Efficacy of an ovine caseous lymphadenitis vaccine formulated using a genetically inactive form of the Corynebacterium pseudotuberculosis phospholipase D

Gaseous lymphadenitis (CLA) is an economically significant disease of sheep caused by the Gram-positive bacterium Corynebacterium pseudotuberculosis. CLA vaccines are currently formulated using Formalin inactivated culture supernatants that are rich in the C. pseudotuberculosis phospholipase D (PLD) exotoxin. One alternative to chemical detoxification is to inactivate the PLD genetically. This procedure not only provides a means to remove an onerous chemical treatment step but also the opportunity to increase gene expression, therefore improve protein yields, Using site-specific mutagenesis the C, pseudotuberculosis PLD was inactivated by substituting a serine residue at histidine 20 within the enzyme active site. CLA vaccine formulated using genetically inactivated PLD protected 44% of sheep against C. pseudotuberculosis challenge compared with 95% protection offered by the formalin inactivated preparation. Since there was no apparent difference in immune response mounted by vaccinated sheep the reason for this variation in vaccine efficacy remains unclear. Although genetic inactivation can be a convenient means to produce toroid vaccines its use to develop a new CLA vaccine provided no net benefit over the conventional formulation. (C) 1999 Elsevier Science Ltd. All rights reserved.