Determination of CD marker expression profiles of S15 vaccine strain Theileria annulata Schizont infected cell line using RT-PCR analysis

Document Type: Original Articles

Author

Department, Razi Vaccine and Serum Research Institute

Abstract

The aim of this study was to identify the cell surface markers “cluster of differentiation” (CD) of Theileria annulata schizont-infected cell lines. CD molecules are very useful for characterization of cells and the different subpopulations of leukocytes. They are usually recognized by specific antibodies by flow cytometry and immunohistochemistry. Here, we emphasize on application of RT-PCR for defining the cell surface markers for S15 vaccine strain T. annulata schizont infected cell line (attenuated) and a new laboratory established T. annulata schizont infected cell line (non-attenuated). The characterization of surface protein profiles of S15 vaccine cell line is important to determine the specific markers that can be used to exclude non attenuated cell lines. RT-PCR was carried out by specific designed primers using a panel of seven bovine CD markers as well as beta-actin as an internal control a house keeping gene. Here, we showed that both of the examined cell lines had a consistent expression for CD4, CD5, CD11a, CD14, CD43 and CD45 markers. But the specific finding in this study was the expression of B cell marker CD79a and CD5 by newly transformed cell line, whereas the CD5 a marker for a B cell subset was expressed by S15 vaccine strain. In conclusion, we consider CD79a surface protein to be a new marker for non-attenuated Theileria annulata schizont infected cell line but not in vaccine strain. In addition, identification of CD marker expression based on the RT-PCR assay might be a good and appropriate alternative technique for flow cytometry.

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