Analysis of Bone Marrow and Blood B Cell Immune Responses to Influenza Vaccination

This is a Phase IV open label and single arm study enrolling up to 55 healthy males and
non-pregnant females in a single site, 18-49 years old, inclusive. The study is designed to
assess the humoral response to influenza vaccination and the longevity of humoral immunity to
influenza vaccination in healthy adults. The laboratory technique will characterize
persistence and clonotype of antigen specific B-cells and plasmablasts in blood and bone
marrow. Enrolled subjects will receive licensed seasonal inactivated influenza vaccine
(administered as a part of the study). The vaccine will be administered according to the
package insert, and study participants will donate serial samples of blood and bone marrow
aspirate for immunology monitoring. Safety will be assessed from the time of study enrollment
through the last study visit, via monitoring of vital signs, change in health status, and
targeted physical exam with safety labs prior to each bone marrow aspirate procedure.
Repeated measurements of humoral immunity will be obtained at 7 days, 14 days, 28 days, 90
days and at one year post vaccination to assess the magnitude, clonal diversity and
persistence of B-cell responses to influenza vaccination. The study duration is up to 4
years. After 1 year, subjects will be offered the opportunity to participate in the study for
up to 3 consecutive years provided eligibility criteria is met each year. Subjects who elect
to continue in the study after first year of participation will be rescreened to verify
continued eligibility and re-consented prior to subsequent participation. Re-enrolling
subjects will receive new subject identifiers and will count towards the total enrollment
number for subsequent years of participation. A separate subject record will be maintained
each year a subject re-enrolls in the study. Enrollment for the next year will begin with the
availability of the seasonal flu vaccine. For subjects who elect to re-enroll in the study,
the Day 365 visit (+/- 3 month window) would also be the Day 0 visit for the subsequent year.
The primary study objective is to investigate the longevity of humoral immunity to influenza
virus in humans. The secondary study objective is the longitudinal tracking of
vaccine-induced B cell responses with special emphasis on broadly neutralizing HA
stem-reactive responses.

Inclusion Criteria:

1. Male or non-pregnant female subjects between 18 and 49 years of age (inclusive).

2. Subjects capable of providing written informed consent prior to initiation of any
study procedures.

3. Subjects able to understand and comply with planned study procedures and be available
for all study visits.

4. Safety labs:

- WBC, within reference range of lower limit of normal of 4,000 / uL, and upper
limit of normal of 10,000 / uL.

- Hemoglobin, within reference range of lower limit of normal of 11.4 gm / dL and
upper limit of normal of 16.1 gm / dL.

- Hematocrit, within reference range of lower limit of normal of 33.3% and an upper
limit of normal of 46.5%.

- Platelet Count, within reference range of lower limit of normal of 150,000 / uL
and upper limit of normal of 400,000 / uL.

- PT / INR, PT below or equal to the upper limit of normal of 13.1 seconds; INR
within normal reference range of less than 1.5 (normal range for
non-anti-coagulated patients).

- Creatinine within reference range of lower limit of normal of 0.4 mg / dL and
upper limit of normal of 1.2 mg / dL.

- Potassium, within reference range of lower limit of normal of 3.6 mM and upper
limit of normal of 5.1 mM.

5. Heart rate > / = 55 to < / = 100 per minute.

6. Systolic blood pressure > / = 90 to < / = 150 mm Hg.

7. Diastolic blood pressure < 90 mm Hg.

8. Oral temperature < 100 degrees Fahrenheit.

9. Respirations even, unlabored, and > 10 / minute to < 20 / minute.

10. Female subjects of child bearing potential must have a negative urine pregnancy test
at the screening visit, enrollment visit and prior to subsequent bone marrow aspirate
procedures. Female subjects of childbearing potential must agree to practice
abstinence, use a barrier method of birth control, or use an FDA approved form of
birth control.

11. Subjects who have not received seasonal flu vaccine for the current year (September -
June).

Exclusion Criteria:

1. If female, active pregnancy or breast-feeding or plans to become pregnant during study
participation.

2. Subject report of having any medical disease or condition that, in the opinion of the
site principal investigator or appropriate sub-investigator, is a contraindication to
study participation. This includes any acute or chronic medical disease or condition,
defined as persisting 3 months (defined as 90 days) or longer, 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.

3. Subject report of taking anticoagulants, or long-term (greater than 14 days) systemic
steroids or other immunosuppressive medications.

4. Subject report of known allergy to lidocaine.

5. Subject report of known hypersensitivity or allergy to eggs, egg or chicken protein,
report of allergy to components of the study vaccine or other components of the study
vaccine.

6. Subject report of known latex allergy.

7. Subject report of a history of severe reactions following previous immunization with
licensed or unlicensed influenza virus vaccines.

8. Subject report of a history of Guillain-Barre syndrome.

9. Subjects who believe they cannot tolerate the bone marrow aspirations without
sedation.

10. Subjects with an active infection or that have an acute illness within 72 hours prior
to study vaccination. Subject having had 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.

11. Subjects who are participating in another clinical trial involving the use of
investigational agents or vaccines.