Journal Article

Cloning, Yeast Expression, Isolation, and Vaccine Testing of Recombinant <i>Ancylostoma</i>-Secreted Protein (ASP)-1 and ASP-2 from <i>Ancylostoma ceylanicum</i>

Gaddam Narsa Goud, Bin Zhan, Kashinath Ghosh, Alex Loukas, John Hawdon, Azra Dobardzic, Vehid Deumic, Sen Liu, Reshad Dobardzic, Bernard C. Zook, Qun Jin, Yueyuan Liu, Laura Hoffman, Sophia Chung Debose, Rachna Patel, Susana Mendez and Peter J. Hotez

in The Journal of Infectious Diseases

Published on behalf of Infectious Diseases Society of America

Volume 189, issue 5, pages 919-929
Published in print March 2004 | ISSN: 0022-1899
Published online March 2004 | e-ISSN: 1537-6613 | DOI: http://dx.doi.org/10.1086/381901
Cloning, Yeast Expression, Isolation, and Vaccine Testing of Recombinant Ancylostoma-Secreted Protein (ASP)-1 and ASP-2 from Ancylostoma ceylanicum

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cDNAs encoding 2 Ancylostoma-secreted proteins (ASPs), Ancylostoma ceylanicum (Ay)-ASP-1 and Ay-ASP-2, were cloned from infective third-stage larvae (L3) of the hookworm A. ceylanicum and were expressed as soluble recombinant fusion proteins secreted by the yeast Pichia pastoris. The recombinant fusion proteins were purified, adjuvant formulated, and injected intramuscularly into hamsters. Hamsters vaccinated either by oral vaccination with irradiated L3 (irL3) or by injections of the adjuvants alone served as positive and negative controls, respectively. Anti-ASP-1 and anti-ASP-2 antibody titers exceeded 1:100,000. Each vaccinated hamster was challenged orally with 100 L3. Two groups of vaccinated hamsters (i.e., those vaccinated with either irL3 or ASP-2 formulated with Quil A) exhibited significant reductions in adult hookworm burdens, compared with control hamsters. The hookworms recovered from the hamsters vaccinated with ASP-2 plus Quil A were reduced in length. Splenomegaly, which was observed in control hamsters, was not seen in hamsters vaccinated with either irL3 or ASP-2 formulated with Quil A. These results indicate that ASP-2 is a promising molecule for the development of a hookworm vaccine.

Journal Article.  6290 words.  Illustrated.

Subjects: Infectious Diseases ; Immunology ; Public Health and Epidemiology ; Microbiology

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