Safety and immunogenicity of the Na-GST-1 hookworm vaccine in Brazilian and American adults

David J Diemert, Janaína Freire, Vanderson Valente, Carlo Geraldo Fraga, Frederico Talles, Shannon Grahek, Doreen Campbell, Amar Jariwala, Maria Victoria Periago, Martin Enk, Maria Flávia Gazzinelli, Maria Elena Bottazzi, Robert Hamilton, Jill Brelsford, Anna Yakovleva, Guangzhao Li, Jin Peng, Rodrigo Correa-Oliveira, Peter Hotez, Jeffrey Bethony, David J Diemert, Janaína Freire, Vanderson Valente, Carlo Geraldo Fraga, Frederico Talles, Shannon Grahek, Doreen Campbell, Amar Jariwala, Maria Victoria Periago, Martin Enk, Maria Flávia Gazzinelli, Maria Elena Bottazzi, Robert Hamilton, Jill Brelsford, Anna Yakovleva, Guangzhao Li, Jin Peng, Rodrigo Correa-Oliveira, Peter Hotez, Jeffrey Bethony

Abstract

Necator americanus Glutathione-S-Transferase-1 (Na-GST-1) plays a role in the digestion of host hemoglobin by adult N. americanus hookworms. Vaccination of laboratory animals with recombinant Na-GST-1 is associated with significant protection from challenge infection. Recombinant Na-GST-1 was expressed in Pichia pastoris and adsorbed to aluminum hydroxide adjuvant (Alhydrogel) according to current Good Manufacturing Practice. Two Phase 1 trials were conducted in 142 healthy adult volunteers in the United States and Brazil, first in hookworm-naïve individuals and then in residents of a N. americanus endemic area in Brazil. Volunteers received one of three doses of recombinant Na-GST-1 (10, 30, or 100 μg) adjuvanted with Alhydrogel, adjuvanted with Alhydrogel and co-administered with an aqueous formulation of Glucopyranosyl Lipid A (GLA-AF), or the hepatitis B vaccine. Vaccinations were administered via intramuscular injection on days 0, 56, and 112. Na-GST-1/Alhydrogel was well tolerated in both hookworm-naïve and hookworm-exposed adults, with the most common adverse events being mild to moderate injection site pain and tenderness, and mild headache and nausea; no vaccine-related severe or serious adverse events were observed. Antigen-specific IgG antibodies were induced in a dose-dependent fashion, with increasing levels observed after each vaccination in both trials. The addition of GLA-AF to Na-GST-1/Alhydrogel did not result in significant increases in specific IgG responses. In both the US and Brazil studies, the predominant IgG subclass induced against Na-GST-1 was IgG1, with lesser amounts of IgG3. Vaccination of both hookworm-naïve and hookworm-exposed adults with recombinant Na-GST-1 was safe, well tolerated, and resulted in significant antigen-specific IgG responses. Based on these results, this vaccine will be advanced into clinical trials in children and eventual efficacy studies.

Trial registration: ClinicalTrials.gov (NCT01261130 for the Brazil trial and NCT01385189 for the US trial).

Conflict of interest statement

ABSVI supports the development and testing of several hookworm vaccines. PH is President of ABSVI and patent holder of the Na-GST-1 hookworm vaccine. All other authors declare that they have no conflicts of interest.

Figures

Fig 1. Study flow diagram for US…
Fig 1. Study flow diagram for US clinical trial.
Fig 2. Study flow diagram for Brazilian…
Fig 2. Study flow diagram for Brazilian clinical trial.
Fig 3. Geometric mean levels of IgG…
Fig 3. Geometric mean levels of IgG against recombinant Na-GST-1 in US participants immunized with Na-GST-1/Alhydrogel (administered with or without GLA-AF) as measured by ELISA.
Bars represent 95% confidence intervals. V1 = first vaccination; V2 = second vaccination; V3 = third vaccination; AU = arbitrary units.
Fig 4. Geometric mean levels of IgG…
Fig 4. Geometric mean levels of IgG against recombinant Na-GST-1 in Brazilian participants immunized with Na-GST-1/Alhydrogel (administered with or without GLA-AF) or the hepatitis B vaccine, as measured by ELISA.
Bars represent 95% confidence intervals. V1 = first vaccination; V2 = second vaccination; V3 = third vaccination; HBV = hepatitis B vaccine; AU = arbitrary units.
Fig 5. Comparison between US and Brazilian…
Fig 5. Comparison between US and Brazilian participants in levels of IgG against recombinant Na-GST-1 as measured by ELISA.
Bars represent 95% confidence intervals. V1 = first vaccination; V2 = second vaccination; V3 = third vaccination; HBV = hepatitis B vaccine; AU = arbitrary units.
Fig 6. Levels of IgG1 (panel A)…
Fig 6. Levels of IgG1 (panel A) and IgG3 (panel B) against recombinant Na-GST-1 in Brazilian participants vaccinated with Na-GST-1/Alhydrogel (administered with or without GLA-AF) or the hepatitis B vaccine, as measured by ELISA.
Bars represent 95% confidence intervals. V1 = first vaccination; V2 = second vaccination; V3 = third vaccination; HBV = hepatitis B vaccine; AU = arbitrary units.

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