Phase I Dose Escalation of T-cell Receptor α/β Depleted Donor Lymphocyte Infusions Following CD34+- Selected Allogeneic Stem Cell Transplantation From Related & Unrelated Donors in Patients With Lymphoid, Myeloid or Plasma Cell Malignancies
Phase: PHASE1
Conditions: Lymphoid Leukemia, Acute, Myeloid Malignancy, Plasma Cell Tumor
Sponsor: Guenther Koehne

Protocol # 2019-KOE-004 
 
Version #3.0 16 SEP 2022 
A Phase I Dose Escalation Trial of T-cell Receptor α/β Depleted Donor Lymphocyte Infusions 
following CD34+-selected Allogeneic Stem Cell Transplantation from Related and Unrelated 
Donors for Patients with Lymphoid, Myeloid or Plasma Cell Malignancies 
 
 
 
Protocol Number: 2019-KOE-004 
Protocol Version: 3.0 
Version Date: 16 SEP 2022 
 
 
 
Principal Investigator 
Guenther Koehne, M.D., Ph.D. 
Deputy Director of Miami Cancer Institute (MCI) 
Miami Cancer Institute  
8900 North Kendall Drive 
Miami, Florida 33176 
Email: GuentherK@baptisthealth.net    
 
 
 
Sponsor 
Miami Cancer Institute at Baptist Health, Inc. 
8900 North Kendall Drive 
Miami, FL 33176 
 
 
 
 
IND # 28075 
 
 
 
 
 
 
 
 
 
 
 
 
 
NCT05350163

Protocol # 2019-KOE-004 
 
Version #3.0 16 SEP 2022 
 
 
1 
CONTENTS 
1 
PROTOCOL SUMMARY ............................................................................................................................... 4 
2 
INTRODUCTION ............................................................................................................................................ 5 
2.1 
Background and Rationale ................................................................................................................. 5 
3 
OBJECTIVES AND SCIENTIFIC AIMS ...................................................................................................... 8 
4 
STUDY DESIGN ............................................................................................................................................. 8 
4.1 
Overall Design ...................................................................................................................................... 8 
5 
STUDY POPULATION .................................................................................................................................. 8 
5.1 
Inclusion Criteria .................................................................................................................................. 8 
5.2 
Exclusion Criteria ................................................................................................................................. 9 
5.3 
Strategies for Recruitment .................................................................................................................. 9 
6 
STUDY INTERVENTION .............................................................................................................................. 9 
6.1 
Study Intervention(s) Administration ............................................................................................... 10 
6.1.1 
Study Intervention Description ..................................................................................... 10 
7 
THERAPEUTIC/DIAGNOSTIC AGENTS ................................................................................................. 11 
7.1.1 
Processing of CD34 negative Fraction for TCR a/b depletion ................................ 11 
7.2 
The CliniMACS System for Positive Selection of TCR α/β CD3+ T-Cells................................. 13 
7.2.1 
The CliniMACS Instrument ........................................................................................... 13 
7.2.2 
CliniMACS Tubing Set ................................................................................................... 13 
7.2.3 
CliniMACS TCR α/β Reagent ....................................................................................... 14 
7.3 
The CliniMACS PBS/EDTA Buffer .................................................................................................. 15 
8 
TREATMENT/INTERVENTION PLAN ...................................................................................................... 15 
8.1 
TCR a/b depleted donor lymphocyte infusions ............................................................................. 15 
8.1.1 
Dosing and time intervals of TCR a/b Depeleted DLI. .............................................. 15 
8.2 
Prophylaxis against acute graft-versus-host disease ................................................................... 16 
9 
EVALUATION post TCR a/b depleted donor lymphocyte infusions ..................................................... 16 
9.1 
Post-infusion Evaluation ................................................................................................................... 16 
10 
TOXICITIES/SIDE EFFECTS ..................................................................................................................... 18 
10.1 
Toxicities/Side Effects of the Transplant and the Combined Effects of the Conditioning 
Regimen and Transplant and TCR a/b depeleted DLI. ................................................................................... 18 
10.1.1 
Infections ......................................................................................................................... 18 
10.1.2 
Graft Failure / Poor Marrow Function .......................................................................... 18 
10.1.3 
Graft-versus-host Disease ............................................................................................ 19 
10.1.4 
Sinusoidal obstruction syndrome (SOS) of the Liver ................................................ 19 
10.1.5 
End Organ Damage ....................................................................................................... 19 
10.1.6 
Disease Relapse ............................................................................................................ 20 
11.3.7 Death 
 
 .......................................................................................................................... 20 
11 
CRITERIA FOR THERAPEUTIC RESPONSE/OUTCOME ASSESSMENT....................................... 20 
11.1 
Disease relapse and recurrence ..................................................................................................... 20 
11.2 
Disease-free Survival ........................................................................................................................ 20 
11.3 
Overall Survival .................................................................................................................................. 20 
12 
STATISTICAL CONSIDERATIONS .......................................................................................................... 20 
12.1 
Study Population definition ............................................................................................................... 22 
12.2 
Subjects Disposition .......................................................................................................................... 22 
12.3 
Efficacy Analysis ................................................................................................................................ 22 

Protocol # 2019-KOE-004 
 
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12.4 
Safety Analysis ................................................................................................................................... 22 
13 
RESEARCH PARTICIPANT REGISTRATION AND RANDOMIZATION PROCEDURES ............... 24 
13.1 
Research Participant Registration ................................................................................................... 24 
13.2 
Randomization ................................................................................................................................... 25 
14 
DATA MANAGEMENT ISSUES ................................................................................................................. 25 
14.1 
Quality Assurance.............................................................................................................................. 25 
14.2 
Data and Safety Monitoring .............................................................................................................. 26 
15 
SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS .............................. 26 
15.1 
Informed Consent Process ............................................................................................................... 26 
15.2 
CRITERIA FOR REMOVAL FROM STUDY .................................................................................. 27 
15.3 
Criteria for suspension or premature termination of the study .................................................... 28 
16 
PROTECTION OF HUMAN SUBJECTS .................................................................................................. 29 
16.1 
Benefits ............................................................................................................................................... 29 
16.2 
Privacy ................................................................................................................................................. 30 
16.3 
Serious Adverse Event (SAE) Reporting ....................................................................................... 30 
16.5 Definition of aN SAE .................................................................................................................................... 32 
17.3.1 IND reporting 
 .......................................................................................................................... 33 
18 PUBLICATION POLICY ..................................................................................................................................... 33 
19 REFERENCES .................................................................................................................................................... 33 
 

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1 
 
PROTOCOL SUMMARY 
 
This is a phase I dose escalating trial designed to identify tolerable, clinically active doses of 
donor-derived TCR α/β depleted donor lymphocyte infusions (DLI) when administered to patients 
with lymphoid or myeloid malignancies following a T-cell depleted allogeneic hematopoietic cell 
transplantation (TCD HSCT). Patients will undergo a preparative regimen with Busulfan, 
Melphalan, Fludarabine, (Bu/Mel/Flu) or Total Body Irradiation (TBI) plus Thiopeta and 
Cyclophosphamide (TBI, Thio, Cy) and anti-thymocyte globulin (ATG), followed by a CD34+-
selected stem cell transplant from a histo-compatible related or unrelated donor. Hematopoietic 
stem cell donors for this trial will include individuals who are 10/10 HLA matched or one antigen 
or allele mismatched at the HLA-A, B, C, DRB1 or DQB1 locus, as defined by high resolution 
methods. Donors who are 8/10 HLA matched with an antigen or allele mismatched at HLA-DQB1 
and at one other locus will also be eligible for the trial. The donor-derived TCR α/β depleted donor 
lymphocyte infusions post transplantation are administered to induce complete remissions in 
patients with residual disease, to decrease the rate of relapse and to reduce viral 
complications/reactivations following allogeneic stem cell transplants. For this study, the TCR α/β 
depleted donor lymphocyte infusions is obtained from the CD34 negative fractions maintained 
after the CD34+ selection of the stem cell product. 
 
The primary objective of this study is to assess the safety of TCR α/β depleted DLI derived from 
the CD34neg fractions from the original transplant donor of the CD34+ selected allogeneic stem 
cell transplant, when administered to patients with ALL, AML, Myelodysplastic Syndrome, 
Myelofibrosis or relapsed/refractory multiple myeloma following TCD HSCT. Secondary 
objectives are: 1) to assess the effects of these adoptively transferred cells depleted of TCR α/β 
T cells on minimal residual disease and the progression of disease, 2) to quantitate levels of TCR 
α/β and TCR γ/δ T cells in addition to reconstitution of NK cells and 3) to determine the 
reconstitution of viral-specific T lymphocytes and associated viral complications following 
adoptive transfer of the T cells. 
 
Patients eligible for this protocol are patients who are undergoing a CD34+-selected allogeneic 
transplant for AML, ALL, MDS, MPD or multiple myeloma as indicated on their respective, primary 
transplant protocols/treatment plans. The patients will be conditioned for transplantation with 
1. Busulfan (Busulfex®) (0.8 mg/Kg/dose Q6H x 10-12 doses), Melphalan (70 mg/m2/day x 2 doses) 
and Fludarabine (25mg/m2/day x 5 doses) or with 2. Hyperfractionated total body irradiation to 
dose of 1375cGy fractions at 4-6 hour intervals three times a day for a total of 11 or 12 doses 
followed by Thiotepa 5mg/kg/day x 2 (or 10mg/kg/day x  1) and cyclophosphamide 60mg/kg/day 
x 2 (or Fludarabine 25mg/m2 x 5 if cyclophosphamide is contraindicated). Patients will also 
receive ATG (Thymoglobulin®) prior to transplant to promote engraftment and to prevent graft-versus 
host disease post transplantation. The preferred source of stem cells will be peripheral blood 
stem cells (PBSC) mobilized by treatment of the donor with G-CSF for 5-6 days. PBSC will be 
isolated, and T-cells depleted by positive selection of CD34+ progenitor cells, using the CliniMACS 
Cell Selection System. The CD34+-selected, T-cell depleted peripheral blood progenitors will then 

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be administered to the patients after they have completed cytoreduction. No drug prophylaxis 
against GvHD will be administered post-transplant. All patients will also receive G-CSF post-
transplant to foster engraftment. 
 
