Tags

Type your tag names separated by a space and hit enter

A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia.
PLoS Negl Trop Dis. 2016 Apr; 10(4):e0004565.PN

Abstract

Snakebite envenomation is a serious medical problem in many tropical developing countries and was considered by WHO as a neglected tropical disease. Antivenom (AV), the rational and most effective treatment modality, is either unaffordable and/or unavailable in many affected countries. Moreover, each AV is specific to only one (monospecific) or a few (polyspecific) snake venoms. This demands that each country to prepare AV against its local snake venoms, which is often not feasible. Preparation of a 'pan-specific' AV against many snakes over a wide geographical area in some countries/regions has not been possible. If a 'pan-specific' AV effective against a variety of snakes from many countries could be prepared, it could be produced economically in large volume for use in many countries and save many lives. The aim of this study was to produce a pan-specific antiserum effective against major medically important elapids in Asia. The strategy was to use toxin fractions (TFs) of the venoms in place of crude venoms in order to reduce the number of antigens the horses were exposed to. This enabled inclusion of a greater variety of elapid venoms in the immunogen mix, thus exposing the horse immune system to a diverse repertoire of toxin epitopes, and gave rise to antiserum with wide paraspecificity against elapid venoms. Twelve venom samples from six medically important elapid snakes (4 Naja spp. and 2 Bungarus spp.) were collected from 12 regions/countries in Asia. Nine of these 12 venoms were ultra-filtered to remove high molecular weight, non-toxic and highly immunogenic proteins. The remaining 3 venoms were not ultra-filtered due to limited amounts available. The 9 toxin fractions (TFs) together with the 3 crude venoms were emulsified in complete Freund's adjuvant and used to immunize 3 horses using a low dose, low volume, multisite immunization protocol. The horse antisera were assayed by ELISA and by in vivo lethality neutralization in mice. The findings were: a) The 9 TFs were shown to contain all of the venom toxins but were devoid of high MW proteins. When these TFs, together with the 3 crude venoms, were used as the immunogen, satisfactory ELISA antibody titers against homologous/heterologous venoms were obtained. b) The horse antiserum immunologically reacted with and neutralized the lethal effects of both the homologous and the 16 heterologous Asian/African elapid venoms tested. Thus, the use of TFs in place of crude venoms and the inclusion of a variety of elapid venoms in the immunogen mix resulted in antiserum with wide paraspecificity against elapid venoms from distant geographic areas. The antivenom prepared from this antiserum would be expected to be pan-specific and effective in treating envenomations by most elapids in many Asian countries. Due to economies of scale, the antivenom could be produced inexpensively and save many lives. This simple strategy and procedure could be readily adapted for the production of pan-specific antisera against elapids of other continents.

Authors+Show Affiliations

Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand. Chulabhorn Graduate Institute, Bangkok, Thailand.Department of Molecular Medicine, University of Malaya, Kuala Lumpur, Malaysia.Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand.Department of Pharmacology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, Thailand.Department of Pre-clinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Salaya, NakornPrathom, Thailand.Department of Pre-clinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Salaya, NakornPrathom, Thailand.The Veterinary and Remount Department, The Royal Thai Army, NakornPrathom, Thailand.Department of Molecular Medicine, University of Malaya, Kuala Lumpur, Malaysia.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

