Determination of CD Markers Profile of the Cell Line Infected by S15 Vaccine Strain of Theileria annulata Schizont Using RT-PCR Analysis

Document Type: Short Communication

Authors

1 Department of wildlife research, Razi vaccine and serum research institute, agriculture research, education, and extension organization (AREEO), Karaj, Iran

2 Department of parasite vaccine research and production, Razi vaccine and serum research institute, agriculture research, education, and extension organization (AREEO), Karaj, Iran

3 Department of parasitology, Veterinary Faculty, Tehran University, Tehran, Iran

4 Department of biotechnology, Razi vaccine and serum research institute, agriculture research, education, and extension organization (AREEO), Karaj, Iran

5 Department of Parasite Vaccine Research and Production, Razi vaccine and serum research institute, agriculture research, education, and extension organization (AREEO), Karaj, Iran

Abstract

The aim of this study was to identify the cell surface cluster of differentiation (CD) markers of the cell lines infected by Theileria annulata schizont. The CD molecules are very useful for the characterization of cells and different subpopulations of leukocytes. They are usually recognized by specific antibodies using flow cytometry and immunohistochemistry. In the current study, we applied reverse transcriptase-polymerase chain reaction (RT-PCR) to define the profile of cell surface markers in a cell line infected by an attenuated S15 vaccine strain of T. annulata schizont and a new laboratory-established cell line infected by a non-attenuated form. In order to determine the specific markers that can be used for excluding the non-attenuated cell lines, the characterization of the surface proteins profile of the S15 vaccine cell line is important. The RT-PCR was carried out by specifically designed primers using a panel of seven bovine CD markers, as well as beta-actin as an internal control house-keeping gene. We showed that both of the examined cell lines had a consistent expression of CD4, CD5, CD11a, CD14, CD43, and CD45 markers. However, the specific finding in this study was the expression of B-cell markers CD79a and CD5 by the newly-transformed cell line. On the other hand, CD5 as a marker for B-cell subset was expressed by S15 vaccine strain. In conclusion, we consider CD79a surface protein as a new marker for the cell lines infected by non-attenuated T. annulata schizont, while the cell lines infected by the vaccine strain do not express this marker. In addition, the identification of CD marker expression based on the RT-PCR assay might be a suitable and appropriate alternative technique for flow cytometry.

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Main Subjects


Article Title [French]

Détermination des Profils d'Expression des Marqueurs CD de la Lignée Cellulaire Infectée par la Souche Vaccinale S15 de Theileria annulata Schizont à l'Aide d'une Analyse RT-PCR

Abstract [French]

Le but de cette étude était d'identifier les marqueurs de surface cellulaire de "cluster de différenciation" (CD) des lignées cellulaires infectées par le Theileria annulata schizont. Les molécules de CD sont très utiles pour la caractérisation des cellules et des différentes sous-populations de leucocytes. Elles sont généralement reconnues par cytométrie en flux et immunohistochimie en utilisant des anticorps spécifiques. Cette étude portait sur l'application de la RT-PCR pour définir les marqueurs de surface cellulaire de la lignée cellulaire infectée par la souche vaccinale S15 de Theileria annulata schizont (atténuée) et une nouvelle lignée produite en laboratoire infectée par T. annulata schizont (non atténuée). La caractérisation des profils protéiques de surface de la lignée cellulaire du vaccin S15 est importante pour déterminer les marqueurs spécifiques pouvant être utilisés pour exclure les lignées cellulaires non atténuées. La RT-PCR a été réalisée à l'aide d'amorces spécifiques conçues à l'aide d'un panel de sept marqueurs CD bovins ainsi que de la bêta-actine en tant que contrôle interne d’un gène domestique (housekeeping gene) . Nos résultats ont montré que les deux lignées cellulaires examinées avaient une expression similaire pour les marqueurs CD4, CD5, CD11a, CD14, CD43 et CD45. Cependant, la co-expression des marqueurs de cellules B CD79a et CD5 s’est avérée être spécifique à la lignée cellulaire nouvellement transformée, alors que le marqueur CD5 d’un sous-ensemble de cellules B était exprimé par la souche de vaccin S15. En conclusion, nous considérons la protéine de surface CD79a comme un nouveau marqueur de la lignée cellulaire infectée par Theileria annulata schizont non atténuée, non-exprimé par la souche vaccinale. L’évaluation de l'expression du marqueur CD par RT-PCR pourrait donc représenter une alternative appropriée à la cytométrie en flux.

Keywords [French]

  • Marqueur de CD
  • CD79a
  • RT-PCR
  • Theileria annulata
  • Vaccin
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