Non-Myeloablative Allogeneic Peripheral Blood Mobilized Hematopoietic Precursor Cell Transplantation for Hematologic Malignancies in High Risk Patients and in Patients With Debilitating Hematologic Diseases
Phase: PHASE2
Conditions: Myeloproliferative Disorders, Acute Myelogenous Leukemia, Chronic Myelogenous Leukemia, Myelodysplastic Syndrome, Acute Lymphoblastic Leukemia
Sponsor: National Heart, Lung, and Blood Institute (NHLBI)

Protocol with SAP for CT.gov posting:  
Official Title: 
Non-myeloablative Allogeneic Peripheral Blood Mobilized Hematopoietic 
Precursor Cell Transplantation for Hematologic Malignancies in High Risk  
Patients and in Patients with Debilitating Hematologic Diseases 
NCT number: 
 
NCT00003838 
Document Type: 
Protocol with Statistical Analysis Plan 
Document Date: 
February 22, 2022 

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99-H-0050
Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
CLINICAL RESEARCH PROTOCOL
Project: #99-H-0050 
Date:
 February 22, 2022
Title: Non-Myeloablative Allogeneic Peripheral Blood Mobilized Hematopoietic Precursor Cell 
Transplantation For Hematologic Malignancies In High Risk Patients and In Patients With Debilitating 
Hematologic Diseases 
Other Underlying Words: Peripheral blood stem cells, non-myeloablative bone marrow transplantation, 
engraftment, graft-versus-host disease, graft-versus-leukemia, graft-versus-tumor,  cyclophosphamide, 
fludarabine, donor apheresis 
Principal Investigator:    
Richard Childs, M.D., CMTB, NHLBI (E)
594-8008
Bldg 10, CRC 3-5332
Medically Responsible Investigator:  
Richard Childs, M.D., CMTB, NHLBI (E)
594-8008
Bldg 10, CRC 3-5332
Subjects of Study
Group
Number
Sex
Age range
Recipients
A
45
either
> 10 and < 75 > 8 and < 65(multiple 
myeloma)
Recipients
B
62
either
> 8 and < 80
Total Recipients
Both A and B
107
either
Donors
-
107
either
  > 2 and < 80
Project Involves Ionizing Radiation? 
Yes (only if medically indicated)
Off Site Project?
No 
Multi-Institutional Project?
No
DSMB Involvement?
Yes
 

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February 22, 2022 (Amendment III)
PRECIS 
Patients with malignant and non-malignant hematologic diseases including severe aplastic anemia (SAA), 
paroxysmal nocturnal hemoglobinuria (PNH), myelodysplastic syndrome (MDS), acute and chronic leukemias, 
Hodgkin's and non-Hodgkin's lymphoma and multiple myeloma (MM) can now be cured by allogeneic bone 
marrow transplantation (BMT).  This curative effect has been ascribed to the use of high dose chemo-
radiotherapy and the anti-tumor or anti-bone marrow effect of the allograft. Dose intensification of conditioning 
regimens in attempts to reduce disease recurrence has been largely unsuccessful because of increased toxicity 
and mortality. Indeed, most evidence now points to donor- derived T-cells as being the principal modality 
leading to the complete eradication of both malignant and non-malignant host hematopoietic cells. 
The assumption that successful allogeneic BMT relies on the myeloablative effect of intensive but hazardous 
chemo-radiotherapy has largely restricted this therapeutic modality to patients with malignant or life-threatening 
hematologic disorders under the age of 55 years. Treatment-related mortality increases substantially with age, 
prior intensive treatment with chemo-radiotherapy, worsening performance status, and co-morbid medical 
conditions. An unacceptable risk of death from conventional BMT renders many patients ineligible for what 
may otherwise be curative therapy. 
Several in vitro studies have demonstrated the existence of donor-derived CD4 and CD8 positive lymphocytes 
with specific reactivity for the patient's leukemia. These cells provide a potent graft-versus- leukemia (GVL) 
effect. This GVL effect is best seen in patients with CML relapsing after BMT, where a single infusion of donor 
lymphocytes has been shown to induce complete remission. In addition to the potent anti-leukemia effect of 
these cells, there is now strong evidence that donor T-cells are capable of completely eradicating residual host 
hematopoietic cells in a non-myeloablative transplant setting (graft-versus-marrow) leading to successful and 
complete donor hematopoietic engraftment. 
Non-myeloablative allogeneic peripheral blood stem cell transplants are currently being investigated in phase 
I/II trials assessing engraftment efficacy and toxicity at a number of transplant centers. Preliminary data, 
including our own experience with >150 patients undergoing this type of procedure, have shown a high rate of 
complete donor engraftment with a low toxicity profile. Two recent studies investigating non-myeloablative 
allo-transplantation in standard risk patients revealed an extremely low rate of transplant- related complications 
and mortality. 
The decreased risk of transplant-related complications associated with non-myeloablative transplants expands 
the eligibility of transplant candidates as well as opens the possibility to evaluate non-myeloablative regimens 
in patients at high risk for complications with standard transplantation. Besides hematologic malignancies, 
allogeneic BMT has been shown to be curative in a number of debilitating hematologic diseases which may 
behave in a relatively indolent fashion, such as paroxysmal nocturnal hemoglobinuria (PNH) and refractory 
anemia (RA) or refractory anemia with ringed sideroblasts (RARS). However, the 30% risk of treatment-related 
mortality (TRM)with standard myeloablative allotransplantation usually precludes these patients from 
potentially curative therapy, because of concerns about shortening life in patients with these disorders. In this 
protocol we investigate non-myeloablative allogeneic peripheral blood stem cell (PBSC) transplantation in two 
groups of subjects where standard allogeneic transplantation is considered to have unacceptable toxicity 
Group A: Subjects with hematologic malignancies with factors putting them at high risk for transplant related 
complications and mortality, including prior intensive chemo-radiotherapy and co-morbid diseases. 
Group B: Subjects with hematologic diseases (both clonal and non-clonal) associated with reasonable longevity 
not currently considered for allogeneic BMT because of prohibitive procedural mortality with conventional 

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February 22, 2022 (Amendment III)
BMT (enrollment closed October 2010).
In this protocol, eligible subjects are treated with an allogeneic PBSC transplant from an HLA identical or 
single HLA antigen-mismatched family donor, using an intensive immunosuppressive regimen without 
myeloablation in an attempt to decrease the transplant related toxicities while preserving the anti-malignancy 
and/or anti-host marrow effect of the graft. The low intensity non-myeloablative conditioning regimen should 
provide adequate immunosuppression to allow stem cell and lymphocyte engraftment. T -cell replete, donor-
derived, granulocyte colony stimulating factor (G-CSF)-mobilized PBSCs will be used to establish 
hematopoietic and lymphoid reconstitution. We will add back lymphocytes in recipients with <100% donor T-
cell chimerism in an attempt to prevent graft rejection and enhance a graft-versus-malignancy effect. 
The primary endpoint of this study is transplant related mortality (200 day survival). Other end points include 
engraftment, degree of donor-host chimerism, incidence of acute and chronic graft versus host disease (GVHD), 
transplant related morbidity, as well as disease-free and overall survival.

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
TABLE OF CONTENTS
 
1.0
 OBJECTIVES.............................................................................................................................................6
2.0 
BACKGROUND.........................................................................................................................................6
3.0
SCIENTIFIC AND CLINICAL JUSTIFICATION ....................................................................................8
4.0
STUDY DESIGN ........................................................................................................................................8
5.0 
ELIGIBILITY ASSESSMENT ...................................................................................................................9
6.0
CLINICAL EVALUATION OF THE RECIPIENT .................................................................................11
7.0 
TREATMENT PLAN................................................................................................................................12
7.7 
GVHD Prophylaxis....................................................................................................................................13
8.0
 DONOR EVALUATION AND PLAN....................................................................................................14
9.0 
MANAGEMENT OF RECIPIENT COMPLICATIONS .........................................................................17
10.0   STUDY MODIFICATIONS .......................................................................................................................18
11.0
ANCILLARY LABORATORY RESEARCH STUDIES.........................................................................18
12.0 
BIOSTATISTIC CONSIDERATIONS.................................................................................................20
13.0
DATA SAFETY AND MONITORING PLAN ........................................................................................24
15.0
PHARMACEUTICALS ............................................................................................................................36
REFERENCES ......................................................................................................................................................39
APPENDIX A
 PREPARATIVE REGIMEN....................................................................................................41
APPENDIX B:  Donor stem cell mobilization and apheresis. ..............................................................................42
APPENDIX C NHLBI LABORATORY RESEARCH STUDIES.......................................................................43
SEE ALSO 
Blood & Marrow Transplant Consortium Supportive Care Guidelines for Allogeneic 
Hematopoietic Stem Cell Transplant Recipients
Available online at:
http://intranet.cc.nih.gov/bmt/clinicalcare/guidelines.shtml
 

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February 22, 2022 (Amendment III)
1.0
 OBJECTIVES 
1.1. To evaluate the safety and toxicity of a low intensity non-myeloablative preparative regimen followed 
by a granulocyte colony stimulating factor (G-CSF) mobilized peripheral blood stem cell (PBSC) 
transplant in a population of subjects with malignant hematologic disorders at increased risk for 
complications with standard myeloablative allo-transplantation, and in subjects with hematologic 
diseases not usually considered for allogeneic bone marrow transplantation (BMT) because of 
prohibitive procedural mortality with conventional BMT . 
1.2. To evaluate engraftment by bone marrow chimerism analysis. 
1.3. To determine the incidence and severity of acute and chronic graft-versus-host-disease (GVHD) 
following the transplant. 
1.4. To examine the possibility of controlling hematologic malignancies by induction of a graft-versus-
malignancy effect (GVM). 
1.5. To determine the disease-free survival, relapse, transplant related mortality (TRM) and death from all 
causes.
2.0 
BACKGROUND 
2.1 
Introduction 
It is now well established that chronic myelogenous leukemia (CML), myelodysplastic syndromes (MDS), 
acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma/disease (HD) 
and non-Hodgkin's lymphoma (NHL) and multiple myeloma (MM) can be cured by allogeneic BMT(1)(2) (23). 
This curative effect has been ascribed to the use of myeloablative chemo-radiotherapy and the antileukemic 
effect of the transplant (the graft-versus-leukemia [GVL] effect) (3). The assumption that the intensive 
myeloablative preparative regimen is essential for the cure of the malignancy went unchallenged until the 
demonstration by Kolb et al, subsequently confirmed by numerous investigators, that donor lymphocytes alone 
exert a powerful antileukemic effect in the context of patients relapsing with myeloid leukemias after BMT 
(4)(5)(6)(7). This observation has important implications. First, it may be possible to cure some hematological 
malignancies with preparative regimens of lower intensity, designed to immunosuppress the recipient to allow 
lymphocyte and stem cell engraftment without major cytoreduction of the malignancy by myeloablation.  
Second, such low-intensity preparative regimens appear to have lower toxicity and may make transplantation 
appropriate in patients where procedural mortality is usually prohibitive, including patients with more indolent 
hematological diseases as well as patients with co-morbid diseases and older patients.  Several groups, 
including our own, have begun to investigate this approach to improve the applicability and outcome following 
allogeneic BMT (10)(17)(20).  Our experience at the NIH has included >150 subjects and has also been quite 
favorable, with a lower incidence of treatment-related complications than is typically observed with a standard 
“myeloablative” transplant approach. 
We now plan to extend the evaluation of low-intensity marrow stem cell transplants to include all subjects at 
high risk for treatment-related complications with standard myeloablative allotransplantation, as well as to 
subjects with a number of debilitating hematologic diseases where allotransplantation has been shown to be 
curative but is usually not performed, due to unacceptable toxicity with standard transplantation (18)(19). The 
design of the study incorporates the following features: 

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
1)
This is a phase II trial to determine the therapeutic potential of a new transplant approach (disease-free 
survival, GVM effect) and to evaluate its toxicity profile (immediate toxicity, graft-vs.-host disease, 
graft rejection, mortality).
2)
Two subject cohorts will be studied:
Group A: Heavily treated and/or debilitated subjects: 
Several groups of patients with hematologic malignancies have been found to be particularly 
poor risks for standard allogeneic transplantation due to extremely high TRM. These include 
patients with a history of previous treatment with dose-intensive chemotherapy (including 
previous allo/auto-transplant) and/or prior radiotherapy, debilitated patients with co-morbid 
diseases, and patients with MM.  The decreased procedure-related toxicity and mortality 
associated with the use of a non-myeloablative preparative regimen should be most evident in 
this high risk group where treatment related mortality is >40% in most studies.  Because the 
transplant does not incorporate intensive chemotherapy and relies instead on a graft-versus-
malignancy (GVM) and graft-versus-marrow effect, which may take months to be effective, 
only subjects in remission or with a stable or slowly proliferating disease will be selected for 
transplant. These diseases include hematologic malignancies such as all acute leukemias in 
complete remission, CML in chronic phase, MDS, CLL, MM in remission or in stable phase 
following chemotherapy, and relapsed HD and NHL.  These disorders are more common in 
older adults and at present there are no other potentially curative options for such patients. 
Group B:Debilitating hematologic disease (closed to enrollment in Oct. 2010): 
Some hematological disorders (both clonal and nonclonal) behave in a relatively indolent fashion but 
nonetheless may be associated with life-threatening neutropenia and/or significant debilitation resulting 
in poor quality of life. Patients with severe aplastic anemia (SAA), acquired or congenital pure red cell 
aplasia (PRCA), refractory anemia (RA) and refractory anemia with ringed sideroblasts (RARS) MDS 
and PNH often have recurrent needs for platelet and/or red blood cell transfusions, and may have 
associated iron overload resulting in end organ damage.  In addition, PNH patients are at increased risk 
for thrombotic events, some of which may be life threatening.  Allogeneic marrow cell transplantation 
has been shown to be curative for these patients but is not attempted in every patient who has a suitable 
donor due to concerns of unacceptable mortality associated with standard myeloablative procedures 
(18)(19). Median survival in these groups without transplantation is variable and ranges from 10 years in 
PNH patients, 7 -8 years in RA and RARS MDS patients, to 12-14 months in SAA patients failing 
immunosuppression.  The low incidence of treatment related complications and mortality associated 
with non-myeloablative PBSC transplantation now makes non-myeloablative transplantation a 
potentially curative therapeutic option with an acceptable risk profile. These disease categories include 
hematopoietic diseases such as PNH, SAA in patients who have failed immunosuppression, and RA and 
RARS MDS. 
3)
Immunosuppression without myeloablation: Subjects will receive chemotherapy sufficient to allow 
donor lympho-hematopoietic engraftment without complete marrow ablation.  If the graft is rejected, 
the subject will reconstitute autologous marrow function to pre- transplantation levels.  We will use a 
combination of two or three agents with known immunosuppressive activity in marrow transplantation: 
cyclophosphamide, which produces transient pancytopenia; fludarabine, which causes less cytopenia 
but equivalent immunosuppression; and in some subjects at increased risk for graft rejection 
antithymocyte globulin (ATG) which is potently immunosuppressive. 
4)
Peripheral blood hematopoietic progenitor cell (PBPC) transplant: An unmanipulated PBSC collection 

