• Evaluations for tumor response and studies to determine whether dendritic cells enhance the infused T cells' anti-tumor activity and their ability to migrate to the tumor site. [ Time Frame: 4 Years ] [ Designated as safety issue: No ]
• Evaluate the characteristics of the infused T cells that correspond with effectiveness in vivo. [ Time Frame: 4 Years ] [ Designated as safety issue: No ]

The primary objective will be to determine whether patients receiving adoptively transferred, tumor antigen-specific T cells in combination with dendritic cells and high dose IL-2 have sustained persistence of infused T cells compared to patients treated with T cells and high dose IL-2 alone. Secondary endpoints will include evaluations for tumor response and studies to determine whether dendritic cells enhance the infused T cells' anti-tumor activity and their ability to migrate to the tumor site. In addition, researchers will evaluate the characteristics of the infused T cells that correspond with effectiveness in vivo.

This study uses cell transfer therapies to treat patients with metastatic melanoma. Immune cells, also called T cells, can recognize and kill melanoma cells. These tumor fighting immune cells are taken and grown in the laboratory and given back to the patient. A vaccine prepared from special blood cells, known as dendritic cells, will be studied to determine whether it will help the immune cells to work better at fighting the melanoma. Patients in this study will also be treated with chemotherapy and high dose IL-2.

A T-cell is a type of lymphocyte. Lymphocytes are a type of white blood cell that protect you from viral infections; help other cells fight bacterial and fungal infections; produce antibodies; fight cancers; and coordinate the activities of other cells in the immune system.

Dendritic cells are cells that specialize in presenting antigens to the T-cells. An antigen is any substance capable of triggering an immune response. The cells in this study capable of presenting the antigens are called dendritic cells, and the T-cells are cells in the immune system responsible for coordinating the destruction of foreign antigens.

Dendritic Cells are able to attach to their surface small portions of the melanoma molecule which are recognized by the immune system and may be responsible for causing lymphocytes to attack the tumor. Besides attaching or binding this part of the antigen, called peptide, to their surface, dendritic cells can, with the help of specialized molecules present on their surface, boost the immune system. The purpose of this experimental protocol is, therefore, to enhance the immune response against the cancer cells in your body.

Chemotherapy is used to greatly reduce the number of normal lymphocytes circulating in your body, so that there will be more "space" for the cancer fighting lymphocytes (T-cells) that will be infused.

IL-2 was recently approved by the FDA for the treatment of metastatic melanoma. IL-2 is a hormone naturally found in the body that boosts the immune system. It is made in the lab, but from a normal human gene.

Before you can start treatment on this study, you will have what are called "screening tests". These tests will help the doctor decide if you are eligible to take part in the study. You will have a complete physical exam, including measurement of height, weight, and blood pressure. You may have an electrocardiogram (ECG - a test that measures the electrical activity of the heart). Blood samples (about 4 tablespoons) will be collected for routine blood tests, and HLA typing. You may also be tested for one or more of the following:

Human Immunodeficiency Virus (HIV) Hepatitis B Virus Hepatitis C Virus Cytomegalovirus (CMV) Herpes Simplex Virus (HSV) Epstein-Barr Virus (EBV) - Chagas Disease Human T cell Lymphotropic Virus Sickle Cell Disease West Nile Virus Pregnancy (within 7 days of treatment) Magnetic Resonance Imaging (MRI)/CT of the brain

If there are any abnormal results, the study doctor or a research staff member will discuss this with you.

A computed tomography (CT) scan of the chest, abdomen (stomach area), and pelvis (waist area) will be done to measure the tumor(s). A PET/CT may also be used. A MRI/CT scan of the brain will also be performed. Women who are able to have children must have a negative blood or urine pregnancy test. Pregnancy testing will be performed within 7 days before treatment. Women must have a documented negative pregnancy test (urine or blood) if they have had menstruation in the past 12 months and who have not been surgically sterilized.

A biopsy procedure of the tumor will be performed. A whole or part of your tumor will be cut and removed using high enough pain medication. This biopsy is being done to help attempt to grow the T-cells necessary for the treatment. Your T-cells will be grown in the laboratory with the help of radiation-exposed cells from other blood donors. These donor cells are tested to be negative for currently screened infectious diseases. Although precautions have been taken to lower the chance of you receiving infectious diseases from the cells, the use of these donor cells may cause an unwanted and unexpected immune response and/or viral infection, which could affect your health and/or well being. If researchers are unable to grow enough cells or if your tumor does not have enough of the necessary components, you will not continue on study and other treatment options will be discussed with you. If the growth of these cells is successful, you will be continued on the study. There is up to a 6 week waiting period between the biopsy and continuing to the next phase of this study. You will be allowed to participate in other treatment plans during this time period.

If eligible to continue on study, you will be randomly assigned (as in the toss of a coin) to one of two groups. Participants in one group will receive chemotherapy and IL-2 plus T-cells. Participants in the other group will receive chemotherapy and IL-2 plus T-cells and the vaccine of dendritic cells.

