Development of a Skin Test for the Na-ASP-2 Hookworm Antigen

Study Overview
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Brief Summary
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Na-ASP-2 is a protein expressed during the larval stage of the N. americanus hookworm life cycle. In a clinical study in previously hookworm-infected adults in Brazil, this protein induced urticarial reactions (rash) in a subset of volunteers. This study will evaluate solutions of varying concentrations of the protein in both a skin prick-puncture and intradermal test that will eventually be used to screen volunteers living in hookworm endemic areas who are being considered as potential participants in vaccine trials, to reduce the potential of inducing allergic reactions in those vaccinated with the Na-ASP-2 Hookworm Vaccine.

Detailed Description
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Study site: George Washington University Medical Center Number of participants: up to 15 Study duration: up to 3 months; each participant will undergo a minimum of one screening visit and one study visit. Each participant will have both skin prick-puncture and intradermal skin tests applied on the same study day. The prick-puncture and intradermal tests will be applied sequentially to the volar aspects of the participant's forearms: Skin prick-puncture: four different concentrations of Na-ASP-2 solution (1, 10, 100, and 1000 μg/mL) will be applied simultaneously in duplicate, to the left forearm. Intradermal: three different concentrations of Na-ASP-2 (10, 100, and 1000 μg/mL) will be injected simultaneously, in duplicate, to the right forearm. Allergen diluent and histamine solutions will be utilized as the negative and positive controls, respectively, for both the skin prick-puncture and intradermal tests. Participants will be observed in the study clinic for at least 30 minutes following application of the skin tests, and will be contacted via telephone 2 days following skin testing. Sizes of the wheal and erythema reactions for each application will be recorded and graded in comparison to the histamine positive control reaction. Each participant will have blood collected via venipuncture (a maximum of 25 mL) immediately prior to application of the skin tests, in order to measure antibodies (particularly IgG and IgE) to Na-ASP-2 using an ELISA procedure.

Official Title
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Development of a Skin Test to Detect Immediate-type Hypersensitivity to the Experimental Na-ASP-2 Hookworm Antigen

Conditions
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Hookworm Infection, Allergy

Intervention / Treatment
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* Biological: Na-ASP-2 Skin Test Reagent


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Males or females between 18 and 45 years of age, inclusive. Good general health as determined by means of the screening procedure. Willingness to participate in the study as evidenced by signing the informed consent document. Exclusion Criteria: History of previous infection with hookworm. Prior extensive and continuous travel, work, or residence (> 1 month) in a hookworm-endemic region. Pregnancy as determined by a positive urine hCG test (if female). Evidence of clinically significant neurologic, cardiac, pulmonary, hepatic, rheumatologic, autoimmune, or renal disease by history or physical examination. Behavioral, cognitive, or psychiatric disease that in the opinion of the investigator affects the ability of the volunteer to understand and cooperate with the study protocol. Other condition that in the opinion of the investigator would jeopardize the safety or rights of a volunteer participating in the trial or would render the subject unable to comply with the protocol. Participation in an investigational vaccine or drug trial within 14 days of starting this study. Volunteer has had medical, occupational, or family problems as a result of alcohol or illicit drug use during the past 12 months. History of a severe allergic reaction or anaphylaxis. Known immunodeficiency syndrome. Use of corticosteroids (excluding topical or nasal) or immunosuppressive drugs within 30 days of starting this study. History of a surgical splenectomy. Extensive dermatitis precluding skin testing Current use of a beta blocker (oral or topical) or anti-histamine medication. A volunteer may participate in the study if they agree to withhold use of an anti-histamine for at least 5 days prior to application of the skin test. Use of a tricyclic anti-depressant within the past month.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 45 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Diagnostic
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Na-ASP-2 Hookworm Antigen Skin Test<br>All participants will have the same number of concentrations of the Na-ASP-2 skin test reagent applied to their arms, using both the prick-puncture and intradermal techniques. | Biological: Na-ASP-2 Skin Test Reagent<br>* Na-ASP-2 Hookworm Skin Test Reagent, 1-1000 mcg/mL solution<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| 100 Mcg/ml Na-ASP-2 Prick-puncture Skin Test | Mean grade of wheal reaction after 2 applications of the skin test reagent per participant. For each skin test reaction, the grade of the test was determined based on the mean of the longest and orthogonal diameters, which was then compared to the equivalent measurements of a histamine solution positive control that was applied at the same time as the test. Grading was as follows: 0 no discernible wheal < ½ histamine diameter ≥ ½ histamine; < histamine diameter = size of histamine control ± 1 mm > histamine diameter; < 2x diameter ≥ 2x histamine control | 15 minutes after skin test application | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| 1000 Mcg/ml Na-ASP-2 Prick-puncture Skin Test | Mean grade of wheal reaction after 2 applications of the skin test reagent per participant. For each skin test reaction, the grade of the test was determined based on the mean of the longest and orthogonal diameters, which was then compared to the equivalent measurements of a histamine solution positive control that was applied at the same time as the test. Grading was as follows: 0 no discernible wheal < ½ histamine diameter ≥ ½ histamine; < histamine diameter = size of histamine control ± 1 mm > histamine diameter; < 2x diameter ≥ 2x histamine control | 15 minutes after skin test application | 
| 100 Mcg/ml Na-ASP-2 Intradermal Skin Test | Mean grade of wheal reaction after 2 applications of the skin test reagent per participant. For each skin test reaction, the grade of the test was determined based on the mean of the longest and orthogonal diameters, which was then compared to the equivalent measurements of a histamine solution positive control that was applied at the same time as the test. Grading was as follows: 0 no discernible wheal < ½ histamine diameter ≥ ½ histamine; < histamine diameter = size of histamine control ± 1 mm > histamine diameter; < 2x diameter ≥ 2x histamine control | 15 minutes after skin test application | 
| 1000 Mcg/ml Na-ASP-2 Intradermal Skin Test | Mean grade of wheal reaction after 2 applications of the skin test reagent per participant. For each skin test reaction, the grade of the test was determined based on the mean of the longest and orthogonal diameters, which was then compared to the equivalent measurements of a histamine solution positive control that was applied at the same time as the test. Grading was as follows: 0 no discernible wheal < ½ histamine diameter ≥ ½ histamine; < histamine diameter = size of histamine control ± 1 mm > histamine diameter; < 2x diameter ≥ 2x histamine control | 15 minutes after skin test application | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Hookworm, Allergy, Vaccine, Skin test