Theileria-infected cell line from an African buffalo (Syncerus caffer)

• DOI: 10.1007/s00436-009-1467-0
• Publication Date: 2009-05-08T07:01:23Z
• Authors: Erich Zweygarth; Otto Koekemoer; Antoinette I. Jos...; Natasha Rambritch; Ronel Pienaar; John Putterill; Abdalla Latif; Fred T. Potgieter
• Keywords: Theileria; Subculture (biology); Giemsa stain

Mononuclear cells were isolated from the peripheral blood of a buffalo infected with a Theileria sp. using density gradient centrifugation, and the cells were put into culture flasks covered by a monolayer of bovine endothelial cells. Twenty days after culture initiation, cells containing macroschizonts were detected in Giemsa-stained smears. The first subculture was carried out on day 45 of culture propagation. Subsequently, infected cells were subcultured twice a week, and each time 1 to 2 x 10(6) per milliliter cells were harvested. DNA was extracted from culture material and a partial polymerase chain reaction amplification of the 18S ribosomal RNA (rRNA) gene was carried out using Theileria genus-specific primers. Sequence data and phylogenetic analysis using the 18S rRNA gene indicated a close relationship to Theileria sp. buffalo, previously described in literature. Here, the first successful attempt to establish a macroschizont-infected lymphoblastoid cell line of Theileria sp. (buffalo) from an African buffalo is described.