If you are randomized to receive dendritic cells, researchers will take some of your blood through a procedure called apheresis. During the apheresis procedure, white blood cells are removed from you using a serum cell separator machine. This requires putting a needle into your arm or a temporary central venous catheter to collect blood to go into the machine. The machine divides whole blood into red cells, plasma (the serum part) and lymphocytes (or white cells). The lymphocytes will be taken out, and the plasma and red cells returned to you through a second needle in your other arm or the central venous catheter. A single apheresis procedure takes about 3-4 hours to complete. If enough cells are not collected, then additional apheresis procedures may need to be performed. The collected cells will be used to make the dendritic cells. A stress cardiac test (stress thallium, stress MUGA, dobutamine echocardiogram) within 6 months of lymphodepletion to rule out cardiac ischemia. A pulmonary function tests, (FEV1 > 65% or FVC >65% of predicted) within 6 months of lymphodepletion. You will also have a CT scan of chest, abdomen and pelvis (a PET/CT may also be used), an MRI/CT of brain, an EKG, and a pregnancy test will be administered.

You are expected to stay in the Houston area for at least 14 days in order to participate in the chemotherapy and cell infusion phase of the study. After researchers have grown your cells from the samples collected, you will be given two chemotherapy medicines to decrease your immune system so the cells given back to you can work without any interference from the cells in your immune system. This chemotherapy will be given for seven days prior to receiving your cells. The two chemotherapy medicines are called cyclophosphamide and fludarabine. The cyclophosphamide will be given for two days (on Days -7 and -6) and the fludarabine for five days (on Days -5 through -1) immediately before the cells are infused. On Days -7 and -6, cyclophosphamide will be given by vein over about 2 hours. A drug called Mesna will also be infused by vein over about 24 hours starting on Day -7. Mesna is given to protect the bladder from side effects of the cyclophosphamide. On Days -5 to -1, fludarabine will be given by vein once a day over about 15-30 minutes.

The chemotherapy treatment will be followed by the infusion of the T-cells (Day 0). A catheter (a thin flexible tube inserted into the body to permit introduction or withdrawal of fluids) will be put in place to give you the cells. The catheter may be placed into a vein in your arm or in a large vein in your neck. If the cells need to be given through a large vein either in your upper chest or in your neck, the area will be numbed with medicine before the catheter is put in. Other catheters may be needed in one or both of your arms for additional routes to give you fluids, medicines, or extra nutrition. If you are assigned to the vaccine group, you will receive the vaccine by vein about 4 hours after the T-cells are given and again on Day 21 if your platelet count is high enough. On Days 1-5 and 22-26 you will receive high dose IL-2 by catheter every 8 hours for up to 15 doses as an inpatient if your platelet count is high enough on Day 21. Each infusion of IL-2 should take about 15 minutes.

You will be monitored closely during this study for any side effects of treatment. During the treatment, you will be monitored with vital signs after cell infusion once an hour for 4 hours after the infusion. Blood samples (about 3 teaspoons each) will be taken for routine blood tests every 1 to 2 days of treatment.

In order to help decrease the risk of infection during this study, you will be given some antibiotics. You will be given levofloxacin once a day by mouth or vein until your white blood cell level returns to an acceptable level. You will also take trimethoprim and sulfamethoxazole (SMX) by mouth twice a day, beginning on Day -7 and continuing for at least 3 months after chemotherapy.

It is possible that you will receive a 2nd treatment (re-treatment) that will be identical to your first schedule of therapy (an entire repeat cycle of chemotherapy, IL-2, and T-cells with or without vaccine). If the disease gets worse, you experience any intolerable side effects, or your doctor feels it is in your best interest, you will be taken off the study and your doctor will discuss other treatment options with you.

You will have blood samples collected to evaluate the activity of the T-cells and their ability to attack the tumor. Blood samples (about 4 tablespoons) will be taken before surgery to remove your tumor or tumor biopsy and again on Days 7, 14, 21, 28, 35, 42 (+/- 7 days) if possible and 70 (+/- 7 days) when possible.

At about 6 weeks (+ 7 days) and at 12 weeks (+ 7 days) after you receive the T-cell and/or vaccine infusion (for each cycle), you will return to the clinic for a physical exam and for repeat scans to evaluate the size of your tumors.

This is an investigational study. The medications being used in this study are FDA approved. The chemotherapy agents and IL-2 are commercially available drugs. However, their use together in this study is experimental. The T-cell and vaccine therapy are authorized for use only in research. A total of up to 83 patients will take part in this study. All will be enrolled at M. D. Anderson.

