Vaccination with a cocktail of Ancylostoma ceylanicum recombinant antigens leads to worm burden reduction in hamsters

Marcin Wiśniewski; Maciej Łapiński; Emilia Daniłowicz-Luebert; Sławomir Jaros; Ewa Długosz; Halina Wędrychowicz

doi:10.1515/ap-2016-0074

Article

Vaccination with a cocktail of Ancylostoma ceylanicum recombinant antigens leads to worm burden reduction in hamsters

Marcin Wiśniewski , Maciej Łapiński , Emilia Daniłowicz-Luebert , Sławomir Jaros , Ewa Długosz and Halina Wędrychowicz

Published/Copyright: July 21, 2016

Published by

Become an author with De Gruyter Brill

Author Information

From the journal Acta Parasitologica Volume 61 Issue 3

Abstract

Hookworms, a group to which Ancylostoma ceylanicum belongs, are gastrointestinal nematodes that infect more than 700 million people around the world. They are a leading cause of anemia in developing countries. In order to effectively prevent hookworm infections research is conducted to develop an effective vaccine using recombinant antigens of the parasite. The aim of this study was to examine the influence of the hosts’ on protection against ancylostomiasis and the shaping of the humoral immune response among Syrian hamsters after immunization with a cocktail of five A. ceylanicum recombinant antigens. Ace-ASP-3, Ace-ASP-4, Ace-APR-1, Ace-MEP-6 and Ace-MEP-7 were obtained in the pET expression system. Immunization with a vaccine cocktail resulted in a 33.5% worm burden reduction. The immunogenicity of the recombinant proteins were determined using ELISA. Statistical analysis showed that vaccinated hamsters developed stronger humoral responses to four of five recombinant antigens (the exception being Ace-ASP-3) compared to hamsters from the control group.

Keywords: Hookworm; Ancylostoma ceylanicum; recombinant antigen; vaccine

Acknowledgements

This research was supported by a grant from the Polish Ministry of Science and Higher Education (Projects No.: 3 P06K 024 24 and NN 308 076 134).

References

Albonico M., Bickle Q., Ramsan M., Montresor A., Savioli L., Taylor M. 2003. Efficacy of mebendazole and levamisole alone or in combination against intestinal nematode infections after repeated targeted mebendazole treatment in Zanzibar. Bulletin of the World Health Organization, 81, 343–352Search in Google Scholar

Albonico M., Smith P.G., Ercole E., Hall A., Chwaya H.M., Alawi K.S., Savioli L. 1995. Rate of reinfection with intestinal nematodes after treatment of children with mebendazole or albendazole in a highly endemic area. Transactions of the Royal Society of Tropical Medicine and Hygiene, 89, 538–34110.1016/0035-9203(95)90101-9Search in Google Scholar

Bąska P., Wisniewski M., Krzyzowska M., Długosz E., Zygner W., Górski P., Wędrychowicz H. 2013. Molecular cloning and characterization of in vitro immune response against astacin-like metalloprotease Ace-MTP-2 from Ancylostoma ceylanicum. Experimental Parasitology, 133, 472–482. DOI: 10.1016/j.exppara.2013.01.006Search in Google Scholar

Bethony J., Loukas A., Smout M., Brooker S., Mendez S., Plieskatt J., Goud G., Bottazzi M.E., Zhan B., Wang Y., Williamson A., Lustigman S., Correa-Oliveira R., Xiao S., Hotez P.J. 2005. Antibodies against a secreted protein from hookworm larvae reduce the intensity of hookworm infection in humans and vaccinated laboratory animals. FASEB Journal, 19, 1743–1745. DOI: 10.1096/fj.05-3936fjeSearch in Google Scholar

Bungiro R.D., Harrison L.M., Cappello M. 2002. Ancylostoma ceylanicum Excretory/Secretory Protein 1: Purification and molecular cloning of a major secretory protein from adult hookworms. Molecular and Biochemical Parasitology, 119, 147–151. DOI:10.1016/S0166-6851(01)00408-XSearch in Google Scholar

Bungiro R.D. Jr, Solis C.V., Harrison L.M., Cappello M. 2004. Purification and molecular cloning of and immunization with Ancylostoma ceylanicum excretory-secretory protein 2, an immunoreactive protein produced by adult hookworms. Infection and Immunity, 72, 2203–2213. DOI: 10.1128/IAI.72.4.2203-2213.2004Search in Google Scholar PubMed PubMed Central

