A Multicenter Phase I-II Study of Tumor Vaccine Following Chemotherapy in Patients With Metastatic Breast Cancer Untreated With Chemo/Radiation in the Previous 18 Months: Vaccine-Induced Bias of T-Cell Repertoire Reconstitution After T-Cell Re-Infusion

Inclusion Criteria

- INCLUSION CRITERIA:

All patients must have a diagnosis of metastatic infiltrating carcinoma of the breast
including hormone receptor testing. At least one site of metastatic disease must have been
confirmed by pathologic or cytologic material. In the choice of a biopsy site, the PI will
weigh the morbidity the diagnostic procedure against the probability of positive yield of
the diagnostic procedure.

All pathologic material must be reviewed by the Pathology Laboratory of the National
Cancer Institute (NCI) before treatment.

The tumor MUST stain positive for CEA, by standard immuno-histochemistry performed at the
Pathology Laboratory of the NCI.

--Method: 5 microM formalin-fixed paraffin-embedded sections are deparaffinized and
blocked with methanol-30% hydrogen peroxide (H2O2). After antigen retrieval by boiling in
citrate buffer, or heating in a microwave oven for 10 minutes, slides are incubated with
monoclonal antibodies anti-CEA (diluted 1/1000 Dako). Then, slides are immunostained with
avidin-biotin-peroxidase complex and developed with diaminobenzidine. Harris' hematoxylin
was used to counter stain the slides. Positivity is defined as greater than 30% of cells
staining.

Patients may be newly diagnosed with metastatic breast carcinoma or known to have breast
carcinoma.

- If newly diagnosed, patients may not have received any chemotherapy for this disease
before entry on study.

- If previously treated for breast cancer, patients may have received chemotherapy or
radiation as adjuvant treatment for non-metastatic disease or metastatic disease but
not in the previous 18 months.

- Patients may have been on hormonal therapy for stage IV disease. Patients with
disease progression on hormonal therapy alone are eligible.

Karnofsky performance status of greater than or equal to 70% (Eastern Cooperative Oncology
Group (ECOG) 0 or 1)

Ejection fraction by multi-gated acquisition scan (MUGA) or 2-dimensional (2-D)
echocardiogram within normal institutional limits. In case of insufficient ejection
fraction, a stress echocardiogram will be performed. In case of an ejection fraction
greater than 35 % but less than 45%, the patient will remain eligible for the study if the
increase of ejection fraction with stress is estimated at 10% or more.

Creatinine clearance greater than or equal to 60 cc/min

Normal urinalysis; if proteinuria is present it must be quantified at less than 1 g / 24 h
on a measured 24 h urine collection

Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) less than 3 times the
upper limit of normal except if believed to be due to tumor involvement of the liver prior
to induction therapy.

Bilirubin less than 1.5 (except if due to tumor involvement prior to induction therapy or
in cases of Gilbert's disease).

Absolute Neutrophil Count greater than l000 / mm^3 and Platelet count greater than 90,000

Corrected carbon monoxide diffusing capacity (DLCO) greater than 50%

No history of abnormal bleeding tendency or predisposition to repeated infections.

Patient must be able to avoid close contact with children under 3 years old, pregnant
women, individuals with eczema or other skin conditions, and immuno-suppressed people for
2 weeks after initial vaccination. (see protocol for specific exclusion criteria for
vaccinia administration). Patients must agree to make specific arrangements, if necessary,
in order to comply and be eligible.

Patients must be able to give informed consent.

EXCLUSION CRITERIA:

Age less than 18 years

Patients in whom an urgent or emergent clinical situation does not safely allow for the
short delay in initiating the Concurrent Therapy (as defined in protocol) necessary for
the pre-treatment immunization and lymphocyte collection (at the discretion of the PI).

Patients requiring chronic immunosuppressive therapy (including corticosteroids) for any
medical condition.

Patients with an autoimmune disease: autoimmune neutropenia, thrombocytopenia, or
hemolytic anemia; Rheumatoid Arthritis, Systemic Lupus Erythematosus, Sjogren syndrome,
Scleroderma, Systemic Sclerosis, Myasthenia Gravis; Multiple sclerosis, Goodpasture
syndrome; Addison's disease, Hashimoto's thyroiditis, or active Graves' disease)

Any abnormality on the following tests suggestive of an autoimmune disease: anti-nuclear
antibody (ANA), anti-deoxyribonucleic acid (DNA), triiodothyronine (T3), thyroxine (T4),
thyroid stimulating hormone (TSH) after review with appropriate consultant. Patients with
endocrine disease that is controlled by replacement therapy including, diabetes, thyroid
and adrenal disease or vitiligo may be enrolled.

Patients with active inflammatory bowel disease

Patients with clinically significant cardiomyopathy requiring treatment or symptomatic
congestive heart failure (CHF), symptomatic arrhythmia that is not controlled by
medication, unstable coronary artery disease (CAD) such as unstable angina who require
active intervention, and patients with a recent infarction or cerebrovascular accident
(CVA) within the past 6 months

Patients testing positive for human immunodeficiency virus (HIV) or hepatitis B or C

Patients known or found to be pregnant or those unwilling to discontinue breastfeeding.
The effects of the chemotherapy, vaccines, and the medications used in this study are
highly likely to be harmful to a fetus. The effects upon breast milk are also unknown and
may be harmful to the infant; therefore, women should not breastfeed while on this study.

Patients of childbearing age who are unwilling to practice an effective form of
contraception. Patients of childbearing potential must use an effective method of
contraception while they are on-study; effective methods include intrauterine device
(IUD), hormonal (birth control pills, injections, or implants), tubal
ligation/hysterectomy (self or partner), partner's vasectomy, or barrier methods (condom,
diaphragm, or cervical cap), or abstinence.

Patients with brain metastases.

Patients with an active second malignancy (excluding treated skin cancers or carcinoma
in-situ) will be ineligible.

Patients with a life expectancy reasonably estimated at less than 6 months.

Patients may be excluded at the discretion of the principal investigator (PI) if it is
deemed that allowing participation would represent an unacceptable medical or psychiatric
risk.

History of splenectomy

Allergy to eggs

Several exclusion criteria are specific to vaccinia administration:

The recombinant vaccinia vaccine should not be administered if the following apply to
either recipients or, for at least two weeks after vaccination, to their close household
contacts (Close household contacts are those who share housing or have close physical
contact):

- Persons with active or a history of eczema or other eczematoid skin disorders

- Persons with other acute, chronic or exfoliative skin conditions (e.g., atopic
dermatitis, burns, impetigo, varicella zoster, severe acne or other open rashes or
wounds) until condition resolves;

- Pregnant or nursing women

- Children under 3 years of age;

- Immunodeficient or immunosuppressed persons by disease or therapy, including HIV
infection.

- History of seizures, encephalitis, or multiple sclerosis

- History of allergy or complications with past vaccinia vaccination.