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Toxins 2019, 11(2), 108; https://doi.org/10.3390/toxins11020108

Design and Production of a Recombinant Hybrid Toxin to Raise Protective Antibodies Against Loxosceles Spider Venom

Laboratory of Immunopathology, Butantan Institute, 05503-900 São Paulo, Brazil
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Received: 16 January 2019 / Revised: 8 February 2019 / Accepted: 10 February 2019 / Published: 12 February 2019
(This article belongs to the Special Issue Arthropod Venom Components and their Potential Usage)
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Abstract

Human accidents with spiders of the genus Loxosceles are an important health problem affecting thousands of people worldwide. Patients evolve to severe local injuries and, in many cases, to systemic disturbances as acute renal failure, in which cases antivenoms are considered to be the most effective treatment. However, for antivenom production, the extraction of the venom used in the immunization process is laborious and the yield is very low. Thus, many groups have been exploring the use of recombinant Loxosceles toxins, particularly phospholipases D (PLDs), to produce the antivenom. Nonetheless, some important venom activities are not neutralized by anti-PLD antibodies. Astacin-like metalloproteases (ALMPs) are the second most expressed toxin acting on the extracellular matrix, indicating the importance of its inclusion in the antigen’s formulation to provide a better antivenom. Here we show the construction of a hybrid recombinant immunogen, called LgRec1ALP1, composed of hydrophilic regions of the PLD and the ALMP toxins from Loxosceles gaucho. Although the LgRec1ALP1 was expressed as inclusion bodies, it resulted in good yields and it was effective to produce neutralizing antibodies in mice. The antiserum neutralized fibrinogenolytic, platelet aggregation and dermonecrotic activities elicited by L. gaucho, L. laeta, and L. intermedia venoms, indicating that the hybrid recombinant antigen may be a valuable source for the production of protective antibodies against Loxosceles ssp. venoms. In addition, the hybrid recombinant toxin approach may enrich and expand the alternative antigens for antisera production for other venoms.
Keywords: phospholipases D; metalloproteases; Loxosceles spp.; recombinant toxins; hybrid immunogen; neutralizing antibodies; antivenoms phospholipases D; metalloproteases; Loxosceles spp.; recombinant toxins; hybrid immunogen; neutralizing antibodies; antivenoms
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Calabria, P.A.; Shimokava-Falcao, L.H.A.; Colombini, M.; Moura-da-Silva, A.M.; Barbaro, K.C.; Faquim-Mauro, E.L.; Magalhaes, G.S. Design and Production of a Recombinant Hybrid Toxin to Raise Protective Antibodies Against Loxosceles Spider Venom. Toxins 2019, 11, 108.

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