Cantacessi C., Campbell B.E., Visser A., Geldhof P., Nolan M., Nisbet A.J., Matthews J.B., Loukas A., Hofmann A., Otranto D., Sternberg P.W., Gasser R.B. 2009. A portrait of the ‘SCP/TAPS’ proteins of eukaryotes – Developing a framework for fundamental research and biotechnological outcomes. Biotechnology Advances, 27, 376–388. DOI: 10.1016/j.biotechadv.2009.02.005Search in Google Scholar PubMed

Chu D., Bungiro R.D., Ibanez M., Harrison L.M., Campodonico E., Jones B.F., Mieszczanek J., Kuzmic P., Cappello M. 2004. Molecular characterization of Ancylostoma ceylanicum Kunitz-type serine protease inhibitor: evidence for a role in hookworm-associated growth delay. Infection and Immunity, 72, 2214–2221. DOI: 10.1128/IAI.72.4.2214-2221.2004Search in Google Scholar PubMed PubMed Central

Długosz E., Wisniewski M., Wójcik L., Wędrychowicz H. 2010. In vitro IL-4, IL-12, and IFN-γ production by splenocytes from Ancylostoma ceylanicum infected hamsters. Bulletin of the Veterinary Institute in Pulawy, 54, 321–325Search in Google Scholar

Fairfax K.C., Vermeire J.J., Harrison L.M., Bungiro R.D., Grant W., Husain S.Z., Cappello M. 2009. Characterisation of a fatty acid and retinol binding protein orthologue from the hookworm Ancylostoma ceylanicum. International Journal for Parasitology, 39, 1561–1571. DOI: 10.1016/j.ijpara.2009.06.005Search in Google Scholar PubMed PubMed Central

Goud G.N., Zhan B., Ghosh K., Loukas A., Hawdon J., Dobardzic A., Deumic V., Liu S., Dobardzic R., Zook B.C., Jin Q., Liu Y., Hoffman L., Chung-Debose S., Patel R., Mendez S., Hotez P.J. 2004. Cloning, yeast expression, isolation, and vaccine testing of recombinant Ancylostoma-secreted protein (ASP)-1 and ASP-2 from Ancylostoma ceylanicum. The Journal of Infectious Diseases, 189, 919–929. DOI: 10.1086/381901Search in Google Scholar PubMed

Hotez P.J., Ashcom J., Zhan B., Bethony J., Loukas A., Hawdon J., Wang Y., Jin Q., Jones K.C., Dobardzic A., Dobardzic R., Bolden J., Essiet I., Brandt W., Russell P.K., Zook B.C., Howard B., Chacon M. 2003. Effect of vaccination with a recombinant fusion protein encoding an astacinlike metalloprotease (MTP-1) secreted by host-stimulated Ancylostoma caninum third-stage infective larvae. Journal of Parasitology, 89, 853–855. DOI:10.1645/GE-46RSearch in Google Scholar PubMed

Hotez P.J., Diemert D., Bacon K.M., Beaumier C., Bethony J.M., Bottazzi M.E., Brooker S., Couto A.R., da Silva Freire M., Homma A., Lee B.Y., Loukas A., Loblack M, Morel C.M., Oliveira R. C., Russell P.K. 2013. The human hookworm vaccine. Vaccine, 31, 227–232. DOI: 10.1016/j.vaccine.2012.11.034Search in Google Scholar

Hotez P.J., Fenwick A., Savioli L., Molyneux D.H. 2009. Rescuing the bottom billion through control of neglected tropical diseases. Lancet, 373, 1570–1575. DOI: 10.1016/S0140-6736(09)60233-6Search in Google Scholar

Loukas A., Bethony J.M., Mendez S., Fujiwara R.T., Goud G.N., Ranjit N., Zhan B., Jones K., Bottazzi M.E., Hotez P.J. 2005. Vaccination with recombinant aspartic hemoglobinase reduces parasite load and blood loss after hookworm infection in dogs. PLoS Medicine, 2, 1008–1017. DOI: 10.1371/journal.pmed.0020295Search in Google Scholar

Mendez S., Zhan B., Goud G., Ghosh K., Dobardzic A., Wu W., Liu S., Deumic V., Dobardzic R., Liu Y., Bethony J., Hotez P.J. 2005. Effect of combining the larval antigens Ancylostoma secreted protein 2 (ASP-2) and metalloprotease 1 (MTP-1) in protecting hamsters against hookworm infection and disease caused by Ancylostoma ceylanicum. Vaccine, 23, 3123–3130. DOI: 10.1016/j.vaccine.2004.12.022Search in Google Scholar

Palmer D.R., Bradley M., Bundy D.A. 1996. IgG4 responses to antigens of adult Necator americanus: potential for use in large-scale epidemiological studies. Bulletin of the World Health Organization, 74, 381–386Search in Google Scholar

