A Phase 1 Trial of CD25/Treg-depleted DLI Plus Ipilimumab for Myeloid Disease Relapse After Matched-HCT
Phase: PHASE1
Conditions: Acute Myeloid Leukemia, Myelodysplastic Syndromes, Myeloproliferative Neoplasms, Chronic Myelomonocytic Leukemia, Myelofibrosis
Sponsor: Dana-Farber Cancer Institute

A Phase 1 Trial of CD25/Treg-depleted DLI Plus Ipilimumab for Myeloid Disease Relapse After 
Matched-HCT 
NCT Number: NCT03912064
IRB Approval Date: Version 5.0 Protocol Version Date 06/26/2023 

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
3
TABLE OF CONTENTS
SCHEMA......................................................................................................................................2
1.
OBJECTIVES....................................................................................................5
1.1
Study Design.............................................................................................5
1.2
Primary Objectives....................................................................................5
1.3
Secondary Objectives................................................................................5
1.4
Correlative Objectives ..............................................................................5
2.
BACKGROUND ...............................................................................................6
2.1
Introduction...............................................................................................6
2.2
IND Agents...............................................................................................7
2.3
Rationale ...................................................................................................9
2.4
Correlative Studies Background ...............................................................9
3.
PARTICIPANT SELECTION.........................................................................10
3.1
Eligibility Criteria ...................................................................................10
3.2
Exclusion Criteria ...................................................................................11
3.3
Inclusion of Women and Minorities .......................................................12
4.
REGISTRATION ............................................................................................12
4.1
General Guidelines for DF/HCC Institutions .........................................12
4.1
Registration Process for DF/HCC Institutions........................................13
4.2
General Guidelines for Other Investigative Sites ...................................13
4.3
Registration Process for Other Investigative Sites..................................13
5.
TREATMENT .................................................................................................13
Overall Treatment Plan...............................................................................................................13
5.1
Treatment Regimen.................................................................................13
5.2
Pre-Treatment Criteria ............................................................................19
5.3
Agent Administration..............................................................................19
5.4
Definition of Dose-Limiting Toxicity (DLT)..........................................20
5.5
General Concomitant Medication and Supportive Care Guidelines.......21
5.6
Duration of Therapy................................................................................21
5.7
Criteria for Taking a Participant Off Protocol Therapy..........................22
5.8
Duration of Follow Up............................................................................22
5.9
End of Study Definition ..........................................................................23
6.
DOSING DELAYS/DOSE MODIFICATIONS..............................................23
7.
ADVERSE EVENTS: LIST AND REPORTING REQUIREMENTS
34
7.1
Definitions...............................................................................................34
7.2
Expected Toxicities.................................................................................36
7.3
Adverse Event Characteristics ................................................................38
7.4
Adverse Event Reporting........................................................................39
7.5
Reporting to the Food and Drug Administration (FDA).........................40

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
4
7.6
Reporting to Hospital Risk Management.....................40
7.7
Routine Adverse Event Reporting ...............................40
8.
PHARMACEUTICAL INFORMATION........................................................40
8.1
Ipilimumab...................................................................40
9.
CORRELATIVE STUDIES ............................................................................45
9.1
Laboratory Correlative Study Sample Collection ...................................45
9.2
Planned Correlative Studies.........................................45
10.
STUDY CALENDAR......................................................................................46
11.
MEASUREMENT OF EFFECT / Clinical response .......................................49
12.
DATA REPORTING / REGULATORY REQUIREMENTS .........................49
12.1
Data Reporting.............................................................50
12.2
Data Safety Monitoring................................................50
12.3
Collaborative Agreements Language...........................50
13.
STATISTICAL CONSIDERATIONS.............................................................50
13.1
Study Design ................................................................50
13.2
Primary endpoints ........................................................51
13.3
Secondary endpoints ....................................................51
13.4
Determining MTD........................................................52
13.5 Sample Size/Accrual Rate................................................53
13.6 Stratification Factors........................................................53
13.7
Analysis plan................................................................53
14.
