H7N9 Vaccination With and Without AS03 and Unadjuvanted H3N2v Vaccination: Standard and Systems Biology Analyses

Study Overview
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Brief Summary
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This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, inclusive, designed to evaluate and compare the immunogenicity between an intramuscular monovalent inactivated influenza A/H7N9 virus vaccine given with and without AS03 adjuvant, and an intramuscular unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine. The primary objectives are (1) assessing the serum anti-HA hemagglutination-inhibition (HAI) response to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v), and (2) identifying differentially expressed genes in human immune cells on Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine with or without AS03) and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 vaccine with or without AS03), compared to baseline assessments performed prior to each study vaccination (Days -7, 1, and 29).

Detailed Description
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This is a single center, randomized, partially-blinded, Phase II, small, targeted, prospective study in approximately 30 healthy male and non-pregnant female subjects aged 18 to 49 years old, inclusive, designed to evaluate and compare the immunogenicity between an intramuscular monovalent inactivated influenza A/H7N9 virus vaccine manufactured by Sanofi Pasteur given with and without AS03 adjuvant manufactured by GlaxoSmithKline, and an intramuscular unadjuvanted monovalent inactivated influenza A/H3N2v virus vaccine manufactured by Sanofi Pasteur. The primary objectives are (1) assessing the serum anti-HA hemagglutination-inhibition (HAI) response to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v), and (2) identifying differentially expressed genes in human immune cells on Days 2, 4, and 29 (following the first study vaccination with A/H7N9 vaccine with or without AS03) and on Days 30, 32, and 36 (following the second study vaccination with A/H7N9 vaccine with or without AS03), compared to baseline assessments performed prior to each study vaccination (Days -7, 1, and 29). The secondary objectives are: (1) compare plasma cytokine and chemokine profiles at specific time points and between treatment arms at post vaccination points, (2) assess the neutralizing antibody responses to influenza A/H7N9 antigen (with and without adjuvant) at Day 57 (approximately one month after the second study vaccination with A/H7N9 vaccine with or without AS03) and influenza A/H3N2v antigen at Day 29 (approximately one month after the study vaccination with A/H3N2v vaccine), and (3) identify differentially expressed genes in human immune cells on Days 2, 4, and 8 following one intramuscular dose of influenza A/H3N2v vaccine compared to baseline assessments performed prior to study vaccination (Day -7 and Day 1).

Official Title
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A Phase II Study to Evaluate and Compare the Immunogenicity of Monovalent Inactivated Influenza A/H7N9 Virus Vaccine Administered With and Without AS03 Adjuvant and Monovalent Inactivated Influenza A/H3N2v Virus Vaccine Administered Without Adjuvant in Healthy Adults Through Standard and Systems Biology Analyses

Conditions
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Avian Influenza, H1N1 Influenza, Influenza

Intervention / Treatment
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* Drug: AS03
* Biological: Influenza Virus Vaccine, Monovalent A/H3N2v A/Minnesota/11/2010 NYMC X-203
* Biological: Monovalent influenza A/H7N9 virus vaccine