The CD34neg cell fraction, usually discarded following the cell processing, containing the CD3+ 
TCR α/β and TCR γ/δ T cells, NK cells and B cells will then be further processed for TCR α/β 
depletion. In this process, the CliniMACS® TCR α/β CD3+ Reagent Kit is used to deplete human 
TCR α/β CD3+ T cells in vitro from heterogeneous hematological cell populations for lymphocyte 
infusions and cell products, depleted of the TCR α/β positive T-cells will be aliquoted for infusions 
post-transplant. Initially, patients in cohort I are treated with single infusions containing 5x10e5 
of TCR γ/δ T cells/kg  at de-escalating time intervals to determine the earliest tolerated time point 
to safely administer these infusions to these patients. Thereafter, starting at the earliest tolerated 
time point of TCR γ/δ T-cell infusions, cohorts of patients are treated with 3 infusions of escalating 
doses of TCR γ/δ T cells/kg at 3-4 week intervals. The dosing of each infusion is adjusted based 
on the donors HLA-match or mismatch, respectively, as outlined in detail in this protocol. 
 
The sample size and duration of the study: 
This is a single center pilot study. 
The duration of this study is estimated to be 18 – 24 months with a total of up to 18 
patients for the primary analysis, 9 -12 patients in  the  HLA-matched group, and 6-9 
patients in the HLA-mismatched group. To account for attrition and feasibility of dosing 
effect on treatment on minimal residual disease and progression of disease, the final 
sample size will be 24 patients. 
 
 
2 
 
INTRODUCTION 
 
2.1 
BACKGROUND AND RATIONALE 
 
During the last 30 years, allogeneic stem cell transplantation has been developed and improved 
for the curative treatment of patients with life-threatening hematological and non-hematological 
malignancies as well as non-malignant diseases. The stem cell transplant is intended to 
reconstitute a healthy and self-sustaining hematopoietic system after eradication of the 
underlying disease by chemo- and/or radiotherapy (conditioning). The success of allogeneic stem 
cell transplantation is limited by the potential development of life-threatening complications 
associated with the procedure. Infection, relapse, and graft-versus-host disease (GVHD) are still 
major causes of mortality and morbidity after allogeneic transplantation. Therefore, the speed 
of reconstitution of T, B, NK and other immune cells is of importance for the reduction of the risk 
for bacterial, viral and fungal infections as well as for mediating anti-tumor effects, especially in 
HLA mismatched and haploidentical transplantation. (1-3) Approaches to reduce the risk of GVHD 
development have been pursued. In particular, GVHD remains a major cause of mortality and 
morbidity after transplantation with allogeneic peripheral blood or bone marrow for both 

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matched and mismatched transplants. (4,5) Strategies of graft engineering have been employed 
with the aim to deplete donor-derived T cells which play a central role in GVHD development 
following an allogeneic stem cell transplant. 
 
Thus, the early morbidity and mortality associated with acute GvHD is a major factor limiting the 
success of transplantation, as is the long-term morbidity associated with chronic GvHD.  The risk 
of acute GvHD following allogeneic bone marrow transplantation (BMT) from HLA-matched 
siblings is 20-60% despite the use of immunosuppressive agents like cyclosporine A (CSA), 
tacrolimus (FK506), methotrexate (MTX), anti-thymocyte globulin (ATG) and corticosteroids, 
alone or in combination (6-9).  Grades II-IV (moderate to severe) acute GvHD are associated with 
an increased risk of transplant-related mortality (10,11).  Mortality rates among patients who 
develop GvHD can be as high as 75% when that disease is unresponsive to therapy (11). 
 
Certain methods of T-cell depletion are effective in reducing or preventing acute GvHD in both 
HLA­matched and HLA-disparate transplant recipients without co-administration of CSA and/or 
MTX. Depletion of T-cells by positive selection of CD34+ hematopoietic cells from GCSF mobilized 
peripheral blood leukocytes (PBSC) and transplantation of these CD34+cell-enriched, T-cell 
depleted cell fractions has been recently studied in the United States, is the CliniMACS system 
(Miltenyi Biotec) (12-14).  In this system, GCSF­ mobilized PBSC are introduced into the closed 
device, which treats the cells with anti CD34 MoAB coated paramagnetic beads.  The CD34+ 
progenitor cells are then separated from other cells by passage through an electromagnetic field, 
and then washed and eluted.  In the large Perugia series, recipients of HLA haplotype disparate 
CD34+ PBSC isolated on the CliniMACS device had only a 10% probability of developing acute or 
chronic GVHD (15).  In a multicenter trial conducted by the BMT Clinical Trials Network under an 
FDA IND, this method yielded a progenitor-enriched cell fraction that provided doses of 
progenitor cells ranging from 2.4-31.3 (med 7.9) x  106  CD34+ cells kg and T-cell doses of 1.1-
84.9 (median 6.6) x 103 CD3+ cells/kg.  HLA matched related HSCT fractionated by this approach 
were administered after cytoreduction with the protocol of hyperfractionated TBI, Thiotepa and 
Cyclophosphamide.  These transplants provided consistent engraftment (no late graft failure in 
44 patients transplanted) and were associated with incidences of 20.5% grade II-IV acute GvHD 
and 7.6% extensive chronic GvHD.  Overall survival for patients with AML transplanted in 1° and 
2° remission was 74% (16).  Overall risk of relapse for patients is 1° or 2° CR was 18%, which 
compares favorably with results of unmodified transplants for these disease indications. 
 
The highly favorable results of the BMT CTN trial published formed the basis for an application 
to the FDA for licensure as a device to prevent GvHD in patients transplanted for AML. Approval 
for use of the CliniMACS CD34 reagent system for processing of hematopoietic progenitor cells 
was obtained for patients with AML when transplanted in 1st morphologic complete remission 
and with CD34+-selected cells from an allogeneic, HLA-identical sibling donor. For other 
indications and for transplants from alternative donors, we performed a clinical trial of T-cell 
depleted CD34+ PBSC transplants fractionated on the CliniMACS which was recently summarized 
in a manuscript submitted. In this trial summary, we describe an excellent outcome of pts 
undergoing CD34+-selected allografts of two chemotherapy-based myeloablative conditioning 

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regimens that secure consistent engraftment of MRD, MUD and single HLA-A, B, C or DR 
mismatched donor-derived CD34+TCDHCTs. Compared to our standard TBI-based regimen the 
incidences of acute, grade 2-4 GVHD and chronic GVHD were significantly lower after Bu/Mel/Flu 
conditioning. Importantly, none of the patients undergoing CD34+-selected allografts on our trial 
have received post transplantation immunosuppressive therapies, which therefore provides an 
ideal platform for the additional administration of post-transplant immunotherapies with donor 
effector cells, such as T-lymphocytes and/or NK cells. 
Unmodified donor lymphocyte infusions (DLI) have been used in T cell depleted and repleted 
allogeneic transplantation in case of decreasing donor chimerism, signs of minimal residual 
disease and therapy refractory viral infections. (17,18) GVHD remains the most common and 
serious adverse event of unmodified DLI after allogeneic transplantation. (19) Different strategies 
have been used with the aim to dissociate the beneficial effects of DLI's such as graft versus 
malignancy activity, from the unwanted, harmful effects, such as GVHD. These approaches 
encompass the use of escalating doses of DLI's, CD8+ T cell depleted DLI's, CD3+ T cell depleted 
DLI's and NK cell infusions. (21, 22) 
In our own studies, we have demonstrated the potential of post-transplant DLI and associated 
complications such as GvHD (23, 24) An important and significant limitation of post-transplant 
donor lymphocyte infusions is the time dependence and amount of donor T-cell infused post allo 
transplant, excluding the safe administration of DLI for prophylaxis within the first 4-6 months 
post allograft. 
To further improve outcomes of CD34+ transplantations and donor lymphocyte infusions, a new 
method for the depletion of unwanted T cells triggering GVHD was developed, namely the 
selective depletion of TCR α/β T cells, which constitute a subpopulation of CD3+ cells. Depletion 
of TCR α/β CD3+ T cells in this setting is very efficient and selective with a 4.5 — 5 log T cell 
reduction. (25) The efficiency of depletion is comparable to that achieved with CD34+ cell 
enrichment. This may allow further reduction of pharmacological immunosuppression post-
transplantation including its numerous negative short- and long-term side effects. This method 
of TCR α/β CD3+ cells depletion leads to lymphocyte compositions, which strongly resemble 
unmanipulated preparations. For example, TCR γ/δ CD3+ T cells and NK cells are preserved. (26)  
In-vitro, when TCR γ/δ T cells were cultivated in co-culture with primary leukemia blasts they 
proliferated and exhibited cytotoxicity against the blasts. (24) In addition, TCR γ/δ+ T cells and 
NK cells have been shown to synergistically enhance their anti-leukemic and anti-viral effector 
cell function in vitro. (27) Furthermore, it has been reported that TCR γ/δ+ T cells seem to exert 
anti-leukemic effects in partially mismatched PBMC in vivo. In a retrospective analysis, patients 
with a high number of TCR γ/δ+ T cells post-transplantation had an improved 5-year survival 
compared to those with normal or low counts (70.8% vs. 19.6%). Therefore, a high content of 
TCR γ/δ +T cells early post T-cell depleted transplants might have a beneficial effect on survival. 
(27)  Since TCR γ/δ+  T cells co-express CD3, but do not present the α/β T cell receptor (TCR α/β), 
they do not provoke GVHD driven by alloantigen T cell receptor activation. Additionally, TCR 
γ/δ+ T cells may support engraftment, may provide protection against infections and bridge the 
time to the immune reconstitution of TCR α/β+ T cells by providing protection against infections 
and they may exert GVT effects. (27-29) In addition, it has been shown, that similar effects are 
exerted by NK cells in the early post-transplant time period. (30 - 33) 

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Taken together, the information suggests that the administration of TCR α/β-depleted donor 
lymphocyte infusions, including functional TCR γ/δ+ T and NK cells at an early time point following  
CD34+-selected allografts provides effector cells capable of enhancing graft-versus-malignancy 
and anti-viral effects, while sparing the induction of GvHD. Most importantly, the CD34 negative 
cell fraction, remaining following a CD34+-cell selection for transplant, is rich of the effector cells, 
including TCR γ/δ+ T and NK cells and is usually discarded after CD34+ processing. In addition, in 
validation trials, we can demonstrate an effective 4-5 log depletion of TCR α/β+ cells from CD34 
depleted cell fractions, leading to high purity of TCR γ/δ+ T cell and NK cell fractions. 
 
3 
OBJECTIVES AND SCIENTIFIC AIMS 
Primary objectives: 
1. To assess the safety of TCR α/β depleted DLI derived from the CD34neg fractions from 
the original transplant donor of the CD34+ selected allogeneic stem cell transplant, when 
administered to patients with ALL, AML, Myelodysplastic Syndrome, Myelofibrosis  or 
relapsed/refractory multiple myeloma following TCD HSCT. 
 
