T-Cell Receptor Alpha Beta+/CD19+ Depletion in Haploidentical Allogeneic Hematopoietic Cell Transplantation (Allo-HCT) for Adult and Pediatric Patients With Hematological Malignancies and Non-malignant Disorders
Phase: NA
Conditions: Acute Lymphoblastic Leukemia in Remission, Acute Myeloid Leukemia in Remission, Myelodysplastic Syndromes, Chronic Myeloid Leukemia, Hemophagocytic Lymphohistiocytoses, Primary Immunodeficiency Diseases, Hemoglobinopathies, Severe Aplastic Anemia, Cytopenia, Bone Marrow Failure Syndrome, Severe Chronic Active Epstein-Barr Virus Infection
Sponsor: Baylor College of Medicine

Coversheet 
Protocol and Statistical Analysis Plan 
Official Study Title 
T-CELL RECEPTOR (TCR) ALPHA BETA+/CD19+ DEPLETION IN 
HAPLOIDENTICAL ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION 
(ALLO-HCT) FOR ADULT AND PEDIATRIC PATIENTS WITH HEMATOLOGICAL 
MALIGNANCIES AND NON-MALIGNANT DISORDERS (HAPLOTAB) 
NCT number 
NCT05236764 
Document Date  
1/23/2025 
 

HAPLOTAB v1.0 (8/25/2021)                 HAPLOTAB v1.4 (02/07/2023) 
HAPLOTAB v1.1 (11/10/2021)               HAPLOTAB v1.5 (05/10/2023) 
HAPLOTAB v1.2 (02/23/2022)               HAPLOTAB v2.0 (07/17/2023) 
HAPLOTAB v1.3 (07/08/2022)               HAPLOTAB v2.1 (01/23/2025) 
 
T-Cell Receptor (TCR) αβ+/CD19+ Depletion in Haploidentical Allogeneic Hematopoietic 
Cell Transplantation (allo-HCT) for Adult and Pediatric Patients with Hematological 
Malignancies and Non-malignant Disorders 
(HAPLOTAB) 
 
Principal Investigators 
Erin Morales, MD 
Baheyeldin Salem, MD 
 
Co-Investigators 
George Carrum, MD 
John Craddock, MD 
Erin Doherty, MD 
Lisa Forbes, MD 
Bambi Grilley, RPh, RAC, CIP, CCRC, CCRP 
LaQuisa Hill, MD 
Rammurti Kamble, MD 
Gabriela Llaurador, MD 
Premal Lulla, MD 
Caridad Martinez, MD 
Carlos Ramos, MD 
Khaled Yassine, MD 
 
Statistician 
Tao Wang, PhD 
 
IND # 
28505 
 
Center for Cell and Gene Therapy 
Dan L. Duncan Comprehensive Cancer Center 
Baylor College of Medicine 
at 
Texas Children's Hospital 
6621 Fannin St. 
Houston, Texas 77030 
and 
Houston Methodist Hospital 
6565 Fannin St. 
Houston, Texas 77030 
 
 

HAPLOTAB v2.0 (07/17/2023) 
2 
 
 
 
Table of Contents 
Checklist Patient Eligibility for Treatment………………………………………………………………………………………………4 
1.0 Objectives ............................................................................................................................................... 6 
Primary Objective ...................................................................................................................................... 6 
    Secondary Objective .................................................................................................................................. 6 
    Exploratory Objective……………………………………………………………………………………………………………………….6 
2.0 Background and Rationale ..................................................................................................................... 6 
2.1 Introduction ........................................................................................................................................ 6 
     2.2 Evolution of the Methodology of T cell depletion Haploidentical HCT .............................................. 7 
     2.3 Role of Natural Killer (NK) cells……………………………………………………………………………………………………10 
     2.4 Current Outcome Data for TCR αβ+ CD19+ Depletion………………………………………….10 
 
2.4.1 Graft Composition…………………………………………………………………………………………………………….10 
  
2.4.2 Engraftment……………………………………………………………………………………………………………………….11 
 
2.4.3 Graft-versus-Host Disease (GVHD)…………………………………………………………………………………….11 
 
2.4.4 Transplant-related Mortality (TRM)……………………………………………………………………………………12 
 
2.4.5 Survival……………………………………………………………………………………………………………………………….12 
 
2.4.6 Immune Reconstitution………………………………………………………………………………………………………12 
 
2.4.7 Infections……………………………………………………………………………………………………………………………13 
3.0 Patient Eligibility…………………………………………………………………………………………………………………………………13 
      3.1 Inclusion Criteria……………………………………………………………………………………………………………………………13 
      3.2 Exclusion Criteria………………………………………………………………………………………………………………………….14 
4.0 Treatment Plan………………………………………………………………………………………………………………………………….15 
      4.1 Pretreatment Procedures…………………………………………………………………………………………………………….15 
       4.2 Donor Selection………………………………………………………………………………………………………………………….15 
       4.3 Donor Collections by Apheresis…………………………………………………………………………………………………15 
       4.4 Donor Stem Cell Apheresis Procedure……………………………………………………………………………………….16 
       4.5 TCR αβ+/CD19+ Depletion………………………………………………………………………………………………………….16 
       4.6 Conditioning Regimens………………………………………………………………………………………………………………16 

HAPLOTAB v2.0 (07/17/2023) 
3 
 
5.0 Evaluations……………………………………………………………………………………………………………………………………….17 
       5.1 Evaluation during Study……………………………………………………………………………………………………………17 
6.0 Criteria for Study Evaluation…………………………………………………………………………………………………………...19 
7.0 Sample Size Considerations……………………………………………………………………………………………………………..19 
      7.1 Study design and sample size………………………………………………………………………………………………………19 
      7.2 Stopping Rules…………………………………………………………………………………………………………………………….20 
      7.3 Statistical Analysis……………………………………………………………………………………………………………………….21 
8.0 Reporting Requirements…………………………………………………………………………………………………………………..21 
      8.1 To register patients……………………………………………………………………………………………………………………..21 
      8.2 Relapse and survival data…………………………………………………………………………………………………………….21 
      8.3 Drug Toxicity and/or Adverse Reactions………………………………………………………………………………………21 
      8.4 Potential Risks and Mitigation Strategies…………………………………………………………………………………….22 
9.0 Off Study Criteria………………………………………………………………………………………………………………………………22 
10.0 Clinical Trial Oversight and Monitoring……………………………………………………………………………………………23 
11.0 Informed Consent…………………………………………………………………………………………………………………………..23 
Appendix A-Lansky and Karnofsky Performance Scale…………………………………………………………………………….24 
Appendix B1-Acute GVHD Staging and Grading………………………………………………………………………………………25 
Appendix B2-Chronic GVHD Staging and Grading (NIH Consensus Grading)…………………………………………..26 
Appendix C-CMV Monitoring and Prophylaxis…………………………………………………………………………………………32 
12.0 References……………………………………………………………………………………………………………………………………..33 
 
 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
4 
 
HAPLOTAB CHECKLIST FOR PATIENT ELIGIBILITY FOR TREATMENT 
 
Patient ID: __________________ 
Patient Name: __________________________ 
 
The following must be answered “YES” for a patient to be eligible to participate on this study: 
YES NO 
VALUE/DATE  
__ 
__ 
 
Age less than or equal to 55 years 
__ 
__ 
 
Lack of suitable conventional donor (as defined in protocol) or 
presence of rapidly progressive disease not permitting time to 
identify an HLA-matched unrelated donor 
__ 
__ 
___/___ 
Lansky/Karnofsky score > 50 
__ 
__ 
___/___ 
Signed written informed consent 
 
__ 
__ 
 
Diagnosis of one of the following: (check one) 
 
 Patient with life threatening hematological malignancy 
including “high-risk” ALL in first complete remission 
(CR1); ALL in second or subsequent remission (≥ CR2); 
high-risk AML in CR1; AML in second or subsequent CR; 
myelodysplastic 
syndromes 
(MDS); 
non-Hodgkin’s 
lymphomas (NHL) in second or subsequent remission (≥ 
CR2); CML 
 
 
 
 Hemophagocytic Lymphohistiocytosis (HLH) including 
familial HLH, relapsed HLH or CNS HLH 
 
 
 
 Primary Immunodeficiency Disorders (PID) 
 
 
 
 Hemoglobinopathies including thalassemia or sickle cell 
disease (SCD) 
 
 
 
 Severe aplastic anemia (SAA) not responding to immune 
suppressive therapy 
 
 
 
 Congenital/hereditary 
cytopenias 
including 
Fanconi 
anemia (FA) without malignant clonal evolution (MDS, 
AML) 
 
 
 
 Other inherited bone marrow failure syndromes (IBMFS)  
 
 
 
 Severe chronic active Epstein Barr virus infection 
(SCAEBV) with predilection for T- or NK-cell malignancy 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
5 
 
 
The following must be answered “NO” for patient to be eligible 
YES NO 
VALUE/DATE  
__ 
__ 
___/___ 
Life expectancy of < 6 weeks limited by disease other than 
leukemia. 
__ 
__ 
___/___ 
Greater than grade II acute graft versus host disease (GVHD) or 
chronic extensive GVHD due to a previous allograft at the time 
of inclusion 
__ 
__ 
___/___ 
Subject receiving an immunosuppressive treatment for GVHD 
treatment due to a previous allograft at the time of inclusion 
__ 
__ 
___/___ 
Symptomatic cardiac disease OR left ventricular shortening 
fraction < 25% or ejection fraction < 40% 
__ 
__ 
___/___ 
Severe renal disease, with creatinine clearance < 40cc/1.73 m2 
 
__ 
__ 
___/___ 
Pre-existing severe restrictive pulmonary disease, FVC < 40% of 
predicted 
 
__ 
__ 
___/___ 
Severe hepatic disease with ALT/AST ≥ x2.5 upper limit of 
normal or bilirubin level ≥ x1.5 upper limit of normal 
__ 
__ 
 
Serious concurrent uncontrolled medical disorder or mental 
illness 
__ 
__ 
 
Pregnant or breastfeeding female subject 
__ 
__ 
 
Current active infectious disease including viral and fungal 
diseases at the time of enrollment; that on evaluation of PI 
precludes ablative chemotherapy or successful transplantation. 
__ 
__ 
 
Active HIV infection 
__ 
__ 
 
Severe personality disorder or mental illness that would preclude 
compliance with the study 
 
 
 
To register patients and check eligibility, call Dr. Morales at 832-826-0860. 
 