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Provide written informed consent prior to initiation of any study procedures. Are able to understand and comply with planned study procedures and be available for all study visits. Are males or non-pregnant females, 18 to 49 years old, inclusive. Are in good health*. *As determined by medical history and targeted physical examination, if indicated based on medical history, to evaluate acute or currently ongoing chronic medical diagnoses or conditions, defined as those that have been present for at least 90 days, that would affect the assessment of the safety of subjects or the immunogenicity of study vaccinations. Chronic medical diagnoses or conditions should be stable for the last 60 days. This includes no change in chronic prescription medication, dose, or frequency as a result of deterioration of the chronic medical diagnosis or condition in the 60 days prior to enrollment. Any prescription change that is due to change of health care provider, insurance company, etc., or that is done for financial reasons, as long as in the same class of medication, will not be considered a deviation of this inclusion criterion. Any change in prescription medication due to improvement of a disease outcome, as determined by the site principal investigator or appropriate sub-investigator, will not be considered a deviation of this inclusion criterion. Subjects may be on chronic or as needed (prn) medications if, in the opinion of the site principal investigator or appropriate sub-investigator, they pose no additional risk to subject safety or assessment of reactogenicity and immunogenicity and do not indicate a worsening of medical diagnosis or condition. Similarly, medication changes subsequent to enrollment and study vaccination are acceptable provided there was no deterioration in the subject's chronic medical condition that necessitated a medication change, and there is no additional risk to the subject or interference with the evaluation of responses to study vaccination. Note: Topical, nasal, and inhaled medications (with the exception of inhaled corticosteroids as outlined in the Subject Exclusion Criteria), herbals, vitamins, and supplements are permitted. Oral temperature is less than 100.4 degrees F. Pulse is 50 to 115 bpm, inclusive. Systolic blood pressure is 85 to 150 mm Hg, inclusive. Diastolic blood pressure is 55 to 95 mm Hg, inclusive. Erythrocyte sedimentation rate (ESR) is less than 30 mm per hour. Women of childbearing potential* must use an acceptable contraception method** from 30 days before first study vaccination until 60 days after last study vaccination. *Not sterilized via tubal ligation, bilateral oophorectomy, hysterectomy or successful Essure (R) placement (permanent, non-surgical, non-hormonal sterilization) with documented radiological confirmation test at least 90 days after the procedure, and still menstruating or <1 year of the last menses if menopausal. **Includes, but is not limited to, non-male sexual relationships abstinence from sexual intercourse with a male partner, monogamous relationship with vasectomized partner who has been vasectomized for 180 days or more prior to the subject receiving the first study vaccination, barrier methods such as condoms or diaphragms with spermicide or foam, effective intrauterine devices, NuvaRing (R), and licensed hormonal methods such as implants, injectables or oral contraceptives (the pill). Women of childbearing potential must have a negative urine or serum pregnancy test within 24 hours prior to study vaccination. Exclusion Criteria: Have an acute illness*, as determined by the site principal investigator or appropriate sub-investigator, within 72 hours prior to study vaccination. *An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion of the site principal investigator or appropriate sub-investigator, the residual symptoms will not interfere with the ability to assess safety parameters as required by the protocol. Have any medical disease or condition that, in the opinion of the site principal investigator or appropriate sub-investigator, is a contraindication to study participation*. *Including acute or chronic medical disease or condition, defined as persisting for at least 90 days that would place the subject at an unacceptable risk of injury, render the subject unable to meet the requirements of the protocol, or may interfere with the evaluation of responses or the subject's successful completion of this study. Have immunosuppression as a result of an underlying illness or treatment, or use of anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study vaccination. Have known active neoplastic disease (excluding non-melanoma skin cancer) or a history of any hematologic malignancy. Have known human immunodeficiency virus (HIV), hepatitis B, or hepatitis C infection. Have known hypersensitivity or allergy to eggs, egg or chicken protein, squalene-based