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
from a G-CSF-stimulated HLA-matched donor should improve the chance of engraftment because of 
the high stem cell dose (>5 x 106/kg CD34+ cells) and the presence of donor lymphocytes. Low-
intensity preparative regimens and PBPC transplants have a low incidence of severe GVHD (20). To 
further reduce the risk of GVHD, subjects will receive cyclosporine (CSA) and methotrexate (MTX) 
after the transplant. Subjects with mixed T-cell chimerism at day 30 will be tapered off CSA over 2 
weeks and subjects with 100% donor T-cell chimerism will begin a CSA taper at day 60 (through day 
100) if no GVHD is present. 
5)
Donor lymphocyte infusion (DLI): Subjects who show progressive disease in the absence of GVHD or 
who are at risk for graft failure due to incomplete donor T-cell engraftment will receive DLI (section 
7.11 ).
3.0
SCIENTIFIC AND CLINICAL JUSTIFICATION 
There is sufficient experimental evidence and clinical data to indicate that the low dose preparative regimen is 
capable of allowing lympho-hematopoietic engraftment with minimal toxicity. Cyclophosphamide alone 
permits engraftment in patients with SAA (9), and fludarabine has been used either alone or in conjunction with 
ATG and other agents to achieve engraftment (10). 
Early data indicate that non-myeloablative stem cell transplants are associated with an excellent transplant 
outcome in younger patients. The presence of high stem cell doses and donor lymphocytes is known to facilitate 
engraftment (11)(12)(13). Preliminary studies, including our own experience with PBPC transplants and low 
intensity preparative regimens, indicate that the risk of transplant related mortality appears to be lower than 
historical controls receiving a myeloablative transplant (10). 
4.0
STUDY DESIGN 
T-cell replete PBPC allograft: Subjects will receive a non-myeloablative preparative regimen of 
cyclophosphamide 60mg/kg/d x 2 days, and fludarabine 25mg/m2 intravenously (IV) over 30 minutes daily x 5 
days followed by a PBPC graft targeted to deliver >5x106 CD34+ cells/kg.  CSA and MTX for GVHD 
prophylaxis will be used initially with target CSA levels in the lower therapeutic range.  The HLA matched 
donor will receive granulocyte colony-stimulating factor (G-CSF) with apheresis collections of PBPC on day 5 
and day 6 if required.    
CSA beginning day -4 and IV MTX  on days +1, +3, and +6 will be given.  Subjects with mixed T-cell 
chimerism on day 30 will begin a CSA taper. Subjects with 100% donor T-cell chimerism by day 30 will be 
tapered off CSA from days 60 through 100 (25% reduction in dose every 10 days-off by day 100).  CSA will 
not be tapered in any subjects with grade > II acute GVHD regardless of chimerism results.   In addition, 
subjects with evidence of disease progression without grade > II GVHD will have CSA discontinued regardless 
of chimerism results.
On day +30, +60 and +100 disease status will be assessed and subjects with disease progression or relapse will 
be treated on protocol as indicated (Section 7.11).  
In addition, on days +30, +60 and +100 the chimeric status of subjects will be assessed by 
microsatellite analysis of the peripheral blood and sometimes bone marrow. Subjects demonstrating less than 
100% donor T-cell chimerism on day 30 will be tapered off CSA over a 12 day period and will be re-evaluated.  

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
Should T-cell chimerism remain less than 100% donor, then infusions of previously collected and 
cryopreserved donor lymphocytes will be given to subjects (in the absence of grade > II GVHD) in an attempt 
to prevent graft rejection. 
5.0 
ELIGIBILITY ASSESSMENT
 
5.1 
Inclusion Criteria-Recipients
5.1.1 
Group A: Subjects at high risk for transplant related complications and mortality as defined below: 
Ages 10 to 75 (both inclusive) with a history of one of the following: 
1)
Treatment with dose intensive chemotherapy and/or radiotherapy 
2) Previous history of allo/auto transplant 
3) History of multiple myeloma or extramedullary plasmacytoma 
4) Chronic disease or co-morbid medical condition including subjects with symptoms or signs of 
significant pulmonary disease, hepatic disease, kidney disease, cardiac disease or disease of other 
organ systems which would result in increased risk of morbidity or death from a standard 
myeloablative transplant. 
Diseases to be included: 
1)   CML, chronic phase 
2) Acute lymphoblastic leukemia (ALL), all subjects in complete or partial remission 
3) AML: AML in first complete or partial remission.  Exceptions: AML with good risk karyotypes: 
AML M3 t(15:17), AML M4Eo (inv. 16), AML t(8;21).  All AML in second or subsequent 
complete remission 
4) MDS: refractory anemia with excess blasts (RAEB), or chronic myelomonocyte leukemia (CMML)  
5) Myeloproliferative diseases associated with either cytopenia or uncontrolled proliferation 
6) CLL or small lymphocytic lymphoma (SLL) with bulky or progressive disease despite prior 
treatment with chemotherapy which includes purine analogs 
7) NHL 
A) Intermediate or high grade relapsed or progressive despite treatment with standard therapy 
ineligible for autologous PBSC transplant
B) NHL intermediate or high grade relapsing despite prior autologous transplant
C) Low grade follicular or small lymphocytic lymphoma: (1) high risk patients who have relapsed 
following conventional chemotherapy, (2) relapsed following autologous marrow or PBSC 
transplant, or (3) chemo-resistant disease
D) Mantle cell lymphoma
E) NHL intermediate or high grade with concurrent BCL2 and MYC translocations who are at high risk 
for relapse and who have low survival with conventional chemotherapy (25-29)8)
HD, relapsed 
after prior autologous transplant or after 2 or more combination chemotherapy regimens and 
ineligible for autologous PBSC transplant
9) EBV driven lymphoproliferative disorders progressing despite standard therapies
10) MM: MM subjects must be between the ages of 8 and 65 (both inclusive) 
11) Mycosis fungoides, which has been shown to be amenable to allogeneic stem cell transplants.
5.1.2 
Group B (closed to enrollment Oct. 2010): Subjects with hematologic diseases associated with 
reasonable longevity, shown to be curable by allogeneic BMT but where concern for a high procedural 
mortality with conventional BMT may delay or prevent such treatment. 
Ages 8 to 80 (both inclusive) with a history of one of the following  

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February 22, 2022 (Amendment III)
1) PNH associated with either life- 
threatening thrombosis, cytopenia, transfusion dependence or recurrent and debilitating hemolytic 
crisis 
2)
Aplastic anemia or PRCA  (acquired or congenital) in subjects associated with transfusion 
dependence and/or neutropenia who are  not candidates for or who have failed immunosuppressive 
therapy
3)
RA or RARS MDS subjects who have associated transfusion dependence and/or neutropenia.  
 
5.1.3
Ability to comprehend the investigational nature of the study and provide informed consent.  The 
procedure will be explained to subjects age 8 -17 years with formal consent being obtained from parents 
or legal guardian.
5.1.4 
Availability of HLA identical or single HLA locus mismatched family donor. 
5.2 
Inclusion Criteria -Donor 
5.2.1
HLA identical or single HLA mismatched family donor.
5.2.2
Age > 2 up to 80 years old. 
5.2.3
Weight > 18 kg
5.2.4 
Ability of donor or guardian of donor to comprehend the investigational nature of the study and provide 
informed consent.
5.3 
Exclusion Criteria-Recipient: any of the following: 
5.3.1 
Pregnant or lactating 
5.3.2     Group A: age < 10 or >75 (multiple myeloma age <8 or > 65)
Group B: Age < 8 or >80 years 
5.3.3     ECOG performance status of 3 or more (See NIH Bone and Marrow Consortium Supportive Care 
Guidelines for Allogeneic Hematopoietic Stem Cell Transplant Recipients -  
http://intranet.cc.nih.gov/bmt/_pdf/ECOG_Karnofsky_Lansky_Scales.pdf)
5.3.4    Psychiatric disorder or mental deficiency severe as to make compliance with the BMT treatment 
unlikely and making informed consent impossible
5.3.5
Major anticipated illness or organ failure incompatible with survival from PBSC transplant
5.3.6
Diffusion capacity of carbon monoxide (DLCO) < 40% predicted 
5.3.7 
Left ventricular ejection fraction: < 30% 
5.3.8 
Serum creatinine > 2.5mg/dl or creatinine clearance < 50 cc/min by 24 hr urine collection 
5.3.9 
Serum bilirubin > 4 mg/dl, transaminases > 5x upper limit of normal
5.3.10 Other malignant diseases liable to relapse or progress within 5 years, with the exception of a separate 
hematologic malignancy where allogeneic stem cell transplant is shown to be potentially curative.
5.4
Exclusion Criteria-Donor: any of the following
 
5.4.1 
Pregnant or lactating 
5.4.2
Donor unfit to receive G-CSF and undergo apheresis (uncontrolled hypertension, history of congestive 
heart failure or unstable angina, thrombocytopenia) 
5.4.3 
HIV positive donor. Donors who are positive for hepatitis B (HBV), hepatitis C (HCV) or human T-cell 
lymphotropic virus (HTLV-I/II) will be used at the discretion of the investigator following counseling 
and approval from the recipient.

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6.0
CLINICAL EVALUATION OF THE RECIPIENT 
-Human biologic materials are collected as follows for the clinical evaluation and management of the subject. 
Bone marrow aspirates will be read by a pathologist.  Samples will be ordered and tracked through the CRIS 
screens.  Should a CRIS screen not be available, the NIH form 2803-1 will be completed and will accompany 
the specimen and be filed in the medical record.
6.1 
Pre-Study Evaluation 
6.1.1 
High resolution molecular HLA- A, B, DR typing 
6.1.2 
Bone marrow aspiration and biopsy with chromosome analysis, PCR, and flow cytometry as 
appropriate to stage and classify underlying disorder.  Myeloma subjects will additionally have an 
SPEP, serum immunofixation, urine protein electrophoresis, urine immunofixation, quantitative 
immunoglobulins and 24 hour urine for creatinine clearance and protein evaluation. PCR to detect 
BCR.ABL when appropriate
6.1.3 
Antibody screen for HBV, HCV, HIV, HTLV-VII, CMV, EBV, toxoplasma, syphilis and VZV 
serology. Consider PPD test for subjects from areas where tuberculosis is prevalent
6.1.4
Coagulation screen, CBC with differential 
6.1.5 
Acute care panel, hepatic panel and mineral panel
6.1.6
24 hour creatinine clearance
6.1.7 
- Pulmonary function testing: vital capacity FEV-1, DLCO 
6.1.8 
Sinus CT scan. Lymphoma subjects will additionally have baseline CT scans of the chest, abdomen, 
pelvis and any other areas of evaluable disease within 30 days of the transplant.MM subjects will, in 
addition, undergo a baseline skeletal series. 
6.1.9   
Cardiac function: EKG, - and baseline echocardiogram for all subjects 
6.1.10
H&P and ECOG status - All subjects age 50 or greater, or age 40 or over, with one of the following 
risk factors: a history of high blood pressure or increased cholesterol, family history of coronary 
disease, smoking or diabetes, will have a baseline cardiac workup, which will include:
1.  Stress nuclear perfusion imagery
2. Cardiac consultation in subjects age 50 and over
6.1.11
Nutritional assessment 
6.1.12 
Dental review 
6.1.13 
Social worker interview and complete durable power of attorney form 
6.1.14 
Ophthalmology consultation 
6.1.15 
Interview with members of primary care team and visit to unit 
6.1.16 
Consent form signed 
-
6.1.18   Pregnancy test for women of child bearing potential
6.2 
In-Patient Monitoring 
Once daily: CBC with differential, acute care, hepatic, and mineral panels, direct bilirubin, temperature, pulse, 
blood pressure, respiratory rate, weight, caloric intake, abdominal girth.
Twice weekly: reticulocytes, pre-albumin, coagulation screen.  
Weekly: 
CMV surveillance, C-reactive protein 
Stool for C difficile when appropriate 
Drug levels when appropriate (e.g., gentamicin, vancomycin, CSA).

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6.3 
Follow Up to Day 100: Out-patient 
At least weekly: CBC, acute care panel, hepatic and mineral screen, temperature, pulse, blood pressure, 
respiratory rate, weight; C-reactive protein: CMV surveillance.  A complete physical exam will be repeated 
weekly. 
Peripheral blood will be drawn and in some recipients a bone marrow aspirate will be taken on days +15,  +30, 
+45, +60 and + 100 to assess for donor-host chimerism in the lymphoid and myeloid cell lines. A sample of 
peripheral blood will also be obtained for chimerism analysis at the time of neutrophil recovery (ANC > 500). 
Day 30, 60, and 100:  Recipients with hematological malignancies or where clinical conditions warrant, will 
have bone marrow aspirate to quantitate engraftment.  All recipients will receive a repeat PFT on day 30.
MM recipients will have staging labs repeated and appropriate radiographic studies. 
Lymphoma recipients (HD, NHL, CLL with adenopathy, and SLL recipients) will have baseline CT scans of 
the chest, abdomen and pelvis performed pre-transplant and on days 30, 60, 100.
6.4 
Beyond Day 100 
At 6, 12, 18, 24, 36, 48 and 60 months (+/- 1 month): CBC, acute care panel, hepatic and mineral screen, 
ferritin, iron, TSH, T4, LH, quantitative immunoglobulins, CD4+ count. Estradiol, testosterone, FSH, chest 
radiograph, bone marrow aspirate or blood sample for morphology, chimerism studies, and karyotype, 
pulmonary function, and an ophthalmology assessment.   
Repeat staging labs in myeloma recipients. 
Lymphoma recipients (HD, NHL, CLL with adenopathy, and SLL recipients) will have CT scans every 3 
months for the first 2 years (after day 100), then at 6 to 12 month intervals until 5 years post transplant.
After 5 years follow up visits are not mandatory, but yearly communication with the recipient and the referring 
physician is continued.
7.0 
TREATMENT PLAN 
7.1 
Pre-transplant Treatment of MDS, Leukemia, MM, and Lymphoma 
Recipients with active disease may receive appropriate chemotherapy according to standard indications to 
control disease prior to the preparative regimen. 
7.2 
Apheresis of Recipient
One collection of 1010 leukocytes by a 10-liter leukapheresis before transplant preparative regimen begins and at 
the time of disease response or acute graft-vs.-host disease for cryopreservation of leukemia cells and/or 
lymphocytes to be used for laboratory investigation of graft-versus-malignancy, graft-versus-marrow, or GVHD 
effectors. 