 
___________________________ 
 
_______________ 
Signature of Enrolling Investigator 
 
 
Date 
 
 

HAPLOTAB v2.0 (07/17/2023) 
6 
 
1.0 OBJECTIVES 
 
1.1. This is a phase I/II study evaluating the safety and feasibility of αβ T cell-depleted (TCD) 
haploidentical (or partially mismatched) hematopoietic cell transplantation (HHCT) by 
negative selection of T cell receptor alpha/beta (TCR αβ+) T cells and CD19+ depletion 
using an automated CliniMACS device in pediatric and adult patients 
 
Primary Endpoints:  
1. Cumulative incidence of both neutrophil and platelet engraftment by day 42 post HCT 
as a composite endpoint. 
2. Cumulative incidence of Grade III or higher acute graft versus-host disease (aGVHD) 
by day 100 post HCT 
Secondary Endpoints: 
1. Cumulative incidence of transplant-related mortality (TRM) at Day 100 and Day 365 
2. Overall survival (OS) up to one year post HCT 
3. Cumulative incidence of chronic graft versus-host disease (GVHD) up to 2-years post-
HCT 
 
 
Exploratory Objectives: 
1. Feasibility of using the automated CliniMACS system to produce a graft with defined 
cell composition  
2. Incidence of viral reactivations – defined as detectable viral load by peripheral blood 
polymerase chain reaction (PCR) – and/or symptomatic infections up to one year post 
HCT 
3. For patients with hematological malignancies, complete donor chimerism within 30 
days from engraftment.  
4. T-, B- and NK-cell subset analysis at Day 30, 60, 100, 180 and 365 post-HCT 
5. Recent Thymic Emigrants (RTE) at Day 100, provided absolute lymphocytes count 
(ALC) is > 200 x10e9/L, then at Day 180 and 365 
6. T-cell repertoire analysis by flow cytometry at Day 180 post-transplant 
7. Proportion of alpha/beta versus gamma/delta T-cell receptor positive cells present at 
Day 30, 60, 100, 180 and 365 post-HCT 
8. NK screening subset at Day 30, 60, 100, 180 and 365 post-HCT 
9. Extended NK cell phenotype at Day 30, 60, 100, 180 and 365 post-HCT 
 
2.0 BACKGROUND AND RATIONALE 
 
2.1. Introduction  
While most hematopoietic cell transplant (HCT) candidates have an acceptable 
donor, regarded as an HLA-matched or minimally mismatched donor, some 
patients lack an optimal donor, that is, donor who is matched at high resolution at 
HLA-A, HLA-B, HLA-C and HLA-DRB1. The likelihood of finding an optimal donor 
varies among racial and ethnic groups with the highest probability among whites 
of European descent, at 75%, and the lowest among Blacks of South or Central 
American descent, at 16%. Likelihoods for other groups are intermediate. Few 

HAPLOTAB v2.0 (07/17/2023) 
7 
 
patients will have an optimal cord-blood unit (CBU) – that is, one matched at the 
antigen level at HLA-A and HLA-B and at high resolution at HLA-DRB1. 1 
 
Allogeneic haploidentical hematopoietic cell transplantation (allo-HHCT) using 
contemporary approaches provides similar outcomes to transplantation from 
matched unrelated donor (MUD) or CBU. 2-4 
 
HHCT platforms based on T cell depletion rely mainly on graft processing to 
achieve an ideal graft composition that prevents rejection, recurrence of leukemia, 
infection and GVHD without requiring post-transplant immune suppressive 
therapy. T cell depletion-based graft require dedicated laboratories and are more 
expensive than conventional unmanipulated HCT, especially if combined with 
adoptive transfer of T cell populations chosen to improve post-transplant immune 
reconstitution. However unlike unmanipulated HHCT, pharmacologic GVHD 
prophylaxis and further treatments are not necessary in T cell depleted 
haploidentical transplants thus reducing the need for and the cost of supportive 
care and post-transplant hospitalizations.  
 
2.2. Evolution of the Methodology of T cell Depletion Haploidentical HCT  
 
Various strategies have been attempted to promote successful utilization of allo-
HHCT. Earlier strategies involved CD34+ selection in the transplanted allo-graft.5,6 
This allowed for the indirect removal of alloreactive T cells responsible for graft-
versus-host disease (GVHD), and for the indirect removal of B cells which can 
cause post-transplant lymphoproliferative disorder (PTLD) and are also thought to 
play a role in the pathogenesis of chronic GVHD (cGVHD), thus removal of B cells 
may reduce the risk of GVHD.7-9 In a landmark phase II study of haploidentical 
transplant using CD34+ immunoselection for patients with high risk acute 
leukemia, the group demonstrated high engraftment rates (93%) and high rate of 
GVHD prevention.5 Even with low rates of GVHD, the leukemia relapse in this, and 
other large studies, was not increased in high-risk leukemia patients. Despite lack 
of a potent T cell mediated graft versus leukemia (GVL) effect with T cell depletion 
(TCD), eradication of residual leukemia cells was still achieved. This may have 
been mediated by, a) the myeloablative nature of the conditioning regimen, b) 
subclinical GVL effect by the residual T cells within the graft being unopposed by 
any post-transplant immunosuppressive therapy, as well and c) T cell independent 
GVL effect of donor NK cells preserved within the graft.  
 
Patients who received TCD HHCT – or similarly with in vivo depletion using 
serotherapy – exhibit a very narrow T cell repertoire leading to prolonged 
susceptibility to serious opportunistic infections. In the study by Aversa and 
colleagues, 27 of 103 patients died from serious infections. Infection-related 
mortality has been the main cause of transplant failure using this approach. 10-12 In 
order to improve immunological reconstitution and minimize risk of severe, life-
threatening infections, successful strategies for adoptive T cell therapy were 
developed; most notably, adoptive transfer of pathogen specific T lymphocytes.13 
Across different studies, donor-derived virus-specific T cells (VSTs) were shown 
to have a response rates in 70-90% of recipients; similarly, in closely matched third 
party VSTs, response rates were between 60-70%.14 Other strategies of delayed 
T cell addback using adoptive T cell therapy included allodepletion and alloanergy 

HAPLOTAB v2.0 (07/17/2023) 
8 
 
induction, immune modulation by co-infusion of regulatory cell populations and the 
use of safety switch gene-modified T cells. 15-20  
 
To improve clinical outcomes, the CD34-positive selection approach was largely 
replaced by negative selection of peripheral blood progenitor cells (PBPCs). This 
approach allows for the preservation of other immune cells – such as Nk cells, 
dentritic cells and monocytes – supporting engraftment and immune recovery.  
One successful application of this strategy involves selective depletion of CD3+ T 
cells (+/- CD19+ B-cells) from the grafts via immunomagnetic beads using the 
automated CliniMACS device (Miltenyi Biotec, Germany).  
 
This method of transplantation was described by Hale et al. in a study of 20 
pediatric patients with acute lymphoblastic leukemia (ALL), acute myeloid 
leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia 
(CML), or non-Hodgkin lymphoma (NHL) using a conditioning regimen of total body 
irradiation (TBI), cyclophosphamide, thiotepa and rabbit antithymocyte globulin 
(ATG).21 Three subsequent trials were completed in patients with various 
hematological malignancies using a reduced intensity conditioning (RIC) regimen 
consisting of fludarabine, melphalan, thiotepa and OKT3.22-24 Rates of engraftment 
were favorable in these studies ranging from 83% to 97%. In the study by Hale et 
al., the rate of acute GVHD (aGVHD) was 16% (all grade I-II) with the rate of 
cGVHD (all limited cases) being 5.2%. 21 The trials using RIC regimens had higher 
rates of aGVHD, though mostly lower grade (grade I-II ranged from 24% to 39%, 
with grade III-IV in only 3%-14%). In these studies, the rate of cGVHD ranged from 
10%-28%. The rate of transplant-related mortality (TRM) in one study using RIC 
regimen was only 2.6%.23 However TRM remained significant in the other studies 
ranging from 20-30%.22  
 
In more recent years, Bader et al. reported on use of grafts depleted of T and B 
cells using CD3- and CD19-coated microbeads and automated CliniMACS device. 
Children with acute leukemia received conditioning consistent of fludarabine, 
thiotepa, melphalan and OKT3 or ATG. Rate of primary engraftment was 88%, 
aGVHD grade II occurred in 20% and grade III-IV in 7%; cGVHD occurred in 21%. 
Non-relapse mortality (NRM) was 8% at 1-year and 20% at 5-years.25 Using the 
same approach, but with RIC regimens in 61 adults, the incidence of grade II-IV 
acute and chronic GVHD was 46% and 18% respectively. Day 100 mortality was 
23% and at 2 years 42%. Relapse rate was 31% and overall survival (OS) at 2 
years was 28%.26 A major concern with this approach was the high incidence of 
GVHD. 
 
To overcome this problem, Chaleff et al. described a large-scale clinical method 
using the Miltenyi Biotec CliniMACS® T-cell receptor alpha/beta (TCR α/β) system 
for the depletion of α/β T lymphocytes from peripheral blood stem cells while 
retaining all other cells. 27 The CliniMACS® TCR α/β system uses murine 
monoclonal antibodies specific for the T cell receptor α/β antigen conjugated to 
biotin in combination with the CliniMACS® anti-Biotin reagent. The pioneering 
experience of the Handgretinger’s group showed that TCR αβ/CD19 depletion 
allows a T cell reduction of 4.5-5 log which is comparable to CD34+ selection 
methods.28,29 It also ensures patients to receive NK cells, monocytes and dendritic 
cells, and most important, the TCR gamma/delta+ (γδ+) T lymphocytes. TCR γδ T 
cells appear to exert anti-leukemic activity since they directly recognize stress-

HAPLOTAB v2.0 (07/17/2023) 
9 
 
induced self-antigens expressed by malignant cells. Strikingly, they do not 
recognize specific processed peptide antigens as presented on major 
histocompatibility (MHC) molecules and so are not expected to induce GVHD.30,31  
 
Early clinical experiences with children transplanted using this approach showed 
excellent full-donor engraftment, a rapid early expansion of donor-derived TCR 
γδ+ T cells that contributed to a very fast immunological reconstitution.28  Using 
the same method of graft engineering, Locatelli and colleagues achieved similar 
results in terms of engraftment, prevention of both acute and chronic GVHD, and 
a rapid recovery of post-transplant immunity in children independently from the 
conditioning regimen, whether TBI based (children with leukemia) or 
chemotherapy based (children with non-malignant disorders).32,33 In 23 children 
with non-malignant disorders, no cases of visceral acute or chronic GVHD were 
observed and survival was 91% at 2 years. 34 The same group in Italy and others 
have tested in phase 1/2 clinical trials, the use of a chimeric gene incorporating the 
death domain of inducible caspase 9 (iC9) in children with either malignant or non-
malignant disorders who receive TCR α/β/CD19 depleted haplo-HCT followed by 
infusion of titrated numbers of iC9 T cells on day 14 +/- 4. The goal was to further 
improve immune reconstitution without increasing the risk of GVHD. These 
modified T cells can be rapidly eliminated by the administration of AP1903 if acute 
GVHD occurs.20,35 TCR γδ+ T cells recovering during the first year after HCT in 
102 patients with acute leukemia correlated with a reduced incidence of infection 
in the study by Perko et al.36 Children with an elevated number of TCR γδ+ T cells 
post engraftment experienced only viral infections, while low/normal TCR γδ+ T 
cell group had viral, bacterial and fungal infections. Enhanced TCR γδ+ T cell 
recovery resulted in higher event-free survival (EFS) rate in 1 year.  
 
