US2022387492A1PendingUtilityA1

Chimeric antigen receptor t cell therapy

Assignee: KITE PHARMA INCPriority: Nov 6, 2019Filed: Nov 6, 2020Published: Dec 8, 2022
Est. expiryNov 6, 2039(~13.3 yrs left)· nominal 20-yr term from priority
G01N 33/57505A61P 35/02C07K 2317/76A61K 35/545C07K 14/70521C07K 14/7051A61K 39/3955G01N 33/505A61K 2039/804C07K 2317/622G01N 2800/52C07K 2319/03A61K 31/675C07K 2319/00C07K 16/2803A61K 38/1774C07K 2319/30A61K 31/7076A61K 2039/545C12N 2510/00C07K 2319/33A61K 9/0019C12N 2501/505A61P 35/00C12N 2506/11A61K 35/17G01N 33/57426A61K 39/001112A61K 2039/5156A61K 40/11A61K 40/31A61K 40/4211A61K 2239/48A61K 2239/38C12N 5/0636
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Claims

Abstract

Provided herein are methods for preparing, producing, processing, culturing, isolating, or making cells suitable for immune or cell therapy, and for their use in cell therapy.

Claims

exact text as granted — not AI-modified
We claim: 
     
         1 . A method for treating mantle cell lymphoma (MCL) or B cell ALL in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a T cell product comprising autologous T cells expressing an anti-CD19 chimeric antigen receptor (CAR). 
     
     
         2 . The method of  claim 1 , wherein the MCL and B cell ALL are relapsed or refractory MCL and B cell ALL, optionally wherein the MCL is classical, blastoid, and pleomorphic MCL. 
     
     
         3 . The method of any one of  claims 1  and  2 , wherein the MCL and B cell ALL is refractory to, or has relapsed following, one or more of chemotherapy, radiotherapy, immunotherapy (including a T cell therapy and/or treatment with an antibody or antibody-drug conjugate), an autologous stem cell transplant, or any combination thereof. 
     
     
         4 . The method of any one of  claims 1  through  3 , wherein the subject has received 1-5 prior treatments, optionally wherein at least one of the prior treatments is selected from autologous SCT, anti-CD20 antibody, anthracycline- or bendamustine-containing chemotherapy, and/or a Bruton Tyrosine Kinase inhibitor (BTKi). 
     
     
         5 . The method of  claim 4 , wherein the BTKi is ibrutinib or acalabrutinib. 
     
     
         6 . The method of any one of  claims 1  through  5 , wherein R/R B cell ALL is defined as refractory to first-line therapy (i.e., primary refractory), relapsed ≤12 months after first remission, relapsed or refractory after ≥2 prior lines of systemic therapy, or relapsed after allogeneic stem cell transplant (SCT), optionally, wherein the subject is required to have ≥5% bone marrow blasts, an Eastern Cooperative Oncology Group performance status of 0 or 1, and/or adequate renal, hepatic, and cardiac function. 
     
     
         7 . The method of any one of  claims 1  through  6 , wherein if the B cell ALL subject has received prior blinatumomab, the subject is required to have leukemic blasts with CD19 expression ≥90%. 
     
     
         8 . The method of any one of  claims 1  through  7 , wherein the subject receives bridging therapy after leukapheresis and before conditioning/lymphodepleting chemotherapy. 
     
     
         9 . The method of any one of  claims 1  through  8 , wherein the MCL subject receives a lymphodepleting chemotherapy regimen of cyclophosphamide 500 mg/m 2  intravenously and fludarabine 30 mg/m 2  intravenously, both given on each of the fifth, fourth, and third days before T cell infusion. 
     
     
         10 . The method of any one of  claims 1  through  9 , wherein the B cell ALL subject receives a lymphodepleting regimen of fludarabine intravenous (IV) 25 mg/m 2 /day on each of the fourth, third, second days before T cell infusion, and cyclophosphamide IV 900 mg/m 2 /day on the second day before infusion. 
     
     
         11 . The method of any one of  claims 8  through  10 , wherein the MCL bridging therapy is selected from dexamethasone (e.g., 20-40 mg or equivalent PO or IV daily for 1-4 days); methylprednisolone, ibrutinib (e.g., 560 mg PO daily), and/or acalabrutinib (e.g, 100 mg PO twice daily); an immunomodulator; R-CHOP, bendamustine; alkylating agents; and/or platinum-based agents, wherein the bridging therapy is administered after leukapheresis and completed in, for example, 5 days or less before conditioning chemotherapy. 
     