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7.3 
Central Venous Line Placement 
A triple lumen Hickman catheter will be placed by an interventional radiologist or a surgeon. 
7.4 
Infection Prophylaxis (See Supportive Care Guidelines)
7.4.1 Strongyloides prophylaxis:  Ivermectin, will be given per NIH Bone and Marrow Consortium Supportive 
Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant Recipients prior to transplant as 
prophylaxis against Strongyloides stercoralis (threadworm), a known human parasite (recipients at risk).
7.5
Preparative Regimen (see Appendix A) 
7.5.1 
Antithymocyte Globulin (ATG or Atgam) (See also NIH Bone and Marrow Consortium Supportive 
Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant Recipients)
Subjects will receive one of two preparative regimens as described below.  Recipients at higher risk for 
graft rejection with this non-myeloablative approach (for example, subjects with aplastic anemia, 
heavily transfused subjects and single HLA locus mismatched subjects) will have ATG added to the 
preparative regimen unless a contraindication exists ( i.e. prior life threatening toxicity associated with 
ATG treatment).
7.5.2 Dose Levels
1) 
Cyclophosphamide 60 mg/kg/d IV day -7, -6 
Fludarabine 25 mg/m2/d IV day -5, -4, -3, -2, -1 
2) 
Aplastic anemia or recipients at increased risk for graft rejection as defined above, 7.5.1 
Cyclophosphamide 60 mg/kg/d IV day -7, -6 
Fludarabine 25 mg/m2/d IV day -5, -4, -3, -2, -1 
Antithymocyte globulin 40mg/kg IV days -5,-4,-3,-2 
7.6
Peripheral Blood Progenitor Cell Transplant (See also Appendix B)
The target for progenitor cells is >5 x 106 CD 34+ cells/kg.
Minimum dose for transplant is 2 x 106 cells/kg.
7.7 
GVHD Prophylaxis 
(For grading - see NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell 
Transplant)
All recipients will receive CSA and MTX as GVHD prophylaxis. CSA (1.5 mg/kg/dose IV BID) will begin on 
day -4. Dosing will be based on actual body weight unless the recipient is “obese” (BMI > 35) when practical 
weight will be used.  Switch to the equivalent oral CSA dose divided bid on day 14 or when able to tolerate PO.   
CSA doses will be adjusted to achieve serum trough levels of 200-400 mcg/mL.  MTX (5 mg/m2/dose) will be 
given IV on days +1, +3 and +6.  Refer to NIH Bone and Marrow Consortium Supportive Care Guidelines for 
Allogeneic Hematopoietic Stem Cell Transplant Recipients for MTX dose adjustments based on serum 
creatinine and total bilirubin values on the day of administration. 
Recipients with <100% donor T-cell chimerism on day 30 will begin a 12  day CSA taper (i.e. 25% reduction in 

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February 22, 2022 (Amendment III)
dose every 3 days). Recipients with 100% donor T-cell chimerism by day 30 will be tapered off CSA from days 
60 through 100 (25% reduction in dose every 10 days-off by day 100).  CSA will not be tapered in any 
recipients with grade > II AGVHD regardless of chimerism results.  In addition, recipients with evidence of 
disease progression without grade > II acute GVHD will have CSA tapered regardless of chimerism results.
7.8 
Transfusion Support 
(See - NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant):  
Filtered and irradiated blood products. CMV negative recipients of CMV negative or positive marrow will 
receive CMV negative blood products. 
7.9
Nutrition: 
Parenteral nutrition will be instituted if daily caloric intake <1000KCal/day in recipients > 18 years. Recipients 
8 to 18 years will start parenteral nutrition if they are unable to meet at least half their daily caloric needs by 
mouth (i.e. 50 KCal/kg/day). 
7.10
Hospital Discharge:  
Recipient will be in the hospital for about 4 weeks and will be discharged when the following criteria are 
fulfilled: 

Recipient afebrile, positive weight balance without requirement for parenteral nutrition

Platelet transfusion requirement absent or less than 2 x weekly

Recipient or family able to care for Hickman line 
7.11
Donor Lymphocyte Transfusions:  
Recipients with disease progression or donor T-cell chimerism of <100% following CSA tapering without grade 
> II GVHD will receive donor lymphocyte infusions (previously collected and cryopreserved) based on the 
following algorithm:
7.11.1  Mixed chimerism without disease progression
 
Monthly increments of DLI 5 x 106, 1 x 107, 5 x 107 CD3+ cells/kg.
7.11.2  Disease progression (independent of chimerism status)
DLI 1 x 107 CD3+ cells/kg and repeat doses depending on response and acute GVHD.
8.0
 DONOR EVALUATION AND PLAN
8. 1 Pre-study Consult and Evaluation 
8.1.1
Pregnancy test for women of child bearing potential
8.1.2
HLA-A,-B,-DR typing 
8.1.3
Confirm HLA identity of donor with recipient 
8.1.4
History and physical examination 
8.1.5
Chest X-ray in donors with underlying pulmonary disease or history of smoking
Hepatitis B, C, HIV, HTLV I/II, CMV antibodies, RPR, VZV serology, anti- T. cruzi, West Nile 
virus NAT (nucleic acid test) and human transmissible bovine spongiform encephalopathy (BSE is 
done by questionnaire of associated risk factors)

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February 22, 2022 (Amendment III)
8.1.6
Red blood cell ABO group, Rh type, antibody screen
8.17.
CBC, coagulation screen, acute care, hepatic, mineral screen 
8.1.8
If eligible: Orientation - visit to Department of Transfusion Medicine inspection of veins to determine 
the need for a central line for apheresis. 
8.2 
Pre-consent Evaluation and Concurrent Care of Minor Donors (donors less than age 18 only) 
For donors less than age 18, a social worker and mental health specialist (psychologist or psychiatrist) will 
meet with the minor prior to the assent process to determine willingness to participate.
Donors who are less than 18 years of age will see a pediatric provider (pediatrician, pediatric nurse 
practitioner or pediatric physician’s assistant) who is separate from the transplant team. This practitioner will 
serve as the donor’s health care provider and advocate during the minor’s participation on the clinical trial.
8.3
Consent to Undergo G-CSF Mobilization and Donate Leukocytes 
8.4 
PBPC Collections (See Appendix B) Algorithm for mobilization, collection, and processing:
8.4.1 Minor donor mobilization with G-CSF (≤ 18 years old)
After medical evaluation and clearance for suitability as an allogeneic donor by the BMT service in consultation 
with DTM, the donor will undergo mobilization with filgrastim (G-CSF) as an outpatient.  G-CSF will be 
administered based on body weight (see below) for at least 5, and up to 7 days, subcutaneously. 
Dosing will be based on actual body weight unless the donor is “obese” (BMI > 35) when practical weight will 
be used.   G-CSF will be administered according to a vial based algorithm to reduce wastage, improve donor 
compliance, and optimize CD34+ yields. The doses for days 1-4 may be given at any time of day, but the doses 
for days 5 (and if necessary, days 6 and 7) must be given very early in the morning, prior to apheresis.  
Predictable side effects of G-CSF including headache, bone pain, and myalgia, will be treated with 
acetaminophen or ibuprofen. Prophylactic treatment of these side effects with the same medications may be 
elected.  Other side effects will be evaluated and treated accordingly.
 
Donor Wt          
Total G-CSF 
Dose (range)
19-30.9 kg
300 mcg  
(10.0 to 15.8 mcg/kg)
31-37.9 kg
480 mcg  
(10.0 to 13.0 mcg/kg) 
38 – 48.9 kg          
600 mcg 
(12.5 to 15.8 mcg/kg)
49 – 56.9 kg           
780 mcg 
(13.9 to 15.9 mcg/kg)
57 – 60.9 kg        
900 mcg 
(15.0 to 15.8 mcg/kg)
61 – 67.9 kg           
960 mcg 
(14.3 to 15.7 mcg/kg)
68 – 108.9 kg         
1080 mcg 
(10.0 to 15.9 mcg/kg)
> 109 kg             
1200 mcg 
(11.0 or less)
8.4.2  Adult donor mobilization with G-CSF (> 18 years old) – per NIH BMT Consortium Supportive Care 
Guidelines for Allogeneic Hematopoietic Stem Cell Transplant Dose algorithms:  Two dosing algorithms are 
recommended for use in the Clinical Center. The standard dose algorithm uses doses in the range of 10 to 12 
mcg/kg /day, with a higher dose given to lighter weight adult donors to improve CD34+ yields. The higher 
dose algorithm is intended for use in (1) adult autologous donors who have received prior myelotoxic agents; 
(2) adult donors whose components will undergo further processing; (3) protocols in which a high transplant 
cell dose is required (CD34+  > 8 x106); and (4) situations with large weight discrepancy between the adult 

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February 22, 2022 (Amendment III)
donor and recipient (recipient > donor) (as assessed by the Principal Investigator). The total dose in both 
regimens is capped at 1200 mcg/day. The Principal Investigator (PI) or his designee will determine the dosing 
algorithm to be utilized.
I. Standard-Dose Filgrastim Algorithm 
Donor Weight 
Total Daily Filgrastim 
Dose (range) 
38 – 60.9 kg 
600 mcg 
(10.0 to 15.8 mcg/kg)
61 – 78.9 kg 
780 mcg 
(10.0 to 12.8 mcg/kg)
79 – 90.9 kg 
900 mcg
(10.0 to 11.4 mcg/kg)
91 – 96.9 kg 
960 mcg 
(10.0 to 10.5 mcg/kg)
97 – 108.9 kg 
1080 mcg 
(10.0 to 11.0 mcg/kg)
 > 109 kg 
1200 mcg 
(11.0 or less) 
II. Higher-Dose Filgrastim Algorithm 
Donor Weight 
Total Daily Filgrastim 
Dose (range) 
38 – 48.9 kg
600 mcg 
(12.5 to 15.8 mcg/kg)
49 – 56.9 kg
780 mcg
(13.9 to 15.9 mcg/kg)
57 – 60.9 kg
900 mcg
(15.0 to 15.8 mcg/kg)
61 – 67.9 kg
960 mcg
(14.3 to 15.7 mcg/kg)
68 - 108 kg
1080 mcg
(10.0 to 15.9 mcg/kg)
 
> 109 kg
1200 mcg
(11.0 or less)
Collection and processing of donor cells
Donors will receive divalent cation prophylaxis to prevent citrate toxicity during apheresis, in accordance with 
standard DTM policies.  The volume processed per apheresis procedure will be determined by DTM medical 
staff on the day of apheresis, based on peak CD34+ cell mobilization response to GCSF and the CD34+ cell 
dose needed, based on kilogram weight of recipient.  This will range from 15 to 35 liters processed per day for 1 
to 3 days, not to exceed a total of 75 liters over 3 days.  In pediatric subjects, defined as less than 40 kg, a 
maximum of 8 blood volumes will be processed per day, for up to 1-3 days.
Obtaining a dose of > 3x106 CD34+ cells/kg will be considered adequate, and no further apheresis will be 
performed. If the target CD34+ cell dose of 3 x 106/kg is not reached, then a 6th (and possibly 7th) dose of G-
CSF will be given; and an apheresis procedure will be performed on day 6 (and possibly day 7).  
If the minimum CD34+ cell dose is not achieved with one mobilization and 3 apheresis collections (i.e.2 x 106 
CD 34+ cells/kg), then the PI will consider options of a bone marrow collection, a second course of 
mobilization and apheresis, or no further donor collections and taking the subject off protocol.
The PBSC collections will not be manipulated except for plasma removal or red blood cell depletion as needed 
for ABO or other red blood cell antigen incompatibility.  PBSC will be either cryopreserved for later thawing 
and infusion, or infused as fresh products within 48 hours of collection.

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February 22, 2022 (Amendment III)
On rare occasions, recipients may develop marrow failure as a consequence of graft-vs.-host hematopoiesis or 
from post transplant drug treatment (i.e. ganciclovir).  When such situations arise, recipients may require a stem 
cell “boost”. Donors will undergo a repeat mobilization with G-SCF (as outlined above) followed by CD34+ 
selection of hematopoietic progenator cells prior to being infused.
8.5 
Donor Lymphocyte Collection 
Prior to undergoing G-CSF mobilization, the donor will undergo a ten liter apheresis for cryopreservation of 
donor lymphocytes to be infused as necessary after transplantation (section 7.11). Aliquots of 5 x 106 CD3+ 
cells/kg, 1 x 107 CD3+cells/kg and 5 x 107 CD3+cells/kg will be cryopreserved. 
9.0 
MANAGEMENT OF RECIPIENT COMPLICATIONS 
The major complications are cytomegalovirus reactivation, acute GVHD, chronic GVHD, relapse of the original 
disease, donor-recipient ABO incompatibility and pulmonary engraftment syndrome.  Recipients with these 
complications will be treated along the following lines: 
9.1 
CMV Reactivation 
(See NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant)  
9.2 
Acute GVHD 
(See NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant)  
 
9.3 
Chronic GVHD 
(See NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell Transplant)
Cyclosporine at standard dose (see Pharmaceuticals, section 15.3). 
Prednisone 20-60 mg daily according to severity. 
Change to alternate day steroid and cyclosporine therapy when response is established. 
Non-responding recipients may be treated with other standard of care therapies such as azathioprine, sirolimus, 
tacrolimus, rituximab, mycophenolate, daclizumab or infliximab,  PUVA, photopheresis, and/or thalidomide at 
the discretion of the attending physician. 
9 .4 Graft Rejection
This transplant protocol uses a non-myeloablative preparative regimen.  Therefore, auto-recovery is 
anticipated in recipients who fail to engraft.  Recipients who fail to demonstrate donor T-cell engraftment will 
be taken off study and referred back to their primary physician for further therapy. 
9.5
Relapse of Hematologic Malignancy:  
Recipients may be treated at the discretion of the PI with donor lymphocyte transfusions, standard of care 
treatment approaches ( i.e. interferon-alpha, GM-CSF, low dose IL-2 by subcutaneous injection, Rituximab, 
chemotherapy, Gleevec)  with or without stem cell rescue. 

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February 22, 2022 (Amendment III)
9.6
Donor-recipient ABO Incompatibility  
(See NIH BMT Consortium Supportive Care Guidelines for Allogeneic Hematopoietic Stem Cell 
Transplant)
9.7
Pulmonary Engraftment Syndrome:  
Recipients who develop pulmonary engraftment syndrome (around 10-14 days post-transplant) will be treated 
with steroids.  
10.0   STUDY MODIFICATIONS 
Graft failures will be monitored throughout the study. Specific need for preparative regimen modifications 
based on the incidence of graft failures will be made as discussed in section 7.5.
11.0
ANCILLARY LABORATORY RESEARCH STUDIES 
11.1 Sample Collection 
During the course of participating on this study, blood and tissue as detailed below will be collected for 
correlative laboratory research studies: 
 11.1.1
Recipient before transplant 
One apheresis collection of approximately 1010 leukocytes in a volume of 200 ml.
11.1.2
Recipient in hospital: Weekly lymphocyte subsets 
11.1.3
Planned outpatient visits: Cytotoxic T-cell precursor frequency (CTLPf) assays and 
lymphocyte subsets on day 60 and 100 then at each scheduled follow-up. 
11.1.4
Donor at time of leukapheresis
Donor leukocytes obtained at leukapheresis will be used for research studies investigating graft-versus-
leukemia effects. The recipient's leukemia cells and normal lymphocytes will serve as targets for all 
subsequent testing of donor-anti host responses. 
11.1.5
Recipient bone marrow samples 
An extra volume (up to 25 ml) of bone marrow aspirate will be collected for research studies at the pre-
transplant evaluation and for monitoring minimal residual disease. The cells will be used to investigate 
lymphocyte interactions with bone marrow progenitor cell proliferation. No marrow aspirates solely for 
research purposes are planned. 
11.1.6    Pulmonary function tests
Pulmonary status at baseline, day 30 and after day 100 will be analyzed to see if reduced intensity 
conditioning (RIC) effects lung function before day 100 or if the decline in lung function is only seen later 
consistent with bronchiolitis obliterans syndrome (BOS).
11.2 Intended Use

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February 22, 2022 (Amendment III)
These specimens will not be read by a pathologist or used for diagnostic purposes.   Studies will not be used 
in assessing the primary endpoint, but are undertaken for descriptive or exploratory ancillary research as 
detailed below: 
11.2.1
GVL and GVHD studies:  Blood and serum from recipients and donors will be obtained by 
leukapheresis to study genomic and proteomic pathways involved in mediating GVHD and in vitro donor 
T-cell responses to recipients leukemia. The cytotoxic T-cell precursor frequency (CTLPf) assay will be 
used to evaluate the donor immune function at various periods after BMT. 
11.2.2
Chimerism studies:   PCR of microsatellites of post-transplant peripheral blood T-lymphocytes, 
B-cells, NK cells and blood myeloid cells will be assessed in serial fashion to determine the chimeric status 
of the recipient as a function of time. These studies will be repeated after lymphocyte infusions to assess 
their effect on host chimeric status as well. The relationship between degrees of donor host chimerism, 
GVHD and tumor response will be analyzed.
11.2.3
Nephrotic syndrome studies:    We have reported an unusually high incidence of nephrotic 
syndrome (6.1% with a median follow up of 2.8 years) in recipients following 
fludarabine/cyclophosphamide conditioning prior to undergoing non-myeloablative stem transplantation. In 
order to understand the pathogenesis of this condition in this recipient population, samples from recipients 
and when available their associated donors may be sent to Pierre Ronco,  M.D., Ph.D., Head, Nephrology 
Department,  Unit 702 At Tenon Hospital, Paris, France  tel : + 33 1 56 01 66 39/38 
pierre.ronco@tnn.aphp.fr  for further evaluation.  We will use a numerical code to indicate if the samples 
were drawn before the onset of or during the nephrotic syndrome.  Samples will be free of any identifying 
information in order to protect the identity of the subjects providing samples.  
Transplant recipients and their associated donor will be asked to sign the revised consent documenting their 
approval to have an anonymized sample of their blood sent to an investigator outside the NIH. See section 
14.5 for telephone consenting procedure to be utilized if recipient is unavailable for an NIH visit.  A 
material transfer agreement (MTA) will be in place prior to transfer of any anonymized samples to this 
external lab.
11.2.4 Ex vivo expanded autologous double negative (DN) T cells:   In order to determine the feasibility and 
safety of using ex vivo expanded autologous double negative (DN) T cells as a novel cell therapy for 
treating leukemia, it is necessary to determine whether DN T cells can be expanded from 
apheresis/peripheral blood mononuclear cells (PBMCs).  It is critical to understand if these ex vivo 
expanded DN T cells kill autologous tumor cells but not normal bone marrow cells.    To address these 
questions, Dr. Zhang at University of Toronto will utilize apheresis/PBMC samples from NIH healthy 
donors to isolate and expand their DN T cells ex vivo.  The expanded DN T cells will then be utilized in an 
assay to determine their ability to kill enriched autologous AC133+ cells (provided by NIH) as well as 
autologous EBV-transformed B cells (provided by NIH).  This will be evaluated both in vitro and in NOD-
SCID mice.  The goals are twofold:
1.  To determine if DN Tcell expansion is feasible from healthy donor apheresis/PBMC.  
2.  Determine DN T cell toxicity to autologous Epstein Barr Virus transformed B cells and normal 
autologous AC133+ cells both in vitro and in a NOD-SCID mouse model.
Other research labs as described on the NHLBI Laboratory Research Studies submitted to and reviewed by 
the IRB (Appendix C).
Etiology of post-transplant syndromes: We will share de-identified blood plasma and/or serum with Dr. 