Outcomes of TCR α/β/CD19 depleted haplo-HCT were evaluated in a cohort of 
children with chemo-refractory AML. The conditioning regimen included 
fludarabine and cytarabine (cytoreduction phase) followed by treosulfan and 
thiotepa (myeloablative phase). Tocilizumab was given instead of ATG in all 
patients, abatacept in 10 patients. Post-transplant CD45RA-depleted donor 
lymphocytes were given prophylactically; with or without a hypomethylating agent. 
Overall results were promising with 95% of patients achieving a complete 
remission, 18% having a grade II-IV acute GVHD and 23% chronic GVHD. At 2 
years, NRM was 9%, relapse rate 42%, EFS and OS were 49% and 53% 
respectively.37 More recently, the advantages of this strategies were demonstrated  
in 20 patients with sickle cell disease (SCD) (children and adults, median age 15 
years). Conditioning consisted of ATG, thiotepa fludarabine, and treosulfan. Two 
patients succumbed to a cytomegalovirus (CMV) pneumonitis and a macrophage 
activation syndrome. One patient required renal replacement therapy for BK virus 
nephritis. None developed grade III-IV GVHD. At a median follow up of 21 months 
(range 9-62 months), overall survival was 90% documenting the feasibility, safety 
and efficacy of TCD haplo-HCT also for advanced stage SCD patients.38  
 
Prezioso et al reported on transplant outcome using this approach in 59 adults 
(median age 48 years, range 19–74) with hematological malignancies.39 All 
patients were conditioned with ATG (median at Day -10), treosulfan, fludarabine, 
and thiotepa. No additional pharmacologic prophylaxis for GvHD was given after 
transplantation. Full donor engraftment was achieved in 56/59 (95%) patients. Two 

HAPLOTAB v2.0 (07/17/2023) 
10 
 
patients developed severe GVHD resulting in death. One of them had received the 
highest dose of ab+ T cells (3.7 × 105/kg). Skin limited grade II acute GvHD was 
observed in 8 patients who responded rapidly to steroids. At the time of this report, 
only two patients have developed chronic GvHD that responded to steroid and 
cyclosporine treatment. Interestingly, naïve and memory T-cell subsets increased 
significantly over the first year after transplantation. Similarly, serum 
immunoglobulin levels normalized within 3 months. In two patients, CMV 
reactivation was associated with a significant expansion of pathogen specific 
CD8+ T cells that contributed to spontaneous elimination of viral load. Disease 
relapse was the main cause of death in 16/59 patients; 15 patients died without 
relapsing, 11 of them from infections.  
 
Similar results reported by a team in Turkey in 34 adult patients with either 
AML(n=24) or ALL (n=10). 40 Conditioning regimen consisted of thiotepa, 
melphalan, fludarabine, and ATG. Full donor chimerism was achieved in 31/34 
patients. Four patients developed severe GvHD (2 acute, 2 chronic). A low NRM 
(11.7%) at day 100 was attributed to a rapid T-cell reconstitution. Relapse was the 
main cause of death (56.3%). DFS at 1 year was 42%. 
 
2.3. Role of Natural Killer (NK) cells 
Earlier investigations in the allo-HCT setting demonstrated several favorable 
actions of NK cells in grafts including facilitating engraftment, graft versus tumor 
(GVT) effects, and activity against opportunistic infections. Tumor cell lysis by NK 
cells can be mediated by mismatch of donor NK cell killer immunoglobulin-like 
receptors (KIRs) and KIR ligands on recipient cells, which facilitates GVL effect.41-
43 Transplantation from NK-alloreactive donors (n= 51) was associated with a 
significantly lower relapse rate in the 61 patients (3 vs. 47%) (P > 0.003) and better 
event-free survival (EFS) (67 vs. 18%, P = 0.02) in a cohort of 112 patients with 
high-risk AML.10 Furthermore, the group from Madrid, Spain reported on 
CD56dim/CD56bright NK cell ratio early post HHCT using TCR α/β/CD19 depleted 
grafts and outcome. In this prospective analysis of 60 patients undergoing 63 
transplants for ALL, AML or other hematological malignancies (23 patients in CR1, 
20 in CR2 and 20 in > CR2), the probability of relapse was 32 +/- 6% and disease 
free survival (DFS) was 52 +/- 6% with a median follow-up of 28 months. A high 
CD56dim/CD56bright NK cell ratio was associated with better DFS (≥3.5; 77 ± 8% vs. 
<3.5; 28 ± 5%; p = 0.001) due to lower relapse incidence (≥3.5; 15 ± 7% vs. <3.5; 
37 ± 9%; p = 0.04). Thus, a rapid expansion of “mature” NK cells post HHCT, and 
lower risk of relapse, further supports an NK cell-mediated GVL effect. 44  
 
2.4. Current Outcome Data for TCR αβ+ CD19+ Depletion  
 
2.4.1. Graft Composition  
Overall, studies have shown good results with respect to graft composition, 
achieving adequate depletion of TCR αβ+ CD19+ cells and good CD34+ 
cell recovery. Several pediatric studies have shown median numbers of 
CD34+ cells per kg body weight in the graft ranging from 7.9 x106 to 18.7 
106. 45-51 Studies conducted have shown comparable median numbers of 
CD34+ cells per kg body weight of 11 x106 to 17.69 x106.  Earlier studies 

HAPLOTAB v2.0 (07/17/2023) 
11 
 
involving CD34+ selection for haploidentical transplant were notable for 
CD34+ cell counts/kg body weight ranging from 1.2 x106 to 13.8 x106. 5,6 
Thus, the cell dose achieved with TCR αβ+ CD19+ depletion is not inferior 
to CD34+ selection. Overall, median numbers of αβ+ T cells and B cells 
per kg body weight ranged from 11 x103 to 55 x103 and 40 x103 to 300 x103, 
respectively, reflecting optimum depletion, typically on the order of 3-4 log 
depletion.34,45,47,48,50 Median numbers of γδ+ T cells and NK cells in the graft 
range from 4 x106 to 11 x106 per kg body weight 34,40,45,47,52  and 22 x106 to 
81.3 x106,34,47-50 respectively. 
 
2.4.2. Engraftment  
Several studies have shown successful primary engraftment rates (83-
100%) of TCR αβ/CD19+ depleted stem cell grafts for HHCT, with average 
absolute neutrophil count (ANC) recovery by day 10-15 and average 
platelet recovery by day 10-17.  Although various conditioning regimens 
have been studied (possibly due to heterogeneity of diseases studied) most 
have commonly included alkylator based chemotherapy, often thiotepa or 
melphalan as well as ATG or radiation. Lang et al reported on 41 pediatric 
patients with both benign and malignant conditions who received a 
conditioning with fludarabine or clofarabine, thiotepa and melphalan in 
combination with either OKT3 or ATG-Fresenius. Five of the 41 patients 
did not engraft, yet all five successfully engrafted on second attempt after 
being reconditioned and infused with grafts from different haploidentical 
donors also depleted of αβ T cells and B cells. 49 
 
2.4.3. Graft-versus-Host Disease (GVHD) 
So far, studies have shown promising results of TCR αβ/CD19+ depleted 
stem cell grafts for allogeneic HHCT with respect to the incidence of GVHD.  
Collectively, grade I-II aGVHD was reported in 3-39% of patients and 
primarily limited to the skin, rather than visceral organs. Across different 
studies, the overall incidence of severe GVHD was low (0-15%). Kaynar et 
al. studied 34 adults with AML or ALL who received a conditioning regimen 
of fludarabine, thiotepa, melphalan and ATG.  Median follow-up was 176 
days. Grade I-II aGVHD was observed in 26% of patients and grade III-IV 
aGVHD was observed in 6%. The rate of cGVHD was 6% with 3% being 
limited and 3% being extensive. The authors reported on at least two 
patients 
treated 
for 
GVHD 
using 
steroids, 
cyclosporine 
and 
mycophenolate. Locatelli et al. studied 80 pediatric patients with AML or 
ALL, each received one of four conditioning regimens, two of which were 
TBI-based. 50 There was no post transplantation GVHD prophylaxis. 
Median follow-up was 46 months. The rate of aGVHD was 30% and all 
were grade I-II. All of these patients responded to treatment with topical or 
systemic steroids. The rate of cGVHD was 5% with all cases limited to the 
skin. Bertaina el al. reported on 23 children with benign disorders who 
received one of four conditioning regimens. 34 There was no post-
transplantation GVHD prophylaxis. Median follow-up was 18 months. 
Grade I-II aGVHD was observed in 23 patients and limited to the skin. No 
patients experienced acute visceral or cGVHD.  
 
NOTE: see appendix B1 and B2 for grading systems 

HAPLOTAB v2.0 (07/17/2023) 
12 
 
 
2.4.4. Transplant-related Mortality (TRM) 
So far, studies have shown promising results of TCR αβ/CD19+ depleted 
stem cell grafts for allogeneic HHCT with respect to TRM.  The reported 
incidence of TRM is variable and ranges from 0-20%. Infection was the 
most common cause of TRM. Lang et al. conducted a study involving 30 
pediatric 
patients 
with 
non-malignant 
conditions, 
hematological 
malignancies and solid tumors who received a conditioning regimen of 
fludarabine, thiotepa and melphalan and either ATG or TBI. TRM in this 
study was 3.3%, with all TRM due to infection. 48 Bhat et al. carried out a 
study involving 22 pediatric patients with both malignant and non-malignant 
hematological conditions who received a conditioning regimen consisting 
of TBI or chemotherapy. TRM in this abstract was 14% with all TRM due to 
infection.45 Overall the rate of TRM in these studies is lower than the rate 
of TRM seen in earlier studies involving transplantation of “megadoses” of 
CD34+ cells or CD3+/CD19+ depleted cells from haploidentical donors (36-
40% and 20-30% respectively).5,6,21,22,24 
 
2.4.5. Survival  
Although survival data might be difficult to interpret from studies given the 
heterogeneity of diseases included, certain studies involving only one or 
two disease types provide insight regarding disease efficacy. 
 
