Novel Mutant Phospholipase D from Hemiscorpius lepturus Acts as A Highly Immunogen in BALB/c Mice Against the Lethality of Scorpion Venom.

Hemiscorpius lepturus (H. lepturus) which belongs to the Scorpionidae family, is the deadliest scorpion in Iran. It causes pathological manifestations like dermonecrosis, hemolysis, renal failure, necrotic ulcers, and in some cases, even death. The venom of this scorpion is well-known for its cytotoxic effects in comparison with the other venomous scorpions which show significant neurotoxic effects. Due to the painless nature of the sting of this scorpion, the clinical symptoms occur in victims 24 to 72 h post-sting. In our previous studies during the last decade, we demonstrated that the medical complications are attributable to the presence of phospholipase D (PLD) as a major toxin in the venom. With the purpose of designing and constructing a vaccine against H. lepturus for humans, animal model experiments were performed. To achieve this goal, non-toxic PLD was developed by mutation of two critical catalytic residues-His12 and His48-into alanines and the product was then denominated mut-rPLD1. The in-vivo tests showed that the mice immunized with interval doses of 10 µg of mut-rPLD1, were completely protected against 10× the LD100 of the venom. In conclusion, this mutant may be an effective vaccine candidate against scorpion envenomation by H. lepturus in future clinical studies.

Vaccination confers significant protection of sheep against infection with a virulent United Kingdom strain of Corynebacterium pseudotuberculosis.

Using a virulent United Kingdom Corynebacterium pseudotuberculosis isolate, an ovine experimental model of caseous lymphadenitis was developed, in which the manifestation of disease was equivalent to the naturally observed infection in this country. Subsequently, the capacity of several experimental vaccines to protect against experimental challenge was determined. Sheep were immunised with a recombinant derivative of phospholipase D, deriving from the virulent UK isolate, a formalin-killed bacterin of the same strain, or a bacterin supplemented with recombinant phospholipase D. Following homologous experimental challenge, the phospholipase D and bacterin vaccines were observed to confer statistically significant protection against infection, and appeared to restrict dissemination of challenge bacteria beyond the inoculation site in the majority of animals. More importantly, the combined vaccine succeeded in providing absolute protection against infection, whereby challenge bacteria were eradicated from all vaccinates. In addition to the experimental vaccines, a commercially available CLA vaccine, unlicensed for use in the European Union, was assessed for its capacity to protect against heterologous challenge. The vaccine conferred significant protection, although the dissemination of infection beyond the inoculation site was not restricted as it had been with the previous vaccines. However, no animals immunised with this vaccine manifested infection within the lungs; thus, a potentially important route of disease transmission was eliminated. The results of this study provide information pertinent to the development of an effective caseous lymphadenitis vaccination strategy in the UK.