Molecular cloning, expression, function and immunoreactivities of members of a gene family of sphingomyelinases from Loxosceles venom glands

Mol Immunol. 2004 Jul;41(8):831-40. doi: 10.1016/j.molimm.2004.03.027.

Abstract

Loxoscelism is the clinical condition produced by the venom of spiders belonging to the genus Loxosceles, which can be observed as two well-defined clinical variants: cutaneous loxoscelism and systemic or viscerocutaneous loxoscelism. We have recently identified, purified and characterised the toxins (sphingomyelinases) from Loxosceles intermedia venom that are responsible for all the local (dermonecrosis) and systemic effects (complement dependent haemolysis) induced by whole venom. In the present study, we have cloned and expressed the two functional sphingomyelinases isoforms, P1 and P2, and shown that the recombinant proteins display all the functional characteristics of whole L. intermedia venom, e.g., dermonecrotic and complement-dependent hemolytic activities and ability of hydrolyzing sphingomyelin. We have also compared the cross-reactivities of antisera raised against the toxins from different Loxosceles species and show here that the cross-reactivity is high when toxins are from the same species (P1 and P2 from L. intermedia) but low when the toxins are from different species (L. intermedia versus L. laeta). These data suggest that in order to obtain a suitable comprehensive neutralizing antiserum using the recombinant toxin as an immunogen, a mixture of the recombinant toxins from the different species has to be used. The use of anti-recombinant toxin antisera may have clinical benefits to those individuals displaying acute loxoscelic lesions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • Cross Reactions
  • Molecular Sequence Data
  • Multigene Family
  • Phosphoric Diester Hydrolases / genetics*
  • Phosphoric Diester Hydrolases / immunology
  • Phosphoric Diester Hydrolases / metabolism
  • Phylogeny
  • Sphingomyelin Phosphodiesterase / genetics*
  • Sphingomyelin Phosphodiesterase / immunology
  • Sphingomyelin Phosphodiesterase / metabolism
  • Spider Venoms / genetics*
  • Spider Venoms / immunology
  • Spider Venoms / metabolism
  • Spiders / genetics*
  • Spiders / immunology
  • Spiders / metabolism

Substances

  • Spider Venoms
  • loxosceles venom
  • Phosphoric Diester Hydrolases
  • Sphingomyelin Phosphodiesterase

Associated data

  • GENBANK/AY304471
  • GENBANK/AY304472