The study conducted by Locatelli et al. in 80 children with acute leukemia 
who received myeloablative conditioning, including ATG, showed OS of 
72% at 5 years and DFS of 71% at 5 years.50 Maschan et al. utilized this 
transplant method in 33 pediatric patients with high-risk AML with 20 
receiving MUD transplants and 13 receiving haploidentical transplants. The 
conditioning regimen consisted of treosulfan, melphalan, fludarabine and 
ATG. OS and DFS at 2 years were 73% and 59% respectively for 
haploidentical cohort. 47 
 
2.4.6. Immune Reconstitution 
Generally, studies evaluating immune reconstitution following HHCT with 
TCR αβ/CD19+ depleted grafts show rapid early reconstitution of γδ T cells 
and NK cells, with more gradual appearance of αβ T cells and B cells over 
3-12 months.33,45,48,50,53 Locatelli and colleagues demonstrated rapid 
recovery of γδ T cells and NK cells with a median of 181 cells/µL (range 1-
1335) and 236 cells/µL (range 47-1813) at one month respectively.50 
Subsequently, αβ reconstitution took place, with a median of 47/µL (range 
1-672) at 1 month and 186/µL (range 12-1340) at 3 months, but with 
recovery to 573/µL (range 135-2146) at 6 months and 1291/µL (range 259-
2795) at 12 months. Similarly, B cell population had a very gradual recovery 
with minimal amounts detected at 1 and 3 months; ultimately, numbers 
reached 160/µL (range 2-1609) and 291/µL (range 40-1616) at 6 and 12 
months respectively.50 Similar results were previously reported by Lang 
and colleagues in pediatric patients where recovery of γδ T cells started as 
early as seven days post-transplant and made up the majority of CD3+ 
cells in the early post-transplant period. NK cells that were co-infused with 
the product were detected in the first week post-transplant followed by early 

HAPLOTAB v2.0 (07/17/2023) 
13 
 
proliferation in week 2. NK cell population reached > 200 cells/µL shortly 
thereafter (with a median of 12.5 days). B cell population recovered by day 
30 and reached normal levels by day 150 post-transplant.49 Other 
advantages of the kinetics of NK cell recovery with HHCT using TCR 
αβ/CD19+ depleted grafted over CD34+ selection approach is that early 
donor derived mature NK cells is often not demonstrated in the CD34+ 
setting until at least 2-3 months post-transplant. 54  Airoldi and colleagues 
performed an in-depth analysis of γδ T cells recovery following TCR 
αβ/CD19+ depletion in 27 pediatric patients with malignant and non-
malignant conditions. Two to three weeks post-transplant, T cell 
populations were predominantly γδ cells (mean 91.5% of CD3+ 
lymphocytes; range 70-100%) and with the αβ T cell population gradually 
increasing over time. Based on previous studies showing that 
aminobisphosphonates can induce activation and proliferation of Vδ2 
subset of γδ population, the authors studied the effect of in vitro exposure 
to zoledronic acid on rate of expansion of γδ cells in subset of 13 patients. 
As expected, exposure to zoledronic acid increased expansion rate of Vδ2 
subset of γδ cells. Similarly, cytotoxic activity of Vδ2 cells was enhanced 
with exposure of leukemia cell lines to zoledronic acid.33  
 
 
2.4.7. Infections  
 
CMV reactivation rates ranged from 30-70% in various studies. However, 
death from CMV occurred in no more than 5%.  Two studies reported on 
BK virus reactivation, ranging from 17 to 25% in their cohorts, but no BK 
virus-related deaths were reported.  Three studies report on adenovirus 
detection, ranging from 5-53%, with two deaths reported. 24,45,48 There were 
no cases of Epstein-Barr virus (EBV)-associated PTLD reported in the five 
studies where it was monitored, and EBV viremia was only very rarely 
detected.  Overall, these data indicate that the risk of viral reactivations 
remains significant in the context of TCR αβ/CD19+ depleted HHCT. 
However, the reported mortality rates associated with these infections are 
very low. Importantly, risk of EBV reactivation and EBV-related PTLD 
appear very low in patients undergoing this type of transplant.  
 
NOTE: See appendix C for CMV monitoring and prophylaxis 
 
 
3.0 PATIENT ELIGIBILITY 
 
 
3.1. Inclusion Criteria 
3.1.1. Age less than or equal to 55 years 
3.1.2. Lack of suitable conventional donor (10/10 HLA matched related or unrelated 
donor) or presence of rapidly progressive disease not permitting time to identify an 
HLA-matched unrelated donor. This does not include cord blood unit (CBU) 
availability. 
3.1.3. Lansky/Karnofsky score > 50 
3.1.4. Signed written informed consent  
3.1.5. Diagnosis of one of the following:  

HAPLOTAB v2.0 (07/17/2023) 
14 
 
a. Patient with life threatening hematological malignancy including “high-risk” 
ALL in first complete remission (CR1); ALL in second or subsequent 
remission (≥ CR2); high-risk AML in CR1; AML in second or subsequent CR; 
myelodysplastic syndromes (MDS); non-Hodgkin’s lymphomas (NHL) in 
second or subsequent remission (≥ CR2); CML 
b. Hemophagocytic Lymphohistiocytosis (HLH) including familial HLH, relapsed 
HLH or central nervous system (CNS) HLH 
c. Primary Immunodeficiency Disorders (PID) 
d. Hemoglobinopathies including thalassemia or sickle cell disease (SCD) 
e. Severe aplastic anemia (SAA) not responding to immune suppressive 
therapy 
f. 
Congenital/hereditary cytopenias including Fanconi anemia (FA) without 
malignant clonal evolution (MDS, AML) 
g. Other inherited bone marrow failure syndromes (IBMFS) 
h. Severe chronic active Epstein Barr virus infection (SCAEBV) with predilection 
for T- or NK-cell malignancy 
 
Note: ‘High risk’ ALL or AML refers to those acute leukemias identified by the presence of specific 
biologic features, which predict high likelihood of failure to conventional chemotherapy. As 
biologic features of high-risk disease evolve with improvement of conventional chemotherapy, it 
is not practical to define this indication with any further specificity. Therefore, high risk AML/ALL 
will be determined by the primary physician. 
 
3.2 
Exclusion Criteria 
3.2.1 Life expectancy of < 6 weeks  
3.2.2 Greater than grade II acute graft versus host disease (GVHD) or chronic 
extensive GVHD due to a previous allograft at the time of inclusion 
3.2.3 Subject receiving an immunosuppressive treatment for GVHD treatment due to a 
previous allograft at the time of inclusion 
3.2.4 Symptomatic cardiac disease or left ventricular shortening fraction < 25% or 
ejection fraction < 40% 
3.2.5 Severe renal disease, with creatinine clearance < 40cc/1.73 m2 
3.2.6 Pre-existing severe restrictive pulmonary disease, FVC < 40% of predicted 
3.2.7 Severe Hepatic Disease with ALT/AST ≥ x2.5 upper limit of normal or bilirubin 
level ≥ x1.5 upper limit of normal 
3.2.8 Serious concurrent uncontrolled medical disorder or mental illness 
3.2.9 Pregnant or breastfeeding female subject 
3.2.10 Current active infectious disease including viral and fungal diseases at the time 
of enrollment; that on evaluation of PI precludes ablative chemotherapy or 
successful transplantation. 
3.2.11 Active HIV infection 
3.2.12 Severe personality disorder or mental illness that would preclude compliance 
with the study. 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
15 
 
4.0 TREATMENT PLAN 
We will use the CliniMACS® TCRαβ-Biotin system, manufactured by Miltenyi Biotech. 
This system will be accessed through three IDE exemption cross references: BB MF 
12011, (Clinimacs® CD19 reagent system), BB MF 12251, (Clinimacs® depletion tubing 
set), and BB MF 15678, Clinimacs® TCR α/β reagent system), all held by the 
manufacturer, Milteny Biotec, Inc., Cambridge, MA. 
 
4.1. Pretreatment Procedures 
 
4.1.1. Patients will have a double lumen central venous catheter placed prior to starting 
conditioning regimen.  
4.1.2. Patients will undergo pre-transplant workup which includes but not limited to 
disease evaluation, organ functioning, infectious disease workup (lab testing and 
radiologic exams) as well as performance scores. This will be performed per our 
institutional standards of practice. 
 
4.2. Donor Selection 
As described in section 3.1.2, the protocol will be open for patients who lack a suitable 
conventional donor or in the presence of a rapidly progressive disease not permitting time 
to identify an unrelated donor. For this protocol, the "best" donor will be defined as a first-
degree haploidentical family member. Matching will be determined by class I and class II 
DNA based HLA typing. The donor should be sufficiently healthy not to be at increased 
risk from the mobilization procedure.  Should more than one "equally" MHC incompatible 
donor be identified, other selection criteria will include: age and size of donor, CMV status 
and sex. The final decision on donor selection will be at the discretion of the primary BMT 
physician and the Principal Investigator (PI) of the study.  
 
4.3. Donor Collections by Apheresis 
 
Donor collection will be accomplished by using leukapheresis whereby the leukocyte 
fraction will be collected using continuous flow centrifugation. Prior to collection, infectious 
disease evaluation will be performed per institutional guidelines and standard of practice. 
The donor will receive recombinant G-CSF (Filgrastim) by daily subcutaneous injections 
(as per institutional SOP), typically 5 days prior to anticipated date of collection, with 
monitoring of CD34+ cell count starting on the fourth day of starting G-CSF. Apheresis will 
typically begin on the fifth day of mobilization. A standard apheresis procedure of 2-3 times 
blood volumes will be performed and processed per institutional practices. The volume 
processed and the duration of the apheresis will be documented and recorded. If less than 
5 x109 peripheral blood mononuclear cells (PBMCs) are collected, the PI or designee 
should be notified immediately.  
 
Every effort will be made to infuse a fresh stem cell product; however, a frozen product 
may be infused when necessary. 
 
All donors will be monitored and PBSC products will be collected according to FACT-
JACIE international standards for cell therapy, product collections, product and 
administration. 
 

HAPLOTAB v2.0 (07/17/2023) 
16 
 
 
4.4. Donor Stem Cell Apheresis Procedure 
Prior to negative depletion of TCR αβ+/CD19+, the target number of CD34+ stem cells by 
apheresis should be ≥ 12-15 x106/kg recipient weight. Accounting for up to 40% loss of 
CD34+ cells during the depletion process, the final product to be infused should contain 
at least 8-10 x106 CD34+ stem cells/kg recipient weight. The actual cell numbers achieved 
will be documented. To ensure achieving the target cell dose, a second successive day of 
apheresis will be allowed. The optimal number of peripheral CD34+ cells is ≥ 0.04 x109/L. 
 
In the event that the CD34+ target cell dose is not achieved with the initial leukapheresis 
collection and, as stated above, the donor is to be collected again for additional CD34+ 
cells (with or without use of plerixafor). 
 