PUBLICATION PLAN....................................................................................53
REFERENCES
55
APPENDIX A
PERFORMANCE STATUS CRITERIA
58
Appendix B: BODY SURFACE AREA AND CREATININE CLEARANCE .59 
Appendix C: ACUTE GVHD GRADING..........................................................60
Appendix D: NCI COMMON TOXICITY CRITERIA .....................................61
Appendix E: AML RESPONSE CRITERIA ......................................................62
Appendix F: MDS/MPN RESPONSE CRITERIA.............................................64
Appendix G: MANAGEMENT OF IMMUNE RELATED AEs DUE TO 
IPILIMUMAB ........................................................................................66

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
5
1. OBJECTIVES
1.1
Study Design:
This is an open-label, single-center Phase 1 study of CD25/Treg-depleted donor 
lymphocyte infusion (DLI) plus Ipilimumab in patients with AML, MDS, MPN who 
relapse after 8/8 HLA-matched hematopoietic cell transplantation (matched-HCT). Four 
dose levels combinations of CD25/Treg-depleted DLI plus Ipilimumab will be 
considered, starting at 3x107 CD3+ cells/kg and 1 mg/kg Ipilimumab. Cells will be infused 
on Day 1 and Ipilimumab will be administered every 3 weeks for 4 doses (days 1, 22, 43, 
and 64). Maintenance Ipilimumab every 3 months for 4 doses will be administered at 
weeks 24, 36, 48 and 60, to patients without disease progression at the day 92 (end of 
week 13) response assessment.
1.2
Primary Objectives
To determine the safety (MTD) of CD25/Treg-depleted DLI plus Ipilimumab in patients 
with myeloid relapse after matched-HCT.
1.3
Secondary Objectives
1. To determine complete remission (CR/CRi) rate at day 43 (6 weeks).
2. To determine the rate of progression-free survival (PFS) and overall survival (OS)at
~3 months (day 92) and ~1 year (week 60) post cell infusion.
3. To determine the ~3 month (day 92) incidence and severity of acute GVHD rates 
after cell infusion.
4. To determine the ~1 year (week 60) incidence and severity of chronic GVHD rates 
after cell infusion.
1.4
Correlative Objectives
1.
To assess the immunologic impact of infusions of CD25/Treg-depleted DLI plus 
Ipilimumab. These include immunophenotypic analysis of T-cell subsets, B and NK 
cells; phenotypic and functional readouts of effector T (Teff) and regulatory T (Treg) 
cell activation status; changes in plasma cytokines and chemokines; tumor 
microenvironment; and tumor mutation profile.
2.
To correlate clinical response with changes in circulating Teff vs. Treg cell counts 
and activation; the persistence of adoptively transferred Teff cells; changes in 
cytokine and chemokine levels; tumor infiltration by activated CD8+ T cells; and 
tumor mutation profile, blast phenotype and tumor microenvironment.
3.
In an exploratory analysis, to perform tumor neoantigen discovery to identify novel 
T cell targets and correlate tumor mutational load with response.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
6
2. BACKGROUND
2.1
Introduction
For many patients with myeloid malignancies (e.g. acute myeloid leukemia (AML), 
myelodysplastic syndromes (MDS), and myeloproliferative neoplasms (MPN)), allogeneic 
hematopoietic cell transplantation (HCT) offers the only opportunity for cure. Unfortunately, 
myeloid disease relapse after transplant is associated with dismal outcomes, and remains a major 
obstacle to HCT success1. Currently, up to 40% of patients with AML/MDS undergoing HLA- 
matched related or unrelated HCT (matched-HCT) relapse2,3. HCT relapses are usually treated 
by withdrawal of IS, chemotherapy for interim disease control, and/or donor lymphocyte 
infusion (DLI) to invoke potentially curative graft-versus-tumor (GVT) immune responses. 