     
         12 . The method of any one of  claims 8  through  10 , wherein the B cell ALL subject may receive any one or more of the following bridging chemotherapy regimens: 
       
         
           
                 
               
                     
                 
                   Predefined Bridging Chemotherapy Regimens 
                 
                     
                 
                     
                 
                 
                 
               
                   Attenuated VAD 
                   Vincristine non-liposomal (1-2 mg IV weekly) or liposomal (2.25 
                 
                     
                   mg/m 2  IV weekly), and dexamethasone 20-40 mg IV or PO daily × 
                 
                     
                   3-4 days per week. Optional doxorubicin 50 mg/m 2  IV × 1 (first 
                 
                     
                   week only) 
                 
                   Mercaptopurine 
                   50-75 mg/m 2 /day by mouth (administer at bedtime on an empty 
                 
                   (6-MP) 
                   stomach to improve absorption) 
                 
                   Hydroxyurea 
                   Doses titrated between 15-50 mg/kg/day (rounded to the nearest 
                 
                     
                   500 mg capsule and given as a single daily oral dose on a 
                 
                     
                   continuous basis) 
                 
                   DOMP 
                   Dexamethasone 6 mg/m 2 /day PO (or IV) divided BID 
                 
                     
                   days 1-5, vincristine 1.5 mg/m 2  (maximum dose 2 mg) IV on 
                 
                     
                   day 1, methotrexate 20 mg/m 2  PO weekly, 6-MP 
                 
                     
                   50-75 mg/m 2 /day PO daily 
                 
                   Attenuated 
                   Fludarabine 30 mg/m 2  IV days 1-2, cytarabine 2 g/m 2  IV days 1-2, 
                 
                   FLAG/FLAG-IDA 
                   G-CSF 5 μg/kg SC or IV starts on day 3 and can continue until 
                 
                     
                   day before the start of conditioning chemotherapy. With or 
                 
                     
                   without idarubicin 6 mg/m 2  IV days 1-2 
                 
                   Mini-hyper CVAD 
                   Course A: Cyclophosphamide 150 mg/m 2  every 12 h × 3 days, 
                 
                   (courses A and/or 
                   dexamethasone 20 mg/d IV or PO daily days 1-4 and 11-14, 
                 
                   B) 
                   vincristine 2 mg IV × 1 
                 
                     
                   Course B: methotrexate 250 mg/m 2  IV over 24 hours on day 1, 
                 
                     
                   cytarabine 0.5 g/m 2  IV every 12 hours x 4 doses on days 2 and 3 
                 
                     
                 
             
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         13 . The method of any one of  claims 1  through  12 , wherein the T cell product comprises CD4+ and CD8+ CAR T cells that are prepared from peripheral blood mononuclear cells (PBMCs) by positive enrichment and consequent partial or complete depletion of circulating cancer cells. 
     
     
         14 . The method of  claim 13 , wherein the PBMC are enriched for T cells by positive selection for CD4+ and CD8+ cells, activated with anti-CD3 and anti-CD28 antibodies in the presence of IL-2, and then transduced with a replication-incompetent viral vector containing FMC63-28Z CAR, a chimeric antigen receptor (CAR) comprising an anti-CD19 single-chain variable fragment (scFv), CD28 and CD3-zeta domains. 
     
     
         15 . The method of any one of  claims 13  and  14 , wherein the T cell product comprises fewer cancer cells than a T cell product comprising T cells from a leukapheresis product that have not been positively selected for CD4+ and CD8+ T cells. 
     
     
         16 . The method of any one of  claims 13  through  15 , wherein the T cell product has other superior product attributes relative to a T cell product comprising T cells from a leukapheresis product that have not been positively selected/enriched for CD4+ and CD8+ T cells. 
     
     
         17 . The method of  claim 16 , wherein the superior product attributes are selected from increased percentage of CDRA45+ CCR7+(naïve-like) T cells, decreased percentage of differentiated T cells, increased percentage of CD3+ cells, decreased IFN-gamma production, decreased percentage of CD3− cells. 
     
     
         18 . The method of any one of  claims 1  through  17 , wherein the MCL subject is administered one or more doses of 1.8×10 6 , 1.9×10 6 , or 2×10 6  CAR positive viable T cells per kg body weight, with a maximum of 2×10 8  CAR positive viable T cells (for patients 100 kg and above) and the B cell ALL subject is administered 0.5×10 6 , 1×10 6 , or 2×10 6  CAR positive viable T cells per kg body weight, with a maximum of 2×10 8  CAR positive viable T cells (for patients 100 kg and above). 
     