Secondary objectives: 
1) To assess the effects of these adoptively transferred cells depleted of TCR α/β T cells 
on minimal residual disease and the progression of disease,  
2) To quantitate levels of TCR α/β and TCR γ/δ T cells in addition to reconstitution of NK 
cells  
3) To determine the development of transplant-associated viral complications following 
adoptive transfer of the T cells. 
4) To correlate the presence and duration of MRD negative status with the duration of 
complete remission of these patients.  
 
4 
STUDY DESIGN  
 
4.1 
OVERALL DESIGN 
 
This is a pilot study designed to identify the safe administration and effects of TCR α/β-depleted 
donor lymphocyte infusions following CD34+-selected allogeneic stem cell transplant from HLA 
compatible related or unrelated donors on minimal residual disease status, infectious 
complication, progression-free and overall survival. 
 
5 
STUDY POPULATION 
 
5.1 
INCLUSION CRITERIA 
 
Subject Inclusion Criteria 

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Patients with hematologic malignancies that are candidates for CD34+ selected, T-cell depleted 
allogeneic hematopoietic stem cell transplantation. 
 
The following inclusion criteria are also required: 
 
• Patient's age includes  ≥18   to < 75 years old. 
• Patients may be of either gender or any ethnic background. 
• Patients must have a Karnofsky (adult) Performance Status of at least 70%. 
• Patients must have adequate organ function measured by: 
 
Cardiac:  asymptomatic or if symptomatic then LVEF at rest must be 50% and must 
improve with exercise. 
 
Hepatic: < 3x ULN AST and: s 1.5 total serum bilirubin, unless there is congenital benign 
hyperbilirubinemia. Patients with higher bilirubin levels due to causes other than active 
liver disease is also eligible with Pl approval e.g. patients with PNH, Gilbert’s disease or 
other hemolytic disorders. 
 
Renal:  serum creatinine: s; 1.2 mg/dl or if serum creatinine is outside the normal range, 
then CrCl > 40 ml/m in (measured or calculated/estimated). 
 
Pulmonary:   asymptomatic or if symptomatic, DLCO 50% of predicted (corrected for 
hemoglobin). 
 
Each patient must be willing to participate as a research subject and must sign an informed 
consent form. 
 
5.2 
EXCLUSION CRITERIA 
 
Subject Exclusion Criteria 
• Patients with active acute GvHD 
 
5.3 
STRATEGIES FOR RECRUITMENT 
 
Patients who fulfill the eligibility criteria as listed in Section 5.0 will be recruited for this study by 
an attending physician of the Hematologic Oncology or BMT Program at Miami Cancer Institute 
(MCI). This protocol will take due notice of NIH/ADAMHA policies concerning inclusion of women 
and minorities in clinical research populations. 
 
6 
STUDY INTERVENTION 
 

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6.1 
STUDY INTERVENTION(S) ADMINISTRATION 
 
6.1.1 STUDY INTERVENTION DESCRIPTION 
 
 
Initially, patients in cohort I will be treated with infusions of TCR γ/δ T cells at time de-escalating 
intervals. The first 3 patients who received a CD34+-selected allograft from a HLA-matched 
related or unrelated donor will obtain an infusion containing 5x10e5/kg TCR γ/δ T cells at 6 -7 
weeks post-transplant, the next 3 patients will be infused with 5x10e5/kg TCR γ/δ T cells at 4-5 
weeks post-transplant and a third group of 3 patients will be treated with 5x10e5/kg TCR γ/δ T 
cells 2-3 weeks post-transplant.   
 
The first 3 patients who received a CD34+-selected allograft from a HLA-mismatched donor will 
be treated with an infusion containing 1x10e5/kg TCR γ/δ T cells at 6 -7 weeks post-transplant, 
the next 3 patients will be infused with 1x10e5/kg TCR γ/δ T cells at 4-5 weeks post-transplant 
and a third group of 3 patients will be treated with 1x10e5/kg TCR γ/δ T cells 2-3 weeks post-
transplant. No immune suppressive therapies will be administered on this trial for these 
infusions.  
 
Subsequently, administration of 3 consecutive infusions of TCR α/β depleted DLI per patient 
cohort will be initiated at the earliest tolerated time point determined through these de-
escalating intervals. Thus, cohort II of 3 patients from matched related or unrelated donors will 
receive the first dose of 5x10e5/kg TCR γ/δ T cells at time point X, followed at 3-4 week intervals 
by a the second and third infusion 1x10e6/kg TCR γ/δ T cells. Cohort III of 3 patients will receive 
the first dose of 5x10e5/kg TCR γ/δ T cells at time point X, followed by an infusion of 1x10e6/kg 
TCR γ/δ T cells at 3-4 weeks later and a third infusion 2x10e6/kg TCR γ/δ T cells 3-4 weeks 
following the second infusion. Three additional patients will then be treated at the maximal 
tolerated dose determined. All patients will be re-assessed for acute GvHD prior to each TCR γ/δ 
T cell infusion, and subsequent infusions will be held if new onset of GvHD is observed.   
 
Cohort II + III of 3 patients from mismatched donors will receive the first dose of 1x10e5/kg TCR 
γ/δ T cells at time point X, followed at 3-4 week intervals by a the second and third infusion of 
5x10e5/kg TCR γ/δ T cells and 1x10e5/kg TCR γ/δ T cells at time point X, followed by an infusion 
of 5x10e5/kg TCR γ/δ T cells at 3-4 weeks later and a third infusion 1x10e6/kg TCR γ/δ T cells 3-4 
weeks following the second infusion, respectively. Three additional patients will then be treated 
at the maximal tolerated dose determined. All patients will be re-assessed for acute GvHD prior 
to each TCR γ/δ T cell infusion, and subsequent infusions will be held if new onset of GvHD is 
observed. A dose variation of ±20% of the indicated cell dose is acceptable for all infusions listed. 
However, the maximum dose of residual TCR α/β T cells contained within each infusion will be 
limited to a maximum of 2x10e4 TCR α/β T cells/kg. 
 
Table 1: Dose Levels of TCR γ/δ T cells following TCR α/β depleted DLI 

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Group Patients (HLA matched) 
TCR γ/δ T cells 
I 
3 per time point 
5 x 10e5/kg at 6-7, 4-5 and 2-3 weeks post-transplant 
II 
3 
5 x 10e5/kg starting time point X, 1 x 10e6/kg 3-4 weeks after first dose, 1 x 
10e6/kg 3-4 weeks after second dose. 
III 
3-6 
5 x 10e5/kg starting time point X, 1 x 10e6/kg 3-4 weeks after first dose, 2 x 
10e6/kg 3-4 weeks after second dose. 
 
 
Group Patients (HLA mismatched) TCR γ/δ T cells 
I 
3 per time point 
1 x 10e5/kg at 6-7, 4-5 and 2-3 weeks post-transplant 
II 
3 
1 x 10e5/kg starting time point X, 5 x 10e5/kg 3-4 weeks after first dose, 5 x 
10e5/kg 3-4 weeks after second dose. 
III 
3-6 
1 x 10e5/kg starting time point X, 5 x 10e5/kg 3-4 weeks after first dose, 1 x 
10e6/kg 3-4 weeks after second dose. 
 
On this trial, enrollment of patients will be staggered to provide time intervals sufficiently enough 
to permit the exclusion of new onset of acute GvHD. We will hold enrollment of the next patient 
for a minimum of 3-4 weeks after the first infusions with TCR γ/δ T cells of the first two patients 
of each cohort and will continue to enroll the following patient after such time period expired in 
the absence of new onset of acute GvHD. 
 
 
 
 
7 
THERAPEUTIC/DIAGNOSTIC AGENTS 
 
7.1.1 PROCESSING OF CD34 NEGATIVE FRACTION FOR TCR A/B DEPLETION 
 
Historically, the CD34 negative cell fraction, remaining following a CD34+-cell selection for 
transplant, has been discarded following the process. However, this cell fraction remains rich of 
the effector cells, including TCR γ/δ+ T and NK cells. The utilization of the CD34 negative cell 
fraction also appears logistically appropriate, as its use avoids another request for effector cell 
infusion from the same stem cell donor. Thus, the infusion of TCR α/β depleted DLI derived from 
the remaining cell fraction offers several advantages. In our initial validation trial, we 
demonstrate an effective 4-5 log depletion of TCR α/β+ cells from CD34 depleted cell fractions, 
leading to high purity of TCR γ/δ+ T cell and NK cell fractions for cryopreservation and planned 
infusion post-transplant. 
For the immunophenotyping of TCR α/β- depleted product, we are using BD FacsCanto II and we 
created our own panel, which include the following CD markers   CD45, CD3, CD4, CD8, CD14, 
CD16, CD19, CD56, TCR α/β and  TCR γ/δ plus 7ADD. Identification  and enumeration of T 
lymphocytes with TCR α/β and  TCR γ/δ  subsets, B lymphocytes and natural killer cells .     

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Viability testing will be performed by flow cytometry and 7-ADD staining. Viability from the TCR 
α/β+ depleted product from the negative fraction has been consistently between 90% to 95% 
and remained 80-85% following cryopreservation and thawing.  
The products will be cryopreserved by controlled rate freezing and stored on vapor phase at <130 
C   with a DMSO concentration of 5 to  7.5% based on total nucleated cells concentration. The 
cell infusions will be prepared in 5% human albumin and plasmalyte to be injectable in humans. 
Supplies and reagents that come into contact with patient product during processing, storage 
and/or administration must be sterile and of appropriate grade for intended use. Reagents 
undergo initial qualification for the intended use.  
Sterility testing is performed on all products prior to cryopreservation. If products have 
undergone manipulation, a post processing sterility testing is also performed. 
Stability program: 
Viability testing will be performed at various test intervals- preprocessing (zero-time point), post 
processing (inclusive of fresh and cryopreserved products). For cryopreserved products (storage 
conditions of ≤ -130°C), post thaw viability will be performed using sample aliquots 
representative of final processed product. This post thaw viability will be performed at time of 
infusion (as applicable), 1 year post storage, and annually, thereafter for the duration of aliquot 
availability. 
Pre and post processing cell counts to determine product identity, quality and strength (potency).  
Pre and post processing sterility testing (gram stain and 14 day culture) to determine purity. 
Post processing endotoxin testing to determine purity. 
For cryopreserved products, container integrity post thaw. 
Potency Program: 
Potency will be developed in an incremental approach based on the biological properties of the 
product. To assess biological function and characterization of product a series of analytical assays 
will be performed to assess potency at the early preclinical phase, as clinical study progress we 
would be able to determine the product effectiveness based on the clinical data.  
A Certificate of Analysis with release specifications will be generated for each product to 
demonstrate product activity, quality and consistency throughout product development. This 
Certificate of Analysis will include the tests used for routine lot release: 
Cell counts for Total nucleated cells  
Flow cytometry analysis for identification and enumeration of T cells, TCR α/β CD3+ T cells, 
γ/δTCR CD3+ T cells, natural killer T cells and B cells 
Sterility: gram stain and cultures 

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Endotoxin 
Viability (by both manual method and flow cytometry) 
 
7.2 
THE CLINIMACS SYSTEM FOR POSITIVE SELECTION OF TCR Α/Β CD3+ T-CELLS. 
 
The CliniMACS System (Miltenyi Biotec, Auburn, CA) including the CliniMACSplus Instrument, a 
CliniMACS Tubing Set, the CliniMACS  TCR α/β Reagent Kit and the CliniMACS PBS/EDT A Buffer 
is intended for the selection and enrichment of human TCR α/β CD3+ positive cells from a 
leukapheresis product. 
 