However, AML/MDS remissions after DLI are infrequent and short-lived, with poor patient 
survivals typically measured in months4,5. Clinical graft-versus-host-disease (GVHD) toxicity is 
common after DLI. Innovative approaches to enhance anti-tumor immune responses are urgently 
needed.
Immune escape (i.e. tumor evasion of the donor immune system) contributes to myeloid disease 
relapse after allogeneic HCT. In HLA-matched HCT, presentation of host hematopoietic and/or 
tumor epitopes within the concordant HLA context can invoke T cell receptor (TCR)-mediated 
recognition and elimination by donor-derived effector T (Teff) cells and induce curative GVT 
responses, a process that may be antagonized by suppression of co-stimulatory immune 
pathways, inhibitory regulatory T (Treg) cells, or by activation of inhibitory immune checkpoint 
pathways6-8.
We therefore hypothesized that Treg cells in the DLI product could suppress GVT, and selective 
depletion of CD25hi cells (including constitutive CD25hi Treg cells) from the DLI product could 
boost its clinical efficacy, if severe clinical GVHD was avoided. In a phase 1 study we 
manufactured CD25/Treg-depleted DLI (via ClinMACS CD25 reagent system, Miltenyi Biotec 
GmbH), documenting its safety and promising efficacy at a dose of 3x107 CD3+ T cells/kg, 
highlighting complete durable remissions in relapsed AML/MDS9.
We additionally hypothesized that blockade of the inhibitory CTLA-4 immune checkpoint with 
Ipilimumab Ipi may induce a GVT effect in patients in relapse after HCT. In a phase 1 trial of 
Ipilimumab monotherapy for patients with relapsed hematologic malignancies including 
AML/MDS after HCT, we documented the feasibility, safety and promising early efficacy of 
CTLA-4 blockade, including some complete durable remissions in patients with myeloid 
disease10. Response was associated with increased tumor infiltration of perforin-expressing Teff 
cells, and fewer circulating Treg cells.
We now propose to leverage our prior experience and combine CD25/Treg-depleted DLIwith 
CTLA-4 blockade via Ipilimumab to maximally augment Teff activity while avoiding infusion 
of suppressive Treg cells. We hypothesize that such combination therapy will enhance the 
efficacy of DLI in patients with myeloid disease relapse after matched-HCT.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
7
2.2
IND Agents
2.2.1
CD25/Treg-depleted DLI for matched HCT relapse
Tregs are CD4+CD25hiFoxp3+ lymphocytes that comprise approximately 5-10% of the 
circulating CD4+ T cell population, that act to dominantly suppress autoreactive lymphocytes, 
and control innate and adaptive immune responses11-18. In murine studies, transfer of CD4+ T 
cells depleted of Tregs induces autoimmunity, while transfer of CD4+ Tregs is protective19,20. 
Patients with systemic lupus erythematosus, rheumatoid arthritis, and multiple sclerosis exhibit 
deficiencies in Treg number and function21-23. The relationship between Tregs and autoimmunity 
prompted interest in their potential role in GVHD. Adoptive transfer of donor Treg at HCT can 
prevent or mitigate GVHD in animal models24-26. Several groups examined the PB of patients at 
various stages after HCT and found that the number of circulating Tregs inversely correlated 
with GVHD27-30. We previously reported that in-vivo Treg enhancement with daily subcutaneous 
(SC) low-dose interleukin-2 (IL-2) therapy can induce clinical responses in steroid-refractory 
chronic GVHD31,32. Tregs can also potentially suppress the GVT response. In vitro Tregs 
suppress proliferation of CD4+ and CD8+ Teff cells after polyclonal or antigenic stimulation17. 
In vivo Tregs diminish immune responses to solid and liquid tumors, including AML33-36. We 
therefore hypothesized that selective depletion of Tregs (by depletion of CD25hi cells) from the 
DLI product could boost its clinical efficacy, if severe clinical GVHD was avoided.