     
         19 . The method of any one of  claims 1  through  17 , wherein if the subject has achieved complete response to the first infusion, the subject may receive a second infusion of anti-CD19 CAR T cells, if progressing following >3 months of remission, provided CD19 expression has been retained and neutralizing antibodies against the CAR are not suspected, wherein response is assessed using the Lugano classification. 
     
     
         20 . The method of any one of  claims 1  through  19 , wherein the subject is monitored for signs and symptoms of cytokine release syndrome (CRS) and neurologic toxicity after T cell administration. 
     
     
         21 . The method of  claim 20 , wherein the subject is monitored daily for at least seven days, preferably for four weeks, following infusion for signs and symptoms of CRS and neurologic toxicity. 
     
     
         22 . The method of any one of  claims 20  and  21 , wherein the signs or symptoms associated with CRS include fever, chills, fatigue, tachycardia, nausea, hypoxia, and hypotension and the signs or symptoms associated with neurologic events include encephalopathy, seizures, changes in level of consciousness, speech disorders, tremors, and confusion. 
     
     
         23 . The method of any one of  claims 20  through  22 , wherein cytokine release syndrome in MCL subjects is managed in accordance with the following protocol: 
       
         
           
                 
                 
                 
               
                     
                 
                   CRS Grade 
                   Tocilizumab 
                   Corticosteroids 
                 
                     
                 
                   Grade 1 
                   If not improving after 24 
                   Not applicable. 
                 
                   Symptoms require 
                   hours, administer tocilizumab 
                     
                 
                   symptomatic treatment only 
                   8 mg/kg intravenously over 1 
                     
                 
                   (e.g., fever, nausea, fatigue, 
                   hour (not to exceed 800 mg). 
                     
                 
                   headache, myalgia, malaise). 
                     
                     
                 
                   Grade 2 
                   Administer tocilizumab 8 
                   Manage per Grade 3 
                 
                   Symptoms require and 
                   mg/kg intravenously over 1 
                   if no improvement 
                 
                   respond to moderate 
                   hour (not to exceed 800 mg). 
                   within 24 hours after 
                 
                   intervention. 
                   Repeat tocilizumab every 8 
                   starting tocilizumab. 
                 
                   Oxygen requirement less than 
                   hours as needed if not 
                   If improving, taper 
                 
                   40% FiO2 or hypotension 
                   responsive to intravenous fluids 
                   corticosteroids. 
                 
                   responsive to fluids or low 
                   or increasing supplemental 
                     
                 
                   dose of one vasopressor or 
                   oxygen. Limit to a maximum of 
                     
                 
                   Grade 2 organ toxicity. 
                   3 doses in a 24-hour period; 
                     
                 
                     
                   maximum total of 4 doses if no 
                     
                 
                     
                   clinical improvement in the 
                     
                 
                     
                   signs and symptoms of CRS. 
                     
                 
                     
                   If improving, 
                     
                 
                     
                   discontinue 
                     
                 
                     
                   tocilizumab. 
                     
                 
                   Grade 3 
                   Per Grade 2 
                   Administer 
                 
                   Symptoms require and 
                     
                   methylprednisolone 1 
                 
                   respond to aggressive 
                     
                   mg/kg intravenously 
                 
                   intervention. 
                     
                   twice daily or 
                 
                   Oxygen requirement greater 
                     
                   equivalent 
                 
                   than or equal to 40% FiO2 or 
                     
                   dexamethasone (e.g., 
                 
                   hypotension requiring high- 
                     
                   10 mg intravenously 
                 
                   dose or multiple vasopressors 
                     
                   every 6 hours) until 
                 
                   or Grade 3 organ toxicity or 
                     
                   Grade 1, then taper 
                 
                   Grade 4 transaminitis. 
                     
                   corticosteroids. 
                 
                     
                     
                   If improving, manage 
                 
                     
                     
                   as Grade 2. 
                 
                     
                     
                   If not improving, 
                 
                     
                     
                   manage as Grade 4. 
                 
                   Grade 4 
                   Per Grade 2 
                   Administer 
                 
                   Life-threatening symptoms. 
                     
                   methylprednisolo 
                 
                   Requirements for ventilator 
                     
                   ne 1000 mg 
                 
                   support or continuous veno- 
                     
                   intravenously per 
                 
                   venous hemodialysis 
                     
                   day for 3 days. 
                 
                   (CVVHD), or Grade 4 organ 
                     
                   If improving, taper 
                 
                   toxicity (excluding 
                     
                   corticosteroids, and 
                 
                   transaminitis). 
                     
                   manage as Grade 3. 
                 