4.5. TCR αβ+/CD19+ Depletion 
The leukapheresis product will undergo negative selection of TCR αβ T cells following the 
standardized protocol in the user’s manual for the CliniMACS (Miltenyi Biotech, Germany).  
TCR αβ+ T-cells are labeled by CliniMACS TCR αβ-Biotin (murine anti-TCR αβ 
monoclonal antibodies conjugated to biotin) which allows the TCR αβ+ T-cells to be 
magnetically labeled with CliniMACS Anti-Biotin reagents (murine anti-biotin monoclonal 
antibodies conjugated to superparamagnetic iron dextran particles) for depletion. The 
CD19+ B cells are labeled by CliniMACS CD19 reagents which allows the CD19+ B cells 
to be magnetically labeled for depletion. All unlabeled cells are selected as target cells 
which should not contain more than the maximum number of TCR αβ+. 
The end points to be achieved for validation of the final product after negative selection 
include: 1) > 80% recovery of CD34+ cells after processing, 2) > 3-log depletion of TCR 
αβ cells after processing with a target infusion of < 1 x105/kg, 3) no growth in sterility 
cultures over 14 days and 4) > 80% viability in CD34+ cells. If at the end of the negative 
selection procedure, the residual number of TCR αβ+ cells is > 1 x105/kg, a residual part 
of the selected graft may be frozen and stored. The number of residual B lymphocytes 
after graft manipulation will be recorded but it will not lead to further graft manipulation or 
influence clinic decision. 
 If the desired CD34+ cell dose is not obtained in the initial product, a second 
leukapheresis collection may be selected for CD34+ cells after the target residual αβ T 
cell number has been reached. The number of residual B lymphocytes after the graft 
manipulation will be recorded but will not influence further graft manipulation or clinical 
decisions.  
All subjects enrolled on the study will receive one dose of Rituximab i.v. at a dose of 200 
mg/m2 once on day -1 regardless of the residual number of CD19 B cells in the final 
product.  
 
4.6. Conditioning Regimens 
Two types of regimens are allowed on the study, myeloablative and reduced intensity. 
Choice of regimen will be made at the discretion of the treating physician, that is deemed 
the most appropriate for each patient and for the condition/disease being treated.  
 

HAPLOTAB v2.0 (07/17/2023) 
17 
 
NOTE: 
- 
Target busulfan area under the curve (AUC) will be between 800-1,200 uM min. 
- 
Dosing of chemotherapy for obese patients will follow the American Society for 
Blood and Marrow Transplantation Practice guideline Committee as described in 
the position statement published by Bubalo et al. in 2014 55. 
 
5.0 EVALUATIONS 
 
5.1. Evaluation During Study 
 
Study evaluations and their specific time points for each are described in details in the 
study calendar; will vary according to the underlying disease and will be performed per 
institutional standard of care.  
 
Data will be collected as per secondary and exploratory objectives (section 1.1). 
 
STUDY CALENDAR: 
 
Baseline 
Evaluations 
(prior to 
initiation of 
conditioning 
regimen) 
Evaluations 
during 
inpatient 
admission  
Weekly 
until Day 
+100 
Day 100 
Evaluations 
Monthly 
through 
Month 11  
Annual 
Evaluations 
Year 1 – 5 
H & P 
  X@ 
See footnote 
2 
X 
X 
X 
X 
Body Weight 
X 
X 
X 
X 
X 
Performance Score 
X 
 Day +30, Day +60 
X 
 
X 
Chest X-ray or CT 
scan* 
X 
 
 
 
 
 
EKG 
X 
 
 
X 
 
X 
ECHO 
X 
 
 
X 
 
X 
PFTs** 
X 
 
 
X 
 
X 
crCl or GFR*** 
X 
 
 
X 
 
 
CBC with diff 
X 
See footnote 
2 
X 
X 
X 
X 
CMP including 
LDH 
X 
X 
X 
X 
X 
Liver panel 
X 
X 
X 
X 
X 
UA 
X 
X 
X 
X 
X 
HLA typing 
X 
 
 
 
 
 
ABO typing 
X 
 
 
X 
 
 
Viral Serologies# 
X 
 
 
 
 
 
Other Serologies# 
X 
 
 
 
 
 
Viral PCR (plasma) 
including CMV, 
EBV and 
adenovirus PCR 
 
See footnote 
2 
X 
X 
X 
X (year 1 
only) 
Aspergillus Antigen 
 
X 
X 
X 
 
Fungitell 
 
X 
X 
X 
 
STR or FISH for 
sex chromosomes 
(if donor and 
recipient opposite 
sex) PB & 
X 
X (every 4 weeks following 
engraftment) 
X 
X 
X 
GVHD grading 
 
X§ 
X 
X 
X 
X 

HAPLOTAB v2.0 (07/17/2023) 
18 
 
 
Baseline 
Evaluations 
(prior to 
initiation of 
conditioning 
regimen) 
Evaluations 
during 
inpatient 
admission  
Weekly 
until Day 
+100 
Day 100 
Evaluations 
Monthly 
through 
Month 11  
Annual 
Evaluations 
Year 1 – 5 
Bone marrow 
aspirate & Biopsy% 
X 
X (Day +28 +/- 3 days) 
X 
month 6 
(as 
indicated) 
X (Day +365 
only +/- 3 
days) 
LP% 
X 
X (Day +28 +/- 3 days) 
X 
 
X (Day +365 
only +/- 3 
days) 
Serum 
immunoglolin G 
 
 
X 
X 
X 
X 
Restaging 
evaluations (as 
indicated)^ 
X 
 
 
X 
 
X (year 1 
only) 
Thyroid Function 
Tests 
 
 
 
 
 
X 
Quantitative serum 
immunoglobulins 
 
 
 
X 
 
X 
T-, B- and NK-cell 
subset analysis a 
 
 
 
See specific time points below (starting at Day +30) 
RTEs b 
T-cell Repertoire 
(flow cytometry) c 
Proportion of 
alpha/beta versus 
gamma/delta T-cell 
receptor d 
NK Screening 
Subset e 
Extended NK cell 
Phenotype f 
 
H&P: history and physical; EKG: electrocardiogram; ECHO: echocardiogram: PFTs: pulmonary function tests; CrCl: creatinine 
clearance; GFR: glomerular filtration rate; CMP: complete metabolic profile; UA: urinalysis; STR: short tandem repeats; FISH: 
fluorescent in-situ hybridization; PB: peripheral blood; LP: lumbar puncture; RTE: recent thymic emigrants 
NOTE:  
1) 
Other disease specific studies will be determined by the treating physician and will be considered standard 
of care clinical evaluations and not research-based studies. 
2) 
Evaluations during inpatient admission will be performed per institutional standard of practice.  
@ complete history and physical including:  
- 
donor/recipient HLA matching Data 
- 
underlying disease with number of remissions and relapses including cytoreductive therapy (as applicable) 
- 
concurrent diseases or therapies 
* CT scan to be done in place of chest X-ray if concern for possible ongoing pulmonary disease or infection 
(particularly fungal infection) or history of the above. Repeat imaging to be done as clinically indicated.  
** May omit in young patient unable to perform testing. Recommended for those ≥ 6 years. 
*** nuclear medicine GFR to replace CrCl (24 urine collection) is history of renal disease or if CrCl less than normal 
range for age 
& Engraftment/Chimerism studies including STR and/or FISH to be done every 4 weeks until Day +100 following 
engraftment but no sooner than Day +28. 

HAPLOTAB v2.0 (07/17/2023) 
19 
 
# Serologies include hepatitis B, hepatitis C antibodies, HIV1/2, HSV, VZV, CMV, EBV, Toxoplasma and RPR. Sent 
to Gulf Coast laboratories  
§ weekly evaluation to begin immediately following engraftment (see section 6.1 for definition) 
% for patients with hematological malignancies, bone marrow aspirate and biopsy and lumbar puncture (as indicated) 
to be done at baseline, around day +28, day +100, day +180 and day +365 (+/- 3 days) post-transplant or as clinically 
indicated. Bone marrow evaluation to include morphology, flow cytometry (or other molecular based disease 
evaluation), Cytogenetics, FISH (as indicated) as well as other genomic molecular testing (as indicated). 
^ restaging evaluations include PET-CT scan or other imaging modality for restaging or malignant disease as 
indicated 
a) 
T-, B- and NK-cell subset analysis at Day 30, 60, 100, 180 and 365 post-HCT 
b) 
Recent Thymic Emigrants (RTE) at Day 100, provided absolute lymphocytes count (ALC) is > 200 x10e9/L, then 
at Day 180 and 365 
c) 
T-cell repertoire analysis by flow cytometry at Day 180 post-transplant 
d) 
Proportion of alpha/beta versus gamma/delta T-cell receptor cell recovery at Day 30, 60, 100, 180 and 365 post-
HCT 
e) 
NK Screening subset at Day 30, 60, 100, 180 and 365 post-HCT 
f) 
Extended NK cell phenotype at Day 30, 60, 100, 180 and 365 post-HCT 
 
 
6.0 CRITERIA FOR STUDY EVALUATION 
 
6.1. Hematopoietic engraftment: Engraftment will be recorded as the first of 3 consecutive 
days where absolute neutrophil count exceeds 0.5 x109/L 
6.2. Toxicity will be graded according to the NCI Common Terminology Criteria for Adverse 
Events (CTCAE) version 5.0. http://ctep.cancer.gov 
6.3. Patients will also be assessed post-transplant at 3, and 12 months by use of the Lansky 
or Karnofsky Performance Scale (see Appendix A) 
6.4. Acute GVHD and chronic GVHD will be graded according to consensus criteria  
6.5. Relapse will be assessed from regular blood and marrow analyses and will be recorded 
by the day of detection 
6.6. Survival will be recorded by the day of death and the cause of death 
6.7. Immune reconstitution will be determined from total lymphocyte count, from T and B cell 
numbers and from T cell subset analyses. Phenotyping will be done at approximately day 
+30, +60, +100, +180 and 365 post-HCT (+/- 7 days). We will also seek evidence of CMV, 
EBV and adenovirus reactivation, and will document all bacterial, viral and fungal 
infections.  
 
7.0 SAMPLE SIZE CONSIDERATIONS 
7.1. study design and sample size 
 
This is a single site and single arm study to evaluate the safety and feasibility of αβ+ T 
cell-depleted haploidentical hematopoietic cell transplantation by negative selection of 
TCR αβ+ and CD19+ cells using automated CliniMACS device in patients with 
hematological malignancies and non-malignant disorders. 
 
The primary safety endpoints are grade III or higher aGVHD by day 100 post HCT. The 
primary efficacy endpoint is a composite endpoint of both neutrophil and platelet 
engraftment by day 42 post HCT, with neutrophil engraftment defined as the first of 3 

HAPLOTAB v2.0 (07/17/2023) 
20 
 
consecutive days with a peripheral blood absolute neutrophil count of ≥ 0.5x109/L and 
platelet engraftment defined as the first day with platelet count of ≥ 20 x109/L without 
transfusion support for 7 consecutive days. Any eligible patients who receive HCT will be 
considered evaluable for both endpoints.  
 