In a phase 1 study for patients with disease 
relapse after matched-HCT, we evaluated the 
feasibility of manufacturing Treg depleted 
DLI products as well as the safety and 
potential efficacy of these cells. Treg depleted 
DLI products were obtained by depleted 
CD25hi cells from donor apheresis products 
using antibody-conjugated magnetic beads 
(Miltenyi Biotech – CliniMacs)9. Sixteen of
21 subjects received prior cytoreductive 
therapy, but only four were in complete 
remission at time of infusion. Two DLI dose 
levels were administered: 1x107 (n=6) and 
3x107 CD3+ cells/kg (n=15). A median 2.3 
log-depletion of CD4+CD25hiFOXP3+ Treg 
cells was achieved in the released product 
(Fig. 1).
Infusion of CD25/Treg-depleted products was not associated with acute toxicity. Seven subjects 
(33%) developed clinically significant GVHD by one year, including one fatal case. At Dose 
Level 1, five treated patients demonstrated progression, while one patient had stable disease. At 
Dose Level 2 however, nine subjects (60%) achieved responses (8 CR, 1 PR), including seven 
who had active disease at time of DLI. One-year survival was 53%. When compared to 
unmodified DLI in contemporaneous patients meeting study eligibility (n=14), CD25/Treg- 
depleted DLI at dose-level 2 was associated with an improved response rate (60% vs. 14%, 
p=0.02) and 1-year event-free survival (EFS, 27% vs. 0%, p=0.0075, Fig. 2), with a reduction in
Fig 1: Flow Cytometric Analysis of CD4+CD25+FoxP3+ 
Cells Confirms Treg Depletion. The initial apheresis 
product (top plots) and CD25-depleted target fraction 
(bottom plots) indicate reduction in CD25hi cells (left) 
nd FoxP3+ c ll (ri ht)

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
9
unpublished data, 5 mg/kg Ipilimumab dose appears less effective than 10 mg/kg, with similar 
toxicity. While promising, these data indicate a need for further enhancement of GVT beyond 
that obtained with Ipilimumab alone.
Exploratory immunohistochemical and gene-expression data indicated that in leukemia cutis, 
Ipilimumab therapy drove infiltration of cytotoxic CD8+ T cells to the site of disease both in 
responders and non-responders, but CD8+ T-cell perforin-1 up-regulation was more apparent in 
two responders compared to non-responders. PB analysis showed decreased circulating CD4+ 
Tregs and increased CD4+ Teff cells in patients who had a response. Moreover, in responders the 
proportion of activated CD4+ Treg cells was less than in non-responders. These observations 
suggest that CTLA-4 blockade may be effective after allogeneic HCT by inducing an 
intratumoral GVT response, but that responses might be blunted by Tregs. Strategies to avoid 
infusing Tregs while de-repressing Teff cells might therefore help to further enhance therapeutic 
GVT responses.
2.3
Rationale
Based on the initial safety and promising efficacy observed in our phase 1 trials of CD25/Treg- 
depleted DLI and Ipilimumab individually, we hypothesize that combination therapy will be safe 
in patients with AML, MDS and MPN who relapse after matched-HCT and provide an enhanced 
curative effect for these patients with otherwise extremely poor prognosis.
2.4
Correlative Studies Background
Clinical trial administration of adoptively transferred CD25/Treg-depleted DLI plus Ipilimumab 
provides a unique opportunity to study their biology within patients. Peripheral blood (PB), bone 
marrow (BM) and accessible extramedullary disease site biopsy (e.g. leukemia cutis, if 
applicable) samples obtained before (at baseline), during, and after therapy (e.g. landmark time 
points like Day +30, Day +100 etc.,) and at the time of relapse (if any) will be obtained to assess 
the immunologic effects of treatment.
