Blood will be collected from the patient to make LMP/BARF1/EBNA1 (NPC)-specific T cells. To grow NPC-specific T cells we have to use special cells called antigen-presenting cells, which train the patient's T cells to be NPC specific. Antigen presenting cells, so called monocytes or dendritic cells, will be grown/isolated from the patient's blood. In addition, we use a cell line called K562 as antigen-presenting cells that has had genes put inside it, which encourage the patient's T cells to grow. K562 cells are cancer cells. As such, if injected they could cause cancer. The cells have been treated with radiation so they cannot grow.
These antigen-presenting cells are coated with a specially produced mixture of LMP, EBNA1 and BARF protein fragments called peptides. These coated antigen-presenting cells are then used to generate the patient's NPC-specific T cells in the presence of growth factors. To get the DNR to attach to the surface of these NPC-specific T Cells, we also inserted the DNR gene into the NPC-specific T cells (DNR.NPC-specific T cells). This is done with a virus called a retrovirus that has been made for this study. This virus will carry the DNR gene into the cells.
Once we have made sufficient numbers of DNR.NPC-specific T cells we will freeze the cells and test them to make sure they recognize EBV proteins present in NPC.
Patients will get treated with 1) either two doses of DNR.NPC-specific T cells (the second dose will be given 2 weeks after the first dose) or 2)cyclophosphamide and fludarabine for 3 days before receiving the DNR.NPC-specific T cells.
The cyclophosphamide and fludarabine will be given through a needle inserted into a vein or patient's port-a-cath).
The T cells will be thawed and infused through a central line, or through a vein in the patient's arm over 1 to 10 minutes. Patients will be followed in the clinic after the infusion for 1 to 4 hours.
All of the treatments will be given by the Center for Cell and Gene Therapy at Texas Children's Hospital or Houston Methodist Hospital.
Medical tests before treatment:
Before being treated, the patient will receive a series of standard medical tests:
Physical exam Imaging Study Blood tests to measure blood cells, kidney and liver function Measurements of the tumor by routine imaging studies. We will use the imaging study that was used before to follow the patient's tumor: Computer Tomogram (CT), Magnetic Resonance Imaging (MRI), or Positron Emission Tomography (PET/CT)
Medical tests during and after treatment:
The patient will receive standard medical tests when they are getting the infusions and after:
Physical exams Blood tests to measure blood cells, kidney and liver function Imaging study 8 weeks after the 1st T-cell infusion(for patients receiving only DNR.NPC-specific T cells ) or Imaging study 6 weeks after the T-cell infusion (for patients receiving fludarabine and cyclophosphamide before DNR.NPC-specific T-cell infusion).
To learn more about the way the DNR.NPC-specific T cells are working and how long they last in the body, an extra 60ml or 3ml/kg of body weight of blood (whichever is less) will be taken pre-chemotherapy (for the patients receiving chemotherapy), on the day of the T-cell infusion (before and at the end of the T-cell infusion), 1, 2, 3, 4. 6, and 8 weeks( the last time point only for patients receiving DNR.NPC-specific T-cells only) after the T-cell infusion and every 3 months for 1 year, every 6 months for 4 years, then yearly for a total of 15 years. One additional blood sample might be drawn 3 to 4 days post the 1st T-cell infusion; this is optional. This volume is considered safe, but may be decreased if the patient is anemic. In addition to the blood tests, the patient will receive 2 imaging studies as stated above.
During the time points listed above, if the DNR.NPC-specific T cells are found in patient's blood at a certain amount an extra 5ml of blood may need to be collected for additional testing.
