Trial Information

Evaluation of Immunologic Response After Pandemic Influenza A (H1N1) Vaccine in Oncologic and Hematologic Patients

Primary objective

1) To assess whether oncologic and hematologic patients develop a protective immunological
response after pandemic Influenza A (H1N1) vaccine Secondary objectives

1. To compare the levels of antibody response against A (H1N1) influenza virus between
oncologic and hematologic patients relative to a cohort of healthy volunteers

2. To assess the incidence of A (H1N1) infection in vaccinated oncologic and hematologic
patients in comparison with a cohort of vaccinated healthy volunteers. To assess the
clinical symptoms attributable to influenza infection in vaccinated oncologic and
hematological patients and healthy volunteers.

3. To compare the levels of antibody response against A (H1N1) influenza virus between the
following subgroups: patients with ongoing chemotherapy; patients who have completed
the chemotherapy treatment; patients treated with autologous or allogeneic peripheral
blood hematopoietic stem cell transplant (PBSCT) Study population and design

The study population consists consecutive patients with oncologic or hematologic diseases
who are planned to receive A (H1N1) influenza vaccine

Study procedures The patients will perform a blood sample collection (serum vial) on day 0
(range: 0- 2 days before vaccination) before the vaccination, a blood sample collection
(serum vial) on day +21 (range: +/- 5 days) after vaccination , a blood sample collection
(serum vial) on day +50 (range: +/- 5 days) and on day +90 range: +/- 5 days) after
vaccination. The samples will be frozen in 500 mcl aliquots at -20°C. At the end of the
collection we will perform immunological test to evaluate the antibody titer and the
cellular response. The serum samples will be stored at the laboratory of Virology of the
University of Milan. The titer of antibodies against the vaccine strain will be measured in
all samples by means of hemagglutination-inhibition (HI) assays with the use of turkey
erythrocytes and according to EMEA guidelines. Response criteria will be the achievement of
a protective title of HI test > 1:40. In addition we will evaluate: geometric mean titers
and a fourfold titer increase compared with prevaccination titers. Cellular-mediated
response will be analysed by incubating CD3+ patients' cells with influenza A Antigens and
evaluation of: 1) cellular expansion by flow-cytometry analysis of dilution of
carboxyfluorescein succinimidyl ester (CFSE); 2) IFN-gamma production by ELISPOT.

A control cohort of healthy volunteers who received A(H1N1) vaccine will perform the same
blood sample collection in order to compare the immunological response between oncologic and
hematologic patients relative to healthy cohort.

Evaluation of clinical response:

Oncologic and hematologic patients will be followed as outpatients or inpatients according
to routine controls for their disease. In case that symptoms of the upper airways or
influenza-like symptoms develop, the symptoms will be recorded in the clinical database,
nasal and pharyngeal swaps will be performed according to the doctor who is taking care of
the patient. In order to evaluate the clinical efficacy of the vaccination, the swaps will
be tested for A (H1N1) influenza virus infection.

No further studies will be performed after 3 months from the vaccination.

Sample size The trial will accrue 25 patients for each subpopulation to be analyzed. This
sample size has a 90% power to evaluate an increase of the probability to have a biological
response from a theoretical value of 20% (low efficacy of the vaccine) to a value of 50%
(target of effectiveness) at the 5% significance (student t test, one tail). The
subpopulations are as follows: patients with ongoing chemotherapy; patients who have
completed the chemotherapy treatment; patients treated with autologous or allogeneic
transplant of hematopoietic stem cell.

We will accrue a "calibration" group comprising at least 100 healthy volunteers. This group
will comprise people working at the National Cancer Institute in Milan who have signed an
informed consent to participate to the study. This size of the sample allows a precision on
the evaluation of the probability of obtaining a biological response (half of the confidence
interval) that is not lower than 10%

Study duration The estimated duration of enrolment is of 6 months. The enrolment of the
hematologic and oncologic patients and of the cohort of healthy volunteers will be performed
in 3 months and the study will be closed on day +90 with a blood sample collection.

Selection criteria Inclusion criteria

- Age ≥18 years

- Oncologic and hematologic patient with the plan of receiving the A (H1N1) vaccine

- Control Group: a silent history for oncologic and hematologic diseases; planned of
receiving the A (H1N1) vaccine

- Written informed consent

Exclusion criteria

- Infusion of human Immunoglobulin ongoing or within prior 30 days

- Therapy with monoclonal or polyclonal antibodies ongoing or within prior 30 days

- Therapy with IL-1 or IL-2 or IFN-gamma ongoing or within prior 30 days

- Autologous PBSCT less than 1 month or Allogeneic PBSCT less than 6 months

- Pregnancy or lactation

- Type I hypersensitivity

- Ongoing Anticoagulant therapy or platelets < 50000/ul

Study Procedures at baseline: Medical history for oncologic and hematologic disease;
gynecologic history for women under 50 years of age

• Serum sample on the day of vaccination or 2 days in advance