The CliniMACS TCR α/β Reagent Kit consists of 1 vial of CliniMACS TCR α/β -Biotin and 2 vials of 
CliniMACS Anti-Biotin Reagent which are used with the CliniMACS System. The CliniMACS TCR 
α/β Reagent Kit is used to deplete human TCR α/β T cells in vitro from heterogeneous 
hematological cell populations for stem cell transplantation and lymphocyte infusions in cases 
where this is clinically indicated. 
Each CliniMACS TCR α/β Reagent Kit is sufficient to label up to 24 x 109 TCR α/β+ T cells out of 60 
x 109 total white blood cells using an indirect labeling strategy. 
 
The components of the CliniMACS System include: 
 
7.2.1 THE CLINIMACS INSTRUMENT 
 
The CliniMACS instrument is a bench-top instrument consisting of a supporting structure to hold 
the column/tubing assembly and various bags, a series of valves through which the tubing set is 
fitted, a magnet between the poles of which the separation column is placed, a peristaltic pump 
through which a section of tubing is placed, software to control the instrument and user interface 
and a computer touchpad with a display window.  The instrument is operated at ambient 
temperature and it is intended to be multi­use item. 
 
The software for the CliniMACS Instrument controls the function of the electromechanical 
components of the instrument and the user interface.  Two separate computers, one a micro-
controller located on a control board of the CliniMACS Instrument and the second a PC 
compatible computer which operates the user interface are incorporated with the instrument. 
Software Version 2.3 1, the current version of software is directly traceable to the version of 
software utilized in pre-clinical testing and European Safety trials and has been inspected and 
approved by TOV product services with the CE Mark. 
 
7.2.2 CLINIMACS TUBING SET 
 

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The CliniMACS Tubing Set consists of a tubing element combined with a pair of proprietary cell 
selection columns.  These form a closed, sterile system for processing the cells.  The separation 
column is a proprietary component of the CliniMACS System consisting of a plastic column 
housing with polypropylene fits in each end.  The interior of the column housing is filled with a 
matrix of submillimeter iron beads coated with a heat-cured biocompatible resin. The columns 
are placed at appropriate locations in the CliniMACS Tubing Set to facilitate the cell selection 
process.  The first column serves as a device to remove components that bind non-specifically to 
the column.  The second column which is placed with in a magnetic field performs the actual cell 
selection.  The columns are incorporated sterile as part of the tubing set and are intended for 
single use only. 
 
The tubing element consists of a series of tubes, connectors, spikes, Luer locks, and collection 
bags. The tubing of the tubing element is comprised of materials that have been qualified for use 
in this application by testing to ISO I 0993. The principal constituents are polyvinyl chloride (PVC) 
and silicone. The connectors are made of various polymers (e.g., ABS and PVC) suitable for use in 
a blood contact environment.  They are solvent bonded to the PVC tubing.  The silicone pump 
tubing is softened with petroleum ether for manufacturing and mechanically fixed   to 
connectors.   The cell wash   bags are composed of PVC. 
 
The CliniMACS Tubing Set is packaged in a thermoformed tray and heat-sealed with a Tyvek® lid. 
The CliniMACS Tubing Set is sterilized by ethylene oxide gas in a validated sterilization cycle and 
supplied as a single-use component for the CliniMACS Instrument. 
 
7.2.3 CLINIMACS TCR Α/Β REAGENT 
 
The CliniMACS TCR α/β -Biotin consists of biotinylated TCR α/β monoclonal antibody (mAb). The 
CliniMACS TCR α/β -Biotin mAb is a mouse IgG2b monoclonal, which is produced by the 
hybridoma cell line clone BMA031.H7. 
 
CliniMACS TCR α/β -Biotin is a sterile solution intended for the indirect magnetic labeling of 
human TCR α/β CD3+ T cells, in combination with the CliniMACS® Anti-Biotin Reagent. It is 
supplied in single use vials which contain nominally 7.5 mL per vial. The vials used are 10 mL glass 
injection vials sealed with a chlorobutyl rubber stopper and flip-off aluminum seal. 
 
The TCR α/β CD3+ T positive cells are specifically labeled for selection by incubation with the 
CliniMACS TCR α/β Reagent. After unbound reagent is washed from the suspension, the cells are 
ready for the automated separation process.  The CliniMACS System passes the antibody-labeled 
suspension; the cells are ready for the automated separation process.  The CliniMACS System 
passes the antibody-labeled suspension through a column in which strong magnetic gradients 
are generated. The Selection Column retains the magnetically labeled TCR α/β positive cells, 
while unlabeled cells flow through the Selection Column and are collected in the Negative 
Fraction Bag.  The cells in the Negative Fraction Bag containing the TCR γ/δ+ T cells will be 

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aliquoted and cryopreserved at doses defined in this protocol for post-transplant infusions. The 
separated TCR α/β positive cells are released from the column by removing the column from the 
magnetic field and clueing the cells into a separate collection bag and will be discarded. 
 
7.3 
THE CLINIMACS PBS/EDTA BUFFER 
 
The CliniMACS PBS/EDTA Buffer is an isotonic and is hydric buffer solution with a pH-value of 7.2 
and osmolality of 290 mosmol/L.  Its formulation is shown in the following table. 
 
Table 1 Formulations of the CliniMACS PBS/EDT A Buffer 
Ingredient 
Compendia 
Amount 
NaCl 
Ph. Eur. 
8.0 g/L 
KCl 
Ph. Eur. 
0.19g/L 
Na2HP04 anhy. 
Ph. Eur. 
1.15g/L 
KH2P04 
Ph. Eur. 
0.19 g/L 
Na2EDTA 
Ph. Eur. 
0.37g/L 
Water for Injection 
Ph. Eur. 
ad IL 
 
The CliniMACS PBS/EDTA Buffer is used as external wash and transport fluid for the in vitro 
preparation of human heterogeneous cell populations intended to be separated with the 
CliniMACS Cell Selection System.   
 
 
8 
TREATMENT/INTERVENTION PLAN 
 
Patients will be treated as inpatient or outpatient on the Miami Cancer Institute Hematologic 
Oncology and Blood & Marrow Transplant Service following their allogeneic stem cell 
transplantation on their respective transplant protocols. The stem cell transplant is considered 
standard of care. The TCR α/β depleted DLI dosing and timing are the elements under 
investigation in this trial. 
 
8.1 
TCR A/B DEPLETED DONOR LYMPHOCYTE INFUSIONS 
 
8.1.1 DOSING AND TIME INTERVALS OF TCR A/B DEPELETED DLI. 
 
The dosing and time intervals of TCR α/β depleted DLI will be adjusted based on the HLA-
match/mismatch of the stem cell donors. 
 
Treatment of the first cohort of patients will be performed to determine the earliest time point 
post allogeneic stem cell transplant tolerating TCR α/β depleted DLI without complications.  

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In this cohort the first 3 patients who received a CD34+-selected allograft from a HLA-matched 
donor will be infused with 5x10e5/kg TCR γ/δ T cells at 6 -7 weeks post-transplant, the next 3 
patients will be treated with 5x10e5/kg TCR γ/δ T cells at 4-5 weeks post-transplant and a third 
group of 3 patients will be treated with 5x10e5/kg TCR γ/δ T cells 2-3 weeks post-transplant.   
The first 3 patients who received a stem cell transplant from a HLA-mismatched donor will be 
treated with an infusion containing 1x10e5/kg TCR γ/δ T cells at 6 -7 weeks post-transplant, the 
next 3 patients will be infused with 1x10e5/kg TCR γ/δ T cells at 4-5 weeks post-transplant and a 
third group of 3 patients will be treated with 1x10e5/kg TCR γ/δ T cells 2-3 weeks post-transplant.  
 
Subsequently, stem cell recipients will receive the first infusion of TCR α/β depleted DLI at the 
earliest tolerated of the patients treated on these de-escalating intervals. Three recipients from 
matched related or unrelated stem cell donors will receive the first dose of 5x10e5/kg TCR γ/δ T 
cells at time point X, followed at 3-4 week intervals by a the second and third infusion 1x10e6/kg 
TCR γ/δ T cells. Recipients who received a stem cell transplant from mismatched donors will 
receive the first dose of 1x10e5/kg TCR γ/δ T cells at time point X, followed at 3-4 week intervals 
by a the second and third infusion of 5x10e5/kg TCR γ/δ T cells.   
 
The final cohort of 3 patients of HLA-matched donors will receive the first dose of 5x10e5/kg TCR 
γ/δ T cells at time point X, followed by an infusion of 1x10e6/kg TCR γ/δ T cells at 3-4 weeks later 
and a third infusion 2x10e6/kg TCR γ/δ T cells 3-4 weeks following the second infusion. In this 
group, three additional patients will then be treated at the maximal tolerated dose determined 
and the final cohort who received a HLA-mismatched transplant will be treated with 1x10e5/kg 
TCR γ/δ T cells at time point X, followed by an infusion of 5x10e5/kg TCR γ/δ T cells at 3-4 weeks 
later and a third infusion 1x10e6/kg TCR γ/δ T cells 3-4 weeks following the second infusion. 
Three additional patients will then be treated at the maximal tolerated dose determined. 
 
8.2 
PROPHYLAXIS AGAINST ACUTE GRAFT-VERSUS-HOST DISEASE 
 
No GvHD prophylaxis will be administered other than the CliniMACS TCR α/β depletion 
performed. 
 
 
9 
EVALUATION POST TCR A/B DEPLETED DONOR LYMPHOCYTE INFUSIONS 
 
9.1 
POST-INFUSION EVALUATION  
 
The chart below shows the approximate dates for tests and procedures performed after TCR α/β 
depleted DLI.  This chart also reflects standard of care transplant follow-up procedures. Certain 
tests may be held or repeated at the discretion of the treating physician if deemed in the best 
clinical interest of the patient. 
 