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February 22, 2022 (Amendment III)
Waldman at NIDDK.  Dr. Waldman will use blood plasma and or serum to conduct studies on the etiology 
of post-transplant syndromes that cause renal failure including nephrotic syndrome, TMA, and 
glomerulonephritis.
We will share the following de-identified samples to Dr. Li Zhang, Senior Scientist, Toronto General 
Research Institute (TGRI) at University Health Network: peripheral blood stem cells (PBSC), peripheral 
blood mononuclear cells (PBC,) coded as TRA-PBMC, GRE-PBMC and WYN-PBMC, and coded Epstein-
Bar-Virus (EBV) transformed lymphoblastoid cell line (LCL), coded as TRA-EBV-LCL, GRE-EBV-LCL 
and WYN-EBV-LCL,  derived from healthy donors enrolled in protocols 97-H-0196 and 99-H-0064 and 
99-H-0050 .
We may share the following coded samples with Dr. Michael Nishimura at Loyola University Chicago, 
Chicago Illinois: frozen PBMC from healthy donors from this protocol to sequence the T cell receptor 
(TCR) genes and clone them into retroviral vectors for the transduction of T cells.
11.3  Tracking, Storage and Disposition of Samples
Storage:  Research samples will be stored coded in the secure laboratory of the PI. 
Tracking:  Samples will be ordered and tracked through the CRIS research screens.  Should a CRIS screen not 
be available, the NIH form 2803-1 will be completed and will accompany the specimen and be filed in the 
medical record.  Specimens will be entered in the NHLBI Biospecimen Inventory System (BSI). Samples will 
not be sent outside NIH without IRB notification and an executed MTA. 
End of study procedures:  Samples from consenting subjects will be stored until they are no longer of 
scientific value or if a subject withdraws consent for their continued use, at which time they will be destroyed. 
Loss or destruction of samples:  Should we become aware that a major breech in our plan for tracking and 
storage of samples has occurred, the IRB will be notified.
11.4  Technology Transfer:
This protocol has the following MTAs
Between NHLBI and Pierre Ronco for the transfer of samples to Pierre Ronco,  MD, PhD, Head, Nephrology 
Department,  Unit 702 At Tenon Hospital, Paris, France  tel : + 33 1 56 01 66 39/38 pierre.ronco@tnn.aphp.fr 
Between NHLBI and Li Zhang, M.D., Ph.D.  Professor Departments of Lab Medicine& Pathobiology, 
Immunology University of Toronto , Toronto General Hospital, Norman Urquhart , 1st Floor Rm 1NU119, 200 
Elizabeth St., Toronto, Ontario,  Canada M5G 2C4.
Between NHLBI and Dr. Michael Nishimura, Loyola University Chicago, Chicago, Illinois.
12.0 
BIOSTATISTIC CONSIDERATIONS 

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
12.1   Design and Sample Size  
This trial is designed to estimate the 200 day survival proportion in each of Groups A and B.  The expected 200 
day survival for Group A (recipients between ages 10 and 75 (MM recipients between 8 and 65)  inclusive who 
are at high risk for transplant-related complications and mortality) is 60% and for Group B (debilitating non-
malignant hematological diseases) is 70%.   Up to forty-five recipients will be enrolled in group A.  Up to 62 
subjects could be enrolled in group B (enrollment closed Oct. 2010 with 57 subjects).   After all recipients have 
been followed for 200 days, a 95% confidence interval for the 200 day survival of each subgroup of will be 
given based on the Kaplan-Meier estimate and Greenwood’s formula for the variance (21).  This sample size will 
assure that the true 200 day survival proportion will be estimated within approximately + 14% with 95% 
confidence if the Kaplan-Meier estimate of 200 day survival is in the range of 60% - 70% (21,22).  If 90% of the 
patients survive beyond 200 days, the 95% confidence interval for survival will be estimated to within 
approximately +/-9%.  
In February 2001, the protocol was amended to add an additional immunosuppressive agent, Mycophenolate 
mofetil (MMF), to the transplant regimen to decrease the incidence of life-threatening GVHD.  This drug has 
been used in combination with CSA by a number of other transplant centers using a similar treatment algorithm, 
and has resulted in a significant reduction in the incidence of life-threatening GVHD (unpublished personal 
communication, Dr. Champlin, M.D. Anderson and Dr. Sandameir, Fred Hutchinson Cancer Center).  Acute 
GVHD has been the major toxicity associated with the procedure and has been fatal in two cases.  The majority 
of recipients suffering severe GVHD has done so within the first 60 days following transplantation and has had 
100% donor T-cell engraftment by chimerism analysis.  Therefore, we wish to cover all recipients with an 
additional immunosuppressive agent from days 0 to at least 30 days post-transplant.  Recipients with mixed T-
cell chimerism on day 30 will be tapered off MMF over the next 2 weeks concomitantly with their CSA taper 
(the group we have previously defined to be at low risk for GVHD).  Recipients with 100% donor T-cell 
chimerism on day 30 (high-risk GVHD group) will remain on MMF until day 60 at which point, they will begin 
a taper over the next 40 days concomitantly with their CSA taper.  Recipients with active GVHD at a time point 
which precedes their planned MMF taper, will remain on this agent, having their MMF taper delayed until 
symptoms of GVHD have resolved.  The addition of MMF to CSA prophylaxis will be incorporated into all the 
non-myeloablative protocols.  In November 2002, a retrospective analysis for all recipients on HB protocols 
undergoing non-myeloablative allogeneic stem cell transplantation following cyclophosphamide/fludarabine 
based conditioning was conducted to determine the impact of adding MMF to CSA as GVHD prophylaxis.  In 
this analysis we evaluated the incidence and severity of acute GVHD in our first 66 consecutive recipients who 
received CSA alone (Group I) versus the next 78 consecutive recipients who received CSA + MMF (Group II).  
We saw no beneficial impact of adding MMF to CSA on the incidence of acute grade II-IV, acute grade III-IV 
GVHD or chronic GVHD.  As a consequence, on 2/7/2003 we amended the protocol to replace MMF with the 
more conventional GVHD prophylactic agent MTX as follows:
1) Discontinued the use of MMF as it has clearly failed to reduce the incidence of acute or chronic GVHD. 
2) Continued CSA as GVHD prophylaxis using the same dose and scheduling regimen that was used for 
recipients in Group I and Group II. 
3) Added MTX to CSA as GVHD prophylaxis in the same fashion and doses as are currently given on protocol 
97-H-0196. CSA and MTX have been used together for over a decade and are considered "standard agents" for 
GVHD prophylaxis. 
1/30/2007:  As a consequence of interim analysis of the first 43 recipients accrued to the Group B arm, an 
amendment was requested to increase accrual to the Group B arm with the following justification:  31 subjects 
had received the CSA + MTX as conditioning regimen and 200 day survival had been approximately 90%.  
Because results with CSA + MTX also appear to decrease GVHD, we would like to extend accrual to 17 

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
additional recipients, making a total of 62 subjects in the Group B arm (50 of whom who received CSA + 
MTX), in order to be able to estimate the 200 day survival for recipients who received CSA + MTX to within 
approximately +/- 8% with 95% confidence.  This will also provide a stronger basis for using this regimen as a 
control group for a future trial which will add an additional agent to the conditioning regimen.
12.2 Endpoints
The primary endpoint of this study is transplant related mortality (200 day survival). 
Secondary end points include engraftment, degree of donor-host chimerism, incidence of acute and chronic 
graft versus host disease (GVHD), transplant related morbidity, death from all causes as well as disease-free 
and overall survival. 
The parameters to be monitored include:
1)
CD34+ cell dose, CD3+ cell dose 
2)
Degrees of donor-recipient lymphoid and myeloid chimerism by microsatellite probe analysis of 
peripheral blood and marrow cells (marrow will be collected on days +30, +60, and +100)
3)
Neutrophil recovery (days to neutrophil count of 0.5 x 109/L and 1.0 x 109/L) 
4)
Platelet recovery (days to platelet count of 50 x 109, days to transfusion independence)
5)
Red cell recovery (days to transfusion independence)
6)
Effect of DLI on donor-host lymphoid and myeloid chimerism measured by microsatellite DNA 
analysis of lymphocytes and myeloid cells
7)
Incidence and severity of acute GVHD
8)
Incidence and severity of chronic GVHD
9)
Non-hematologic effects attributable to the preparative regimen
10) Relapse of malignancy or disease control 
11) TRM by day 200
12) Disease-free survival and overall survival  
12.3  Other Analyses
12.3.1. The proportion of graft failures will continue to be monitored.  If there are 6 among the first 42 
recipients, we will reassess our preparative regimen because such results are inconsistent with prior 
beliefs that the graft failure rate is most likely to be 5% and unlikely to be greater than 15%.  A beta 
distribution with parameters 2 and 38 was used as the prior distribution for this calculation.
12.3.2. The proportion of recipients with Grade 3 or higher acute (day 100 or less) and chronic graft 
versus host disease will also be estimated with 95% confidence intervals.
12.3.3 Although the primary endpoint for this protocol occurs at 200 days, recipients will be followed 
up for 60 months so that long-term survival can be estimated.
12.4 Stopping Rules for Safety 
12.4.1 Stopping rule for primary endpoint
We wish to stop this trial if the 200 day survival proportion for either Group A or B is lower than the 
expected 60 and70% respectively with conventional transplantation.  Stopping rules will be applied to 
Group A and Group B separately.  For each, an interim analysis is planned at 200 days after the 15th and 

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30th recipients in each group receive their transplant.  The final analysis will be undertaken 200 days after 
all recipients in each group receive their transplant.  At each interim analysis, the proportion of 200 day 
survivors will be estimated and a 2-sided 99.9% confidence interval given. If the upper bound of the 
confidence interval lies below 60% this suggests that that part of the trial should stop for lack of efficacy.  
This procedure assures that the 95% confidence interval specified in section 12.1. for the final analysis in 
each group can be used with approximately 95% confidence. Operationally, the Z-value 3.29 is used to 
compute the confidence interval at the two interim looks and the Z-value 1.96 is used for the final 
confidence interval.  
This results in the following stopping rule: 
1st interim analysis (n1 = 15): Stop for failure if 0, 1 or 2 are alive at day 200.
2nd interim analysis (n2 = 30): Stop for failure if 9 or fewer are alive at day 200. 
It should be noted that the boundaries may be crossed even if all recipients are not 200 days post-transplant.  
  
12.5 Off Study Criteria
12.5.1 Withdrawal by the recipient from the transplant procedure 
Recipients and their donors will be given ample opportunity to withdraw from the study prior to admission 
for transplant.  Thereafter, the nature of the procedure does not permit safe withdrawal from the protocol. 
The recipient and donor have the right, at any time to elect not to participate in the research aspects of the 
protocol (donation of blood and bone marrow for non-routine tests). 
12.5.2 Withdrawal by the physician from experimental protocol 
Recipients with disease relapse will be taken off protocol and the attending physician may carry out other 
treatments than described in this protocol if they are considered to be in the best interests of the recipient. 
Recipients who fail to achieve allogeneic marrow engraftment (donor chimerism or mixed chimerism) will 
be removed from study after full autologous marrow recovery is achieved.
Recipients with disease progression after 6 month restaging who have active grade >II GVHD or who have 
failed to respond to a previous lymphocyte transfusion and/or to interferon-alpha or IL-2 may be removed 
from study.
Recipients with disease progression associated with a significant decline in performance status which 
negates further treatment on protocol (i.e. DLI) will be removed from study.
12.6 Data Management
The PI will be responsible for overseeing entry of data into an in-house password protected electronic 
system and ensuring data accuracy, consistency and timeliness. The PI, associate investigators (AI) 
/research nurses and/or a contracted data manager will assist with the data management efforts.  All human 
subjects personally identifiable information (PII) as defined in accordance to the Health Insurance 
Portability and Accountability Act,  eligibility and consent verification will  be recorded.  Primary data 
obtained during the conduct of the protocol will be kept in secure network drives or in approved alternative 
sites that comply with NIH security standards.  Primary and final analyzed data will have identifiers so that 
research data can be attributed to an individual human subject participant, e.g., study-specific identifying 

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number (SSPIN) or other unique code or minimum PII required for subject identification.
Laboratory values from referring home physicians may also be entered into the system. 
CIBMTR: For the purposes of quality assurance (i.e. accreditation of the Transplant program), anonymized 
data will be released to the Center for International Blood and Marrow Transplant Research (CIBMTR) 
according to federally mandated policies and procedures. 
End of study procedures:  Data will be stored in locked cabinets and in a password protected database 
until it is no longer of scientific value. 
Loss or destruction of data:  Should we become aware that a major breech in the plan to protect subject 
confidentiality and trial data has occurred, the IRB will be notified.
Publication policy: Given the research mandate of the NIH, subject data including the results of testing and 
responses to treatment will be entered into an NIH-authorized and controlled research database. Any future 
research use will occur only after appropriate human subject protection institutional approval such as 
prospective NIH Intramural IRB review and approval or an exemption from the NIH Office of Human 
Subjects Research (OHSR).
13.0
DATA SAFETY AND MONITORING PLAN 
13.1 Safety Monitoring
Principal Investigator: Accrual, efficacy and safety data will be monitored by the PI.  The study will be 
continuously evaluated for any unusual or unpredicted complications with the aim of detecting and preventing 
unacceptable increase in morbidity and mortality over that anticipated from unmanipulated bone marrow 
transplantation.
IRB:  Accrual and safety data will be monitored reviewed annually by the Institutional Review Board (IRB). 
Prior to implementation of this study, the protocol and the proposed subject consent and assent forms will be 
reviewed and approved by the properly constituted Institutional Review Board (IRB) operating according to 
the 45 CFR 46 Code of Federal Regulations.  This committee will also approve all amendments to the 
protocol or informed consent, and conduct continuing annual review so long as the protocol is open to accrual 
or follow up of subjects. 
DSMB: The NHLBI Data Safety and Monitoring Board will review the protocol at six-twelve month 
intervals.  A progress report will be forwarded to the DSMB at these times and their recommendations will be 
expeditiously implemented. The DSMB may recommend early termination of the study for considerations of 
safety and efficacy.  
13.2
Definitions
Adverse Event (AE):  Any untoward medical occurrence in a human subject, including any abnormal sign (e.g., 
abnormal physical exam or laboratory finding), symptom, or disease, temporally associated with the subject’s 
participation in the research, whether or not considered related to the subject’s participation in the research.
An abnormal laboratory value will be considered an AE if the laboratory abnormality is characterized by any of 

