Safety Trial of Live Attenuated Influenza (H7N3) Vaccine
Safety Trial of Live Attenuated Influenza (H7N3) Vaccine
The study hypothesis is that two doses of cold-adapted, live monovalent
 A/17/mallard/Netherlands/00/95 (H7N3) influenza vaccine will be safe and immunogenic in
 healthy adults.
This is a phase I, double-blind, individually-randomized (3:1, vaccine:placebo), controlled
 trial with two groups, LAIV H7N3 and matched placebo. Healthy male and female adults 18
 through 49 years of age will be invited to participate. For feasibility reasons and in order
 for an independent Safety Monitoring Committee (SMC) to review safety data in a small group
 of subjects initially, the total cohort of 40 subjects will be enrolled in two sub-cohorts:
 one cohort of 12 subjects, randomized at 3:1 (9 vaccine and 3 placebo), followed two weeks
 later by a second cohort of 28 subjects randomized at 3:1 (21 vaccine and 7 placebo). After
 all 12 volunteers of the first sub-cohort have been observed for the first isolation period
 (Day 1 to Day 7), an interim safety review will be performed by the SMC. The SMC will review
 all adverse events (AEs), including clinical laboratory evaluations (pre- and
 post-vaccination) and shedding data, for all subjects and will advise if the volunteers of
 the first sub-cohort may receive dose two of study vaccine or placebo and if the additional
 28 volunteers of the second sub-cohort may be enrolled into the study. As for the first
 sub-cohort, the SMC will also review all safety data for the second sub-cohort and for the
 entire participant population of the trial. For each sub-cohort, the procedures and timelines
 are here summarized.

 On the day of first screening, about 7 days (between 4 and 14 days) prior to administration
 of dose one of study vaccine or placebo, subjects will be screened for eligibility through
 medical history review, physical examination, testing for serologic evidence of chronic viral
 infection [human immunodeficiency virus (HIV), hepatitis B virus (HBV) or hepatitis C virus
 (HCV), with proper pre- and post-test counseling], routine biochemical and hematological
 blood tests and urinalysis by dipstick.

 Subject screening for eligibility will continue and be completed on the second screening day
 (S2). This second screening day will occur the same day as scheduled admission to the
 isolation unit and administration of study vaccine or placebo (Day 0). Women will undergo
 pregnancy tests using urine samples. All subjects will undergo an ear, nose and throat (ENT)
 examination. Fully eligible subjects will be admitted to the isolation unit. At that time,
 nasal swab, nasal wick, and blood specimens will be collected for virologic and immunological
 testing prior to administration of study vaccine or placebo. Blood and urine specimens will
 be again collected for routine biochemical and hematological blood tests and urinalysis by
 dipstick; these results will serve to define baseline status for subject prior to receipt of
 study vaccine or placebo but will not be used for screening purposes. Subjects will be
 unaware of which allocation, LAIV H7N3 or matched placebo, is received; study vaccine and
 placebo will be masked. Subjects will be carefully monitored for adverse reactions while in
 the isolation unit.

 All subjects will remain in the isolation unit for at least 7 days after receipt of study
 vaccine or placebo. Nasal swabs will be collected daily while subjects are in isolation to
 test for presence of influenza virus shed in the nasal passage. Any subject exhibiting
 conjunctivitis will also have a conjunctival swab collected on the day of appearance of the
 sign. Any subject exhibiting influenza A virus shedding, as determined by real-time reverse
 transcriptase polymerase chain reaction (RT-PCR) positivity on a nasal swab specimen, in the
 2 days prior to each planned discharge day after each dose (Days 6 or 7 or Days 34 or 35)
 will be kept in the isolation until PCR-diagnosis results confirm that no influenza virus is
 present in a tested clinical specimen for at least two consecutive days. Any subject still
 exhibiting evidence of influenza virus shedding in a nasal swab on Days 6 or 7 or Days 34 or
 35 post-administration with each dose might be placed on influenza antiviral (oseltamivir)
 treatment at the standard dose for treatment of 75 milligrams (mg) twice a day for a course
 of 5 days.

 After discharge from the isolation unit, subjects will complete diary cards for AEs and use
 of concomitant medications. Subjects will return to the isolation unit at four weeks (Day 28)
 after administration of dose one of study vaccine or placebo. At that time, similar
 procedures will be used for admittance to the isolation unit, for receipt of dose two of
 study vaccine or placebo and for isolation and follow-up, with the additional procedure of
 review of interim histories (and diary cards) since first discharge after dose one.

 After second discharge from the isolation unit, subjects will again complete diary cards for
 AEs and use of concomitant medications. Subjects will then return to the study center at four
 weeks (Day 56) after administration of dose two of study vaccine or placebo for their final
 study visit. Interim histories (and diary cards) will again be reviewed and final blood and
 nasal wick specimens will be collected. Women will also undergo a final pregnancy screen.
 Subjects will complete the study at this time.