                     
                     
                   If not 
                 
                     
                     
                   improving, 
                 
                     
                     
                   consider 
                 
                     
                     
                   alternate 
                 
                     
                     
                   immunosuppres 
                 
                     
                     
                   sants. 
                 
                     
                 
             
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         24 . The method of any one of  claims 20  through  23 , wherein neurologic toxicity in MCL subjects is managed in accordance with the following protocol: 
       
         
           
                 
                 
                 
               
                     
                 
                   Grading 
                     
                     
                 
                   Assessment 
                   Concurrent CRS 
                   No Concurrent CRS 
                 
                     
                 
                   Grade 2 
                   Administer tocilizumab per claim 
                   Administer dexamethasone 10 
                 
                     
                   15 for management of Grade 2 
                   mg intravenously every 6 
                 
                     
                   CRS. 
                   hours until the event is Grade 
                 
                     
                   If not improving within 24 hours after 
                   1 or less, then taper 
                 
                     
                   starting tocilizumab, administer 
                   corticosteroids. 
                 
                     
                   dexamethasone 10 mg intravenously 
                     
                 
                     
                   every 6 hours until the event is Grade 1 
                     
                 
                     
                   or less, then taper corticosteroids. 
                     
                 
                     
                   If still not improving, manage as Grade 3. 
                     
                 
                 
                 
               
                     
                   Consider non-sedating anti-seizure medicines (e.g., levetiracetam) for 
                 
                     
                   seizure prophylaxis. 
                 
                 
                 
                 
               
                   Grade 3 
                   Administer tocilizumab per claim 
                   Administer 
                 
                     
                   15 for management of Grade 2 
                   dexamethasone 10 
                 
                     
                   CRS. 
                   mg intravenously 
                 
                     
                   In addition, administer dexamethasone 10 
                   every 6 hours. 
                 
                     
                   mg intravenously with the first dose of 
                   Continue dexamethasone use 
                 
                     
                   tocilizumab and repeat dexamethasone 
                   until the event is Grade 1 or 
                 
                     
                   dose every 6 hours. Continue 
                   less, then taper corticosteroids. 
                 
                     
                   dexamethasone use until the event is 
                   If not improving, manage as 
                 
                     
                   Grade 1 or less, then taper 
                   Grade 4. 
                 
                     
                   corticosteroids. 
                     
                 
                     
                   If improving, discontinue 
                     
                 
                     
                   tocilizumab and manage as Grade 2. 
                     
                 
                     
                   If still not improving, manage as 
                     
                 
                     
                   Grade 4 (below). 
                     
                 
                 
                 
               
                     
                   Consider non-sedating anti-seizure medicines (e.g., levetiracetam) for 
                 
                     
                   seizure prophylaxis. 
                 
                 
                 
                 
               
                   Grade 4 
                   Administer tocilizumab per claim 
                   Administer 
                 
                     
                   15 for management of Grade 2 
                   methylprednisolone 1000 
                 
                     
                   CRS. 
                   mg intravenously per day 
                 
                     
                   Administer methylprednisolone 1000 mg 
                   for 3 days. 
                 
                     
                   intravenously per day with first dose of 
                   If improving, then manage as 
                 
                     
                   tocilizumab and continue 
                   Grade 3. 
                 
                     
                   methylprednisolone 1000 mg 
                   If not improving, 
                 
                     
                   intravenously per day for 2 more days. 
                   consider alternate 
                 
                     
                   If improving, then manage as Grade 3. 
                   immunosuppressants. 
                 
                     
                   If not improving, consider 
                     
                 
                     
                   alternate immunosuppressants. 
                     
                 
                 
                 
               
                     
                   Consider non-sedating anti-seizure medicines (e.g., levetiracetam) for 
                 
                     
                   seizure prophylaxis. 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
               
            
             
                
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
             
                
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
               
            
             
                
                
                
               
            
           
         
       
     
     
         25 . The method of any one of  claims 1  through  24 , wherein the MCL subject is a high-risk patient as determined by a Ki-67 tumor proliferation index ≥50% and/or presence of a TP53 mutation. 
     
     
         26 . The method of any one of  claims 20  through  22 , wherein CRS in a B cell ALL subject is managed according to the following protocol: 
       
         
           
                 
                 
                 
               
                     
                 
                   CRS Grade 
                   Tocilizumab 
                   Corticosteroids 
                 
                     
                 
                   Grade 1 
                   If not improving after 24 
                   Not applicable. 
                 
