Evaluation of Pertussis Toxin Expression in B2 and THIJS Media

Document Type: Original Articles

Authors

1 Department of Microbiology and Virology, Faculty of Medical Sciences, Zanjan University of Medical Sciences, Zanjan, Iran

2 Department of Human Bacterial Vaccines Research and Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran

Abstract

Whole-cell pertussis vaccine (wP) has been imperative and highly effective in preventing childhood deaths due to pertussis. Pertussis toxin is one of the virulence factors of Bordetella pertussis in all available pertussis vaccines. wP production in Razi Vaccine and Serum Research Institute is according to bioreactor culture of B. pertussis strains in B2 medium. The aim of this study was to evaluate B. pertussis strain 509 PT production in B2 and Thalen-IJssel (THIJS) media by Chinese Hamster Ovary (CHO) cell and enzyme-linked immunosorbent assay methods (ELISA). In the current study, B. pertussis strain 509 was cultured in B2 and THIJS media. Six samples were taken during the log growth phase within 2-3 h intervals (triplicate). The growth rate was calculated using opacity and the quantification of cell-associated and released PT measured by ELISA and CHO cell assays. THIJS medium was significantly showed an increase in the bacterial growth rate. During the first 29 h, bacterial concentrations in B2 and THIJS culture medium were 19 and 29 IOU, respectively. In THIJS medium, greater amount of pertussistoxin production was cell-associated. In B2 medium, maximum cell-associated toxin by ELISA and CHO cell assays were in the ODs of 1.1 and 0.9 and for THIJS medium in the ODs of 1.6 and 1.1, respectively. B. pertussis strain 509 in THIJS medium produced higher cell mass and cell-associated pertussis toxin than that of B2. It can be used for the production of whole-cell vaccine with higher pertussis toxin and accordingly using lower biomass per dose leading to the reduction of vaccine toxicity.

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Main Subjects


Article Title [French]

Évaluation de l'expression de la toxine de coqueluche dans les milieux B2 et THIJS

Abstract [French]

Le vaccin anticoquelucheux à germes entièrs a été impératif et très efficace dans la prévention des décès d'enfants dus à la coqueluche. La toxine de la coqueluche est l’un des facteurs de virulence de Bordetella pertussis dans tous les vaccins anticoquelucheux disponibles. La production de wP à l'Institut de recherche sur les vaccins et les sérums de Razi est menée en conformité à la culture en bioréacteur des souches de B. pertussis dans un milieu B2. Le but de cette étude était d’évaluer la production de la souche 509 PT de B. pertussis dans les milieux B2 et Thalen-IJssel (THIJS) par dosage sur la cellule d'ovaire de hamster chinois (CHO) et par ELISA. Dans cetteétude, la souche 509 de B. pertussis a été cultivée dans les milieux B2 et THIJS. Six échantillons ont été prélevés au cours de la phase de croissance logarithmique dans des intervalles de 2 à 3 heures (en triple exemplaire). Le taux de croissance a été calculé en utilisant l'opacité et la quantification du PT libéré et associé aux cellules mesuré par les tests ELISA et cellulaire CHO. Le milieu THIJS a montré de manière significative une augmentation du taux de croissance bactérienne. Au cours des 29 premières heures, les concentrations bactériennes dans les milieux de culture B2 et THIJS étaient respectivement de 19 et 29 IOU.Dans le milieu THIJS, une plus grande quantité de production de toxine de coqueluche était associée aux cellules. Dans le milieu B2, les concentrations maximales de toxines obtenues par les tests ELISA et CHO étaient dans les ODs de 1,1 et 0,9 et dans le milieu THIJS dans les ODs de 1,6 et 1,1, respectivement. La cuture de la souche 509 de B. pertussis dans le milieu THIJS a produit un taux de toxines de pertussis par cellule et par masse cellulaire supérieure à celui obtenu en mileieu B2. Ce dernier peut être utilisé pour la production de vaccins à germes entiers contenant une concentration plus importante de toxine anticoqueluche et, par conséquent, d’utiliser une biomasse moins importante de microorganism par dose entraînant une réduction de la toxicité du vaccin.

Keywords [French]

  • Bordetella pertussis
  • vaccin anticoquelucheux
  • ELISA
  • cellules de CHO
  • toxine de coqueluche
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