Pearson M.S., Bethony J.M., Pickering D.A., de Oliveira L.M., Jariwala A., Santiago H., Miles A.P., Zhan B., Jiang D., Ranjit N., Mulvenna J., Tribolet L., Plieskatt J., Smith T., Bottazzi M.E., Jones K., Keegan B., Hotez P.J., Loukas A. 2009. An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection. FASEB Journal, 23, 3007–3019. DOI: 10.1096/fj.09-131433Search in Google Scholar

Pearson M.S., Jariwala A.R., Abbenante G., Plieskatt J., Wilson D., Bottazzi M.E., Hotez P.J., Keegan B., Bethony J.M., Loukas A. 2015. New tools for NTD vaccines: A case study of quality control assays for product development of the human hookworm vaccine Na-APR-1M74. Human Vaccines and Immunotherapeutics, 11, 1251–1257. DOI: 10.4161/21645515.2014.980199Search in Google Scholar

de Silva N.R. 2003. Impact of mass chemotherapy on the morbidity due to soil-transmitted nematodes. Acta Tropica, 86, 197–214. DOI: 10.1016/S0001-706X(03)00035-4Search in Google Scholar

Siwińska A.M., Bąska P., Daniłowicz-Luebert E., Januszkiewicz K., Długosz E., Wędrychowicz H., Cappello M., Wisniewski M. 2013. Cloning and molecular characterization of cDNAs encoding three Ancylostoma ceylanicum secreted proteins. Acta Parasitologica, 58, 112–118. DOI: 10.2478/s11686-013-0116-zSearch in Google Scholar PubMed

Stoltzfus R.J., Dreyfuss M.L., Chwaya H.M., Albonico M. 1997. Hookworm control as a strategy to prevent iron deficiency. Nutrition Reviews, 55, 223–232. DOI: 10.1111/j.1753-4887.1997.tb01609.xSearch in Google Scholar PubMed

Xiao S., Zhan B., Xue J., Goud G.N., Loukas A., Liu Y., Williamson A., Liu S., Deumic V., Hotez P. 2008. The evaluation of recombinant hookworm antigens as vaccines in hamsters (Mesocricetus auratus) challenged with human hookworm, Necator americanus. Experimental Parasitology, 118, 32–40. DOI: 10.1016/j.exppara.2007.05.010Search in Google Scholar PubMed

Wasyl K., Zawistowska-Deniziak A., Bąska P., Wędrychowicz H., Wisniewski M. 2013. Molecular cloning and expression of the cDNA sequence encoding a novel aspartic protease from Uncinaria stenocephala. Experimental Parasitology, 134, 220–227. DOI: 10.1016/j.exppara.2013.03.013Search in Google Scholar

Wędrychowicz H. 2015. Antiparasitic DNA vaccines in 21st century. 2015. Acta Parasitologica, 60, 179–189. DOI: 10.1515/ap-2015-0026Search in Google Scholar

Wiśniewski M., Jaros S., Bąska P., Cappello M., Długosz E., Wędrychowicz H. 2016. Molecular cloning and analysis of Acemep-7 cDNA vaccine efficiency on the laboratory model Ancylostoma ceylanicum – Syrian hamster. Experimental Parasitology, 163, 1–7. DOI: 10.1016/j.exppara.2016.01.006Search in Google Scholar

Wiśniewski M., Jaros S., Bąska P., Cappello M., Wędrychowicz H. 2013. Ancylostoma ceylanicum metalloproteases 6 DNA vaccination induces partial protection against hookworm challenge infection. Acta Parasitologica, 58, 367–383. DOI: 10.2478/s11686-013-0151-9Search in Google Scholar

Wiśniewski M., Lapiński M., Zdziarska A.M., Długosz E., Bąska P. 2014. Molecular cloning and analysis of Ancylostoma ceylanicum glutamate-cysteine ligase. Molecular and Biochemical Parasitology, 196, 12–20. DOI: 10.1016/j.molbiopara.2014.07.003Search in Google Scholar

Zhan B., Liu Y., Badamchian M., Williamson A., Feng J., Loukas A., Hawdon J.M., Hotez P.J. 2003. Molecular characterisation of the Ancylostoma-secreted protein family from the adult stage of Ancylostoma caninum. International Journal for Parasitology, 33, 897–907. DOI: 10.1016/S0020-7519(03)00111-5Search in Google Scholar

Received: 2016-1-4

Revised: 2015-3-16

Accepted: 2016-3-23

Published Online: 2016-7-21

Published in Print: 2016-9-1

You are currently not able to access this content.

Articles in the same Issue

https://doi.org/10.1515/ap-2016-0074

Keywords for this article

Hookworm; Ancylostoma ceylanicum; recombinant antigen; vaccine