• Biological: Dendritic Cell Immunization
• Drug: Cyclophosphamide
• Drug: Fludarabine
• Biological: T Cells
• Biological: Interleukin-2

Inclusion Criteria:

1. Patients must have metastatic melanoma or stage III in-transit or regional nodal disease. (Turnstile I)
2. Patients must receive an MRI/CT of the brain. If new lesions are present, patient must have definitive treatment. PI or his designee should make final determination regarding enrollment. (Turnstile I)
3. Age greater than or equal to 12 years. (Turnstile I)
4. Clinical performance status of ECOG 0-2. (Turnstile I)
5. Patients previously treated with immunotherapy, targeted therapy, or no therapy will be eligible. Patients receiving cytotoxic agents will be evaluated by the PI or his designee as to suitable eligibility. (Turnstile I)
6. Patients with a negative pregnancy test (urine or serum) must be documented at screening for women of childbearing potential. (Turnstile I)
7. Patients must be HLA-A2 for cohort A. (Turnstile II-Chemotherapy/Cell Infusion-Inclusion Criteria)
8. Patients must have adequate TIL available as described in Appendix B. (Turnstile II)
9. Patients must have measurable metastatic melanoma. (Turnstile II - Chemotherapy/Cell Infusion -Inclusion Criteria).
10. Patients may have a maximum of 2 brain lesions which measure </= 1cm each. (Turnstile II - Chemotherapy/Cell Infusion- Inclusion Criteria).
11. Patients of both genders must practice birth control for four months after receiving the preparative regimen. Patients must have a documented negative pregnancy test (urine or serum) for women who have menstruation in the past 12 months and without sterilization surgery. (Turnstile II)
12. Unless surgically sterile by bilateral tubal ligation or vasectomy of partner(s), the patient agrees to continue to use a barrier method of contraception throughout the study such as: condom, diaphragm, hormonal, IUD, or sponge plus spermicide. Abstinence is an acceptable form of birth control. (Turnstile II)
13. Clinical performance status of ECOG 0 - 2 at the time of chemotherapy infusion. (Turnstile II - Chemotherapy/Cell Infusion-Inclusion Criteria).
14. Absolute neutrophil count greater than or equal to 1000/mm3. (Turnstile II - Chemotherapy/Cell Infusion- Inclusion Criteria).
15. Platelet count greater than or equal to 100,000/mm3. (Turnstile II - Chemotherapy/Cell Infusion- Inclusion Criteria).
16. Hemoglobin greater than or equal to 8.0 g/dl. (Turnstile II - Chemotherapy/Cell Infusion).
17. Serum ALT less than three times the upper limit of normal. (Turnstile II - Chemotherapy/Cell Infusion- Inclusion Criteria).
18. Serum creatinine less than or equal to 1.6 mg/dl. (Turnstile II - Chemotherapy/Cell Infusion-Inclusion Criteria).
19. Total bilirubin less than or equal to 2.0 mg/dl, except in patients with Gilbert's Syndrome who must have a total bilirubin less than 3.0 mg/dl. (Turnstile II - Chemotherapy/Cell Infusion - Inclusion Criteria).
20. Patients randomized to receive the DC arm of Cohort A will have the following tests: Donor infectious disease panel must be performed which includes the following tests: Hepatitis B surface antigen (HBsAg), Anti-Hepatitis B core antibody (HBC Abs),Anti-Hepatitis C Virus antibody (HCV Ab), Anti-Human Immunodeficiency Virus (HIV) antibody (HIV ½ Ab), Anti-Human T cell lymphotrophic Virus (HTLV) antibody (HTLV I/II Ab), Rapid Plasma Reagen (RPR), Cytomegalovirus antibody (CMV), HCV/HIV Nucleic Acid Test, West Nile Virus Nucleic Acid Test, Chagas Disease, and Sickledex.
21. A stress cardiac test (stress thallium, stress MUGA, dobutamine echocardiogram or other stress test that will rule out cardiac ischemia) within 6 months of lymphodepletion. (Turnstile II - Chemotherapy/Cell Infusion-Inclusion Criteria).
22. Pulmonary function tests (FEV1>65% or FVC>65%of predicted) within 6 months of lymphodepletion. (Turnstile II - Chemotherapy/Cell Infusion - Inclusion Criteria).

Exclusion Criteria:

1. Active systemic infections requiring intravenous antibiotics, coagulation disorders or other major medical illnesses of the cardiovascular, respiratory or immune system. PI or designee shall make the final determination regarding appropriateness of enrollment. (Turnstile I - Screening Exclusion Criteria).
2. Patients who are pregnant or nursing. (Turnstile I)
3. Has had prior systemic cancer therapy within the past four weeks at the time of the start of the lymphodepletion regimen. (Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)
4. Women who are pregnant will be excluded because of the potentially dangerous effects of the preparative chemotherapy on the fetus. (Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)
5. Any active systemic infections requiring intravenous antibiotics, coagulation disorders or other major medical illnesses of the cardiovascular, respiratory or immune system, such as abnormal stress test and/or abnormal PFT. PI or his designee shall make the final determination regarding appropriateness of enrollment.(Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)
6. Any form of primary or secondary immunodeficiency. Must have recovered immune competence after chemotherapy or radiation therapy as evidenced by lymphocyte counts (> 500/mm3), WBC (> 3,000/mm3) or absence of opportunistic infections. (Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)
7. Require steroid therapy or steroid-containing compounds, or have used systemic steroids in the past 30 days, or have used topical or inhalational steroids in the past 2 weeks prior to lymphodepletion. (Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)
8. Presence of a significant psychiatric disease, which in the opinion of the principal investigator or his designee, would prevent adequate informed consent or render immunotherapy unsafe or contraindicated. (Turnstile II - Chemotherapy/Cell Infusion Exclusion Criteria)