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Activity 
Post TCR α/β 
Depleted DLI until 
discharge. 
DISCHARGE TO 
DAY 100 
 
Day 100 
6 
Months 
12 
Months 
24 
Months 
Windows 
See below for 
windows 
See below for 
windows 
+/- 4 days 
+/- 2 
weeks 
+/- 1 month +/- 1 month 
Blood counts and 
chemistry (CBC, 
Comprehensive 
Metabolic Panel) 
CBC: Daily 
+/- 12 hours 
CMP: 2 times a 
week 
CBC/CMP: 
Weekly, 2 times 
a week  
 
 
Every 2-4 weeks 
 
x 
 
x 
 
Physical exam for  
GvHD evaluation 
Monitored 
routinely as per 
inpatient 
transplant 
guidelines 
 
 
 
Weekly 
 
 
 
Every 2-4 weeks 
 
x 
 
x 
Disease evaluation 
 
BMA 1 ,2 
 
30 - 100 days after transplant, if clinically 
indicated 
 
x 
 
x 
 
x 
T-cell 
Chimerism2 
N/A 
30 days after 
transplant 
x 
x 
x 
x 
Peripheral blood 
lymphocyte (PBL) 
phenotyping3 
 
 
Every 2 weeks 
 
 
Monthly 
 
x 
 
x 
 
x 
 
x 
 Viral PCR 
analyses (CMV, 
EBV, HHV-6)4 
 
 
 Twice weekly 
 
 
Weekly  
 
x 
 
x 
 
x 
 
x 
Adverse Event 
Collection 
x  
On infusion day 1 
and discharge 
 
x 
On day 100  
x 
 
 

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Concomitant 
Medications 
x 
On infusion day 1 
 
x 
x 
x 
x 
 
1) This should apply to patients with malignant disease in which the BM was previously 
involved. BM Chimeras should also be assessed whenever possible.  BMA may be delayed 
if patient is clinically unstable. 
2) May be done more frequently if clinically indicated.  Tests need not be done if patient has 
a low number of peripheral white cells. 
3) PBL phenotyping should be performed until CD4 counts is >200. 
4) Will be performed as per standard transplant guidelines. 
 
10 TOXICITIES/SIDE EFFECTS  
 
Patients recruited to this trial are individuals who are either referred by physicians or self- 
referred for peripheral blood stem cell transplantation as a potentially curative treatment for 
their malignancy or other life-threatening disorders. All patients will undergo a detailed 
consultations discussing the risks and potential benefits of an allogeneic stem cell transplantation 
as well as risks and benefits of the infusions of TCR α/β depleted DLI post transplantation. The 
risks and potential benefits of the procedure, as well as the participation in any given research, 
experimental, or therapeutic protocol are also discussed. 
 
 
10.1 TOXICITIES/SIDE EFFECTS OF THE TRANSPLANT AND THE COMBINED EFFECTS OF THE 
CONDITIONING REGIMEN AND TRANSPLANT AND TCR A/B DEPELETED DLI. 
 
 
10.1.1 INFECTIONS 
 
Transplantation puts the patient at higher risk for bacterial, viral, or fungal infections, which are 
potentially life threatening. These risks are potentially higher with T-cell depleted (TCD) 
transplants.  Prophylaxis will be initiated and patients will be closely monitored for signs of 
infections and will receive early and appropriate treatment 
 
10.1.2 GRAFT FAILURE / POOR MARROW FUNCTION 
 
T-cell depletion of donor cells is associated with an increased incidence of graft failure in 
allogeneic transplant recipients. After allogeneic transplantation, the recipient's marrow function  
may  be poor,  and  leukopenia, anemia, or thrombocytopenia may result from many causes 
including graft rejection induced by surviving host immune T-cells, or ongoing suppression of 

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engrafted donor blood-forming cells by GvHD,  infection  or marrow suppression or 
immunosuppressive drugs and other medications. Graft failure may result in death if not 
reversed.   In patients with immune rejection second transplants can be administered   with 
immunosuppressive therapy, including non-myeloablative conditioning regimens.  For patients 
who are engrafted with donor cells  but have severe cytopenia affecting one or more blood cell 
lineages, secondary transplants of CliniMACS fractionated CD34+ T-cell depleted PBSCs may be 
administered to booster and replenish donor hematopoietic cells without conditioning or after 
treatment with anti-thymocyte globulin. 
 
10.1.3 GRAFT-VERSUS-HOST DISEASE 
 
Acute or chronic GvHD may develop after allogeneic transplantation that can be disabling and 
can lead to death.  GvHD is thought to be initiated by T-cells contained in the PBSC graft.  CD34+ 
selection and CD3+ depletion reduces the number of T-cells in the PBSC but GvHD can occur after 
TCD transplants as well as after TCR α/β depleted DLI.   Acute and/or chronic GvHD will be treated 
with immunosuppressive drugs as per the transplant service guidelines. 
 
10.1.4 SINUSOIDAL OBSTRUCTION SYNDROME (SOS) OF THE LIVER 
 
SOS is a manifestation of damage to the liver by the conditioning regimen that usually develops 
within two weeks after allogeneic transplant and is characterized by at least two of the following: 
• Hyperbilirubinemia (total bilirubin > 2 mg/dL) 
• Hepatomegaly or right upper quadrant pain, or 
• Rapid weight gain (> 5% above baseline) 
 
TCD is not expected to affect the risk of SOS. However, the effect of TCR α/β depleted DLI on the 
development of SOS is unknown. 
Recipients developing SOS will be monitored closely and will receive appropriate supportive care 
and careful fluid management. 
 
10.1.5 END ORGAN DAMAGE 
 
End organ damage of all or any of the major organs, including the brain, may occur as a result of 
cumulative toxicity from anti-neoplastic therapy, reactions to other drugs, and as a result of 
destructive processes (e.g., infection, GvHD, etc.) and may have a fatal outcome.  Toxicities may 
occur in any individual patient due to multiple events and cumulative effects that may involve 
any and all organs, including the brain. Brain damage can result in severe loss of cognitive or 
neurologic function.  Data from previous studies do not suggest that the risk of end organ damage 
is appreciably affected by TCD or by TCR α/β depleted DLI or the preparative regimens to be used 
in this study. However, the effect of TCR α/β depleted DLI on the development of end organ 
damage is unknown. 
 

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Recipients developing end organ damage will be monitored closely and will receive appropriate 
supportive care and careful fluid management. 
 
 
10.1.6 DISEASE RELAPSE 
 
Allogeneic transplantation using T-cell depleted peripheral blood stem cells has, in some cases, 
been associated with an increased incidence of leukemic relapse in patients with chronic 
myelogenous leukemia, compared to recipients who receive unmanipulated donor cells. The risk 
of relapse has not been increased in patients with acute leukemia. Nevertheless, despite 
cytoreduction, the transplant and TCR α/β depleted DLI, relapse may occur. 
 
10.1.7 DEATH 
 
There is an approximately 5-10% risk of treatment related mortality within the first month of 
transplant due to the risk of severe regimen related toxicity, hemorrhage, opportunistic infection, 
or other complications.  It is not expected that the regimens to be used in this protocol or TCR 
α/β depleted DLI will increase this risk. 
 
11 CRITERIA FOR THERAPEUTIC RESPONSE/OUTCOME ASSESSMENT 
 
 
11.1 DISEASE RELAPSE AND RECURRENCE 
 
Relapse of leukemia is achieved if one or more of the following criteria are met: greater than 
5% circulating leukemic blasts in the marrow or peripheral blood; the presence of blasts in 
any extra medullary site or; disease determined by clinical assessment. Cytogenetic and/or 
Molecular analysis of the marrow and/or peripheral blood will also be obtained for the 
diagnosis of relapse. 
 
11.2 DISEASE-FREE SURVIVAL 
 
DFS is defined as the minimum time interval of times to relapse/recurrence, to death or to 
the last follow- up, from the time of transplant. 
 
11.3 OVERALL SURVIVAL 
Overall survival is defined as time from transplant to death or last follow-up. 
 
12 STATISTICAL CONSIDERATIONS  
 

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As the study is confirmatory in nature, the sample size is assessed pragmatically.  Because of 
the small sample size and multiple strata there can be no inferential statistical analysis. As each 
strata has less than 10 subjects the sample size is too small for any analysis other than 
descriptive statistics. As such, within each cohort, the percentage of patients who have 
achieved feasibility will be reported.  There are two strata of patients: HLA matched and HLA 
mismatched (Tables 1 and 2).  Within each strata there are three cohorts. For the HLA matched 
strata (cohort 1), 3 patients will receive infusions containing 5x10e5/kg TCR γ/δ T cells at 6-7 
weeks post-transplant.  The next three patients will be given the same dose at 4-5 weeks post-
transplant. A third group of three patients will be treated at the same dose at 2-3 weeks post-
transplant.  The same timing procedure is applied to those in the HLA mismatched group, the 
only difference being the infusion dosage which is 1x10e5/kg TCR γ/δ T cells. 
 
In both strata after cohort I, the earliest tolerated time point will be selected according to pre-
specified criteria. For HLA matched cohort II, 3 patients will receive an infusion using 5x10e5/kg 
TCR γ/δ T cells at the time point post-transplant chosen from cohort I. After this, they will 
receive 2 1x10e6/kg TCR γ/δ T cells infusion at 3-4 week intervals after the first dose.  In the HLA 
mismatched group, 3 patients will receive an infusion using 1x10e5/kg TCR γ/δ T cells at the 
ideal time point chosen from cohort I. Then they will receive 2 5x10e5/kg TCR γ/δ T cells 
infusion at 3-4 week intervals after the first dose. 
 
In cohort III, for HLA matched patients, 3 to 6 patients will receive an infusion using 5x10e5/kg 
TCR γ/δ T cells at the ideal time point chosen from cohort I. Then they will receive a 1x10e6/kg 
TCR γ/δ T cells infusion at 3-4 week intervals after the first dose.  The third dose will be 
2x10e6/kg TCR γ/δ T cells infusion 3-4 weeks after the second dose. 
 
In cohort III, HLA mismatched patients, 3 to 6 patients will receive an infusion using 1x10e5/kg 
TCR γ/δ T cells at the ideal time point chosen from cohort I. Then they will receive a 5x10e5/kg 
TCR γ/δ T cells infusion at 3-4 week intervals after the first dose.  The third dose will be 
1x10e6/kg TCR γ/δ T cells infusion 3-4 weeks after the second dose.  
 