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the following:

Results in discontinuation from the study

Is associated with clinical signs or symptoms

Requires treatment or any other therapeutic intervention

Is associated with death or another serious adverse event, including hospitalization

Is judged by the Investigator to be of significant clinical impact

If any abnormal laboratory result is considered clinically significant, the investigator will provide details 
about the action taken with respect to the test drug and about the patient’s outcome.
Serious Adverse Event (SAE):  A serious adverse event that:

results in death;

is life-threatening (places the subject at immediate risk of death from the event as it occurred);

results in in-patient hospitalization or prolongation of existing hospitalization;

results in a persistent or significant incapacity;

results in a congenital anomaly/birth defect; or

based upon appropriate medical judgment, may jeopardize the subject’s health and may require medical 
or surgical intervention to prevent one of the other outcomes listed in this definition.
Suspected adverse reaction: Suspected adverse reaction means any adverse event for which there is a 
reasonable possibility that the drug caused the adverse event. For the purposes of IND safety reporting, 
‘reasonable possibility’ means there is evidence to suggest a causal relationship between the drug and the 
adverse event. A suspected adverse reaction implies a lesser degree of certainty about causality than adverse 
reaction, which means any adverse event caused by a drug.
Serious event: An event is serious if it meets the definition of a serious adverse event (above) or if it requires 
immediate corrective action by a PI and/or IRB to protect the safety, welfare or rights of subjects.
Unexpected adverse reaction: An adverse event or suspected adverse reaction is considered “unexpected” if it is 
not listed in the investigator brochure or is not listed at the specificity or severity that has been observed; or, if 
an investigator brochure is not required or available, is not consistent with the risk information described in the 
general investigational plan or elsewhere in the current application. "Unexpected”, also refers to adverse events 
or suspected adverse reactions that are mentioned in the investigator brochure as occurring with a class of drugs 
or as anticipated from the pharmacological properties of the drug, but are not specifically mentioned as 
occurring with the particular drug under investigation.
Unanticipated Problem (UP):  Any incident, experience, or outcome that meets all of the following criteria:
1.
unexpected in terms of nature, severity, or frequency in relation to
a.
the research risks that are described in the IRB-approved research protocol and informed consent 
document; Investigator’s Brochure or other study documents; and
b.
the characteristics of the subject population being studied; and
2.
related or possibly related to participation in the research; and
3.
places subjects or others at a greater risk of harm (including physical, psychological, economic, or 
social harm) than was previously known or recognized.
Unanticipated Problem that is not an Adverse Event:  An unanticipated problem that does not fit the definition 
of an adverse event, but which may, in the opinion of the investigator, involves risk to the subject, affect others 
in the research study, or significantly impact the integrity of research data.  For example, report occurrences of 

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breaches of confidentiality, accidental destruction of study records, or unaccounted-for study drug.
Protocol Deviation (PD):  Any change, divergence, or departure from the IRB approved research protocol.
Non-compliance: The failure to comply with applicable NIH HRPP policies, IRB requirements, or regulatory 
requirements for the protection of human research. Noncompliance may be further characterized as:
1. Serious non-compliance: Non-compliance that:
a. Increases risks, or causes harm, to participants.
b. Decreases potential benefits to participants.
c. Compromises the integrity of the NIH HRPP.
d. Invalidates the study data.
2. Continuing non-compliance: Non-compliance that is recurring. An example may be a pattern of non-
compliance that suggests a likelihood that, absent an intervention, non-compliance will continue. 
Continuing noncompliance could also include a failure to respond to IRB requests to resolve previous 
allegations of non-compliance.
3. Minor (non-serious) non-compliance: Non-compliance that, is neither serious nor continuing.
13.2.1 Event Characterization and Reporting to the IRB and Clinical Director (CD) 
All reportable events will be reported to the NIH intramural IRB in accordance with the Policy 801.
The PI or designee will refer to NHLBI DIR Policy to determine Clinical Director reporting requirements and 
timelines.
13.3 Adverse Events 
Adverse events used to evaluate the safety of this protocol regimen will be collected to include any 
unfavorable and unintended signs (including abnormal laboratory findings), symptoms or diseases (i.e. 
incidence of GVHD, graft failure, regimen related toxicities, or infectious complications) which either occur 
during the study, having been absent at baseline or if present at baseline, appear to worsen and  are 
determined to be possibly, probably or definitely related to this investigational treatment (see section 12.2 
parameters to be monitored). 
The following are expected outcomes and will not be reported to the IRB (per review and approval by the 
IRB) unless they meet the criteria for an SAE (life threatening):

Renal insufficiency

Hepatic insufficiency

Transient cardiac arrhythmias

Transient cardiac insufficiency

Pulmonary insufficiency

Neutropenia and its complications

Thrombocytopenia and its complications

Anemia and its complications

Transfusion reactions

Treatable infections from bacteria, viruses, protozoa and fungi

Late effects of transplant regimens including: cataracts, infertility, growth impairment, hypothyroidism, 
and dental caries

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
Headache, insomnia, psychosis, mood changes, disorientation, seizures from metabolic imbalance

Nausea, vomiting, diarrhea, mucositis, weight loss, dry mouth, hiccoughs, constipation

Well-characterized drug reactions - allergic manifestations, "red man" syndrome

Well-characterized drug side effects from drugs used routinely in transplant recipients (e.g.; preparative 
regimen chemotherapy, immunosuppressive drugs, antimicrobials) 

Common side effects of antiemetics, analgesics, anti-inflammatory agents and known complications of 
steroid therapy

Complications from intravenous catheters, thrombotic occlusion, infection, local reactions, cardiac 
arrhythmia 
The following are expected outcomes and will not be reported to the IRB at each occurrence unless they meet 
the criteria of an SAE. They will be reported in summary form at the time of continuing review and at 
termination of the protocol.  The PI will incorporate these events into the protocol and consent as appropriate:

Acute graft-versus-host disease

Chronic graft-versus-host disease

Graft failure / graft rejection

Veno-occlusive disease

Hemorrhagic cystitis

Nephrotic syndrome

Regimen-related toxicity 

Cytomegalovirus reactivation and disease

Disease relapse or progression
Adverse events that are deemed possibly, probably or definitely related to the recipient's underlying 
disease(s), prior therapies, or concurrent non-protocol treatments will not be collected or reported.  
13.4 Adverse Events - Donors
The following are expected outcomes for the donor that will be listed in the protocol and informed consent 
but will not be reported to the IRB unless they meet the criteria of an SAE:

Common side effects of G-CSF administration (bone pain, fatigue, arthralgias, headache, insomnia, fever, 
worsening of pre-existing skin rashes, increases of alkaline phosphatase, lactate dehydrogenase and/or 
uric acid levels, elevated blood leukocyte count, or thrombocytopenia)

Hypotension during apheresis
The following expected outcomes for the donor would not be reported to IRB at each occurrence unless they 
meet the criteria of an SAE. The PI will incorporate these events into the protocol and consent as appropriate. 
They will be reported in summary form at the time of continuing review and at termination of the protocol:

Ischemic chest pain during G-CSF administration

Splenic enlargement

Cutaneous vasculitis

Bone pain, muscle aches or headaches  not controlled with non-narcotic analgesics
13.5 Reporting of SAEs
PI:  All serious adverse events will be reported to Dr. Childs, PI of this study 

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Richard W. Childs, M.D.
Bldg 10, Room 3-5332
Phone: 301-594-8008
Pager:
104-4125-7
E-Mail: childsr@nhlbi.nih.gov 
DSMB:  All SAEs will be included for review by the DSMB at scheduled DSMB meetings.  If the serious 
adverse event is thought to be due to the experimental component of the protocol, accession to the protocol 
will be stopped until a full discussion with the IRB has been held. A stopping rule for the primary endpoint 
was added to this protocol in February, 2001 at the DSMB’s request.
14.0 HUMAN SUBJECT PROTECTIONS 
As of June 2018, this study is no longer recruiting patients. The study status has been changed to follow-up 
only.
14.1  Rationale for Subject Selection 
All subjects with eligible diseases itemized in section 5.0 will be considered for the protocol. Gender, ethnic 
background or race will not be taken into consideration. 
Strategies for subject recruitment: Hematologists and oncologists throughout the country will be informed of 
this protocol by letter. Information about the protocol will be posted on the NIH web page. The protocol will 
also be listed in the physician's data query (PDQ) and clinicaltrials.gov.
Reimbursement for protocol participation, travel, food, and lodging will be consistent with NIH 
guidelines.  In determining reimbursement, the following factors are considered applicable to this protocol: 
the subjects are diagnosed with a rare disease; the subject population is sick; the protocol offers the potential 
for direct benefit; the protocol regimen is demanding; and in order to complete accrual in a reasonable 
timeframe a geographically dispersed participant population is required. 
Payment for participation:  $0.
14.2 Participation of Children
Participation as a transplant recipient:
We are limiting the protocol to recipients over the age of 8 years as the PI does not have expertise in the care 
of younger children undergoing allogeneic transplantation.  The procedure will be explained to recipients 
between the ages of 8 -17 years with formal consent being obtained from parents or legal guardians. 
Participation as a stem cell donor:
Allogeneic bone marrow (and peripheral blood) transplantation (BMT) provides an option for cure for 

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subjects accrued to this protocol and is therefore is considered an accepted standard clinical intervention for 
all diseases accrued to the protocol (ALL, AML, CML, CLL, NHL, MM, SAA, PRCA, PNH, MDS).  The 
donor would be donating stem cells to his/her family member regardless of the objectives of this research 
protocol.  
We are, however, excluding from participation as donors, children who weigh <18 kg and are < 2 years of 
age. The risks of the apheresis procedure are related to the weight of the child, more precisely his/her 
extracorporeal volume, which is weight-dependent.  The risks have to do with (1) need for a central line, (2) 
need for an allogeneic red cell prime, (3) need for systemic heparinization because subject is too small to get 
citrate.  
> 25 kg:  The procedure and associated risk is the same as that in an adult, however a central line is almost 
always needed.   
19 to 25 kg:  A central line is usually required.  Donors may or may not need a central line (at the discretion 
of the Apheresis department). With concurrent magnesium and calcium infusion, children may be safely anti-
coagulated with citrate.
For donors less than age 18, a social worker and mental health specialist (psychologist or psychiatrist) will 
meet with the minor prior to the assent process to determine willingness to participate.  Donors who are less 
than 18 years of age will see a pediatric provider (pediatrician, pediatric nurse practitioner or pediatric 
physician’s assistant) who is separate from the transplant team. This practitioner will serve as the donor’s 
health care provider and advocate during the minor’s participation on the clinical trial.
Donors, less than 18 years of age  will be included in the discussion of the research when formal consent is 
obtained  We will encourage pediatric donors to ask questions about any of the tests or treatments and to sign 
an assent document.  Donors less than age of assent will be included in the discussion of the research, 
however they will not sign assent.  Rather, the research team will rely on the determination of the mental 
health specialist as to the young subject’s willingness to participate.
As participants in laboratory research studies:  Pediatric subjects may participate only in those laboratory 
studies that the IRB categorizes as “does not involve greater than minimal risk to children” provided that 
adequate provisions are made for soliciting the assent of the children and the permission of their parents or 
guardians (see section 14.4.5). 
14.3 Hazards and Discomforts 
14.3.1  The recipient 
Related to the transplant: 
The mortality from conventional BMT may be as high as 40%. Although our data as well as that of 
others suggest a significant reduction in TRM with non-myeloablative PBSC transplantation, the 
procedure nevertheless carries risk.  It is therefore only appropriate to carry out this experimental 
procedure in the context of debilitating or life- threatening conditions and with full informed consent 
from the recipient, donor and/or immediate family. The specific hazards of this study using a non-
myeloablative preparative regimen and high PBPC content graft are graft rejection, GVHD and disease 
relapse. The major discomforts are those of nausea, mucositis, anorexia, diarrhea, fever and malaise, 
and intolerance of the isolation period.  

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Related to the common drugs used in BMT:
Antithymocyte Globulin: Febrile reactions, allergic symptoms including rash, wheezing, anaphylaxis, 
cytopenias, serum sickness, nausea, vomiting, diarrhea, arthralgias, hypotension, tachycardia, 
bradycardia, fever, chills, myelosuppression. Several cases of a severe lung injury related to ATG 
treatment have been reported. Although this side effect appears extremely rare, it is serious and can be 
fatal. There is no information about the mechanism or specific treatment for this condition. A few patients 
recovered after intensive medical support including use of a breathing machine. 
Cyclosporine: CSA is metabolized primarily in liver but the major toxicity is renal. Side effects include renal 
impairment, reversible renal insufficiency, hemolytic uremic syndrome, elevated bilirubin and 
transaminases that normalize with continued administration or reduced dose, hypertrichosis, headaches, 
nausea, gingival hypertrophy, parasthesias (painful hands and feet), hypertension, hypomagnesium, 
bilirubinemia, hypertrichosis, nausea, tremor, and seizure.  An extremely rare complication of 
cyclosporine is blindness, which may be irreversible.  Posterior reversible encephalopathy syndrome 
(PRES) is an increasingly recognized neurologic disorder seen in 6% of patients on cyclosporine which 
manifest with acute to subacute hypertension and/or seizures.  In the event of hypertension, subjects will 
be prescribed 1 or more medications to control blood pressure in an effort to decrease the risk of this 
complication.
Cyclophosphamide: Immediate: tingling and metallic taste, nausea and vomiting, ADH-like effect, 
cardiotoxic at high doses (>70 mg/kg). Rare:   pulmonary toxicity, urticaria and flushing, mucositis 
Delayed: marrow suppression, nausea, mucositis, rash, hemorrhagic cystitis, myocardial damage, alopecia, 
infertility.
Fludarabine: Myelosuppression, fever and chills, nausea and vomiting, malaise, fatigue, anorexia, 
weakness, and rarely hemolysis and pulmonary toxicity, hemolytic anemia and interstitial pneumonitis. 
Serious opportunistic infections have occurred in CLL patients treated with fludarabine.  Methotrexate:  
Mucositis, nausea, dizziness, neutropenia, thrombocytopenia, malaise, fatigue, fever, melena, headaches, 
blurred vision, rashes, alopecia, elevated liver functions. 
Hematologic: Myelosupression (leukopenia [nadir 7 days] thrombocytopenia, anemia). 
Hepatic:  Acute (elevated transaminases) and chronic (fibrosis and cirrhosis) hepatic toxicity. Chronic 
toxicity has generally occurred after prolonged use (generally 2 years or more) and after a total dose of at 
least 1.5 grams.
Urogenital:  Severe nephropathy or renal failure, azotemia, cystitis, hematuria, defective oogenesis or 
spermatogenesis, transient oligospermia, menstrual dysfunction and vaginal discharge, infertility, 
abortion, fetal defects. With high doses of methotrexate, close attention to renal function including 
adequate hydration, and urine alkanization are essential for safe administration. 
Gastrointestinal:  Gingivitis, pharyngitis, stomatitis, anorexia, nausea, vomiting, diarrhea, hematemesis, 
melena, gastrointestinal ulceration and bleeding, enteritis. Should be used with extreme caution in the 
presence of peptic ulcer disease or ulcerative colitis. Therapy may be discontinued if ulcerative stomatitis 
or other severe GI adverse reactions occur.
Pulmonary:  Interstitial pneumonitis deaths have been reported, and chronic interstitial obstructive 
pulmonary disease has occasionally occurred. Pulmonary symptoms or a nonspecific pneumonitis may be 
indicative of a potentially dangerous lesion and require interruption of treatment and careful investigation; 
infection needs to be excluded. This lesion can occur at all dosages.
Skin: Erythematous rashes, pruritis, urticaria, photosensistivty, pigmentary changes, alopecia, 
ecchymosis, telangiectasia, acne, furunculosis.