Viable cells as well as plasma will be isolated and cryopreserved at each time point using 
validated lab SOPs. Since all patients will have active relapse at the initiation of therapy, baseline 
samples will have tumor cells as well as normal donor immune cells. We will conduct detailed 
immunologic and tumor assessments, characterizing immune cells pre- and post-treatment, 
evaluating their functional activity, and assessing the tumor microenvironment to identify 
biologic predictors of response. We hypothesize that infusion of DLI preferentially depleted of 
Treg cells in combination with Ipilimumab will result in enhanced GVT mediated by donor Teff 
cells. To examine this hypothesis, correlative studies will use multiple orthogonal approaches to 
monitor changes in T cell activation, T cell specificity and T cell function in responders and non- 
responders. Specifically, we will seek to correlate clinical response with 1) changes in circulating 
Teff vs. Treg cell counts and activation status via flow cytometry and mass cytometric analysis;
2) the persistence of adoptively transferred Teff cells via TCR sequencing;
3) changes in cytokine and chemokine levels; 4) tumor infiltration by activated CD8+ T cells; 
and 5) tumor mutation profile, blast phenotype and tumor microenvironment via unbiased single- 
cell expression profiling of the immune microenvironment (e.g. CD8+ T, CD4+ Teff, CD4+ Treg, 
NK, B, DC, Macrophage, MDSC, blast cells) using specialized microfluidic platforms (e.g.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
10
inDrop and 10x Genomics) we have optimized for use in cryopreserved BM, PB, and other single 
cell suspensions. In an exploratory analysis, we will perform tumor neoantigen discovery to 
identify novel T cell targets and correlate tumor mutational load with response.
3. PARTICIPANT SELECTION
3.1
Eligibility Criteria
3.1.1
Histologically or cytologically or molecularly confirmed relapse of AML, MDS or MPN 
including CMML, myelofibrosis or MDS/MPN. Disease relapse includes patients with myeloid 
blasts by flow and/or morphology in marrow or extramedullary disease sites; or persistence 
and/or recurrence of disease defining mutations.
3.1.2
Relapse at ≥2 months after any 8/8 or better HLA-matched HCT
3.1.3
Available original stem cell donor.
3.1.4
Age ≥18 years. Because no dosing or adverse event data are currently available on the 
use of Ipilimumab in participants <18 years of age, children are excluded from this study.
3.1.5
ECOG performance status ≤2 (Karnofsky performance status ≥60, see Appendix A).
3.1.6
Recipient donor T cell chimerism 20% within 4 weeks prior to cell infusion.
3.1.7
<50% bone marrow involvement within 4 weeks prior to cell infusion.
3.1.8
No systemic corticosteroid therapy for GVHD (≤5 mg of prednisone or equivalent doses 
of other systemic steroids for non-GVHD, non-autoimmune indications for at least 4 
weeks prior to cell infusion).
3.1.9
No other systemic medications/treatments (e.g. ECP) for GVHD for at least 4 weeks prior 
to cell infusion.
3.1.10 Ability to understand and willingness to sign written informed consents.
3.1.11 Adequate organ function as defined below:

Total bilirubin: ≤1.5 x institutional upper limit of normal (ULN) (except Gilbert’s
or disease-related hemolysis, then < 3 x ULN)

AST(SGOT)/ALT(SGPT): ≤3 x institutional ULN

creatinine clearance: ≤1.5 x institutional ULN

O2 saturation: ≥90% on room air

LVEF >40%

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
11
3.1.12 The effects of CD25/Treg-depleted DLI and Ipilimumab on the developing human fetus 
are unknown. For this reason and because immunomodulatory agents are known to be 
teratogenic, women of child-bearing potential and men must agree to use adequate 
contraception (hormonal or barrier method of birth control; abstinence) prior to study 
entry and for the duration of study participation. Should a woman become pregnant or 
suspect she is pregnant while participating in this study or within 23 weeks after the last 
dose of study drug, she should inform her treating physician immediately. Men treated 
or enrolled on this protocol must also agree to use adequate contraception prior to the 
study, for the duration of study participation, and for at least 31 weeks after completion 
of Ipilimumab administration.
3.1.13 Negative pregnancy test for females of childbearing potential only
3.2
Exclusion Criteria
3.1.14 Extramedullary relapse involving immuno-privileged sites (e.g. CNS, testes, eyes). Other 
sites of extramedullary relapse (e.g. leukemia cutis, granulocytic sarcoma) are acceptable.
