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. 2019 Aug 13;10(1):3642.
doi: 10.1038/s41467-019-11639-2.

Horse immunization with short-chain consensus α-neurotoxin generates antibodies against broad spectrum of elapid venomous species

Guillermo de la Rosa  1   2 , Felipe Olvera  3 , Irving G Archundia  3 , Bruno Lomonte  4 , Alejandro Alagón  3 , Gerardo Corzo  5
Affiliations

Horse immunization with short-chain consensus α-neurotoxin generates antibodies against broad spectrum of elapid venomous species

Guillermo de la Rosa et al. Nat Commun. .

Abstract

Antivenoms are fundamental in the therapy for snakebites. In elapid venoms, there are toxins, e.g. short-chain α-neurotoxins, which are quite abundant, highly toxic, and consequently play a major role in envenomation processes. The core problem is that such α-neurotoxins are weakly immunogenic, and many current elapid antivenoms show low reactivity towards them. We have previously developed a recombinant consensus short-chain α-neurotoxin (ScNtx) based on sequences from the most lethal elapid venoms from America, Africa, Asia, and Oceania. Here we report that an antivenom generated by immunizing horses with ScNtx can successfully neutralize the lethality of pure recombinant and native short-chain α-neurotoxins, as well as whole neurotoxic elapid venoms from diverse genera such as Micrurus, Dendroaspis, Naja, Walterinnesia, Ophiophagus and Hydrophis. These results provide a proof-of-principle for using recombinant proteins with rationally designed consensus sequences as universal immunogens for developing next-generation antivenoms with higher effectiveness and broader neutralizing capacity.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Time course of the specific antibody response of horses against the consensus short-chain α-neurotoxin (ScNtx). Horse hyperimmune sera raised against the ScNtx were titrated by enzyme-linked immunosorbent assay (ELISA). Dashed lines indicate samples used for neutralization tests, corresponding to days 98 (blue), 147 (red), and 205 (orange) of the immunization. Points represent mean ± SD of triplicate wells of the ELISA
Fig. 2
Fig. 2
Neutralization potencies, plotted as median effective dose (ED50). Serum samples were obtained from individual horses immunized with the consensus short-chain α-neurotoxin (ScNtx) at days 98, 147, and 205. ED50 values are expressed in µL/mouse (volume of anti-ScNtx able to neutralize 3 × LD50 (lethal dose, 50%) of purified short-chain neurotoxins ScNtx, MlatA1 or rD.H.). Values were estimated in groups of five mice, injected by the intravenous route. Error bars represent the 95% confidence limits for the ED50 value. The data obtained from the groups of “Horse” and “Day” of immunization were analyzed statistically by two-way analysis of variance (ANOVA) followed by paired Student’s t test. A p value of <0.05 was considered significant, as indicated by *. Horse 2 produces antibodies with the best neutralization potencies from after the 147th day of immunization
Fig. 3
Fig. 3
Multiple sequence alignment of the most lethal short-chain α-neurotoxins. ScNtx: Synthetic consensus α-neurotoxin; rD.H: Micrurus diastema (recombinant); MlatA1: Micrurus laticollaris (recombinant); P01426: Naja spp.; P80548: Micrurus nigrocinctus; Atratoxin: Naja atra; MS1: Micrurus surinamensis; P01424: Naja melanoleuca; pelamitoxin: Hydrophis platura; W-III: Walterinnesia aegyptia. Differences from the ScNtx are indicated by bold letters; cysteines and disulfide bridge arrangements are colored in blue
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