adjuvants, or other components of the study vaccines. Have a history of severe reactions following previous immunization with licensed or unlicensed influenza virus vaccines. Have a personal or family history of narcolepsy. Have a history of Guillain-Barré syndrome. Have a history of convulsions or encephalomyelitis within 90 days prior to study vaccination. Have a history of a potentially immune-mediated medical condition. Have a history of alcohol or drug abuse within 5 years prior to study vaccination. Have any diagnosis, current or past, of schizophrenia, bipolar disease, or other psychiatric diagnosis that may interfere with subject compliance or safety evaluations. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for danger to self or others within 10 years prior to study vaccination. Have taken oral or parenteral (including intra-articular) corticosteroids of any dose within 30 days prior to study vaccination. Have taken high-dose inhaled corticosteroids within 30 days prior to study vaccination. High-dose defined as >840 mcg/day of beclomethasone dipropionate CFC or equivalent. Received licensed live vaccine within 30 days prior to the first study vaccination, or plans to receive licensed live vaccine within 30 days before or after each study vaccination. Received licensed inactivated vaccine within 14 days prior to the first study vaccination, or plans to receive licensed inactivated vaccine within 14 days before or after each study vaccination. Received immunoglobulin or other blood products (with exception of Rho D immunoglobulin) within 90 days prior to study vaccination. Received an experimental agent* within 30 days prior to the first study vaccination, or expects to receive an experimental agent** during the 13-month study-reporting period. *Including vaccine, drug, biologic, device, blood product, or medication. **Other than from participation in this study. Are participating or plan to participate in another clinical trial with an interventional agent* that will be received during the 13-month study-reporting period. *Including licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication. Prior participation in a clinical trial of influenza A/H7 vaccine* or have a history of influenza A/H7 virus actual or potential exposure or infection prior to the first study vaccination. *And assigned to a group receiving influenza A/H7 vaccine, does not apply to documented placebo recipients. Prior participation in a clinical trial of influenza A/H3N2v vaccine* or have a history of influenza A/H3N2v virus actual or potential exposure or infection prior to the first study vaccination. *And assigned to a group receiving influenza A/H3N2v vaccine, does not apply to documented placebo recipients. Occupational exposure to or substantial direct physical contact* with birds in the past year or during the 28 days after each study vaccination. *Casual contact with birds at petting zoos or county or state fairs or having pet birds does not exclude subjects from study participation. Occupational exposure to or substantial direct physical contact* with pigs in the past year or during the 28 days after each study vaccination. *Casual contact with pigs at petting zoos or county or state fairs does not exclude subjects from study participation. Female subjects who are breastfeeding or plan to breastfeed at any given time from the first study vaccination until 30 days after the last study vaccination. Plan to travel outside the US (continental US, Hawaii, and Alaska) within 28 days after each study vaccination. Blood donation or planned blood donation within 30 days before enrollment until 30 days after the last blood draw for this study.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 49 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: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Arm 1<br>10 Subjects: one dose of 15 µg of A/H3N2v, at Day 1 | Biological: Influenza Virus Vaccine, Monovalent A/H3N2v A/Minnesota/11/2010 NYMC X-203<br>* Inactivated monovalent subvirion H3N2v vaccine containing hemagglutinin (HA) of A/Minnesota/11/2010 NYMC X-203 virus.<br>|
| Experimental: Arm 2<br>10 Subjects: two doses of 3.75 µg of AH7N9 AS03,at Day 1 and at Day 29 | Drug: AS03<br>* AS03 oil-in-water emulsion adjuvant.<br>Biological: Monovalent influenza A/H7N9 virus vaccine<br>* Monovalent influenza A/H7N9 virus vaccine.<br>|