Sequential boundaries will be used to monitor dose-limiting toxicity rate. An SAE will be 
considered as a dose limiting toxicity.  The accrual will be halted if excessive numbers of dose-
limiting toxicities are seen, that is, if the number of dose-limiting toxicities is equal to or 
exceeds b out of n patients with full follow-up (see Table 3).  
 
This is a Pocock-type stopping boundary that yields the probability of crossing the boundary at 
most 0.05when the rate of dose-limiting toxicity is equal to the acceptable rate [event 
probability of 0.1].  SAEs, particularly GVHD, will be assessed subjectively to determine if the 
cause is the infusion of cells or the actual transplant.   
 
 
The main objective of this study is to assess the safety of TCR α/β depleted DLI derived from the 
CD34neg fractions from the original transplant donor of the CD34+ selected allogeneic stem cell 

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transplant, when administered to patients with ALL, AML, Myelodysplastic Syndrome, 
Myelofibrosis  or relapsed/refractory multiple myeloma following TCD HSCT. Due to the study 
design, the analysis will be descriptive. In each cohort, percentages and number experiencing a 
dose limiting toxicity will be reported. As the secondary objectives are all qualitative in nature, 
no statistical analysis will be performed 
 
There is no formal interim analysis planned for this study.  
 
12.1 STUDY POPULATION DEFINITION 
 
• Safety Population: All subjects who received at least one dose of TCR α/β depleted 
DLI. 
• Efficacy Evaluable Population: All subjects who meet all eligibility criteria and received 
at least one cycles of intervention. 
 
12.2 SUBJECTS DISPOSITION 
 
Subject disposition (analysis population allocation, entered, discontinued, along with primary 
reason for discontinuation) will be summarized using frequency and percent for both treatment 
and follow-up phases. A summary of subjects enrolled by site will be provided. Protocol 
deviations will be summarized using frequency tabulations. 
 
12.3 EFFICACY ANALYSIS 
 
If study compliance becomes a problem efficacy analysis will not be performed. Data for those 
who completed the study according to protocol will be used. Sample size permitting, all analyses 
previously specified will be applied to this subgroup. 
 
12.4 SAFETY ANALYSIS 
 
The safety population will be included in the safety analyses.  
 
Safety and tolerability will be monitored through continuous reporting of adverse events and 
serious adverse events, laboratory abnormalities, and incidence of subjects experiencing dose 
modifications, dose interruptions, and/or premature discontinuation. Data from all subjects 
who receive one or more doses will be included in the safety analyses. Adverse events, physical 
examinations (including vital sign measurements), clinical laboratory information, and 
concomitant medications/procedures will be tabulated and summarized. Descriptive statistics 
will be presented. 
 

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For both AEs and SAEs, the PI must assess the severity / intensity of the event. The severity / 
intensity of AEs will be graded based upon the subject’s symptoms according to the current 
active minor version of the CTCAE, Version 4.0;  
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm#ctc_40   
AEs that are not defined in the CTCAE should be evaluated for severity / intensity according to 
the following scale:   
• Grade 1 = Mild – transient or mild discomfort; no limitation in activity; no medical 
intervention/therapy required; 
• Grade 2 = Moderate – mild to moderate limitation in activity, some assistance may be 
needed; no or minimal medical intervention/therapy required; 
• Grade 3 = Severe – marked limitation in activity, some assistance usually required; 
medical intervention/therapy required, hospitalization is possible;  
• Grade 4 = Life threatening – extreme limitation in activity, significant assistance required; 
significant medical intervention/therapy required, hospitalization or hospice care 
probable;  
• Grade 5 = Death - the event results in death. 
 
The term “severe” is often used to describe the intensity of a specific event (as in mild, 
moderate or severe myocardial infarction); the event itself, however, may be of relatively minor 
medical significance (such as severe headache). This criterion is not the same as “serious” which 
is based on subject/event outcome or action criteria associated with events that pose a threat 
to a subject’s life or functioning. Seriousness, not severity, serves as a guide for defining 
regulatory obligations. 
 
Table 2. Infusion Regimen for HLA matched donors 
Cohort 
Number of patients 
Infusion: TCR γ/δ T cells 
I  
 
 
 
3 
5x10e5/kg at 6-7 weeks post-
transplant 
 
3 
5*10e5/kg at 4-5 weeks post-
transplant 
 
3 
5*10e5/kg at 2-3 weeks post-
transplant 
II 
3 
5*10e5/kg at X* weeks post-
transplant,  1*10e6/kg at 3-4 
weeks post-first dose, 
1*10e6/kg at 3-4 weeks post-
second dose 
III 
3-6 
5*10e5/kg at X* weeks post-
transplant,  1*10e6/kg at 3-4 
weeks post-first dose, 

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2*10e6/kg at 3-4 weeks post-
second dose 
 
*X represents the time which is chosen from cohort one 
based on pre-specified criteria. 
 
Table 2. Infusion Regimen for HLA mismatched donors 
Cohort 
Number of patients 
Infusion: TCR γ/δ T cells 
I  
 
 
 
3 
1*10e5/kg at 6-7 weeks post-
transplant 
 
3 
1*10e5/kg at 4-5 weeks post-
transplant 
 
3 
1*10e5/kg at 2-3 weeks post-
transplant 
II 
3 
1*10e5/kg at X* weeks post-
transplant,  5*10e5/kg at 3-4 
weeks post-first dose, 
5*10e5/kg at 3-4 weeks post-
second dose 
III 
3-6 
1*10e5/kg at X* weeks post-
transplant,  5*10e5/kg at 3-4 
weeks post-first dose, 
1*10e6/kg at 3-4 weeks post-
second dose 
 
*X represents the time which is chosen from cohort one 
based on pre-specified criteria. 
 
 
Table 3.  Table of stopping boundaries  
Number of patients (n) 
1 2 3 4 5 6 7 8 9 
1
0 
1
1 
1
2 
1
3 
1
4 
1
5 
1
6 
1
7 
1
8 
Boundary (Any SAE) (b) 
- 
2 3 3 3 3 4 4 4 
4 
4 
4 
5 
5 
5 
5 
5 
6 
 
 
 
 
 
13 RESEARCH PARTICIPANT REGISTRATION AND RANDOMIZATION PROCEDURES 
 
13.1 RESEARCH PARTICIPANT REGISTRATION 
 
Confirm eligibility as defined in the section entitled Study Population. 

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Obtain informed consent by following procedures defined in section entitled Informed Consent 
Process. 
 
During the registration process, registering individuals will be required to complete a protocol 
specific Eligibility Checklist. 
 
All participants must be registered through the Clinical Research Office (CRO) at MCI. The signed 
consent form and other relevant documents will be uploaded into OnCore, MCI’s Clinical Trial 
Management System (CTMS) using the electronic database (eDC) and electronic case report form 
(eCRF). 
 
13.2 RANDOMIZATION 
This research study does not require a randomization. 
 
13.3 ENROLLMENT INTO COHORTS 
 
Enrollment into each cohort will follow the regimen outlined in Table 2 above according to 
whether the patient is receiving either a matched or mismatched donor cells. The cohorts will be 
filled sequentially. 
 
14 DATA MANAGEMENT ISSUES 
 
A Clinical Research Coordinator (CRC) will be assigned to the study and will be responsible for  
adult accruals. The responsibilities of the CRC and principal  investigator  include project 
compliance,  data collection,  abstraction  and  entry,  data reporting,  regulatory  monitoring,  
problem resolution  and  prioritization,  and coordinate  the activities  of the protocol study team.  
The data collected for this study will be entered into a secure database.   Source documentation 
will be available to support the computerized patient record. 
 
14.1 QUALITY ASSURANCE 
 
Real-time quality control activities will be conducted to assess missing data and inconsistencies.   
Accrual  rates  and extent and accuracy  of evaluations  and follow-up will be monitored 
periodically  throughout  the study period  and potential  problems will be brought  to the 
attention  of the study team for discussion and action. All necessary precautions will be taken to 
avoid missing data due to small sample size. In addition, imputation of missing data will be done 
after assessing for feasibility. 
 
Random-sample data quality and protocol compliance audits will be conducted by the study 
team. 

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14.2 DATA AND SAFETY MONITORING 
 
The MCI Data Safety and Monitoring Committee (DSMC) will monitor this clinical trial according 
to the MCI Data and Safety Monitoring Plan (DSMP). In its oversight capacity, the MCI DSMC 
bears responsibility for suspending or recommending this study. 
 
DSMC oversight of study conduct includes ongoing review of adverse event data, and periodic 
review of viral complications associated with transplant.  The guidelines appearing in Section 16.3 
are offered for DSMC consideration in assessing adverse events.  In addition, the DSMC will 
review reports from all audits, site visits, or study reviews pertaining to this clinical trial and take 
appropriate action. 
Risk Based Monitoring 
Routine monitoring or audit activities for this study will be conducted by the MCI OCR Office of 
Research Integrity authorized personnel in accordance with current FDA Regulations, ICH GCP 
guidelines, site’s Standard Operating Procedures (SOPs), IRB and the respective local and national 
government regulations. The general scope of such visits would be to inspect study data including 
but not limited to, regulatory requirements, source documentation, original medical records/files 
and eCRF completion, as applicable, following a risk-based monitoring approach. 
The study will be monitored based on procedures established in the Risk-Based Monitoring Plan 
at the time of protocol activation. A series of monitoring forms, tools and templates including 
MCI Clinical Research Monitoring/Audit Deficiencies Guidance and Summary Monitoring Letters 
are utilized to ensure standard and consistent monitoring practices. The monitoring frequency 
may also be adjusted as per the monitor’s discretion based on accrual rate, trial complexity, 
major deficiency findings, visit rating and safety reporting.  
To ensure compliance with current federal regulations and the ICH GCP guidelines, data 
generated by this study must be available for inspection upon request by representatives of the 
FDA, national and local health authorities, and duly authorized representatives of any entity 
providing support for this trial. 
 
15 SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS 
 
15.1 INFORMED CONSENT PROCESS 
 
Before protocol-specified procedures are carried out, consenting professionals will explain full 
details of the protocol and study procedures as well as the risks involved to participants prior to 
their Inclusion in the study.  Participants will also be informed that they are free to withdraw 

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from the study at any time.  All participants must sign an IRB -approved consent form indicating 
their consent to participate.  This consent form meets the requirements of the Code of Federal 
Regulations and the Institutional Review Board of this Center. The consent form will include the 
following: 
 
1. The nature and objectives, potential risks and benefits of the intended study. 
2. The length of study and the likely follow-up required. 
3. Alternatives to the proposed study. (This will include available standard and 
investigational therapies.  In addition, patients will be offered an option of supportive 
care for therapeutic studies.) 
4. The name of the investigator(s) responsible for the protocol. 
5. The right of the participant to accept or refuse study interventions/interactions and to 
withdraw from participation at any time. 
 