 For assessment of safety, subjects will be observed for two hours after each administration
 of study vaccine or placebo. Twice daily (early morning and late afternoon) examination will
 be also used to assess reactions for 7 days after each administration of study vaccine or
 placebo. ENT examination will also occur once per day on Days 7, 28, 35 and 56. Subjects will
 complete diary cards for unsolicited AEs from the day of each discharge until return to the
 isolation unit for dose two (at Day 28) or until return to the study center for the final
 study visit at four weeks post dose two (at Day 56). To assess safety, blood and urine
 specimens will also be collected on days 7, 28 (prior to administration of dose two of study
 vaccine or placebo), 35 and 56, for testing by routine biochemical and hematological blood
 tests and by urinalysis by dipstick.

 For the evaluation of mucosal immunoglobulin A (IgA) antibody, nasal wick specimens will be
 collected on Day 0 (prior to administration of dose one of study vaccine or placebo), on Day
 28 (prior to administration of dose two of study vaccine or placebo) and on Day 56. For the
 evaluation of serum antibodies (by hemagglutination inhibition [HAI], microneutralization and
 IgA and immunoglobulin G [IgG] EIA), serum specimens will be collected on Day 0 (prior to
 administration of dose one of study vaccine or placebo), on Day 28 (prior to administration
 of dose two of study vaccine or placebo) and on Day 56. To study virus infectivity (by
 isolation in chicken embryos) and stability (by molecular sequencing of any isolated virus),
 nasal swab specimens will be taken on Days 1, 2, 3, 5, 7, 29, and 31. To assess priming and
 stimulation of cytotoxic T lymphocytes and other cytokine indicators, whole blood for
 isolation of peripheral blood mononuclear cells (PBMCs) will be collected on Days 0 (prior to
 administration of dose one of study vaccine or placebo), on Day 28 (prior to administration
 of dose two of study vaccine or placebo) and on Day 56.
Inclusion Criteria:

 -  Legal male or female adult 18 through 49 years of age at the enrollment visit.

 -  Literate and willing to provide written informed consent.

 -  Free of obvious health problems, as established by the medical history and screening
 evaluations, including physical examination.

 -  Capable and willing to complete diary cards and willing to return for all follow-up
 visits

 -  Willing to comply with the rules of the isolation unit (including willing and able to
 take oseltamivir influenza antiviral medication, should that be recommended by a study
 physician).

 -  For females, willing to take reliable birth control measures throughout the entire
 period of participation in the study.

 Exclusion Criteria:

 -  Participation in another clinical trial involving any therapy within the previous
 three months or planned enrollment in such a trial during the period of this study.

 -  Receipt of any non-study vaccine within four weeks prior to enrollment or refusal to
 postpone receipt of such vaccines until four weeks after study completion.

 -  Practice of nasal irrigation on a regular basis within the past six months or has
 engaged in nasal irrigation within two weeks prior to enrollment.

 -  Recent history of frequent nose bleeds (>5 within the past year).

 -  Clinically relevant abnormal paranasal anatomy.

 -  Recent history (within the past month) of rhino or sinus surgery, or surgery for any
 traumatic injury of the nose.

 -  Current or recent (within two weeks of enrollment) acute respiratory illness with or
 without fever.

 -  Other acute illness at the time of study enrollment.

 -  Receipt of immune globulin or other blood products within three months prior to study
 enrollment or planned receipt of such products during the period of subject
 participation in the study.

 -  Chronic administration (defined as more than 14 consecutively-prescribed days) of
 immunosuppressants or other immune-modulating therapy within six months prior to study
 enrollment. (For corticosteroids, this means prednisone or equivalent, >=0.5 mg per kg
 per day; topical steroids are allowed, exclusive of nasal.)

 -  Participation in any previous trial of any H5 or H7 containing influenza vaccine.

 -  History of asthma.

 -  Hypersensitivity after previous administration of any influenza vaccine.

 -  History of wheezing after past receipt of any live influenza vaccine.

 -  Other AE following immunization, at least possibly related to previous receipt of any
 influenza vaccine.

 -  Suspected or known hypersensitivity to any of the study vaccine components, including
 chicken or egg protein.

 -  Seasonal (autumnal) hypersensitivity to the natural environment.

 -  Acute or chronic clinically significant pulmonary, cardiovascular, hepatic, metabolic,
 neurologic, psychiatric or renal functional abnormality, as determined by medical
 history, physical examination or clinical laboratory screening tests, which in the
 opinion of the investigator, might interfere with the study objectives. Subjects with
 physical examination findings or clinical laboratory screening results which would be
 graded 2 or higher on the AE severity grading scale will be excluded from entry into
 the study and will be excluded from receipt of dose two of study vaccine or placebo.

 -  History of leukemia or any other blood or solid organ cancer.

 -  History of thrombocytopenic purpura or known bleeding disorder.

 -  History of seizures.

 -  Known or suspected immunosuppressive or immunodeficient condition of any kind,
 including HIV infection.

 -  Known chronic HBV or HCV infection.

 -  Known tuberculosis infection or evidence of previous tuberculosis exposure.

 -  History of chronic alcohol abuse and/or illegal drug use.

 -  Claustrophobia or sociophobia.

 -  Pregnancy or lactation. (A negative pregnancy test will be required before
 administration of study vaccine or placebo for all women of childbearing potential.)

 -  Any condition that, in the opinion of the investigator, would increase the health risk
 to the subject if he/she participates in the study or would interfere with the
 evaluation of the study objectives.