                   Symptoms require 
                   hours, administer tocilizumab 
                     
                 
                   symptomatic treatment only 
                   8 mg/kg intravenously over 1 
                     
                 
                   (e.g., fever, nausea, fatigue, 
                   hour (not to exceed 800 mg). 
                     
                 
                   headache, myalgia, malaise). 
                     
                     
                 
                   Grade 2 
                   Administer tocilizumab 8 
                   Manage per Grade 3 
                 
                   Symptoms require and 
                   mg/kg intravenously over 1 
                   if no improvement 
                 
                   respond to moderate 
                   hour (not to exceed 800 mg). 
                   within 24 hours after 
                 
                   intervention. 
                   Repeat tocilizumab every 8 
                   starting tocilizumab. 
                 
                   Oxygen requirement less than 
                   hours as needed if not 
                   If improving, taper 
                 
                   40% FiO2 or hypotension 
                   responsive to intravenous fluids 
                   corticosteroids. 
                 
                   responsive to fluids or low 
                   or increasing supplemental 
                     
                 
                   dose of one vasopressor or 
                   oxygen. Limit to a maximum of 
                     
                 
                   Grade 2 organ toxicity. 
                   3 doses in a 24-hour period; 
                     
                 
                     
                   maximum total of 4 doses if no 
                     
                 
                     
                   clinical improvement in the 
                     
                 
                     
                   signs and symptoms of CRS. 
                     
                 
                     
                   If improving, 
                     
                 
                     
                   discontinue 
                     
                 
                     
                   tocilizumab. 
                     
                 
                   Grade 3 
                   Per Grade 2 
                   Administer 
                 
                   Symptoms require and 
                     
                   methylprednisolone 1 
                 
                   respond to aggressive 
                     
                   mg/kg intravenously 
                 
                   intervention. 
                     
                   twice daily or 
                 
                   Oxygen requirement greater 
                     
                   equivalent 
                 
                   than or equal to 40% FiO2 or 
                     
                   dexamethasone (e.g., 
                 
                   hypotension requiring high- 
                     
                   10 mg intravenously 
                 
                   dose or multiple vasopressors 
                     
                   every 6 hours) until 
                 
                   or Grade 3 organ toxicity or 
                     
                   Grade 1, then taper 
                 
                   Grade 4 transaminitis. 
                     
                   corticosteroids. 
                 
                     
                     
                   If improving, manage 
                 
                     
                     
                   as Grade 2. 
                 
                     
                     
                   If not improving, 
                 
                     
                     
                   manage as Grade 4. 
                 
                   Grade 4 
                   Per Grade 2 
                   Administer 
                 
                   Life-threatening symptoms. 
                     
                   methylprednisolo 
                 
                   Requirements for ventilator 
                     
                   ne 1000 mg 
                 
                   support or continuous veno- 
                     
                   intravenously per 
                 
                   venous hemodialysis 
                     
                   day for 3 days. 
                 
                   (CVVHD), or Grade 4 organ 
                     
                   If improving, taper 
                 
                   toxicity (excluding 
                     
                   corticosteroids, and 
                 
                   transaminitis). 
                     
                   manage as Grade 3. 
                 
                     
                     
                   If not 
                 
                     
                     
                   improving, 
                 
                     
                     
                   consider 
                 
                     
                     
                   alternate 
                 
                     
                     
                   immunosuppres 
                 
                     
                     
                   sants. 
                 
                     
                 
             
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         27 . The method of any one of  claims 20  through  22  and  26 , wherein neurologic toxicity in a B cell ALL subject is managed in accordance with one of the following two protocols: 
       
         
           
                 
                 
                 
               
                     
                 
                   NE 
                     
                   Revised Management 
                 
                   Grade 
                   Original Management Guidelines 
                   Guidelines 
                 
                     
                 
                   Grade 1 
                   Supportive care 
                   Supportive care 
                 
                     
                   Neurological examination and 
                   Closely monitor neurologic 
                 
                     
                   additional work-up as clinically 
                   status 
                 
                     
                   indicated 
                   Consider prophylactic 
                 
                     
                     
                   antiepileptic 
                 
                   Grade 2 
                   Supportive Care and Evaluation 
                   Supportive Care and Evaluation 
                 
                     
                   Neurological examination, brain MRI, 
                   Continuous cardiac telemetry 
                 
                     
                   and evaluation of CSF; consider EEG as 
                   and pulse oximetry as 
                 
                     
                   clinically indicated 
                   indicated 
                 
                     
                   Consider prophylactic antiepileptic 
                   Serial neurological 
                 
                     
                     
                   examinations to include 
                 
                     
                     
                   fundoscopy and Glasgow 
                 
                     
                     
