Mitchell’s laboratory for RNA isolation in accordance with SOP -UFBTIP -116 and autologous tumor cell targets for immunologic monitoring . Prior to leukapheresis • Tests as required by the Apheresis Center (total estimated blood volume requi red for these evaluations is 10- 20 mLs based on patient weight per institutional guidelines) o β-HCG if female and greater than 12 years of age o Infectious disease testing to include, at a minimum, HIV, Hepatitis B, Hepatitis C and CMV o CBC with differential, Complete or Basic Metabolic panel (per institutional standard) , T4/T8 ratio o To prevent the development of hypocalcemia from the citrate used for leukapheresis, all patients over age 3 will be given oral Tums, 2 tablets, b.i.d for 2 days prior to the leukapheresis procedure if standard of care. Patients who have lowered levels of Ca will be treated per Apheresis lab standard. Prior to DC + xALT Therapy • At the time of first vaccination, all patients will receive a standard 0.5 ml Td (2 Lf tetanus and 2 Lf diphtheria toxoids) booster intramuscularly. The rationale for this vaccination is to enhance DC trafficking to vaccine -site draining lymph nodes (VDLNs). • Additionally, 24 hours prior to first immunization, blood will be drawn for the following purposes: o CBC with differential, T4/T8 ratio, and research blood. Total estimated blood volume required for these evaluations is 12 mL. During and After Immunizations • Clinical evaluations with each vaccine will include a general physical examination, complete neurologic examination, and KPS or LPS rating. • Radiographic evaluations consisting of a contrast enhanced brain CT or MRI scan will be obtained every 8 + 4 weeks. Blood will be obtained for T4/T8 ratio, C BC with manual differential and for Immunologic Monitoring prior to non-mobilized leukapheresis , prior to immunotherapy , 1 day post Vaccine #1, 4 (+/ - 1) days post Vaccine #1 , weekly post V accine #1 for six weeks , and bimonthly , if feasible, until progress ion. Weekly blood draws should be drawn prior to subsequent vaccination. Blood will be processed in accordance with SOP -UFBTIP - 126. Total estimated volume of blood required over the six week period for investigational purposes will be 80 mLs. Re-MATC H Protocol Page 81 of 106 Version: 20180628 STUDY ROAD MAP : Timepoint Date Study Requirements Baseline Inclusion & exclusion criteria Tissue c onsent Medical h istory , physical & neurological exam MRI brain & spine Performance Status: KPS or LPS CBC w/ differential , PT, PTT, comprehensive metabolic panel β-HCG pregnancy test Type & Screen if clinically indicated Surgery Resection, biopsy or cytology collection Tumor tissue for extraction of total tumor RNA Pathology slides for research: 1 H&E & 19 unstained slides MRI brain with & without gadolinium post -surg ery Non -Mobilized Leukapheresis Treatment consent CBC w/ differential , comprehensive or basic metabolic panel (per institutional standard) , T4/T8 ratio , research labs** β-HCG pregnancy test Infectious disease testing to include, at a minimum, HIV, Hepatitis B, Hepatitis C & CMV Oral tums if standard of care Induction or Salvage Chemo Cycle 1 (28 days) Group A: Cyclophosphamide , Mesna, GCSF Group B: Etop oside, Temozolomide, GCSF Cycle 2 (28 days) Group A: Cyclophosphamide , Mesna, GCSF Group B: Etop oside, Temozolomide, GCSF Cycle 3 (28 days) Group A: Etopo side, Temozolomide Group B: Etoposide , Temozolomide Cycle 4 (28 days) Group A : Etoposide ,Temozolomide Group B: Etoposide , Temozolomide Mobilized Leukapheresis CBC w/ differential , comprehensive or basic metabolic panel (per institutional standard) , T4/T8 ratio β-HCG pregnancy test Infectious disease testing to include, at a minimum, HIV, Hepatitis B, Hepatitis C & CMV Oral tums if standard of care Re-MATC H Protocol Page 82 of 106 Version: 20180628 Timepoint Date Study Requirements Pre Consolidation or NMA Chemo Medical history, physical exam MRI brain (spine if indicated) CBC w/ differential , comprehensive metabolic panel , GFR β-HCG pregnancy test ECHO or MUGA EKG (Group A only) DLCO, FEV1 & FVC or Crying Vital Capacity (Group A only) CXR & Pulse Oximetry (Group B & Group A subjects who cannot complete PFTs) Consolidation or NMA Chemo Group A : Carboplatin, Etoposide, Thiotepa, PBSCs , G-CSF Group B: Cyclophosphamide, Fludarabine , PBSCs, G-CSF Prior to Immunotherapy Td booster ( 4 to 24 hours prior) CBC w/ diff, T4/T8 ratio, research labs** Immunotherapy Vaccine #1 Group A: Day +1 Group B: 96 hours post Fludarabine Physical & neurological exam Performance Status: KPS or LPS Tylenol, Benadryl, Intravenous TTRNA -xALT EMLA , Intradermal TTRNA -DC 1 day post: CBC w/ diff, T4/T8 ratio, research labs ** 4 (+/-1) days post: CBC w/ diff, T4/T8 ratio, research labs ** Week 1 post: CBC w/ diff, T4/T8 ratio, research labs ** Week 2 post: CBC w/ diff, T4/T8 ratio, research labs ** Week 3 post: CBC w/ diff, T4/T8 ratio, research labs ** Week 4 post: CBC w/ diff, T4/T8 ratio, research labs ** Week 5 post: CBC w/ diff, T4/T8 ratio, research labs ** Week 6 post: CBC w/ diff, T4/T8 ratio, research labs ** Vaccine #2 14 days post Vaccine #1 Physical & neurological exam Performance Status: KPS or LPS EMLA , Intradermal TTRNA -DC Vaccine #3 14 days post Vaccine #2 Physical & neurological exam Performance Status: KPS or LPS EMLA , Intradermal TTRNA -DC Bi-Monthly follow -up Physical & neurological exam Contrast -enhanced Brain CT or MRI scan CBC w/ diff , T4/T8 ratio, research labs , if feasible ** **2 yellow ( ACD -A) & 1 red ( non-SST) 20.3 Laboratory and Clinical Evaluation Facilities UF Health Clinical Laboratories Re-MATC H Protocol Page 83 of 106 Version: 20180628 UF Health Clinical Laboratories, comply with federal regulations defined in the Clinical Laboratory Improvement Amendments of 1988 (CLIA’88) and the national accreditation standards defined by the College of American Pathologists (CAP). UF Health Clinical Laboratories are accredited by the Colle ge of American Pathologist’s Laboratory Accreditation Program.