| Experimental: Arm 3<br>10 Subjects: two doses of 3.75 µg of A/H7N9, at Day 1 and Day 29 | Biological: Monovalent influenza A/H7N9 virus vaccine<br>* Monovalent influenza A/H7N9 virus vaccine.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Percentage of Participants Achieving Seroconversion Based on HAI Titer (A/H7N9 Vaccine, With and Without Adjuvant) | Seroconversion is defined as either a pre-vaccination HAI titer <1:10 and a post-vaccination HAI titer =>1:40 or a pre-vaccination HAI titer =>1:10 and a minimum four-fold rise in post-vaccination HAI antibody titer. Venous blood samples for serum were used for this assay. HAI based on the H7N9 strain A/Shanghai/02/2013xPR8 were assessed. | Day 29 Post-Vaccination 2 (Day 57) | 
| Percentage of Participants Achieving Seroconversion Based on HAI Titer (A/H3N2v Vaccine) | Seroconversion is defined as either a pre-vaccination HAI titer <1:10 and a post-vaccination HAI titer =>1:40 or a pre-vaccination HAI titer =>1:10 and a minimum four-fold rise in post-vaccination HAI antibody titer. Venous blood samples for serum were used for this assay. HAI based on the H3N2 strain A/Minnesota/11/2010 were assessed. | Day 29 Post-Vaccination 1 (Day 29) | 
| Number of Differentially Expressed Genes Based on RNA Expression in Human Immune Cells (A/H7N9 Vaccine, With and Without Adjuvant) | RNA expression levels (read counts) for each gene were determined by RNA-Sequencing of RNA extracted from six separated cell types: monocytes , dendritic cells, neutrophils, NK cells, B cells, and T cells. Post-vaccination gene expression levels were compared to pre-vaccination gene expression levels (combined summed read counts for Day -7 and Day 1 samples for post-vaccination 1 timepoints and Day 29 read counts for post-vaccination 2 timepoints) using a negative binomial model to identify differentially expressed (DE) genes (FDR-adjusted p-value < 0.05 and mean fold change => 1.5 in either direction). | Days 2, 4, and 29 Post-Vaccination 1 (Day 2, 4, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of Differentially Expressed Genes Based on RNA Expression in Human Immune Cells (A/H3N2v Vaccine) | RNA expression levels (read counts) for each gene were determined by RNA-Sequencing of RNA extracted from six separated cell types: monocytes, dendritic cells, neutrophils, NK cells, B cells, and T cells. Post-vaccination gene expression levels were compared to pre-vaccination gene expression levels (combined summed read counts for Day -7 and Day 1 samples for post-vaccination 1 timepoints) using a negative binomial model to identify differentially expressed (DE) genes (FDR-adjusted p-value < 0.05 and mean fold change => 1.5 in either direction). | Days 2, 4, and 8 Post-Vaccination 1 (Day 2, 4, and 8, respectively) | 
| Cytokine and Chemokine Concentration by Study Arm and Study Visit Day - Part 1 | Venous blood samples for plasma were used for this assay. A Luminex assay was used to measure concentrations in ng/L. Concentrations for 11 of the total 24 cytokines and chemokines were measured: Fractalkine, Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF), Interferon-Gamma-Induced Protein-10(IP-10), Interferon-Alpha (IFNa), Interferon-Beta (IFNb), Interferon-Gamma (IFNy), Interferon-inducible T Cell Alpha Chemoattractant (ITAC), Interleukin 1 Beta (IL-1b), Interleukin 10 (IL-10), Interleukin 13 (IL-13), and Interleukin 2 (IL-2). If no cytokine or chemokine measurements were collected, number of participants is reported as 0. | Day 1 Pre-Vaccination 1; Days 2, 4, 8, and 29 Post-Vaccination 1 (Day 2, 4, 8, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 
| Cytokine and Chemokine Concentration by Study Arm and Study Visit Day - Part 2 | Venous blood samples for plasma were used for this assay. A Luminex assay was used to measure concentrations in ng/L. Concentrations for 9 of the total 24 cytokines and chemokines were measured: Interleukin 21 (IL-21), Interleukin 23 (IL-23), Interleukin 4 (IL-4), Interleukin 5 (IL-5), Interleukin 6 (IL-6), Interleukin 7 (IL-7), Interleukin 8 (IL-8), Interleukin 12, p70 (IL-12p70), and Interleukin 17A (IL-17A). If no cytokine or chemokine measurements were collected, number of participants is reported as 0. | Day 1 Pre-Vaccination 1; Days 2, 4, 8, and 29 Post-Vaccination 1 (Day 2, 4, 8, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 