Based on previous experiences, we expect the engraftment rate is at least 85% and the 
aGVHD ≥ grade 3 rate is at most 10%. For the purpose of this protocol, the treatment 
regimen is deemed useless if the engraftment rate is only 70% or the aGVHD ≥ grade 3 
rate is more than 30%. A Bryant and Day56 two-stage optimal design will be used in order 
to allow early stopping for lack of satisfactory or excessive aGVHD. Although GVHD only 
occurs after engraftment, we anticipate all patients will achieve engraftment so the GVHD 
rate among engrafted patients after HCT will be very similar with that among patients who 
receive HCT. Therefore, an acceptable aGVHD rate of 10% and an unacceptable rate of 
30% are still used for the Bryant and Day design. The study will enroll a total of 47 (18+29) 
evaluable patients in two stages, with a one-sided significance level of 10% for efficacy, a 
one-sided significance level of 5% for toxicity, and 80% Power. This design results in an 
expected maximum sample size of 28 if engraftment rate and/or aGVHD rate are 
unacceptable. The probability of stopping early due to low engraftment and excessive 
aGVHD is 89%; due to excessive aGVHD, but with acceptable engraftment rate is 71%; 
and due to low engraftment rate with acceptable aGVHD rate is 68%. 
      7.2. Stopping Rules 
 
Based on the Bryant and Day two-stage design, at the end of the first stage, if the number 
of engraftment is less than or equal to 13 or if the number of aGVHD ≥ grade 3 is more 
than or equal to 5 out of 18 patients, the study will be stopped early due to either low 
efficacy or high toxicity. If we observe an engraftment rate <=90% after the first stage, we 
will report the aGVHD rate among the engrafted patients and consult with the FDA. 
Otherwise if not stopping at the first stage,  the study proceeds to the second stage and 
enrolls 29 more patients. At the end of study, if 36 or fewer patients have engrafted or 10 
or more patients experience aGVHD ≥ grade 3, the treatment regimen will be considered 
to demonstrate low efficacy or high toxicity.  
 
Additional accrual halting rules for the study to mitigate risks for patients will be applied in 
any of the following settings: 
• 
any treatment-related death within 100 days of HSCT, not related to disease 
progression; 
• 
occurrence of any case of post transplantation lymphoproliferative disorder 
(PTLD); 
• 
excessive incidence of failure to engraftment is ≥30% when the number of patients 
who failed engraftment is equal or greater than the boundary shown in the table below; 
• 
excessive incidence of grade 3 or higher acute or chronic GVHD is ≥30% when the 
number of patients who experience grade 3 or higher acute or chronic GVHD is equal or 
greater than the boundary shown in the table below.  
 
Number of treated patients (n) 
6 
12 
24 
30 
36 
42 
Boundary (x) 
2 
4 
8 
9 
11 
13 
 
 

HAPLOTAB v2.0 (07/17/2023) 
21 
 
Once halted, the study team will review the outcomes of the patients and report to 
regulatory authorities for review. The study will not resume until approvals to continue are 
granted by the FDA.  
 
7.3.Statistical Analysis 
 
Incidences of neutrophil and platelet engraftment by day 42 post HCT from enrolled 
evaluable patients will be estimated along with the associated 95% confidence intervals.  
 
Incidence of acute GVHD and chronic GVHD by grades among patients who achieve 
engraftment will be reported as rate and its associated 95% confidence interval. Other 
adverse events will also be summarized descriptively as frequency tables. Listings of all 
adverse events by patients will include the time to onset, the duration of each event, the 
severity of each event, and the relationship of the event to study therapy, whether it was 
a serious event, and whether it caused withdrawal.  
 
Feasibility of the automated CliniMACS system to produce a graft with defined cell 
composition will be reported as proportion of patients with success in using CliniMACS® 
System for processing hematopoietic stem and progenitor cells collected from an 
allogeneic, HLA-haplotype donors to obtain a CD34+ cell-enriched population with the 
target dose of CD34+ cells of 10-12 x106/kg (minimum of ≥2.5x106/kg) and a target dose 
of T-cell receptor (TCR) alpha/beta CD3+ cells is ≤1x105/kg. 
 
Cumulative incidence of transplant-related mortality (TRM) at Day 100 and Day 365 
among all evaluable patients will be reported with frequencies and proportions. OS will 
be summarized with survival curves using Kaplan-Meier method. OS is defined as the 
time from HCT to death. Alive patients will be censored at the last follow-up. 
 
Incidences of viral reactivations, defined as detectable viral load by peripheral blood PCR, 
and symptomatic infections up to one year post HCT will be summarized as proportions 
among all evaluable patients. 
 
 For patients with hematological malignancies, complete donor chimerism by day 30 post-
transplant will be summarized descriptively. Lab data including lymphocyte, T cell, B cell, 
and NK cell at each time point post HSC will be summarized appropriately and presented 
in plots. Testing of trends over time will be performed with paired statistical methods but 
the analyses will be considered exploratory.     
 
8.0 REPORTING REQUIREMENTS 
 
8.1. To register patients, contact Dr. Erin Morales at 832-826-0860 at Texas Children’s 
Hospital or Dr. George Carrum at 713-441-1450 at Houston Methodist Hospital. After 
entry, the following data will be collected: 
8.1.1. Eligibility (at enrollment) 
8.1.2. On study  
8.1.3. Concomitant Medications 
8.1.4. Adverse Events 
8.1.5. Off study  

HAPLOTAB v2.0 (07/17/2023) 
22 
 
8.1.6. Death information (if applicable) 
8.1.7. Other transplant related data is collected on CIBMTR forms as required 
 
8.2. Relapse and survival data will be collected until one year post-transplant 
 
8.3. Drug Toxicity and/or Adverse Reactions 
8.3.1. Adverse events will be collected as per SOP J02.05.XX and SOP J02.78.XX 
8.3.2. Serious adverse events will be collected and reported as per SOP J02.06.XX 
which includes the requirement that serious adverse events, particularly 
Suspected Unexpected Serious Adverse Reactions (SUSARs) will be reported to 
the FDA per IND safety reporting requirement: 21 CFR 312.32 
 
8.4. Potential Risks and Mitigation Strategies 
Adverse events (including grade 3 or higher) are possible and expected toxicities associated 
with any transplant procedure regardless of the graft processing approach, donor type or 
whether transplant is autologous or allogeneic. These are primarily attributable to conditioning 
regimens including chemotherapy and/or radiation therapy, immunosuppressive medications 
(which will not be a standard of care on this protocol) as well as other supportive interventions 
(including antimicrobials, diuretics, among others). Accordingly, the occurrence of AEs (other 
than GVHD exceeding the acceptable rates specified in the study protocol) should not result 
in early termination of the study or pause in study accrual.  
Management of potential post-transplant complications will be done per institutional guidelines 
and as outlined in the SOPs below: 
 
F05.07.XX Treatment of Regimen-related Complications 
 
F07.04.XX Treatment of Infections 
 
F08.03.XX Acute GVHD Treatment 
 
F08.04.XX Chronic GVHD Treatment 
 
F05.05.XX Adverse Reactions to Transplantation 
 
F05.02.XX Post-Transplant Evaluation 
 
F05.15.XX Recipient Care 
 
Subjects with engraftment failure (as defined in the study protocol), will be taken off study, will 
be evaluated for a second allogeneic HCT using a standard of care approach and per our 
institutional guidelines.  
 
9.0 OFF STUDY CRITERIA 
 
- 
Refusal of further study follow-up by patient or legal guardian 
- 
Treatment with hematopoietic cell product, relating to patients receiving additional 
stem and progenitor cell products 
- 
Completion of study specified procedures  
- 
Relapse 
- 
Lost to follow-up 
- 
Death  
 

HAPLOTAB v2.0 (07/17/2023) 
23 
 
 
10.0 
CLINICAL TRIAL OVERSIGHT AND MONITORING 
 
This protocol will be conducted in accordance with the Cell/Gene Therapy Monitoring Plan 
on file with the FDA.   
 
This protocol will be monitored in accordance with the current Data Safety Monitoring Plan 
of the Dan L Duncan Cancer Center at Baylor College of Medicine. 
 
The conduct of this clinical trial will be evaluated in accordance with the Texas Children's 
Cancer Center and Center for Cell and Gene Therapy Quality Assurance Policy and 
Procedure Plan and Quality Control Policy and Procedure Plan. 
 
 
11.0 
INFORMED CONSENT 
All patients and/or their legal guardian must sign a document of informed consent 
consistent with local institutional and federal guidelines stating that they are aware of the 
investigational nature of this protocol and of the possible side effects of treatment.  
Further, patients must be informed that no efficacy of this therapy is guaranteed, and 
that unforeseen toxicities may occur.  Patients have the right to withdraw from this 
protocol at any time.  No patient will be accepted for treatment without such a document 
signed by the patient or their legal guardian. Full confidentiality of patients and patient 
records will be provided according to institutional guidelines. 
 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
24 
 
Appendix A: Lansky and Karnofsky Performance Scale 
 
Karnofsky Scale (recipient age ≥ 16 years) 
 
Lansky Scale (recipient age ≥ 1 year and 
<16 years) 
 
Able to carry on normal activity; no special 
care is needed 
 
Able to carry on normal activity; no special 
care is needed 
 
100 Normal, no complaints, no evidence of 
disease 
 
100 Fully active 
 
90 Able to carry on normal activity 
 
90 Minor restriction in physically strenuous 
play 
 
80 Normal activity with effort 
 
80 Restricted in strenuous play, tires more 
easily, otherwise active 
 
Unable to work, able to live at home, cares 
for most personal needs, a varying amount 
of assistance is needed 
 
Mild to moderate restriction 
 
70 Cares for self, unable to carry on normal 
activity or to do active work 
 
70 Both greater restrictions of, and less time 
spent in active play 
 
60 Requires occasional assistance but is able 
to care for most needs 
 
60 Ambulatory up to 50% of time, limited 
active play with assistance/supervision 
 
50 Requires considerable assistance and 
frequent medical care 
 
50 Considerable assistance required for any 
active play, fully able to engage in quiet play 
 
Unable to care for self, requires equivalent 
of institutional or hospital care, disease 
may be progressing rapidly 
 
Moderate to severe restriction 
 
40 Disabled, 
requires 
special 
care 
and 
assistance 
 
40 Able to initiate quite activities 
 
30 Severely 
disabled, 
hospitalization 
indicated, although death not imminent 
 
30 Needs considerable assistance for quiet 
activity 
 
20 Very sick, hospitalization necessary 
 
20 Limited to very passive activity initiated by 
others (e.g., TV) 
 
10 Moribund, fatal process progressing rapidly 
 
10 Completely disabled, not even passive play 
 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
25 
 
Appendix B1: Acute GVHD Staging and Grading (MAGIC Consortium)  
 