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Central nervous system: Headaches, drowsiness, blurred vision. There have been reports of 
leukoencephalopathy following intravenous administration of methotrexate to patients who have 
craniospinal irradiation. Aphasia, hemiparesis, paresis, and convulsions have also occurred following 
administration of methotrexate. Following low doses, occasional patients reported transient subtle 
cognitive dysfunction, mood alteration, or unusual cranial sensations.
Other:  Opportunistic infections, arthralgia/myalgia, loss of libido/impotence, diabetes, and osteoporosis. 
A few cases of anaphylactoid reactions have been reported.  
Mycophenolate mofetil (use stopped in 2003): Nausea, diarrhea, leukopenia, headache, weakness, urinary 
frequency, leg cramps or pain, liver function test abnormalities, infection related to immunosuppression,and 
skin rash. Hemorrhagic gastritis and pancreatitis have been reported rarely. Malignant neoplasms have been 
reported in psoriasis patients treated with oral mycophenolic acid, although a definite cause-effect 
relationship has not been established.  
Antimicrobials in general: Allergic reactions, renal impairment (gentamicin, vancomycin, amphotericin, 
acyclovir), red man" syndrome -vancomycin, hepatic damage (acyclovir, rifampicin). 
Risks related to pregnancy and breastfeeding: Pregnant and nursing women are excluded from accrual. 
Subjects are required to use effective contraceptive measures to prevent pregnancy during protocol 
participation to eliminate the possibility of drug effects on a developing fetus. Following transplantation, 
woman of childbearing age will be counseled regarding when it is safe to consider becoming pregnant 
should they so choose.  Female subjects will be advised that should pregnancy occur the research team must 
be informed immediately.  An SAE will be filed on any pregnancy, and this report will be followed up with 
information on the outcome of the pregnancy and complications of (should there be any).  
14.3.2  The donor 
Related to filgrastim G-CSF:  
The hazard to the donor is low. The discomfort from G-CSF mobilization and leukapheresis for collection 
of PBPC are probably lower than those associated with marrow harvesting. G-CSF has been given to large 
numbers of donors without major side effects or long term consequences. The side effects of G-CSF are 
bone pain, fatigue, insomnia, myalgia, headache insomnia, chills, low grade fever, rash, injection site 
irritation, exacerbation of preexisting inflammatory conditions, splenomegaly, reversible elevation in uric 
acid, LDH and leucocyte alkaline phosphatase which are usually mild and self limiting.   Reversible 
thrombocytopenia (mild to moderate decreases in platelet number), with platelet counts falling to the range 
of 100,000 / mm3 is common. Two patients have been reported to experience non-fatal splenic rupture after 
more prolonged treatment with higher doses of G-CSF.  One of these two patients had concurrent 
mononucleosis, a second cause for splenic rupture. Patients with ongoing ischemic heart disease have been 
reported to have angina seemingly temporally-related to G-CSF administration and apheresis. In addition, a 
rare occurrence of pulmonary hemorrhage has been reported in a healthy donor who was a cigarette smoker 
and had underlying pulmonary disease (24). 
Leukapheresis and apheresis of stem cells:  
The apheresis procedures will be performed in accordance with standard apheresis donation policies and 
procedures operative in the Dept. of Transfusion Medicine and will be in compliance with the Blood Donor 
Standards of the American Association of Blood Banks and the rules and regulations of the Food and Drug 
Administration. Adverse reactions to apheresis procedures are rare, but include:

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Pain and hematoma at the needle placement site.
Vasovagal episodes, characterized by transient hypotension, dizziness, nausea and rarely, syncope are seen 
in less than 2% of the procedures. Hypotension secondary to volume depletion is known for the rare 
potential for a cerebrovascular or cardiovascular event. 
Cutaneous or circumoral parasthesias, chills, nausea, heartburn and rarely muscle spasms may result from 
the use of citrate anticoagulant used to prevent clotting in the extracorporeal circuit. Citrate reactions are 
usually relieved by slowing the rate of the anticoagulant infusion and by administering oral calcium 
carbonate tablets or with intravenous calcium gluconate.
Donation of PBPC on three successive days significantly increases the risk of thrombocytopenia 
(<100,000/ul).  However, thrombocytopenia is transient and unlikely to cause clinical sequelae.  
Prior to each apheresis, the potential risks associated with the procedure will be explained to the subject and 
a separate informed consent obtained.
Bone marrow harvest:  
In exceptional instances the donor may be required to donate bone marrow.  There is no additional risk to 
the donor giving marrow after PBPC donation (than would normally be associated with bone marrow 
harvesting). No major risks are involved with bone marrow harvest, other than a small risk of infections, 
pain, bleeding, and hematoma formation at the site of the aspiration exists with the procedure.
Related to central line placement:  
It is estimated that less than 50% of the donors will require intravenous central line placement (femoral 
vein) to successfully complete apheresis.  Intravenous line placement in the femoral vein using a temporary 
double-lumen Arrow catheter carries a small risk of bleeding, bruising or pain and a very low risk of 
accidental injury to the adjacent artery and nerve. Some subjects may experience a vaso-vagal reaction 
(lightheadedness, or, rarely, fainting due to temporary lowering of blood pressure). These risks are 
minimized by using only trained experienced MICU staff for the procedure. Central line placement in a 
minor may be done under conscious sedation.   3 of 10 minor donors (4/15/2009) required femoral lines, 
one of which required conscious sedation.
14.4 Risks in Relation to Benefit 
14.4.1 For adult transplant subjects.   
Clinically the approach is ethically acceptable because we are offering adult subjects with debilitating and 
often lethal hematological diseases incurable with conventional treatments other than allogeneic BMT, an 
alternative to symptomatic therapy.   The protocol aims to decrease the risk of TRM, thus making more 
patients candidates for potentially curative therapy. The risk of death due to complication from the 
allogeneic BMT procedure is justified by the anticipated benefit of potentially eradicating the underlying 
hematological diseases of the subjects, and the relation of the anticipated benefit to the risk is at least as 
favorable to the subjects as the risk presented by available non-transplantation therapies.
Therefore, for adult transplant recipients on this protocol, the research involves greater than minimal risk to 
subjects with the prospect of direct benefit (45 CFR 46.102).

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14.4.2 For pediatric transplant subjects.   
The inclusion of children satisfies the criteria set forth in 45 Code of Federal Regulations 46, Subpart D: 
46.405 as follows:
(a) the risk is justified by the anticipated benefit to the subjects: Clinically the approach is ethically 
acceptable because we are offering a child with debilitating and often lethal hematological diseases 
incurable with conventional treatments other than allogeneic BMT, an alternative to symptomatic 
therapy
(b) the relation of the anticipated benefit to the risk is at least as favorable to the subjects as that 
presented by available alternative approaches. The protocol aims to decrease the risk of transplant-
related mortality, thus making more patients candidates for potentially curative therapy 
(c) the relation of the anticipated benefit to the risk is at least as favorable to the subjects as that 
presented by available alternative approaches. The protocol aims to decrease the risk of transplant-
related mortality, thus making more patients candidates for potentially curative therapy
 and
 (d) adequate provisions are made for soliciting the assent of the children and permission of their parents 
or guardians, as set forth in 46.408.
Therefore, for pediatric transplant recipients on this protocol, the research involves greater than minimal risk 
but presenting the prospect of direct benefit to the individual subjects (45 CFR 46.405).
14.4.3  For adult donors  
Normal identical or single HLA locus mismatched family members will be co-enrolled into this study as stem 
cell donors. The stem cell collection aspect of this protocol is not investigational.  Despite the risks associated 
with this procedure (section 14.3.2), potential benefit does exist for family donors. The donor derives 
psychosocial benefit from donating stem cells both at the time of donation and possibly into the future, 
especially in view of the reduced life expectancy due to this disease in a family member.  Other potential 
benefits include detection of illnesses, determination of blood cell counts, and evaluation of kidney and liver 
function in the potential donor at the time of screening. 
Therefore, for adults participating as stem cell donors, the research involves greater than minimal risk but 
presents the prospect of direct benefit to the individual subject   (45 CFR 46.102).  
14.4.4 For pediatric donors
The inclusion of children as donors satisfies the criteria set forth in 45 Code of Federal Regulations 46, Subpart 
D as follows:
Stem cell transplant is an accepted standard clinical intervention for the diseases under investigation. The donor 
would be donating stem cells to his/her family member regardless of the objectives of this research protocol. 
The stem cell collection procedure is not considered part of the research for the donors, the risks of the stem cell 
collection procedure would not be considered risks of the research for the pediatric donors.
Therefore participation as a stem cell donor on this protocol is considered exempt from the criteria set forth in 
45 CFR 46, Subpart D.

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
14.4.5 For pediatric donors – healthy volunteers- involved in laboratory research studies
 The inclusion of children satisfies the criteria set forth in 45 Code of Federal Regulations 46, Subpart D: 
46.404 as follows:
(a) The research does not involve greater than minimal risk.  Blood and bone marrow specimens for 
research are obtained concurrently with clinically indicated sampling.  Therefore, there is no risk 
associated with sample collection for research because research will only be performed on material 
obtained during standard clinical intervention.
(b) Research specimens will be stored in the Dr. Child’s laboratory. Samples will never be labeled with 
the child’s name. Samples will be assigned a unique code known only to the principal investigator, 
which will serve as a link to the child’s clinical information collected as part of this research 
protocol.  No samples will be provided to investigators outside the Branch. Therefore 
confidentiality is protected.
(c) Only those laboratory tests approved by the IRB and involving not greater than minimal risk will be 
conducted (See Appendix C, laboratory studies D1-D6 are determined to be greater than minimal 
risk).  Research will not include genetic testing.  Therefore, there is no genetic testing associated 
risk. 
(d)  adequate provisions are made for soliciting the assent of the children and permission of their 
parents or guardians, as set forth in 46.408.
Therefore, for pediatric donors – healthy volunteers- participating in laboratory research studies on this 
protocol, the research does not involve greater than minimal risk  (45 CFR 46.404).
14.5 Informed Consent
The investigational nature and research objectives of this trial, the procedure and its attendant risks and 
discomforts will be carefully explained to the subject and a signed informed consent document will be 
obtained prior to entry onto this study.  Investigators names marked with an asterisk on the cover sheet of this 
protocol (Drs. Childs, Srinivasan, Barrett, Gormley or Young) will lead this discussion. 
If the subject is a minor, the parent who signs the consent for the minor must be a legally recognized parent or 
guardian.  Where deemed appropriate by the clinician, and the child's parent or guardian, the child will also be 
included in all discussions about the trial and a minor's assent will be obtained.   The parent or guardian will 
sign on the designated line on the informed consent attesting to the fact that the child had given assent.
If the donor is a minor, assent will not be sought until an evaluation by a social worker and mental health 
specialist (psychologist or psychiatrist) is completed to determine the minor donor’s willingness to 
participate.   As detailed above, the parent who signs the consent for the minor must be a legally recognized 
parent or guardian.  Where deemed appropriate by the clinician, and the child's parent or guardian, the child 
will also be included in all discussions about the trial and a minor donor’s assent will be obtained.   The 
parent or guardian will sign on the designated line on the informed consent attesting to the fact that the child 
had given assent.
We will inform minors during the assent process that for safety, we need to do a pregnancy test.  Subject will 
also be told that if it is positive, we will counsel her and help her tell her parents.  If the minor does not want 
to proceed she will be advised not to sign the assent, and her enrollment on this screening protocol will end.

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At any time during participation in the protocol that new information becomes available relating to risks, 
adverse events, or toxicities, this information will be provided orally or in writing to all enrolled or 
prospective patient participants.  Documentation will be provided to the IRB and if necessary the informed 
consent amended to reflect relevant information.
Reconsenting for nephritic syndrome testing: Subjects already consented on a stem cell transplant protocol 
and not available for a NIH clinic visit:  The revised consent will be sent to the transplant recipient or donor to 
be executed by telephone.  The investigational nature of the testing will be carefully explained to the subject.  
The PI, or an AI will lead this discussion (Drs. Childs, Srinivasan, Gormley or nurse Cho, Cook or Ramos).  
If the subject chooses to participate he/she will be directed to sign and date the revised consent and have 
someone witness and date his/her signing of the document. The informed consent document will then be 
mailed to the PI or AI who led the discussion, who will sign and date and mail back a fully executed copy for 
the subject’s records.   The informed consent process will be documented on a progress note and a copy of 
said note and the original informed consent will be filed in the subject’s medical record.   
(**See Clinical Center Policy and Communication Bulletin M77-2 Dated 30 May 2001. Subject: Informed 
Consent, Section IX. Consent from someone not at the clinical center:  “For research protocols or any 
procedures performed for the purposes of research that involve obtaining consent via technology and/or 
electronic process, rather than in person, the procedures for obtaining consent, including how information will 
be transmitted and documented and by whom, shall be detailed in the written protocol.  Review and approval 
must first be obtained from the institute clinical director and the relevant IRB)
Re-Consent for Minors when they reach the age of majority:  When a pediatric subject reaches age 18, 
continued participation will require re-consenting of the now adult with the standard protocol consent document 
to ensure legally effective informed consent has been obtained. Should sample or data analysis continue 
following completion of active participation and the subject has reached 18 years of age, we will attempt to 
contact the subject using the last known contact information to obtain consent for continued use of data or 
samples  collected during their prior visit. Given the length of time that may have transpired for some of the 
subjects since their last visit for this study, we request waiver of informed consent for those individuals who 
after good faith efforts to contact them, we are unable to contact.
Requirements for Waiver of Consent consistent with 45 CFR 46.116 (d), each of which must be addressed in 
relation to the protocol:
(1) The research involves no more than minimal risk to the subjects.
a.
Analysis of samples and data from this study  involves no additional risks to subjects.
(2) The waiver or alteration will not adversely affect the rights and welfare of the subjects.
a.
Retention of these samples or data in secure locations does not affect the welfare of 
subjects. 
(3) The research could not practicably be carried out without the waiver or alteration.
a.
Considering the length of time between a minor’s enrollment and their age of majority, it is 
possible that more than a few subjects may be lost to follow up.  A significant reduction in 
the number of samples analyzed could impact the quality of the research. 
(4) Whenever appropriate, the subjects will be provided with additional pertinent information after 
participation.
a.
We only plan to request a waiver of reconsent for those subjects who have been lost to 
follow-up.  