Before any protocol-specific procedures can be carried out, the consenting professional will fully 
explain the aspects of patient privacy concerning research specific information.   In addition to 
signing the IRB Informed Consent, all patients must agree to the Research Authorization 
component of the informed consent form. 
 
Each participant and consenting professional will sign the consent form. The participant will 
receive a copy of the signed informed consent form. 
 
15.2 CRITERIA FOR REMOVAL FROM STUDY 
 
Research participants may be removed from the study if requested by the research participant.  
Management will depend on where they are in their treatment course. Such research 
participants will receive appropriate supportive care.  Patients may also be removed from the 
study at any point deemed appropriate by the principal investigator. 
 
Patients may be removed from study, and not followed further, if one or more of the following 
events occur: 
 
• Significant noncompliance on the part of the patient 
• Refusal of the patient to continue treatment or observations 
• Progressive disease or relapse 
• Decision by the investigator that termination is in the patient’s best medical interest 
• Unrelated medical illness or complication 
• Lost to follow-up 
 
A participant will be considered lost to follow-up if he or she fails to return for no more than two 
scheduled visits and is unable to be contacted by the study site staff.  
 

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The following actions must be taken if a participant fails to return to the clinic for a required study 
visit: 
 
• 
The site will attempt to contact the participant and reschedule the missed visit within 3 
business days and counsel the participant on the importance of maintaining the assigned visit 
schedule and ascertain if the participant wishes to and/or should continue in the study. 
• 
Before a participant is deemed lost to follow-up, the investigator or designee will make 
every effort to regain contact with the participant (where possible, 3 telephone calls and, if 
necessary, a certified letter to the participant’s last known mailing address or local equivalent 
methods). These contact attempts should be documented in the participant’s medical record or 
study file.  
• 
Should the participant continue to be unreachable, he or she will be considered to have 
withdrawn from the study with a primary reason of lost to follow-up. 
 
 
 
15.3 CRITERIA FOR SUSPENSION OR PREMATURE TERMINATION OF THE STUDY 
 
 
Miami Cancer Institute (MCI) at Baptist Health, Inc., Principal Investigator (PI) or regulatory 
authority may suspend or prematurely terminate, at the investigation sites for which they are 
responsible, the clinical investigation for significant and documented reasons. If suspicion of an 
unacceptable risk to subjects arises during the clinical investigation, or when so instructed by the 
regulatory authorities, MCI shall suspend the clinical investigation while the risk is assessed. MCI 
shall terminate the clinical investigation if an unacceptable risk is confirmed. 
 
MCI shall consider terminating or suspending the participation of a particular investigation site 
or investigator in the clinical investigation if monitoring or auditing identifies serious or repeated 
deviation. If suspension or premature termination occurs, the terminating party shall justify its 
decision in writing and promptly inform the other parties with whom they are in direct 
communication. The Principal Investigator and MCI shall keep each other informed of any 
communication received from the regulatory authority. 
 
If, for any reason, MCI suspends or prematurely terminates the investigation at an individual 
investigation site, MCI shall inform the responsible regulatory authority as appropriate. If the 
suspension or premature termination was in the interest of safety, MCI shall inform all other 
Principal Investigators. 
 
If suspension or premature termination occurs, 
 
• MCI shall remain responsible for providing resources to fulfill the obligations from the 
protocol and existing agreements for following up the subjects enrolled in the clinical 
investigation, and 

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• The Principal Investigator or authorized designee shall promptly inform the enrolled 
subjects at his/her investigation site, if appropriate. 
 
When MCI concludes an analysis of the reason for the suspension, implements the necessary 
corrective actions, decides to lift the temporary suspension, MCI shall inform the Principal 
Investigator, and, where appropriate, the regulatory authority of the rationale, and provide them 
with the relevant data supporting this decision. The regulatory authorities shall also provide 
confirmation of information before the clinical investigation resumes. If subjects have been 
informed of the suspension, the Principal Investigator or authorized designee shall inform them 
of the reasons for resumption.  
 
16 PROTECTION OF HUMAN SUBJECTS 
 
The risks associated with a T-cell depleted transplant followed by TCR α/β depleted DLI on this 
study are those associated with the toxicities, as detailed in Section 10, as well as the risks of an 
allogeneic transplant, particularly graft failure, or graft vs. host disease, as also detailed in Section 
10. 
 
To protect against the toxicities, the patient will be transplanted in a single room, HEPA filtered 
environment.  Organ toxicities such as mucositis, enteritis and hepatic dysfunction as well as 
infectious complications will be treated by standard procedures developed for transplantation to 
support our patients. Blood and platelet counts will be supported by transfusion. Graft failure 
might necessitate a second transplant, after additional conditioning.  Approaches to the diagnosis 
and treatment of graft failure that secure consistent engraftment have been developed by the 
Transplantation Services. Similarly, advanced treatments will be instituted in the event the 
patient develops graft vs. host disease. 
 
Despite a transplant and TCR α/β depleted DLI, the patient's disease may recur.   In this case, 
standard and/or experimental therapies, such as phase I /II drugs, antibodies or cell therapies, 
will be available to the patient for consideration as treatment options. 
 
16.1 BENEFITS 
 
A transplant is administered with curative intent. The integration of post-transplant TCR α/β 
depleted DLI is intended to improve the outcome. The approaches being evaluated may achieve 
this goal and may also be effective in preventing infections, acute and chronic graft vs. host 
disease and disease relapse. 
 
The results of this study will also define risks and benefits of T-cell depleted grafts and TCR α/β 
depleted DLI and will greatly accelerate further development of graft manipulations employing 
these approaches. 
 

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Consent Process: Participation in this study is voluntary.  All patients will be required to sign a 
statement of informed consent, which must conform to MCI IRB guidelines. 
 
Alternatives:   Enrollment in this study is voluntary.  Alternative treatment options will be 
presented to the patient prior to taking part in this study. Alternative  treatment  options may  
include getting a transplant  from  a volunteer  unrelated  donor, if one is  available;  getting 
treatment for the cancer with either  chemotherapy  or a transplant without being on  a study; 
taking  part in  another study; or getting no  treatment. 
 
Costs: The patient's health plan/insurance company will need to pay for all of the costs of 
treatment in this study. The patient will be responsible for the costs of standard medical care, all 
hospitalizations and any transplant complications. Pre-authorization for the transplant will be 
cleared with the health plan/insurance company prior to admission. Patients will not be paid for 
taking part in this study. 
 
Research tests will be done at no cost to the patient. 
 
Confidentiality: Every effort will be made to maintain patient confidentiality. Research and 
hospital records are confidential. 
 
16.2 PRIVACY 
 
MCI’s Privacy Office may allow the use and disclosure of protected health information pursuant 
to a completed and signed Research Authorization form. The use and disclosure of protected 
health information will be limited to the individuals described in the Research Authorization 
form. A Research Authorization form must be completed by the Principal Investigator and 
approved by the IRB (IRB). 
 
16.3 SERIOUS ADVERSE EVENT (SAE) REPORTING 
 
An adverse event is considered serious if it results in ANY of the following outcomes:  
• 
Death  
• 
A life-threatening adverse event  
• 
An adverse event that results in inpatient hospitalization or prolongation of existing 
hospitalization 
• 
A persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions 
• 
A congenital anomaly/birth defect 
• 
Important Medical Events (IME) that may not result in death, be life threatening, or 
require hospitalization may be considered serious when, based upon medical judgment, they 
may jeopardize the patient or participant and may require medical or surgical intervention to 
prevent one of the outcomes listed in this definition 

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Note: Hospital admission for a planned procedure/disease treatment is not considered an SAE.  
 
SAE reporting is required as soon as the participant starts investigational treatment/intervention.  
SAE reporting is required for the participant’s duration of the study in accordance with the study 
calendar (Section 9). 
 
Please note: Any SAE that occurs prior to the start of investigational treatment/intervention and 
is related to a screening test or procedure (i.e., a screening biopsy) must be reported.  
All SAEs must be submitted to the research team at MCI.  
 
The report should contain the following information: 
• 
The date the adverse event occurred 
• 
The adverse event 
• 
The grade of the event  
• 
Relationship of the adverse event to the treatment(s)  
• 
If the AE was expected 
• 
Detailed text that includes the following 
o 
An explanation of how the AE was handled 
o 
A description of the participant’s condition 
o 
Indication if the participant remains on the study 
• 
If an amendment will need to be made to the protocol and/or consent form 
• 
If the SAE is an Unanticipated Problem  
 
All SAEs will be reviewed by the PI and entered in OnCore within 48 business hours of awareness 
of the event. A follow up SAE report must be entered in OnCore within 5 business days for all 
inpatient hospitalizations graded as 4 and 5. 
 
For all inpatient hospitalizations graded as 1, 2, and 3, a follow up SAE report must entered in 
OnCore within 10 business days. 
 
All SAEs will be reported to the IRB of record in accordance with its policies and procedures. 
 
The IRB requires a SAE report be submitted electronically to the IRB Office. The report should 
contain the following information: 
 
• Subject ID 
• Disease/histology (if applicable) 
• Protocol number and title 
• The date the adverse event occurred 
• The adverse event 
• Relationship of the adverse event to the treatment (drug, device, or intervention) 

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• If the AE was expected 
• The severity of the AE 
• The intervention 
• Detailed text that includes the following 
o An explanation of how the AE was handled  
o A description of the subject's condition 
o Indication if the subject remains on the study 
o If an amendment will need to be made to the protocol and/or consent form. 
 
The Pl's signature and the date it was signed are required on the completed report.  
 
16.5 DEFINITION OF AN SAE 
 
 
An SAE is an undesirable experience that meets one of the follow in criteria: 
• ls fatal or life-threatening 
• Is disabling 
• Results in a congenital anomaly or occurrence of malignancy 
• Important medical event that jeopardizes the participant AND requires medical or surgical 
intervention to prevent one of the outcomes above 
Note: Hospital admission for a planned procedure/disease treatment is not considered an SAE. 
 
Attribution: 
• Unrelated : The AE is clearly NOT related to the TCR α/β depleted DLI   
• Unlikely: The AE is doubtfully related to the TCR α/β depleted DLI  
• Possible: The AE may be related to the TCR α/β depleted DLI  
• Probable: The AE is likely related to the TCR α/β depleted DLI  
• Definite: The AE is clearly related to the TCR α/β depleted DLI  
 
Expected and Unexpected Event: 
• Expected: Any experience previously reported (in nature, severity, or incidence) in the in 
the safety information of the study drugs general investigational plan. 
• Unexpected: Any experience not previously reported (in nature, severity, or incidence) in 
the safety information of the study drugs general investigational plan. 
 