Stage 
Skin (active erythema only) 
Liver (bilirubin) 
Upper GI 
Lower GI (stool output/day) 
0 
No active (erythematous) GVHD 
rash 
< 2 mg/dl 
No or intermittent 
nausea, vomiting or 
anorexia 
Adult: < 500 ml/day or < 3 episodes/day 
Child: < 10 ml/kg/day or < 4 episodes/day 
1 
Maculopapular rash < 25% BSA 
2-3 mg/dl 
Persistent nausea, 
vomiting or anorexia 
Adult: 500-999 ml/day or 3-4 episodes/day 
Child: 10-19.9 ml/kg/day or 4-6 episodes/day 
2 
Maculopapular rash 25-50% BSA 
3.1-6 mg/dl 
-  
Adult: 1000-1500 ml/day or 5-7 episodes/day 
Child: 20-30 ml/kg/day or 7-10 episodes/day 
3 
Maculopapular rash > 50% BSA 
6.1-15 mg/dl 
- 
Adult: > 1500 ml/day or > 7 episodes/day 
Child: > 30 ml/kg/day or > 10 episodes/day 
4 
Generalized erythroderma (> 50% 
BSA) plus bullous formation and 
desquamation > 5% BSA 
> 15 mg/dl 
-  
Severe abdominal pain with or without ileus, or 
grossly bloody stools (regardless of stool 
volume) 
 
Overall clinical grade (based upon most severe target organ involvement):  
 
 
Grade 0: No stage 1-4 of any organ  
 
Grade I: Stage 1-2 skin without liver, upper GI or lower GI involvement  
 
Grade II: Stage 3 rash and/or stage 1 liver and/or stage 1 upper GI and/or stage 1 lower GI 
 
Grade III: stage 2-3 liver and/or stage 2-3 lower GI, with stage 0-3 skin and/or stage 0-1 upper GI 
 
Grade IV: stage 4 skin, liver or lower GI involvement, with stage 0-1 upper GI 
 
Abbreviations: BSA = body surface area; GI = gastrointestinal  
 
 

HAPLOTAB v2.0 (07/17/2023) 
26 
 
 
Appendix B2: Chronic GVHD Staging and Grading (NIH Consensus Grading) 
 
Chronic GVHD will be assessed at least every 3 months using the NIH Consensus staging and 
grading (see table below from Jagasia et al).  Every attempt to diagnose patients with physical 
criteria should be made.  Histologic criteria should be used if needed. 
 
Utilize chronic GVHD organ scoring system (0-3) from Jagasia et al on the following pages 
The organ and site-specific scores are then combined for a global assessment of chronic 
GVHD severity (mild, moderate, or severe).* (Table adapted from Jagasia et al.) 
 
 
 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
27 
 
 
 
 

HAPLOTAB v2.0 (07/17/2023) 
28 
 
 

HAPLOTAB v2.0 (07/17/2023) 
29 
 
 

HAPLOTAB v2.0 (07/17/2023) 
30 
 
 
*Adapted from Lee et al, 2015 
 

HAPLOTAB v2.0 (07/17/2023) 
31 
 
 
 
*Adapted from Lee et al, 2015 
 
 
Jagasia MH, Greinix HT, Arora M, Williams KM, Wolff D, Cowen EW, Palmer J, Weisdorf D, Treister NS, Cheng GS, Kerr H, Stratton P, Duarte RF, 
McDonald GB, Inamoto Y, Vigorito A, Arai S, Datiles MB, Jacobsohn D, Heller T, Kitko CL, Mitchell SA, Martin PJ, Shulman H, Wu RS, Cutler CS, 
Vogelsang GB, Lee SJ, Pavletic SZ, Flowers ME.  National Institutes of Health Consensus Development Project on Criteria for Clinical Trials in Chronic 
Graft-versus-Host Disease: I. The 2014 Diagnosis and Staging Working Group report.  Biol Blood Marrow Transplant. 2015 Mar;21(3):389-401.e1. doi: 
10.1016/j.bbmt.2014.12.001. Epub 2014 Dec 18. 
Lee SJ, Wolff D, Kitko C, Koreth J, Inamoto Y, Jagasia M, Pidala J, Olivieri A, Martin PJ, Przepiorka D, Pusic I, Dignan F, Mitchell SA, Lawitschka A, 
Jacobsohn D, Hall AM, Flowers ME, Schultz KR, Vogelsang G, Pavletic S. Measuring Therapeutic Response in Chronic Graft-versus-Host Disease. 
National Institutes of Health Consensus Development Project on Criteria for Clinical Trials in Chronic Graft-versus-Host Disease: IV. The 2014 Response 
Criteria Working Group Report.  Biol Blood Marrow Transplant. 2015 Mar 19. pii: S1083-8791(15)00155-X. doi: 10.1016/j.bbmt.2015.02.025.  

HAPLOTAB v2.0 (07/17/2023) 
32 
 
Appendix C: CMV monitoring and Prophylaxis:  
CMV prophylaxis (CMV seropositive recipients) with letermovir 480 mg daily beginning 
between day 0 and day 28; to continue until at least day 100; if letermovir is not approved 
for use in age range, consider on an individual basis after consultation with pharmacist 
and infectious diseases. 
OR  
For CMV-seronegative recipients receiving CMV-seropositive donor product can consider 
prophylaxis with ganciclovir or valganciclovir (BID for 2 weeks, then daily Monday-Friday) 
from engraftment to at least day 100. 

HAPLOTAB v2.0 (07/17/2023) 
33 
 
12.0 
REFERENCES  
1. Gragert L, Eapen M, Williams E, et al. HLA match likelihoods for hematopoietic stem-cell 
grafts in the U.S. registry. N Engl J Med. 2014;371(4):339-348. doi: 10.1056/NEJMsa1311707 
[doi]. 
2. Sun Y, Beohou E, Labopin M, et al. Unmanipulated haploidentical versus matched unrelated 
donor allogeneic stem cell transplantation in adult patients with acute myelogenous leukemia in 
first remission: A retrospective pair-matched comparative study of the beijing approach with the 
EBMT database. Haematologica. 2016;101(8):e352-4. doi: 10.3324/haematol.2015.140509 
[doi]. 
3. Bertaina A, Zecca M, Buldini B, et al. Unrelated donor vs HLA-haploidentical α/β T-cell- and 
B-cell-depleted HSCT in children with acute leukemia. Blood. 2018;132(24):2594-2607. doi: 
10.1182/blood-2018-07-861575 [doi]. 
4. Ruggeri A, Labopin M, Sanz G, et al. Comparison of outcomes after unrelated cord blood and 
unmanipulated haploidentical stem cell transplantation in adults with acute leukemia. Leukemia. 
2015;29(9):1891-1900. doi: 10.1038/leu.2015.98 [doi]. 
5. Aversa F, Terenzi A, Tabilio A, et al. Full haplotype-mismatched hematopoietic stem-cell 
transplantation: A phase II study in patients with acute leukemia at high risk of relapse. J Clin 
Oncol. 2005;23(15):3447-3454. doi: JCO.2005.09.117 [pii]. 
6. Aversa F, Tabilio A, Velardi A, et al. Treatment of high-risk acute leukemia with T-cell-
depleted stem cells from related donors with one fully mismatched HLA haplotype. N Engl J 
Med. 1998;339(17):1186-1193. doi: 10.1056/NEJM199810223391702 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
34 
 
7. Ratanatharathorn V, Logan B, Wang D, et al. Prior rituximab correlates with less acute graft-
versus-host disease and better survival in B-cell lymphoma patients who received allogeneic 
peripheral blood stem cell transplantation. Br J Haematol. 2009;145(6):816-824. doi: 
10.1111/j.1365-2141.2009.07674.x [doi]. 
8. Arai S, Sahaf B, Narasimhan B, et al. Prophylactic rituximab after allogeneic transplantation 
decreases B-cell alloimmunity with low chronic GVHD incidence. Blood. 2012;119(25):6145-
6154. doi: 10.1182/blood-2011-12-395970 [doi]. 
9. Iori AP, Torelli GF, De Propris MS, et al. B-cell concentration in the apheretic product predicts 
acute graft-versus-host disease and treatment-related mortality of allogeneic peripheral blood 
stem cell transplantation. Transplantation. 2008;85(3):386-390. doi: 
10.1097/TP.0b013e3181622e36 [doi]. 
10. Ruggeri L, Mancusi A, Capanni M, et al. Donor natural killer cell allorecognition of missing 
self in haploidentical hematopoietic transplantation for acute myeloid leukemia: Challenging its 
predictive value. Blood. 2007;110(1):433-440. doi: blood-2006-07-038687 [pii]. 
11. Sairafi D, Mattsson J, Uhlin M, Uzunel M. Thymic function after allogeneic stem cell 
transplantation is dependent on graft source and predictive of long term survival. Clin Immunol. 
2012;142(3):343-350. doi: 10.1016/j.clim.2011.12.001 [doi]. 
12. Waller EK, Langston AA, Lonial S, et al. Pharmacokinetics and pharmacodynamics of anti-
thymocyte globulin in recipients of partially HLA-matched blood hematopoietic progenitor cell 
transplantation. Biol Blood Marrow Transplant. 2003;9(7):460-471. doi: S1083879103001277 
[pii]. 

HAPLOTAB v2.0 (07/17/2023) 
35 
 
13. Perruccio K, Tosti A, Burchielli E, et al. Transferring functional immune responses to 
pathogens after haploidentical hematopoietic transplantation. Blood. 2005;106(13):4397-4406. 
doi: 2005-05-1775 [pii]. 
14. Bollard CM, Heslop HE. T cells for viral infections after allogeneic hematopoietic stem cell 
transplant. Blood. 2016;127(26):3331-3340. doi: 10.1182/blood-2016-01-628982 [doi]. 
15. André-Schmutz I, Le Deist F, Hacein-Bey-Abina S, et al. Immune reconstitution without 
graft-versus-host disease after haemopoietic stem-cell transplantation: A phase 1/2 study. 
Lancet. 2002;360(9327):130-137. doi: S0140-6736(02)09413-8 [pii]. 
16. Amrolia PJ, Muccioli-Casadei G, Huls H, et al. Adoptive immunotherapy with allodepleted 
donor T-cells improves immune reconstitution after haploidentical stem cell transplantation. 
Blood. 2006;108(6):1797-1808. doi: blood-2006-02-001909 [pii]. 
17. Guinan EC, Boussiotis VA, Neuberg D, et al. Transplantation of anergic histoincompatible 
bone marrow allografts. N Engl J Med. 1999;340(22):1704-1714. doi: 
10.1056/NEJM199906033402202 [doi]. 
18. Di Ianni M, Falzetti F, Carotti A, et al. Tregs prevent GVHD and promote immune 
reconstitution in HLA-haploidentical transplantation. Blood. 2011;117(14):3921-3928. doi: 
10.1182/blood-2010-10-311894 [doi]. 
19. Bacchetta R, Lucarelli B, Sartirana C, et al. Immunological outcome in haploidentical-HSC 
transplanted patients treated with IL-10-anergized donor T cells. Front Immunol. 2014;5:16. doi: 
10.3389/fimmu.2014.00016 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
36 
 