| Cytokine and Chemokine Concentration by Study Arm and Study Visit Day - Part 3 | Venous blood samples for plasma were used for this assay. A Luminex assay was used to measure concentrations in ng/L. Concentrations for 4 of the total 24 cytokines and chemokines were measured: Macrophage Inflammatory Protein-1 Alpha (MIP1a), Macrophage Inflammatory Protein-1 Beta (MIP1b), Macrophage Inflammatory Protein-3 Alpha (MIP3a), and Tumor Necrosis Factor Alpha (TNF-a). If no cytokine or chemokine measurements were collected, number of participants is reported as 0. | Day 1 Pre-Vaccination 1; Days 2, 4, 8, and 29 Post-Vaccination 1 (Day 2, 4, 8, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 
| Fold Change in Cytokine and Chemokine Concentrations From Pre-vaccination - Part 1 | Venous blood samples for plasma were used for this assay. A Luminex assay was used to measure concentrations in ng/L. Concentrations for 7 of the total 24 cytokines and chemokines were measured: Fractalkine, GM-CSF, IP-10, IFNa, IFNb, IFNy, and ITAC. If no cytokine or chemokine measurements were collected, number of participants is reported as 0. | Days 2, 4, 8, and 29 Post-Vaccination 1 (Day 2, 4, 8, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 
| Fold Change in Cytokine and Chemokine Concentrations From Pre-vaccination - Part 2 | Venous blood samples for plasma were used for this assay. A Luminex assay was used to measure concentrations in ng/L. Concentrations for 17 of the total 24 cytokines and chemokines were measured: IL-1b, IL-10, IL-13, IL-2, IL-21, IL-23, IL-4, IL-5, IL-6, IL-7, IL-8, IL-12p70, IL-17A, MIP1a, MIP1b, MIP3a, and TNF-a. If no cytokine or chemokine measurements were collected, number of participants is reported as 0. | Days 2, 4, 8, and 29 Post-Vaccination 1 (Day 2, 4, 8, and 29, respectively); Days 2, 4, and 8 Post-Vaccination 2 (Day 30, 32, and 36, respectively) | 
| Percentage of Participants Achieving Seroconversion Based on Neut Titer (A/H7N9 Vaccine, With and Without Adjuvant) | Seroconversion is defined as either a pre-vaccination Neut titer <1:10 and a post-vaccination Neut titer =>1:40 or a pre-vaccination Neut titer =>1:10 and a minimum four-fold rise in post-vaccination Neut titer. Venous blood samples for serum were used for this assay. Neut based on the H7N9 strain A/Shanghai/02/2013xPR8 were assessed. | Day 29 Post-Vaccination 2 (Day 57) | 
| Percentage of Participants Achieving Seroconversion Based on Neut Titer (A/H3N2v Vaccine) | Seroconversion is defined as either a pre-vaccination Neut titer <1:10 and a post-vaccination Neut titer =>1:40 or a pre-vaccination Neut titer =>1:10 and a minimum four-fold rise in post-vaccination Neut titer. Venous blood samples for serum were used for this assay. Neut based on the H3N2 strain A/Minnesota/11/2010 were assessed. | Day 29 Post-Vaccination 1 (Day 29) | 
| Geometric Mean Titers of Serum HAI Antibody (A/H7N9 Vaccine, With and Without Adjuvant) | Venous blood samples for serum were used for this assay. HAI based on the H7N9 strain A/Shanghai/02/2013xPR8 were assessed. | Day 1 Pre-Vaccination 1, Day 29 Post-Vaccination 1 (Day 29), and Day 29 Post-Vaccination 2 (Day 57) | 
| Geometric Mean Titers of Serum HAI Antibody (A/H3N2v Vaccine) | Venous blood samples for serum were used for this assay. HAI based on the H3N2 strain A/Minnesota/11/2010 were assessed. | Day 1 Pre-Vaccination 1 (Day 1), Day 29 Post-Vaccination 1 (Day 29) | 
| Geometric Mean Titers of Serum Neut Antibody (A/H7N9 Vaccine, With and Without Adjuvant) | Venous blood samples for serum were used for this assay. Neut based on the H7N9 strain A/Shanghai/02/2013xPR8 were assessed. | Day 1 Pre-Vaccination 1 (Day 1), Day 29 Post-Vaccination 1 (Day 29), and Day 29 Post-Vaccination 2 (Day 57) | 
| Geometric Mean Titers of Serum Neut Antibody (A/H3N2v Vaccine) | Venous blood samples for serum were used for this assay. Neut based on the H3N2 strain A/Minnesota/11/2010 were assessed. | Day 1 Pre-Vaccination 1 (Day 1), Day 29 Post-Vaccination 1 (Day 29) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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A/H5N1, A/H7N9, ASO3 Adjuvant, influenza, monovalent, pH1N1, vaccines