINAL

Application of new assays for rapid confirmation and genotyping of isolates of Rubella virus

“…Although virus isolation is time‐consuming, labor‐intensive, and not particularly sensitive, it is still recommended by the WHO [2007] for studying the etiology and epidemiology of rubella. RV can be cultured in a variety of cell lines, such as RK13, SIRC, BHK, and Vero (Vero/SLAM) cells, but serial propagation can be established only in a limited number of cell lines [Figueiredo et al, 2004]. Previous studies have shown that specific CPE may appear in RK13 cells under controlled conditions, or in RK13 and SIRC cells after one or two passages in Vero cells [Figueiredo et al, 2000; Best et al, 2005].…”