                   Coma Score, brain MRI, 
                 
                     
                     
                   evaluation of CSF, EEG; 
                 
                     
                     
                   consider neurology consult 
                 
                     
                     
                   Administer antiepileptics for 
                 
                     
                     
                   patients with seizures 
                 
                     
                   Tocilizumab 
                   Tocilizumab 
                 
                     
                   Consider tocilizumab 8 mg/kg IV over 1 
                   For patients with 
                 
                     
                   hour (not to exceed 800 mg) for patients 
                   concurrent CRS, administer 
                 
                     
                   with comorbid conditions (eg, grade >2 
                   tocilizumab 8 mg/kg IV over 
                 
                     
                   CRS) 
                   1 hour (not to exceed 800 
                 
                     
                     
                   mg); repeat every 4-6 hours 
                 
                     
                     
                   as needed if not responsive to 
                 
                     
                     
                   IV fluids or increasing 
                 
                     
                     
                   supplemental oxygen, for a 
                 
                     
                     
                   maximum of 3 doses in 24 
                 
                     
                     
                   hours 
                 
                     
                     
                   Discontinue tocilizumab if 
                 
                     
                     
                   patient improves 
                 
                     
                   Corticosteroids 
                   Corticosteroids 
                 
                     
                   N/A 
                   For patients without 
                 
                     
                     
                   concurrent CRS, administer 
                 
                     
                     
                   dexamethasone 10 mg IV 
                 
                     
                     
                   every 6 hours 
                 
                     
                     
                   For patients with concurrent 
                 
                     
                     
                   CRS, if no improvement 
                 
                     
                     
                   within 24 hours after starting 
                 
                     
                     
                   tocilizumab, administer 
                 
                     
                     
                   dexamethasone 10 mg IV 
                 
                     
                     
                   every 6 hours 
                 
                     
                     
                   Taper corticosteroids if 
                 
                     
                     
                   patient improves 
                 
                   Grade 3 
                   Supportive Care and Evaluation 
                   Supportive Care and Evaluation 
                 
                     
                   Per grade 2 
                   Manage in monitored care or 
                 
                     
                   Monitor with continuous cardiac 
                   ICU 
                 
                     
                   telemetry and pulse oximetry 
                     
                 
                     
                   Tocilizumab 
                   Tocilizumab 
                 
                     
                   Consider tocilizumab 8 mg/kg IV over 1 
                   Per grade 2 
                 
                     
                   hour (not to exceed 800 mg); repeat 
                   Discontinue tocilizumab if 
                 
                     
                   every 4-6 hours if symptoms have not 
                   patient improves 
                 
                     
                   stabilized or improved 
                     
                 
                     
                   Corticosteroids 
                   Corticosteroids 
                 
                     
                   Consider corticosteroids (eg, 
                   Administer dexamethasone 10 
                 
                     
                   dexamethasone 10 mg IV every 6 hours 
                   mg IV every 6 hours 
                 
                     
                   or methylprednisolone l mg/kg BID) 
                   Taper corticosteroids if 
                 
                     
                   for worsening symptoms despite 
                   patient improves 
                 
                     
                   tocilizumab 
                     
                 
                   Grade  
                   Supportive Care and Evaluation 
                   Supportive Care and Evaluation 
                 
                     
                   Per grade 2 
                   Per grade 3 
                 
                     
                   Monitor with continuous cardiac 
                   Mechanical ventilation may 
                 
                     
                   telemetry and pulse oximetry 
                   be required 
                 
                     
                     
                   Administer 
                 
                     
                     
                   immunosuppresants if patient 
                 
                     
                     
                   does not improve 
                 
                     
                   Tocilizumab 
                   Tocilizumab 
                 
                     
                   Administer tocilizumab per grade 3 if 
                   Per grade 2 
                 
                     
                   not previously administered 
                     
                 
                     
                   Corticosteroids 
                   Corticosteroids 
                 
                     
                   Administer corticosteroids (eg, 
                   Administer high-dose 
                 
                     
                   methylprednisolone ig/d × 3 days, 
                   corticosteroids (eg, 
                 
                     
                   followed by 250 mg BID × 2 days, then 
                   methylprednisone ig/d × 3 
                 
                     
                   125 mg BID × 2 days, then 60 mg BID × 
                   days) 
                 
                     
                   2 days) 
                   Taper corticosteroids if 
                 
                     
                     
                   patient improves 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         28 . The method of any one of  claims 1  through  27 , wherein the B cell ALL subject may receive any one or more of the following bridging chemotherapy regimens: 
       