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February 22, 2022 (Amendment III)
14.6 Conflict of Interest  and Tech Transfer Agreements
The PI assured that each AI listed on the protocol title page received a copy of the NIH’s Guide to preventing 
conflict of interest.  Investigators added subsequent to the initial circulation were provided a copy of the 
document when they were added. Copies of the Conflict of Interest Statement were forwarded to the Clinical 
Director.  No initial or subsequent members of the research team reported a potential conflict of interest.  
This protocol has no associated patents, CRADAs, or CTAs.  
15.0
PHARMACEUTICALS
15.1 ANTI-THYMOCYTE GLOBULIN, Equine (Atgam, lymphocyte immune globulin, horse [equine] 
ATG)
Supply: Commercially available.
Product description: Anti-thymocyte globulin (equine) is the purified, concentrated, and sterile gamma globulin, 
primarily monomeric IgG, from hyperimmune serum of horses immunized with human thymus lymphocytes. It 
is available in 5 mL (250 mg) ampules containing 50 mg of horse gamma globulin/mL stabilized in 0.3 molar 
glycine to a pH of approximately 6.8.
Preparation: Dilute anti-thymocyte globulin (ATG) for intravenous infusion in an inverted bottle of sterile 
vehicle so the undiluted drug does not contact the air inside. Dilute the total daily dose of ATG (equine) in a 
compatible diluent sterile solution (0.9% sodium chloride injection, 5% dextrose and 0.225% sodium chloride 
injection, or 5% dextrose and 0.45% sodium chloride injection) to a final concentration that does not exceed 4 
mg/mL. Do not dilute anti-thymocyte globulin in dextrose injection (e.g. 5% dextrose injection) as low salt 
concentrations can cause precipitation. 
Storage and stability: Vials of ATG (equine) should be stored in a refrigerator at 2– 8 oC (36 – 46 oF). Once 
diluted for intravenous administration, ATG (equine) is physically and chemically stable for up to 24 hours at 
concentrations up to 4 mg/mL.  It is recommended that diluted ATG (equine) be stored in a refrigerator if it is 
prepared prior to the time of infusion. Even if it is stored in a refrigerator, the total time in dilution should not 
exceed 24 hours (including infusion time).
Route of administration: ATG (equine) diluted for intravenous administration should be infused preferentially 
into a central venous catheter through an in-line filter with a pore size of 0.2 to 1 micron. The use of high-flow 
veins will minimize the occurrence of phlebitis and thrombosis. Do not infuse a dose of ATG (equine) in less 
than 4 hours. Follow nursing care SOP guidelines for monitoring during the infusion.
15.2 CYCLOPHOSPHAMIDE (Cytoxan, Neosar)
Supply: Commercially available.
Product description: Cyclophosphamide is available as a lyophilized powder for injection in multiple vial sizes. 
Preparation: Cyclophosphamide powder for injection should be reconstituted with sterile water for injection to 
yield a concentration of 20 mg/mL as described in the product labeling. Once reconstituted, the prescribed dose 
will be further diluted in 250 mL of 0.9% sodium chloride injection or 5% dextrose in water for intravenous 
administration over 60 minutes.
Storage and stability: Vials of cyclophosphamide are stored at room temperature. Once reconstituted as 
directed, solutions of cyclophosphamide are stable for 24 hours at room temperature, or 6 days when 
refrigerated at 2-8o C. 
Route of administration: The prescribed dose of cyclophosphamide will be diluted in an additional 250 mL of 
0.9% sodium chloride injection or 5% dextrose in water for intravenous administration over 60 minutes.

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
15.3 CYCLOSPORINE (Gengraf, Sandimmune, Neoral)
Supply/product description: Cyclosporine will be obtained by the NIH Clinical Center Pharmacy Department 
from commercial sources and is available in capsules (25 mg and 100 mg), USP [MODIFIED], oral solution 
(100 mg/ml), USP [MODIFIED], and as a parenteral concentrate for injection (50 mg/ml).  When oral capsules 
are prescribed for this protocol, the cyclosporine capsules, USP [NON-MODIFIED] should NOT be used.
Preparation: For parenteral doses, each milliliter of concentrate (50mg/ml) should be diluted in 20 to 100 ml of 
dextrose 5% in water or sodium chloride 0.9%.  Parenteral doses of cyclosporine will be prepared in non-PVC 
containers and infused with non-PVC administration sets/tubing.  Oral cyclosporine solution may be mixed in 
orange juice or other beverages, but not milk.
Storage and Stability: Capsules, oral solution, and ampules of parenteral concentrate bear expiration dates and 
are stored at room temperature and protected from light.  Cyclosporine concentrate for injection that has been 
diluted to a final concentration of approximately 2mg/ml is stable for 24 hours in 5% dextrose or 0.9% sodium 
chloride injection in glass, PVC or non-PVC plastic containers. To minimize the potential for sorption to PVC 
plastic bags and tubing as well the leaching of phthalate plasticizer (DEHP) into the solution, only non-PVC 
plastic bags and intravenous administration sets should be utilized.
Administration: Cyclosporine may be given intravenously or orally.
15.4     FILGRASTIM (G-CSF, Neupogen)
Supply: Commercially available.
Product description: Filgrastim injection is available in a concentration of 300mcg/ml in 1ml (300mcg) and 
1.6ml (480mcg) vials.
Preparation: For subcutaneous administration, the appropriate prescribed dose is drawn up from the vial with no 
further dilution prior to administration. For intravenous administration, the commercial solution for injection 
should be diluted prior to administration. It is recommended that the prescribed dose be diluted with dextrose 
5% in water (DO NOT DILUTE WITH NORMAL SALINE) to a concentration greater than 5mcg/ml. 
Filgrastim diluted to concentrations between 5 and 15mcg/ml should be protected from absorption to plastic 
materials by the addition of Albumin (human) to a final concentration of 2mg/ml. When diluted in 5% dextrose 
or 5% dextrose plus Albumin (human), filgrastim is compatible with glass bottles, PVC and polyolefin IV bags, 
and polypropylene syringes.  
Storage and stability:  Filgrastim for injection should be stored in the refrigerator at 2 to 8oC (36 to 46oF). Avoid 
shaking.
Route of administration: Subcutaneous injection or intravenous infusion over 15-30 minutes.
15.5 FLUDARABINE PHOSPHATE (Fludara)
Supply: Commercially available.
Product description: Fludarabine phosphate is commercially available as both a lyophilized powder for injection 
in vials containing 50 mg of fludarabine phosphate with mannitol 50 mg and sodium hydroxide for pH 
adjustment and a solution for injection in 2 mL vials containing 50 mg of fludarabine phosphate (25 mg/mL of 
fludarabine) with 25 mg/mL mannitol and sodium hydroxide for pH adjustment.
Preparation: Fludarabine lyophilized powder for injection should be reconstituted with 2 mL of sterile water for 
injection, USP to a concentration of 25 mg/mL. The prescribed dose of fludarabine should be diluted in 100 mL 
of either 0.9% sodium chloride or 5% dextrose in water for intravenous administration over 30 minutes.
Storage and stability: Fludarabine vials should be stored under refrigeration between 2-8 oC (36- 46 
oF).  Reconstituted fludarabine phosphate is chemically and physically stable for 24 hours at room temperature 
or for 48 hours if refrigerated. The manufacturer recommends use of either the reconstituted powder for 

37
99-H-0050
Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
injection or the solution for injection (once diluted for administration) within 8 hours because neither product 
contains an antimicrobial preservative.
Administration: The prescribed dose of fludarabine should be diluted in 100 mL of either 0.9% sodium chloride 
or 5% dextrose in water for intravenous administration over 30 minutes.
15.6  METHOTREXATE (Methotrexate sodium)
Supply:  Commercially available.
Product description: Methotrexate is commercially available as a 25 mg/mL preservative-free isotonic solution 
for injection.
Preparation: The desired dose will be diluted in 5% dextrose in water or 0.9% sodium chloride.
Storage and stability: Methotrexate should be stored at room temperature and protected from light. Once the 
prepared dose is diluted for intravenous administration, the solution is stable for 24 hours refrigerated or at 
room temperature when protected from light.
Administration: The prescribed dose of methotrexate will be given as an intravenous infusion over 15 minutes. 
Refer to BMT Supportive Care Guidelines for dose adjustments based on serum creatinine and total bilirubin.
15.7 MYCOPHENOLATE MOFETIL (MMF)—not used after February 2003
Generic:  Mycophenolate mofetil.
Other:  CellCept, Roche Laboratories.
Classification:  Immunosuppressive agent.
Dose: 1 gram p.o. twice daily given to some recipients with steroid resistant acute GVHD or steroid dependent 
chronic GVHD.
Action: Mycophenolic acid produces potent, noncompetitive inhibition of inosine monophosphate 
dehydrogenase, thus blocking de novo synthesis of guanosine nucleotides; as lymphocytes depend upon the de 
novo pathway for purine synthesis, lymphocyte proliferation is inhibited. In vitro and in vivo studies have 
demonstrated the ability of mycophenolic acid to block proliferative responses of T and B lymphocytes, and 
inhibit antibody formation and the generation of cytotoxic T-cells.
Metabolism: Mycophenolate mofetil is rapidly converted to its active metabolite, mycophenolic acid in the 
liver.  Mycophenolic acid is subsequently metabolized to mycophenolic acid glucuronide which is inactive and 
is excreted via the urine and bile.  Enterohepatic recirculation of mycophenolic acid may occur.  The 
elimination half-life of mycophenolic acid is approximately 11 to 18 hours.  

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Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
REFERENCES 
1 
The clinical practice of stem cell transplantation. Barrett AJ and Treleavan JG (Eds) Isis Medical Media 
Oxford 1998. 
2 
Armitage JO. Bone marrow transplantation. N Eng J Med 330: 827-38, 1994 
3 
Weiden PL, Sullivan KM, Flournoy N, Storb R, Thomas ED, The Seattle Marrow Transplant Team: 
Antileukemoc effect of chronic graft-versus-host disease. Contribution to improved survival after allogeneic 
marrow transplantation. N Engl. J Med 304: 1529-1533, 1981 
4 
Kolb HJ, Mittermuller J, Clemm C et al. Donor leukocyte transfusions for treatment of recurrent chronic 
myelogenous leukemia in marrow transplant recipients. Blood 76:2462-2465, 1990. 
5 
Porter DL, Aoth MS, McGarigle C, Ferrara JLM, Antin JH. Induction of graft-versus-host disease as 
immunotherapy for relapsed chronic myeloid leukemia. N Engl J Med 330: 100-106' 1994 
6 
Kolb HJ, Schattenberg A, Goldman JM, et al. Graft-versus-Ieukemia effect of donor lymphocyte 
transfusion in marrow grafted patients. Blood 86: 2041-2050, 1995. 
7 
Khouri 1, Keating MJ, Przepiorka D, et al. Stem cell transplantation (SCT) for chronic lymphocytic 
leukemia (CLL): graft-versus-Ieukemia (GVL) without acute graft-versus-host disease (GVHD). Blood 86: 
457a, 1995. 
8 
Khouri 1, Keating MJ, Przepioka et el. Engraftment and induction of GVL with fludarabine (FAMP)- j 
based non-myeloablative preparative regimen in patients with CLL and lymphoma. Blood : 
88;10:suppl,310a,1996. 
9 
Huss R, Deeg HJ, Gooley T, Bryant E, Leinsenring W, Clift A, Buckner CD, Martin P, Appelbaum FA. 
Effect of mixed chimerism on graft-versus-host disease, disease recurrence and survival after HLA-identical 
marrow transplantation for aplastic anemia or chronic myelogenous leukemia. Bone Marrow 
Transplantation 1996 Oct;18(4):767-76 
10 Giralt S, Estey E, Albitar M, van Besienk K: Engraftment of allogeneic hematopoietic progenitor cells with 
purine analog containing chemotherapy. Blood 1997, Jun 15;89 (12)4531-6 
11 Schmitz N, Greger P, Suttorp Met al. Primary transplantation of allogeneic peripheral blood progenitor cells 
mobilized by Filgastrim {G-CSF) Blood 1995, 85: 1666-1674. 
12 Goodman JM. Peripheral blood stem cells for allografting. Blood 1995, 85: 1413-1415. 
1 3 Korbling M and Champlin R. Peripheral blood progenitor cell transplantation: A replacement for marrow 
auto- or allografts. Stem Cells, 1996; 14: 164-176. 
14 Fleming TA: One-sample multiple testing procedure for phase II trials. Biometrics 1982; 38: 143-151 . 
15 Jennison C and Turnbull B. Confidence intervals for a binomial parameter following a multistage test with 
application to MIL-STD 105D and medical trials. Technometrics 1983;25:49-50
16 Kaplan EL and Meier P. Nonparametric estimation from incomplete observations. J Am Statist Assoc 1958; 
53:457-481. 
17 Khouli IF, Keating M, Korbling Met al. Transplant lite: induction ot graft-versus-malignancy using 
tludarabine-based nonablative chemotherapy and allogeneic blood progenitor-cell transplantation as 
treatment tor lymphoid malignancies. J Clin Oncol, 1998:16;8, 2817-24 - 
18 Storb R, Evans R, Thomas D, et al. Paroxysmal Nocturnal Haemoglobinuria and refractory marrow failure 
treated by marrow transplantation. Brit J Haem 1973;24:743-750 
19 Kolb H.J., Holler E, Bender-Gotze Ch et al. Myeloablative conditioning for marrow transplantation in 
myelodysplastic syndromes and paroxysmal nocturnal haemoglobinuria. Bone Marrow Transplantation 
1989; 4: 29-34 
20 Slavin S, Nagler A, Naparstek E et al. Nonmyeloablative stem cell transplantation and cell therapy as an 
alternative to conventional bone marrow transplantation with lethal cytoreduction for the treatment of 
malignant and nonmalignant hematologic diseases. Rapid Communication Blood 1998;91:756-763 
21 Kaplan EL, Meier P. Nonparametric estimation from incomplete observations. J Am Statist Assoc 1958; 
53:457-481. 

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February 22, 2022 (Amendment III)
22. Simon, R. and Lee Y.J. Confidence limits for survival probabilities and median survival time. Cancer 
Treatment Reports 1982; 66: 37-42. 
23. Maloney DG, Molina AJ, Sahebi F, Stockeri-Goldstein KE, Sandmaier BM, Bensinger W, Storer B, 
Hegenbart U, Somio G, Chauncey T, Bruno B, Applebaum FR, Blume KG, Forman SJ, McSweeney P and 
Storb R.  Allografting with nonmyeloablative conditioning following cytoreductive autografts for the 
treatment of patients with multiple myeloma.  Blood 2003: 102: 3447-3454.
24. Kopp HG, Horger M, Faul C, Hartmann JT, Kanz L, Lang P, Vogel W,  Granulocyte colony-stimulating 
factor induced pulmonary hemorrhage in a healthy stem cell donor. Granulocyte colony-stimulating factor 
induced pulmonary hemorrhage in a healthy stem cell donor. J Clin Oncol. 2007 Jul 20;25(21):3174-5.
25. Johnson NA, Savage KJ, Ludkovski O, Ben-Neriah S, Woods R, Steidl C, Dyer MJ, Siebert R, Kuruvilla J, 
Klasa R, Connors JM, Gascoyne RD, Horsman DE. Lymphomas with concurrent BCL2 and MYC 
translocations: the critical factors associated with survival. Blood. 2009 Sep 10;114(11):2273-9.
26. Aukema SM, Siebert R, Schuuring E, van Imhoff GW, Kluin-Nelemans HC, Boerma EJ, Kluin PM., 
Double-hit B-cell lymphomas. Blood. 2011 Feb 24;117(8):2319-31.
27. Tomita N. BCL2 and MYC dual-hit lymphoma/leukemia. J Clin Exp Hematop. 2011;51(1):7-12.
28. Tomita N, Tokunaka M, Nakamura N, Takeuchi K, Koike J, Motomura S, Miyamoto K, Kikuchi A, Hyo R, 
Yakushijin Y, Masaki Y, Fujii S, Hayashi T, Ishigatsubo Y, Miura I. Clinicopathological features of 
lymphoma/leukemia patients carrying both BCL2 and MYC translocations. Haematologica. 2009 
Jul;94(7):935-43.
29. Snuderl M, Kolman OK, Chen YB, Hsu JJ, Ackerman AM, Dal Cin P, Ferry JA, Harris NL, Hasserjian RP, 
Zukerberg LR, Abramson JS, Hochberg EP, Lee H, Lee AI, Toomey CE, Sohani AR. B-cell lymphomas 
with concurrent IGH-BCL2 and MYC rearrangements are aggressive neoplasms with clinical and 
pathologic features distinct from Burkitt lymphoma and diffuse large B-cell lymphoma. Am J Surg Pathol. 
2010 Mar;34(3):327-40.