20. Zhou X, Dotti G, Krance RA, et al. Inducible caspase-9 suicide gene controls adverse 
effects from alloreplete T cells after haploidentical stem cell transplantation. Blood. 
2015;125(26):4103-4113. doi: 10.1182/blood-2015-02-628354 [doi]. 
21. Hale GA KK. Haploidentical stem cell transplantation with CD3 depleted mobilized 
peripheral blood stem cell grafts for children with hematologic malignancies. Blood. 
2005;106(11):2910. 
22. Hale GA KK. Mismatched family member donor transplantation for patients with refractory 
hematologic malignancies: Long-term followup of a prospective clinical trial. Blood. 
2006;108(11):3137. 
23. Handgretinger R, Chen X, Pfeiffer M, et al. Feasibility and outcome of reduced-intensity 
conditioning in haploidentical transplantation. Ann N Y Acad Sci. 2007;1106:279-289. doi: 
annals.1392.022 [pii]. 
24. Bethge WA, Faul C, Bornhäuser M, et al. Haploidentical allogeneic hematopoietic cell 
transplantation in adults using CD3/CD19 depletion and reduced intensity conditioning: An 
update. Blood Cells Mol Dis. 2008;40(1):13-19. doi: S1079-9796(07)00132-5 [pii]. 
25. Bader P, Soerensen J, Jarisch A, et al. Rapid immune recovery and low TRM in 
haploidentical stem cell transplantation in children and adolescence using CD3/CD19-depleted 
stem cells. Best Pract Res Clin Haematol. 2011;24(3):331-337. doi: 10.1016/j.beha.2011.04.005 
[doi]. 
26. Federmann B, Bornhauser M, Meisner C, et al. Haploidentical allogeneic hematopoietic cell 
transplantation in adults using CD3/CD19 depletion and reduced intensity conditioning: A phase 
II study. Haematologica. 2012;97(10):1523-1531. doi: 10.3324/haematol.2011.059378 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
37 
 
27. Chaleff S, Otto M, Barfield RC, et al. A large-scale method for the selective depletion of 
alphabeta T lymphocytes from PBSC for allogeneic transplantation. Cytotherapy. 2007;9(8):746-
754. doi: 782850553 [pii]. 
28. Handgretinger R. Negative depletion of CD3(+) and TcRαβ(+) T cells. Curr Opin Hematol. 
2012;19(6):434-439. doi: 10.1097/MOH.0b013e3283582340 [doi]. 
29. Handgretinger R. New approaches to graft engineering for haploidentical bone marrow 
transplantation. Semin Oncol. 2012;39(6):664-673. doi: S0093-7754(12)00189-3 [pii]. 
30. Carding SR, Egan PJ. Gammadelta T cells: Functional plasticity and heterogeneity. Nat Rev 
Immunol. 2002;2(5):336-345. doi: 10.1038/nri797 [doi]. 
31. Lamb LS, Lopez RD. Gammadelta T cells: A new frontier for immunotherapy? Biol Blood 
Marrow Transplant. 2005;11(3):161-168. doi: S108387910400597X [pii]. 
32. Locatelli F, Bauquet A, Palumbo G, Moretta F, Bertaina A. Negative depletion of α/β+ T cells 
and of CD19+ B lymphocytes: A novel frontier to optimize the effect of innate immunity in HLA-
mismatched hematopoietic stem cell transplantation. Immunol Lett. 2013;155(1-2):21-23. doi: 
S0165-2478(13)00150-8 [pii]. 
33. Airoldi I, Bertaina A, Prigione I, et al. Γδ T-cell reconstitution after HLA-haploidentical 
hematopoietic transplantation depleted of TCR-αβ+/CD19+ lymphocytes. Blood. 
2015;125(15):2349-2358. doi: 10.1182/blood-2014-09-599423 [doi]. 
34. Bertaina A, Merli P, Rutella S, et al. HLA-haploidentical stem cell transplantation after 
removal of αβ+ T and B cells in children with nonmalignant disorders. Blood. 2014;124(5):822-
826. doi: 10.1182/blood-2014-03-563817 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
38 
 
35. Lucarelli B MP. BPX-501 cells (donor T cells transduced with iC9 suicide gene) are able to 
clear life-threatening viral infections in children with primary immune deficiencies given 
alpha/beta T-cell depleted HLA-haploidentical hematopoietic stem cell transplantation (haplo-
HSCT). Blood. 2015;126(23):4299. 
36. Perko R, Kang G, Sunkara A, Leung W, Thomas PG, Dallas MH. Gamma delta T cell 
reconstitution is associated with fewer infections and improved event-free survival after 
hematopoietic stem cell transplantation for pediatric leukemia. Biol Blood Marrow Transplant. 
2015;21(1):130-136. doi: S1083-8791(14)00608-9 [pii]. 
37. Shelikhova L, Ilushina M, Shekhovtsova Z, et al. Αβ T cell-depleted haploidentical 
hematopoietic stem cell transplantation without antithymocyte globulin in children with 
chemorefractory acute myelogenous leukemia. Biol Blood Marrow Transplant. 2019;25(5):e179-
e182. doi: S1083-8791(19)30076-X [pii]. 
38. Foell J, Schulte JH, Pfirstinger B, et al. Haploidentical CD3 or α/β T-cell depleted HSCT in 
advanced stage sickle cell disease. Bone Marrow Transplant. 2019;54(11):1859-1867. doi: 
10.1038/s41409-019-0550-0 [doi]. 
39. Prezioso L, Manfra I, Bonomini S, et al. Haploidentical hematopoietic stem cell 
transplantation in adults using the αβTCR/CD19-based depletion of G-CSF-mobilized peripheral 
blood progenitor cells. Bone Marrow Transplant. 2019;54(Suppl 2):698-702. doi: 
10.1038/s41409-019-0608-z [doi]. 
40. Kaynar L, Demir K, Turak EE, et al. TcRαβ-depleted haploidentical transplantation results in 
adult acute leukemia patients. Hematology. 2017;22(3):136-144. doi: 
10.1080/10245332.2016.1238182 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
39 
 
41. Ruggeri L, Capanni M, Urbani E, et al. Effectiveness of donor natural killer cell alloreactivity 
in mismatched hematopoietic transplants. Science. 2002;295(5562):2097-2100. doi: 
295/5562/2097 [pii]. 
42. Ruggeri L, Aversa F, Martelli MF, Velardi A. Allogeneic hematopoietic transplantation and 
natural killer cell recognition of missing self. Immunol Rev. 2006;214:202-218. doi: MR455 [pii]. 
43. Velardi A, Ruggeri L, Mancusi A, Aversa F, Christiansen FT. Natural killer cell 
allorecognition of missing self in allogeneic hematopoietic transplantation: A tool for 
immunotherapy of leukemia. Curr Opin Immunol. 2009;21(5):525-530. doi: 
10.1016/j.coi.2009.07.015 [doi]. 
44. Diaz MA, Zubicaray J, Molina B, et al. Haploidentical stem cell transplantation in children 
with hematological malignancies using αβ(+) T-cell receptor and CD19(+) cell depleted grafts: 
High CD56(dim)/CD56(bright) NK cell ratio early following transplantation is associated with 
lower relapse incidence and better outcome. Front Immunol. 2019;10:2504. doi: 
10.3389/fimmu.2019.02504 [doi]. 
45. Bhat S NS. Outcomes of myeloablative haplo-identical hematopoietic stem cell transplant in 
pediatric patients with TCR α/β; and CD 19 depletion 
. Biol Blood Marrow Transplant. 2017;23(3, Supplement):S191-S192. 
46. Shelikhova L SZ. Tcrαβ+/CD19+-depletion in hematopoietic stem cells transplantation from 
matched unrelated and haploidentical donors following treosulfan or TBI-based conditioning in 
pediatric acute lymphoblastic leukemia patients. Blood. 2016;128(22):4672. 
47. Maschan M, Shelikhova L, Ilushina M, et al. TCR-alpha/beta and CD19 depletion and 
treosulfan-based conditioning regimen in unrelated and haploidentical transplantation in children 

HAPLOTAB v2.0 (07/17/2023) 
40 
 
with acute myeloid leukemia. Bone Marrow Transplant. 2016;51(5):668-674. doi: 
10.1038/bmt.2015.343 [doi]. 
48. Lang PJ SP. TCR-alpha/beta and CD19 depleted haploidentical stem cell transplantation 
following reduced intensity conditioning in children: First results of a prospective multicenter 
phase I/II clinical trial. Blood. 2016;128(22):389. 
49. Lang P, Feuchtinger T, Teltschik HM, et al. Improved immune recovery after transplantation 
of TCRαβ/CD19-depleted allografts from haploidentical donors in pediatric patients. Bone 
Marrow Transplant. 2015;50 Suppl 2:S6-10. doi: 10.1038/bmt.2015.87 [doi]. 
50. Locatelli F, Merli P, Pagliara D, et al. Outcome of children with acute leukemia given HLA-
haploidentical HSCT after αβ T-cell and B-cell depletion. Blood. 2017;130(5):677-685. doi: 
10.1182/blood-2017-04-779769 [doi]. 
51. Li Pira G, Malaspina D, Girolami E, et al. Selective depletion of αβ T cells and B cells for 
human leukocyte antigen-haploidentical hematopoietic stem cell transplantation. A three-year 
follow-up of procedure efficiency. Biol Blood Marrow Transplant. 2016;22(11):2056-2064. doi: 
S1083-8791(16)30281-6 [pii]. 
52. Schumm M, Lang P, Bethge W, et al. Depletion of T-cell receptor alpha/beta and CD19 
positive cells from apheresis products with the CliniMACS device. Cytotherapy. 
2013;15(10):1253-1258. doi: S1465-3249(13)00546-X [pii]. 
53. Park M, Im HJ, Lee YJ, et al. Reconstitution of T and NK cells after haploidentical 
hematopoietic cell transplantation using αβ T cell-depleted grafts and the clinical implication of 
γδ T cells. Clin Transplant. 2018;32(1):10.1111/ctr.13147. Epub 2017 Dec 8. doi: 
10.1111/ctr.13147 [doi]. 

HAPLOTAB v2.0 (07/17/2023) 
41 
 
54. Pende D, Marcenaro S, Falco M, et al. Anti-leukemia activity of alloreactive NK cells in KIR 
ligand-mismatched haploidentical HSCT for pediatric patients: Evaluation of the functional role 
of activating KIR and redefinition of inhibitory KIR specificity. Blood. 2009;113(13):3119-3129. 
doi: 10.1182/blood-2008-06-164103 [doi]. 
55. Bubalo J, Carpenter PA, Majhail N, et al. Conditioning chemotherapy dose adjustment in 
obese patients: A review and position statement by the american society for blood and marrow 
transplantation practice guideline committee. Biol Blood Marrow Transplant. 2014;20(5):600-
616. doi: S1083-8791(14)00050-0 [pii]. 
56. Bryant J, Day R. Incorporating toxicity considerations into the design of two-stage phase II 
clinical trials. Biometrics. 1995;51(4):1372-1383.