3.1.15 Participants who had cytotoxic chemotherapy or other investigational agents within 
4 weeks prior to cell infusion (6 weeks for nitrosoureas or mitomycin C), or 
immunotherapy within 8 weeks prior, or those who have not recovered from adverse 
events due to agents administered more than 4 weeks prior. Use of hydroxyurea, 
HMAs (e.g. azacytidine, decitabine) and/or approved novel agents (e.g. venetoclax, 
FLT3 inhibitors, IDH1/2 inhibitors etc.) to control counts within 4 weeks prior to 
cell infusion is permitted.
3.1.16 Prior history of DLI
3.1.17 Prior history of treatment with anti-CTLA-4 or anti-PD-1 pathway therapy, or CD137 
agonist therapy.
3.1.18 Prior history of severe (grade 3 or 4) acute GVHD, or ongoing active GVHD requiring 
systemic treatment.
3.1.19 Organ transplant (allograft) recipient.
3.1.20 History of allergic reactions attributed to compounds of similar chemical or biologic 
composition to Ipilimumab or other agents used in study.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
12
3.1.21 Autoimmune disease: Patients with a history of inflammatory bowel disease, including 
ulcerative colitis and Crohn’s Disease, are excluded from this study, as are patients with 
a history of symptomatic disease (e.g., rheumatoid arthritis, systemic progressive 
sclerosis [scleroderma], systemic lupus erythematosus, autoimmune vasculitis [e.g., 
Wegener’s Granulomatosis]) and motor neuropathy considered of autoimmune origin 
(e.g. Guillain-Barre Syndrome and Myasthenia Gravis). Patients with Hashimoto’s 
thyroiditis are eligible to go on study.
3.1.22 Uncontrolled intercurrent illness including, but not limited to, ongoing or active 
infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac 
arrhythmia, or psychiatric illness/social situations that would limit compliance with 
study requirements.
3.1.23 Pregnant women are excluded from this study because of the unknown teratogenic risk. 
Because there is an unknown but potential risk for adverse events in nursing infants 
secondary to treatment of the mother, breastfeeding should be discontinued if the 
mother is treated on this study.
3.1.24 HIV-positive participants on combination antiretroviral therapy are ineligible because of 
the potential for pharmacokinetic interactions with agents used in this study. In addition, 
these participants are at increased risk of lethal infections when treated with marrow- 
suppressive therapy. Appropriate studies will be undertaken in participants receiving 
combination antiretroviral therapy when indicated.
3.1.25 Individuals with active uncontrolled hepatitis B or C are ineligible as they are at high 
risk of lethal treatment-related hepatotoxicity after HCT.
3.3
Inclusion of Women and Minorities
Both men and women of all races and ethnic groups are eligible for this trial.
4. REGISTRATION
4.1
General Guidelines for DF/HCC Institutions
Institutions will register eligible participants in the Clinical Trials Management System 
(CTMS) OnCore. Registrations must occur prior to the initiation of any protocol-specific 
therapy or intervention. Any participant not registered to the protocol before protocol-specific 
therapy or intervention begins will be considered ineligible and registration will be denied.
An investigator will confirm eligibility criteria and a member of the study team will complete 
the protocol-specific eligibility checklist.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
13
Following registration, participants may begin protocol-specific therapy and/or intervention. 
Issues that would cause treatment delays should be discussed with the Overall Principal 
Investigator (PI). If the subject does not receive protocol therapy following registration, the 
subject must be taken off-study in the CTMS (OnCore) with an appropriate date and reason 
entered.
4.2
Registration Process for DF/HCC Institutions
Applicable DF/HCC policy (REGIST-101) must be followed.