         
           
                 
               
                     
                 
                   Predefined Bridging Chemotherapy Regimens 
                 
                     
                 
                     
                 
                 
                 
               
                   Attenuated VAD 
                   Vincristine non-liposomal (1-2 mg IV weekly) or liposomal (2.25 
                 
                     
                   mg/m 2  IV weekly), and dexamethasone 20-40 mg IV or PO daily 
                 
                     
                   x 3-4 days per week. Optional doxorubicin 50 mg/m 2  IV × 1 (first 
                 
                     
                   week only) 
                 
                   Mercaptopurine (6- 
                   50-75 mg/m 2 /day by mouth (administer at bedtime on an empty 
                 
                   MP) 
                   stomach to improve absorption) 
                 
                   Hydroxyurea 
                   Doses titrated between 15-50 mg/kg/day (rounded to the nearest 
                 
                     
                   500 mg capsule and given as a single daily oral dose on a 
                 
                     
                   continuous basis) 
                 
                   DOMP 
                   Dexamethasone 6 mg/m 2 /day PO (or IV) divided BID 
                 
                     
                   days 1-5, vincristine 1.5 mg/m 2  (maximum dose 2 mg) IV on 
                 
                     
                   day 1, methotrexate 20 mg/m 2  PO weekly, 6-MP 
                 
                     
                   50-75 mg/m 2 /day PO daily 
                 
                   Attenuated 
                   Fludarabine 30 mg/m 2  IV days 1-2, cytarabine 2 g/m 2  IV days 1-2, 
                 
                   FLAG/FLAG-IDA 
                   G-CSF 5 μg/kg SC or IV starts on day 3 and can continue until 
                 
                     
                   day before the start of conditioning chemotherapy. With or 
                 
                     
                   without idarubicin 6 mg/m 2  IV days 1-2 
                 
                   Mini-hyper CVAD 
                   Course A: Cyclophosphamide 150 mg/m 2  every 12 h × 3 days, 
                 
                   (courses A and/or 
                   dexamethasone 20 mg/d IV or PO daily days 1-4 and 11-14, 
                 
                   B) 
                   vincristine 2 mg IV × 1 
                 
                     
                   Course B: methotrexate 250 mg/m 2  IV over 24 hours on day 1, 
                 
                     
                   cytarabine 0.5 g/m 2  IV every 12 hours × 4 doses on days 2 and 3 
                 
                     
                 
             
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         29 . Autologous T cells expressing an anti-CD19 CAR for use in a method for treating mantle cell lymphoma (MCL) or B cell ALL according to any one of  claims 1  through  28 . 
     
     
         30 . Use of autologous T cells expressing an anti-CD19 CAR in the manufacturing of a medicament for treating mantle cell lymphoma (MCL) or B cell ALL according to any one of  claims 1  through  28 . 
     
