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. 2020 Jan 31:26:e20190048.
doi: 10.1590/1678-9199-JVATITD-2019-0048. eCollection 2020.

Proteomics and antivenom immunoprofiling of Russell's viper (Daboia siamensis) venoms from Thailand and Indonesia

Thava Malar Changra Lingam  1 , Kae Yi Tan  1 , Choo Hock Tan  2
Affiliations

Proteomics and antivenom immunoprofiling of Russell's viper (Daboia siamensis) venoms from Thailand and Indonesia

Thava Malar Changra Lingam et al. J Venom Anim Toxins Incl Trop Dis. .

Abstract

The Eastern Russell's viper, Daboia siamensis, is a WHO Category 1 medically important venomous snake. It has a wide but disjunct distribution in Southeast Asia. The specific antivenom, D. siamensis Monovalent Antivenom (DsMAV-Thailand) is produced in Thailand but not available in Indonesia, where a heterologous trivalent antivenom, Serum Anti Bisa Ular (SABU), is used instead. This study aimed to investigate the geographical venom variation of D. siamensis from Thailand (Ds-Thailand) and Indonesia (Ds-Indonesia), and the immunorecognition of the venom proteins by antivenoms.

Methods: The venom proteins were decomplexed with reverse-phase high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by in-solution tryptic digestion, nano-liquid chromatography-tandem mass spectrometry and protein identification. The efficacies of DsMAV-Thailand and SABU in binding the various venom fractions were assessed using an enzyme-linked immunosorbent assay optimized for immunorecognition profiling.

Results: The two most abundant protein families in Ds-Thailand venom are phospholipase A2 (PLA2) and Kunitz-type serine protease inhibitor (KSPI). Those abundant in Ds-Indonesia venom are PLA2 and serine protease. KSPI and vascular endothelial growth factor were detected in Ds-Thailand venom, whereas L-amino acid oxidase and disintegrin were present in Ds-Indonesia venom. Common proteins shared between the two included snaclecs, serine proteases, metalloproteinases, phosphodiesterases, 5'nucleotidases and nerve growth factors at varying abundances. DsMAV-Thailand exhibited strong immunorecognition of the major protein fractions in both venoms, but low immunoreactivity toward the low molecular weight proteins e.g. KSPI and disintegrins. On the other hand, SABU was virtually ineffective in binding all fractionated venom proteins.

Conclusion: D. siamensis venoms from Thailand and Indonesia varied geographically in the protein subtypes and abundances. The venoms, nevertheless, shared conserved antigenicity that allowed effective immunorecognition by DsMAV-Thailand but not by SABU, consistent with the neutralization efficacy of the antivenoms. A specific, appropriate antivenom is needed in Indonesia to treat Russell's viper envenomation.

Keywords: Antivenomics; Eastern Russell's viper; Neutralization; Venomics.

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Conflict of interest statement

Competing interests: The authors declare no conflict of interest.

Figures

Figure 1.
Figure 1.. Protein decomplexation of Daboia siamensis venom from (A) Thailand and (B) Indonesia. Upper panel: C18 reverse-phase high-performance liquid chromatography (HPLC) profile of the venom. Lower panel: 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the protein fractions, under reducing conditions. Lane C: whole venom.
Figure 2.
Figure 2.. Venom proteomes of Daboia siamensis from (A) Thailand and (B) Indonesia. KSPI: Kunitz-type serine protease inhibitor; PLA2: phospholipase A2; snaclec: snake venom C-type lectin/lectin-like protein; SVSP: snake venom serine protease; SVMP: snake venom metalloproteinase; LAAO: L-amino acid oxidase; VEGF: snake venom vascular endothelial growth factor; VNGF: snake venom nerve growth factor; 5′NUC: 5′-nucleotidase; PDE: phosphodiesterase; Dis: disintegrin.
Figure 3.
Figure 3.. Immunoreactivity of Thai Daboia siamensis monovalent antivenom (DsMAV-Thailand) toward HPLC venom protein fractions of D. siamensis from (A) Thailand and (B) Indonesia. Naja sputatrix venom was used as negative control. Absorbance values were obtained by indirect ELISA and expressed as mean ± S.E.M. from three independent experiments.
Figure 4.
Figure 4.. Immunoreactivity of Indonesian Serum Anti Bisa Ular (SABU) toward HPLC venom protein fractions of D. siamensis from (A) Thailand and (B) Indonesia. Naja sputatrix venom was used as positive control. Absorbance values were obtained by indirect ELISA and expressed as mean ± S.E.M. from three independent experiments.
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