REPORTABLE EVENTS: 
• Grades 1-2: Adverse Event Reporting NOT required. 
• Grades  3-4: Possible,  Probable  or  Definitely  attributed  to  the study drug   (as detailed  
as 1 1.2.1), will be reported *. 
• Grades 5: Regardless of Attribution will be reported*. 
 

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*Reportable events are those, which occur within 30 days of the last dose of treatment on 
protocol. Events beyond 30 days will be reported at the discretion of the PI. 
 
For IND protocols: 
The AE report should be completed as above, and the FDA assigned IND number written at the 
top of the report. If appropriate, the report will be forwarded to the FDA by the SAE staff through 
the Office of Clinical Research (OCR). 
 
17.3.1 IND REPORTING  
 
This protocol has an IND; therefore, the SAE will also be reported to the FDA through the OCR 
and the report will include the FDA assigned IND number and name. 
 
All FDA correspondence will occur in accordance with the requirements as set forth in 21CFR Part 
312. 
 
18 PUBLICATION POLICY 
 
The results of the clinical investigation will be made publicly available through clinicaltrials.gov 
or via other publishing bodies, in case of positive or negative results upon study completion. 
 
19 REFERENCES  
 
1. Seggewiss R, Einsele H: Immune reconstitution after allogeneic transplantation and 
expanding options for immunomodulation: an update. Blood, May 2010; 115: 3861 – 
3868.   
2. Lang P, Handgretinger R. Haploidentical SCT in children: an update and future 
perspectives. Bone Marrow Transplant. (2008) 42 Suppl 2:S54-9..  
3. Reisner Y, Hagin D, Martelli MF. Haploidentical hematopoietic transplantation: current 
status and future perspectives. Blood (2011) 118(23):6006-17   
4. Madan Jagasia, Mukta Arora, Mary E. D. Flowers, et. al. Risk factors for acute GVHD and 
survival after hematopoietic cell transplantation Blood, Jan 2012; 119: 296 – 307.   
5. Mukta Arora, John P. Klein, Daniel J. Weisdorf, et. al. Chronic GVHD risk score: a Center 
for International Blood and Marrow Transplant Research analysis Blood, Jun 2011; 117: 
6714 - 6720.   
6. Chao, N. J., G. M. Schmidt, et al. (1993). "Cyclosporine, methotrexate, and prednisone 
compared with cyclosporine and prednisone for prophylaxis of acute graft-versus-host 
disease." N Engl J Med 329(1 7): 1225-1230. 
7. Nash, R. A., J.  H. Antin, et al. (2000). "Phase 3 study comparing methotrexate and 
tacrolimus with methotrexate and cyclosporine for prophylaxis of acute graft-versus-
host disease after marrow transplantation from unrelated donors."  Blood 96(6): 2062-
2068. 

Protocol # 2019-KOE-004 
 
Version 3.0 10 JAN 2022  
 
Page 34 of 35 
 
8. Ratanatharathorn, V., R A. Nash, et al. (1 998). "Phase l II study com paring 
methotrexate and tacrolimus (prograf, FK506) with methotrexate and cyclosporine for 
graft-versus-host disease prophylaxis after HLA- identical sibling bone marrow 
transplantation." Blood 92(7): 2303-2314. 
9. Gratwohl, A., R. Brand, et al. (2002). "Graft-versus-host disease and outcome in HLA- 
identical sibling transplantations for chronic myeloid leukemia." Blood 100 (12): 3877-
3886.  
10. Martin, P. J., G. Schoch, et al. (1990). "A retrospective analysis of therapy for acute graft-
versus-host disease: initial treatment." Blood 76(8): 1464-1472.  
11. Reisner  Y,  Kapoor N,  Kirkpatrick D, Pollack  MS,  Dupont B, Good RA,  O'Reilly RJ. 
Transplantation for acute leukaemia with HLA-A and B non identical parental marrow 
cells fractionated with soybean agglutinin and sheep red blood cells.Lancet. 1981 Aug 
15; 2(8242):327-31 
12. Handgretinger, R., T. Kl ingebiel, et al. (2001). "Mega dose transplantation of purified 
peripheral blood CD34(+) progenitor cells from HLA-rnismatched parental donors in 
children." Bone Marrow Trans plant 27(8): 777- 783.  
13. Jakubowski,  A. A., T. N.  Small, et al. (2007).  "T cell depleted stem-cell transplantation 
for adults w ith hematologic  malignancies:  sustained  engraftment  of  HLA-matched  
related  donor  grafts without the use of antithymocyte globulin." Blood 1 10( 13): 4552-
4559. 
14. Urbano-lspizua, A., C. Rozman, et al. (2002). "Risk factors for acute graft-versus-host 
disease in patients undergoing transplantation with CD34+ selected blood cells from 
HLA-identical siblings."  Blood 100(2): 724-727. 
15. Aversa, F., A. Terenzi, et al. (2005). "Full haplotype-mismatched hematopoietic stern-cell 
transplantation: a phase II study in patients with acute leukemia at high risk of relapse." 
J Clin Oncol 23(15):  3447-3454. 
16. Devine S, Soiffer R, Pasquini M, Carter S, Hari P, DeVore S, Stein A, Lazarus  H, Linker C, 
Stadtrnauer E, Keever-Taylor  C, O'Reil ly  RJ . HLA-identical sibling matched,  CD34+ 
selected, T cell depleted  peripheral  blood  stem cells  following rnyeloablative 
conditioning  for first or second remission acute rnyeloid  leukemia (AM L): Results of 
blood  and marrow  trans plant clinical trials network (BMT CTN) Protocol 0303. Blood, 1 
14 (22), 273. 
17. Karl S. Peggs, Kirsty Thomson, Daniel P. Hart, Johanne Geary, Emma C. Morris, Kwee 
Yong, Anthony H. Goldstone, David C. Linch, and Stephen Mackinnon Blood, Susan K. 
Fautsch, Roby Nicklow, Todd DeFor, and Bruce R. Blazar: Lymphodepletion followed by 
donor lymphocyte infusion (DLI) causes significantly more acute graft-versus-host 
disease Feb 2004; 103: 1548 – 1556.   
18. Wolfgang A. Bethge, U., Hegenbart, Monic J. Stuart, Barry E. Storer, Michael B. Maris, 
Mary E. D. Flowers, David G. Maloney, Thomas Chauncey, Benedetto Bruno, Ed Agura, 
Stephen J. Forman, Karl G. Blume, Dietger Niederwieser, Rainer Storb, and Brenda M. 
Sandmaier: Adoptive immunotherapy with donor lymphocyte infusions after allogeneic 
hematopoietic cell transplantation following nonmyeloablative conditioning; Blood, Feb 
2004; 103: 790 – 795.   

Protocol # 2019-KOE-004 
 
Version 3.0 10 JAN 2022  
 
Page 35 of 35 
 
19. Frey NV and Porter DL: Graft versus host disease after donor leukocyte infusion: 
presentation and management. Best Pract Res Clin Haematol. 2008, 21: 205-222.   
20. Meyer RG, Britten CM, Wehler D et al. Prophylactic transfer of CD8-depleted donor 
lymphocytes after T-cell-depleted reduced-intensity transplantation. Blood 2007; 
109:374-382.   
21. S Farhan, DA Lee, RE Champlin, and SO Ciurea: NK cell therapy: targeting disease relapse 
after hematopoietic stem cell transplantation, Immunotherapy, Mar 2012; 4(3): 305-13.  
22.  Handgretinger R: Negative depletion of CD3+ and TCRα/β+ T cells. Curr Opin Hematol, 
Nov 2012; 19(6): 434-9. 
23. Tyler, E, Jungbluth,A, O’Reilly, R, Koehne, G. WT1- specific T-cell responses in high- risk 
multiple myeloma patients undergoing allogeneic T cell-depleted hematopoietic stem 
cell transplantation and donor lymphocyte infusions. Blood. 2013 Jan 10; 121(2): 308-
317 
24. O’Reilly,R, Dao, Tao, Koehne, G, Scheinberg,D, Doubrovina, E. Adoptive Transfer of 
Unselected or Leukemia-Reative T-cells in the Treatment of Relapse Following 
Allogeneic Hematopoietic Cell Transplantation. Semin Immunol.2010 Jun;22(3): 162-172 
25. Handgretinger R: Negative depletion of CD3+ and TCRα/β+ T cells. Curr Opin Hematol, 
Nov 2012; 19(6): 434-9. 
26. Schumm M, Lang P, Bethge W, Vogel W, Faul C, Feuchtinger T, Pfeiffer M, Handgretinger 
R. Depletion of TcRab+ and CD19-positive cells from leukapheresis products with the 
CliniMACS device. BMT. 2011Apr; 46 Suppl1:P1093.   
27. Munchel AT, Kasamon YL, Fuchs EJ. Treatment of hematological malignancies with 
nonmyeloablative, HLA-haploidentical bone marrow transplantation and high dose, 
post-transplantation cyclophosphamide. Best Pract Res ClinHaematol (2011) 
Sep;24(3):359-68.   
28. Maniar A, Zhang X, Lin W et al. Human γ/δ T lymphocytes induce robust NK cell mediated 
antitumor cytotoxicity through CD137 engagement. Blood (2010)116(10):1726-33.   
29. Godder KT, Henslee-Downey PJ, Mehta J et al. Long term disease-free survival in acute 
leukemia patients recovering with increased γ/δ T-cells after partially mismatched 
related donor bone marrow transplantation. BMT (2007) 39(12):751-757.   
30. Blazar BR, Taylor PA, Bluestone JA et al. Murine γδ T-cells affect allo-engraftment via the 
recognition of nonclassical major histocompatibility complex class Ib antigens. Blood 
(1996) 109:2646–2656.   
31. Bonneville M, Scotet E. Vγ9Vδ2 T cells: promising new leads for immunotherapy of 
infections and tumors. CurrOpinImmunol (2006) 18:1–8.   
32. Grimes HL, Schanie CL, Huang Y et al. Graft facilitating cells are derived from 
hematopoietic stem cells and functionally require CD3, but are distinct from T 
lymphocytes. ExpHematol (2004) 32:946–954.   
33. Leung, W. Use of NK cell activity in cure by transplant. British Journal of Haematology 
(2011), 155(1): 14-29.   
34. Ivanova, A., Qaqish BF, Schell, MJ. Continuous toxicity monitoring in phase II trials in 
oncology. Biometrics (2005) 61: 540-545.