     
         31 . A method of predicting:
 (i) objective response of a subject to a CAR T cell treatment (optionally, according to the method of any one of  claims 1  through  28 ) comprising measuring peak CAR T cell levels and comparing them to a reference standard, wherein objective response is positively associated with peak CAR T cell levels, wherein objective response includes both complete response and partial response, and wherein all responses are assessed using the Lugano classification.   (xii) minimal residual disease (e.g., at week 4) in response to a CAR T cell treatment (optionally, according to the method of any one of  claims 1  through  28 ) comprising measuring peak CAR T cell levels and comparing them to a reference standard, wherein negative minimal residual disease is associated with higher peak CAR T cell levels.   (xiii) grade ≥3 CRS and/or grade ≥3 neurologic events (NE) in a subject receiving CAR T cell treatment (optionally, according to a method of any one of  claims 1  through  28 ) comprising measuring peak CAR T cell expansion after treatment and comparing the levels to a reference value, wherein the higher the CAR T cell expansion, the higher the chance for grade ≥3 CRS and/or grade ≥3 NE events.   (xiv) grade ≥3 CRS and/or grade ≥3 NE comprising measuring the peak levels of GM-CSF and IL-6 post-CAR T cell treatment (optionally, according to the method of any one of  claims 1  through  28 ) and comparing them to a reference level, wherein the higher the peak level of these cytokines, the higher the chance for grade ≥3 CRS and/or grade ≥3 NE.   (xv) grade ≥3 CRS in a subject receiving CAR T cell treatment (optionally, according to a method of any one of  claims 1  through  28 ) comprising measuring the peak level of serum ferritin post-CAR T cell treatment and comparing it to a reference level, wherein the higher the peak level of ferritin, the higher the chance for grade ≥3 CRS.   (xvi) grade ≥3 CRS comprising measuring the peak levels of serum IL-2 and IFN-gamma post-CAR T cell treatment (optionally, of any one of  claims 1  through  28 ) and comparing them to a reference level, wherein the higher the peak level of IL-2 and IFN-gamma, the higher the chance for grade ≥3 NE.   (xvii) grade ≥3 CRS comprising measuring the cerebrospinal fluid levels of C-reactive protein, ferritin, IL-6, IL-8, and/or vascular cell adhesion molecule (VCAM) post-CAR T cell treatment (optionally, of any one of  claims 1  through  28 ) and comparing them to a reference level, wherein the higher the cerebrospinal fluid levels of C-reactive protein, ferritin, IL-6, IL-8, and/or vascular cell adhesion molecule (VCAM), the higher the chance for grade ≥3 NE   (xviii) grade ≥3 CRS post-CAR T cell treatment (optionally, according to a method of any one of  claims 1  through  28 ) comprising measuring peak serum levels of IL-15, IL-2 Rα, IL-6, TNFα, GM-CSF, ferritin, IL-10, IL-8, MIP-1a, MIP-1b, granzyme A, granzyme B, and/or perforin after anti-CD19 CAR T treatment and comparing the levels to reference levels, wherein the peak serum levels of IL-15, IL-2 Rα, IL-6, TNFα, GM-CSF, ferritin, IL-10, IL-8, MIP-1a, MIP-1b, granzyme A, granzyme B, and/or perforin associate positively with grade ≥3 CRS.   (xix) grade ≥3 CRS post-CAR T cell treatment of B cell ALL (optionally, according to a method of any one of  claims 1  through  28 ) comprising measuring peak serum level of IL-15 after anti-CD19 CAR T treatment and comparing the levels to reference levels, wherein the peak serum level of IL-15 associates negatively with grade ≥3 CRS.   (xx) grade ≥3 CRS and/or grade ≥3 NE post-CAR T cell treatment (optionally, according to a method of any one of  claims 1  through  28 ) comprising measuring peak serum levels of IL-6, TNFα, GM-CSF, IL-10, MIP-1b, and granzyme B after anti-CD19 CAR T treatment and comparing the levels to reference levels, wherein peak serum levels of IL-6, TNFα, GM-CSF, IL-10, MIP-1b, and granzyme B associate positively with grade ≥3 CRS and grade ≥3 NE.   (xxi) whether a patient is going to be MRD (10 −5  sensitivity) negative at 4 weeks/one month post-CAR T cell treatment (optionally, of any one of  claims 1  through  28 ), comprising measuring peak serum levels of IFN-γ, IL-6, and/or IL-2 after treatment and comparing the level to a reference standard, wherein peak serum levels of IFN-γ, IL-6, and/or IL-2 associate positively with being MRD negative at one month.   
     
     
         32 . The method of any one of  claims 20  through  24 ,  26 ,  27 , and  30  through  31 , wherein CRS and NE are graded by the method described in Lee et al., Blood 2014; 124: 188-195. 
     
     
         33 . The method of  claim 31 , wherein the reference standard is established by any method generally used in the biomarker arts, such as quartile analysis of patient populations with known responses, grades of toxicity, and MRD levels. 
     
     
         34 . The method of  claim 31 , wherein CAR T cell levels are measured by CAR gene copies per microgram of DNA in blood. 
     
     
         35 . The method of any one of  claims 1  through  34 , further comprising reducing the levels/activity of the cytokines that associate positively with grade ≥3 CRS and/or grade ≥3 NE post CAR T cell infusion to reduce grade ≥3 CRS and/or grade ≥3 NE. 
     
     
         36 . A method of improving the effectiveness of CAR T cell treatment (e.g., of classical, blastoid, and pleomorphic MCL, and B cell ALL), in a subject in need thereof, comprising manipulating the T cell phenotype of the T cell product administered to the subject, optionally wherein the manipulation comprises increasing the number of CD3+ T cells, decreasing the number of CD3− cells, increasing the number/percentage of CDRA45+ CCR7+(naïve-like) T cells and/or decreasing the number/percentage of differentiated cells in the T cell product during production, decreasing the levels of IFN-gamma production by the T cells, wherein the improvement is observed relative to the effectiveness of a T cell product that is prepared without any intentional manipulation of the number/percentage of CDRA45+ CCR7+ (naïve-like) T cells and/or the number/percentage of differentiated cells in the T cell product.

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