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APPENDIX A
PREPARATIVE REGIMEN
HLA IDENTICAL SIBLING REGIMEN
Day
Hour
Regimen
0 
(midnight)
Hydrate: NS 2.6ml/kg/hr 10 mEq/l KCl (starting 12 hours
pre-cyclophosphamide and continue hydration until 24 hours after last 
cyclophosphamide infusion)
11
Ondansetron 8 mg IV q12 hrs (or equivalent serotonin antagonist) + decadron 
8 mg PO q12hrs x 1 day
Mesna (per Supportive Care Guidelines) 
Furosemide 20 mg IV
-7
12
Cyclophosphamide 60 mg/kg IV in 250 ml D5 in 1 hr
Mesna (per Supportive Care Guidelines)
-6
11
Ondansetron 8 mg IV q12 hrs (or equivalent serotonin antagonist)+  decadron  
8 mg PO q12 hrs x 1 day
Furosemide 20 mg IV
12
Cyclophosphamide 60 mg/kg I.V. in 250 ml D5 I.V. over 1 hr
13
Mesna (per supportive care guidelines)
Day -5
Stop IV hydration (24 hours after last cyclophosphamide dose)
Monitor electrolytes and fluid balance while receiving hydration for 
cyclophosphamide 
Day -5 to Day -1
9
Fludarabine 25 mg/m2 IVBP over 30 minutes for 5 days
SINGLE HLA ANTIGEN MIS MATCH REGIMEN
Same as HLA identical sibling regimen except some subjects will receive ATG at a dose of 40 mg/kg/d for 4 
consecutive days on day -5 to day –2 (see section 7.5 preparative regimen).

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Richard W. Childs, M.D.
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APPENDIX B:  Donor stem cell mobilization and apheresis.
I.
Donor stem cell mobilization with G-CSF and donor stem cell apheresis
After medical evaluation and clearance for suitability as an allogeneic donor, each donor will undergo 
mobilization with G-CSF as an outpatient. The donor will receive 10 mcg /kg G-CSF subcutaneously daily 
starting day –4 before apheresis. On day 0, a large volume leukapheresis of 15- 25 liters will be performed. The 
volume of the leukapheresis procedure will be targeted to obtain a progenitor cell dose of 5x106 CD34+ 
cells/kg. Obtaining a dose of  > 3x106 CD34+ cells/kg will be considered adequate and no further apheresis will 
be performed. If after the first day of apheresis, the product contains < 3x106/kg CD34+ cells, a decision 
regarding further apheresis will be made at the discretion of the PI, in consultation with Department of 
Transfusion Medicine. The minimum acceptable CD34+ cell dose to proceed with the protocol is 2x106/kg. 
Predictable side effects of G-CSF, including headache, bone pain, and myalgia, will be treated with 
acetaminophen or ibuprofen.  Prophylactic treatment of these side effects may also be carried out with the same 
medications. Other side effects will be evaluated and treated accordingly.
Leukapheresis procedures will be carried out in the Apheresis Unit, Department of Transfusion Medicine. These 
procedures will be done via a 2-armed approach or by temporary central venous catheter in the femoral vein, 
using the Baxter CS3000Plus instrument or the Cobe Spectra.  These procedures will use ACD-A anticoagulant, 
routinely used in normal donors.  If the donor is very small or relatively intolerant to ACD-A, and the adverse 
effects cannot be controlled by usual means, consideration will be given to using partial anticoagulation with 
heparin.
II.      Leukapheresis PBPC product
The processing goals for this protocol are to provide a product with a maximum dose of CD34+ cells. ABO or 
other erythrocyte incompatibility is not an issue because the resulting number of contaminating erythrocytes is 
well below 1 ml using this system. The final cell suspension will be prepared for infusion by transferring the 
cells into a 50-150 ml volume of an infusible solution (Plasmalyte A with 1% human serum albumin), suitable 
for intravenous infusion over 15 minutes to 1 hour, depending on clinical circumstances.

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February 22, 2022 (Amendment III)
APPENDIX C NHLBI LABORATORY RESEARCH STUDIES
IRB APPROVED NHLBI LABORATORY RESEARCH STUDIES v. 2.5.13
DESCRIPTION OF LABORATORY STUDY BY BRANCH SECTION
Does this test 
pose a greater 
than minimal 
risk to 
pediatric 
subjects per 45 
CFR 46.404?
Does this test 
pose a 
greater than 
minimal risk 
to healthy 
pediatric 
donors per 45 
CFR 46.404?
A
Stem Cell Allotransplantation Section (Dr. A. John Barrett) (No longer active)
B
Molecular Hematopoiesis  Section  (Dr. Cynthia Dunbar) 
B.1
Flow cytometric analysis of cell surface and cytoplasmic proteins, including cell 
adhesion molecules, putative retroviral receptors, and markers of differentiation, 
using bone marrow and mobilized peripheral blood cells.
No
No
B.2
Hematopoietic progenitor-derived colony ascertainment in vitro (as described above), 
and engraftment of immunodeficient mice for detection of human stem cell number 
and function.
No
No
B.3
Testing ability of hematopoietic progenitor cells to be transduced with retroviral, 
lentiviral, and novel gene transfer vectors in vitro.
No
No
B.4
Reprogramming of adult mature cells, including skin fibroblasts and blood cells, into 
induced pluripotent stem cells in vitro.
No
No
C
Cell Biology Section (Dr. Neal Young) 
C.1
Studies of blood and bone marrow hematopoietic progenitor numbers, including early 
and late erythroid progenitors, myelomonocytic progenitors, and multi-potential 
progenitor cells.  In addition, bone marrow may be placed in long-term bone marrow 
culture to assess the function of stroma and stem cells and to assay more primitive 
progenitors, as well as organelle culture.  Whole or selected bone marrow 
populations are cultured short-term for CD34 cell expansion.
No
No
C.2
Assays of apoptosis in hematopoietic cells and their progeny, using flow cytometric 
methods such as annexin and caspase-3 staining, propidium iodide uptake, and 
mitochondrial permeability tests.
No
No
C.3
Separation and functional study of cell populations characteristic of paroxysmal 
nocturnal hemoglobinuria, identified by absence of glycosylphosphatidylinositol 
anchored proteins.
No
No
C.4
Studies of mutation rates in hematopoietic cells and in buccal mucosa cells, using 
conventional hypoxanthine phosphoribosyltransferase activity functional assays, 
sequencing of mitochondrial DNA after specific gene amplification, and 
measurement of GPI-anchored deficient cells in blood and bone marrow.
No
No

43
99-H-0050
Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
C.5
Assays of immune function of T-cells, including intracellular cytokine staining, 
ELISPOT, semiquantitative gene amplification for gamma-interferon, tumor necrosis 
factor, interleukin-2, and other cytokines, and functional assessment in co-culture 
using specific neutralizing monoclonal antibodies.  In addition, peripheral blood 
lymphocytes are subjected to spectratyping for CDR3 size distribution as well as 
nucleotide sequence of CDR3 peaks obtained.
No
No
C.6
Studies of engraftment of human normal and diseased bone marrow and peripheral 
blood in immunodeficient mice in order to determine the presence of hematopoietic 
repopulating stem cells as well as functional differences among selected populations.
No
No
C.7
Flow cytometric analysis of blood and bone marrow for lymphocyte phenotype, 
especially for evidence of activation of lymphocytes, for markers of apoptosis, and 
for antigens associated with primitive and mature hematopoietic cell populations.
No
No
C.8
Flow cytometric analysis of blood and bone marrow for hematopoietic stem cell 
progenitors and CD34 positive cells.
No
No
C.9
Studies of chromosomal instability in myelopdysplastic syndromes including BM cell 
and CD34 cell response to PAS crosslinking and examination of the cytotoxic effect 
of lymphocytes to the abnormal clone of cells.
No
No
C.10
Surface Enhanced Laser/Desorption Ionization (SELDI)  time-of-flight mass 
spectrometry (Ciphergen)  (proteomics methodology).
No
No
C.11
Mitochondrial DNA (mtDNA) sequence heterogeneity.
No
No
C.12
Measurement of EBV viral load.
No
No
C.13
Measurement of EBV LMP-1 via RT-PCR for LMP-1 RNA or flow cytometry for 
LMP-1.
No
No
C.14
Outgrowth assay of EBV transformed B cells.
No
No
C.15
Quantification of serumchemokines and cytokines (e.g. SDF-1, IL-10, IL-6, CXCR4, 
CXCL12).
No
No
C.16
Quantification of EBV cytotoxic T cells (tetramerstaining).
No
No
C.17
Telomere length measurement by Southern blot, Q-PCR, flow-fish, in situ 
hybridization and STELA
No
No
C.18
Telomere repair complex gene mutations by nucleotide sequencing of some or all of 
the following:  DKC1 ,TERC, TERT, SBDS, NOp10 , NHP2.
No
No
C.19
Analysis of  inflammatory markers and/or bacterial, viral, fungal or protozoal 
elements in plasma or serum using molecular, colorimetric, enzymatic, flow 
cytometric or other assays in subjects receiving immunosuppressive therapy, 
chemotherapy and/or bone marrow transplantation.
No
No
C.20
Confocal microscopic imaging of bone marrow.
No
No
C.21
Characterization of intracellular signaling proteins by cell permeabilization and flow 
cytometry, and quantitative immunoblots.
No
No
C.22
Assays for chromosomal aneuploidy by florescence in situ hybridization (FISH) and 
other molecular techniques.
No
No
C.23
Conversion of human dermal fibroblasts into hematopoietic progenitors using Oct4 
transfection.
No
No
C.24
Quantification of gene expression with RNA-seq
No
No
C.25
Characterization of chromatin and promoter/enhancer landscapes with ATAC-seq
No
No
C.26
Measurement of protein markers with SomaLogic's SOMAscan assay
No
No
D
Virus Discovery Section (Dr. Neal Young) THESE ASSAYS WILL NOT BE 
PERFORMED ON SAMPLES FROM HEALTHY PEDIATRIC DONORS

44
99-H-0050
Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
D.1
Assays of serum, blood cells, and bone marrow cells for B19 parvovirus and possible 
B19  variants using gene amplification, cell culture, and hematopoietic colony 
inhibition assays.
No
N/A
D.2
Assays of blood, bone marrow, liver, and other tissues for potentially novel viruses, 
using a variety of techniques including RNA and DNA assays, differential display, 
gene amplification with conserved and random primers, cell culture assays, 
immunohistochemical methods, and inocculation of mice, rabbits, and monkeys, as 
well as antibody measurements.
No
N/A
D.3
Assays of blood, bone marrow, and liver for known viruses, including herpesviruses 
such as cytomegalovirus, human herpesviruses 6, 7, and 8, enteric viruses such as A-
6, circiviruses, and parvoviruses, using assays as in (2).
No
N/A
D.4
Spectra-typing of blood cells to determine response to known or putative viral 
infections.
No
N/A
D.5
HLA typing or subtyping to determine risk factors/determinants for hepatitis-AA 
studies.
No
N/A
D.6
Cytotoxic lymphocyte assays with intracellular cytokine measurement for 
determining anti-viral response and lymphocyte cloning to obtain clones with 
specific antiviral activity.
No
N/A
E
Solid Tumor Section (Dr. Richard Childs)
E.1
Cr51 cytotoxicity assay to evaluating killing of patient tumor cells by patient NK cell 
clones and T-cells.
No
No
E.2
ELISA for IL-12  maturity of DC's made from subjects monocytes.
No
No
E.3
ELISA for IFN ã to evaluate specificity of CTL clones.
No
No
E.4
H thymidine uptake to evaluate proliferation potential of antigen specific T-cells.
No
No
E.5
PCR of STR to assess chimerism status of cellular subsets grown in-vitro or retrieved 
from subjects post-transplant.
No
No
E.6
Flow sorting of PBL and/or tissue samples to evaluate chimerism of different subsets.
No
No
E.7
Surface marker analysis of peripheral blood mononuclear cells using flow cytometry.
No
No
E.8
cDNA expression arrays to evaluate T-cells expression/gene patterns in subjects with 
GVHD and a GVT effect.
No
No
E.9
Geno typing of tumor or tissue samples by high density cDNA arrays.
No
No
E.10
VHL mutation analysis on kidney cancer tissue.
No
No
E.11
Transduction of dendritic and tissue cells with tumor antigens using plasmids, viral 
vectors and hybrid fusions.
No
No
E.12
Lasar capture microdisection of cells from tumor biopsies and tissue samples  to 
determine origin (donor vs patient).
No
No
E.13
Quantification of polyoma virus BK exposure by serology and PCR in stem cell 
transplant donors and recipients from blood and urine samples.
No
No
E.14
Quantification of polyoma virus BK specific T cells in stem cell transplant donors 
and recipients from peripheral blood samples.
No
No
E.15
Determination of origin of neovasculature endothelial cells in tumor and tissue 
samples obtained from subjects post transplant.
No
No
E.16
Quantification of lymphocyte subsets CD34 progenitors and endovasculator 
progenitors in G-CSF mobilized peripheral cell allografts.
No
No
E.17
Testing for polyoma virus BK latency in CD34 progenitors, B cells and T cells in the 
G-CSF mobilized peripheral cell allografts.
No
No
E.18
Determination of etiology of membraneous nephropathy using serum from subjects.
No
No

45
99-H-0050
Richard W. Childs, M.D.
February 22, 2022 (Amendment III)
E.19
Serum Proteomic patterns analysis to diagnose complications related to allogeneic 
transplantation.
No
No
E.20
Determine cell origin (donor vs patient) of tissue samples using IHC, IF, sorting, and 
FISH.
No
No
F
Lymphoid Malignancies Section (Dr. Adrian Wiestner)
F.1
Culture of cells from research subjects to investigate molecular disease mechanisms, 
model host tumor interactions, and to test effect of drugs on cell survival and cellular 
functions.
No
No
F.2
Generation of stable cell lines for the study of hematologic malignancies.
No
No
F.3
Modifications of cells using standard expression systems or biologic molecules, e.g. 
interfering RNA, to investigate the effects of candidate genes on cellular functions.
F.4
Identification and monitoring of B or T cell populations as identified by flow 
cytometry and by their B cell or T cell receptor expression.
No
No
F.5
Measurement of gene expression in cells or tissues. Techniques frequently used 
include gene expression profiling on microarrays, quantitative RT-PCR, Western 
blotting, flow cytometry and ELISA assays.
No
No
F.6
Analysis of chromosomal abnormalities or mutations in malignant cells and non-
malignant cells including FISH technology and DNA sequencing.
No
No
F.7
Assays of immune function of B-cells and T-cells, including intracellular cytokine 
staining, ELISPOT, quantitative RT-PCR for cytokines or other immune regulatory 
genes.
No
No
F.8
Analysis of antibody specificities in serum and antigen specificity of the B-cell 
receptor on cells. Techniques may include expression of antibodies in phage display 
systems, generation of antibodies in cell culture systems and use of such antibodies to 
screen for cognate antigens.
No
No
F.9
Transplantation of human cells into mice (xenograft model) to study disease biology 
and to investigate the effect of experimental therapy.
No
No
F.10
Measurements of drug concentrations, biologic molecules and disease markers in 
blood, serum, and plasma.
No
No