27058956

Citation

Ratanabanangkoon, Kavi, et al. "A Simple and Novel Strategy for the Production of a Pan-specific Antiserum Against Elapid Snakes of Asia." PLoS Neglected Tropical Diseases, vol. 10, no. 4, 2016, pp. e0004565.
Ratanabanangkoon K, Tan KY, Eursakun S, et al. A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia. PLoS Negl Trop Dis. 2016;10(4):e0004565.
Ratanabanangkoon, K., Tan, K. Y., Eursakun, S., Tan, C. H., Simsiriwong, P., Pamornsakda, T., Wiriyarat, W., Klinpayom, C., & Tan, N. H. (2016). A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia. PLoS Neglected Tropical Diseases, 10(4), e0004565. https://doi.org/10.1371/journal.pntd.0004565
Ratanabanangkoon K, et al. A Simple and Novel Strategy for the Production of a Pan-specific Antiserum Against Elapid Snakes of Asia. PLoS Negl Trop Dis. 2016;10(4):e0004565. PubMed PMID: 27058956.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia. AU - Ratanabanangkoon,Kavi, AU - Tan,Kae Yi, AU - Eursakun,Sukanya, AU - Tan,Choo Hock, AU - Simsiriwong,Pavinee, AU - Pamornsakda,Teeraporn, AU - Wiriyarat,Witthawat, AU - Klinpayom,Chaiya, AU - Tan,Nget Hong, Y1 - 2016/04/08/ PY - 2015/12/15/received PY - 2016/03/01/accepted PY - 2016/4/9/entrez PY - 2016/4/9/pubmed PY - 2016/8/12/medline SP - e0004565 EP - e0004565 JF - PLoS neglected tropical diseases JO - PLoS Negl Trop Dis VL - 10 IS - 4 N2 - Snakebite envenomation is a serious medical problem in many tropical developing countries and was considered by WHO as a neglected tropical disease. Antivenom (AV), the rational and most effective treatment modality, is either unaffordable and/or unavailable in many affected countries. Moreover, each AV is specific to only one (monospecific) or a few (polyspecific) snake venoms. This demands that each country to prepare AV against its local snake venoms, which is often not feasible. Preparation of a 'pan-specific' AV against many snakes over a wide geographical area in some countries/regions has not been possible. If a 'pan-specific' AV effective against a variety of snakes from many countries could be prepared, it could be produced economically in large volume for use in many countries and save many lives. The aim of this study was to produce a pan-specific antiserum effective against major medically important elapids in Asia. The strategy was to use toxin fractions (TFs) of the venoms in place of crude venoms in order to reduce the number of antigens the horses were exposed to. This enabled inclusion of a greater variety of elapid venoms in the immunogen mix, thus exposing the horse immune system to a diverse repertoire of toxin epitopes, and gave rise to antiserum with wide paraspecificity against elapid venoms. Twelve venom samples from six medically important elapid snakes (4 Naja spp. and 2 Bungarus spp.) were collected from 12 regions/countries in Asia. Nine of these 12 venoms were ultra-filtered to remove high molecular weight, non-toxic and highly immunogenic proteins. The remaining 3 venoms were not ultra-filtered due to limited amounts available. The 9 toxin fractions (TFs) together with the 3 crude venoms were emulsified in complete Freund's adjuvant and used to immunize 3 horses using a low dose, low volume, multisite immunization protocol. The horse antisera were assayed by ELISA and by in vivo lethality neutralization in mice. The findings were: a) The 9 TFs were shown to contain all of the venom toxins but were devoid of high MW proteins. When these TFs, together with the 3 crude venoms, were used as the immunogen, satisfactory ELISA antibody titers against homologous/heterologous venoms were obtained. b) The horse antiserum immunologically reacted with and neutralized the lethal effects of both the homologous and the 16 heterologous Asian/African elapid venoms tested. Thus, the use of TFs in place of crude venoms and the inclusion of a variety of elapid venoms in the immunogen mix resulted in antiserum with wide paraspecificity against elapid venoms from distant geographic areas. The antivenom prepared from this antiserum would be expected to be pan-specific and effective in treating envenomations by most elapids in many Asian countries. Due to economies of scale, the antivenom could be produced inexpensively and save many lives. This simple strategy and procedure could be readily adapted for the production of pan-specific antisera against elapids of other continents. SN - 1935-2735 UR - https://www.unboundmedicine.com/medline/citation/27058956/A_Simple_and_Novel_Strategy_for_the_Production_of_a_Pan_specific_Antiserum_against_Elapid_Snakes_of_Asia_ L2 - http://dx.plos.org/10.1371/journal.pntd.0004565 DB - PRIME DP - Unbound Medicine ER -