4.3
General Guidelines for Other Investigative Sites
N/A
4.4
Registration Process for Other Investigative Sites
N/A
5. TREATMENT
OVERALL TREATMENT PLAN
This is a Phase 1 study of CD25/Treg-depleted DLI plus Ipilimumab for eligible patients with 
relapsed myeloid malignancies after 8/8 HLA-matched donor bone marrow or peripheral blood 
stem cell transplantation. Donor CD25hi cells will be depleted ex-vivo from peripheral blood 
mononuclear cells prior to infusion, and Ipilimumab will be administered after DLI. The study 
will follow a Phase 1 design in which matched-HCT recipients will receive escalating or de- 
escalating doses of CD25/Treg depleted donor lymphocytes and Ipilimumab. Patients will 
receive maintenance Ipilimumab per their assigned dose level.
5.1
Treatment Regimen
5.1.1
Patient preparation:
Prior to enrollment in the study, all patients will undergo a BM aspirate and biopsy (and/or biopsy 
of accessible extra-medullary disease sites, e.g. leukemia cutis) to confirm histologic or cytologic 
relapse. Patients may receive combination chemotherapy to control relapsed disease and/or re- 
induce remission if clinically indicated prior to DLI. Patients must have all cytotoxic or cytostatic 
agents (except hydroxyurea) discontinued at least 4 weeks prior to DLI (6 weeks for nitrosoureas 
or mitomycin C). Hydroxyurea may be instituted before or after the CD25/Treg-depleted donor 
lymphocyte infusion, if needed to temporarily control peripheral blood counts, after approval by 
the Study PI.
5.1.2 Donor Leukapheresis.
Prior stem cell donors will undergo apheresis to obtain donor lymphocytes. When possible, this 

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
14
will be performed via peripheral intravenous access. Donor evaluation, consents and apheresis 
may be performed at other centers (including international sites), and will require completion of 
appropriate consents and medical evaluations, per each donor center’s standard operating 
procedures (SOPs). Apheresis for this study does not entail any added risk to the donor compared 
with unmodified DLI.
We anticipate that sufficient numbers of CD3+ donor lymphocytes will be collected from a single 
apheresis. After apheresis, an aliquot of the product will undergo analysis to determine the dose 
of CD3+ cells. Dosing will be determined by the CD3+ cells/kg as defined in Table 2. An aliquot 
will also be removed for microbiologic/sterility testing. If the collected product does not contain 
sufficient CD3+ cells (i.e. at least 1.5X the number of CD3+ cells/kg planned for infusion) the 
unmanipulated product will be administered off-study, dosed per the providers preference, and 
another patient will be recruited instead. The product, if adequate, will be processed per CMCF 
standard operating procedure and infused within the stated product expiration time.
5.1.3 Preparation and Infusion of CD25/Treg-depleted cells.
CD25/Treg depletion: All cell processing will be performed at the DFCI cGMP CMCF (Connell 
O’Reilly Cell Manipulation Core Facility). Upon receipt of sufficient donor leukapheresis 
product from the donor center, the product will be selectively depleted of CD25hi Treg cells using 
the CliniMACS® CD25 Reagent System according to manufacturer’s instructions. A detailed 
protocol for the preparation and use of the CliniMACS® CD25 reagent is provided to the clinical 
site in the CliniMACS® User Manual. Samples of the donor leukocyte preparation will be 
removed prior to processing and after completion of the CD25hi cell depletion. These samples 
will be used to determine cell viability, and enumerate the CD3+, CD4+, CD8+, CD14+, 
CD20+, CD25+, CD45+, CD56+, CD127+, and FOXP3+ cells (in combination as 
necessary, e.g. CD4+CD25hiFOXP3+ cells) before and/or after processing. Gram stain, 
endotoxin and sterility testing will also be performed on the donor leukocyte preparation. The 
CD25hi depleted product will be denoted as ‘TC-T Reg (CD25hi) reduced cells’. Release criteria 
are indicated in Table
1. Product will be infused even if the final CD3+ cell dose/kg is below target at the maximum 
number of cells available after requisite QC samples obtained.
If the total product collected contains <1.5-fold excess of CD3+ cells/kg per planned infusion 
dose, the product will NOT undergo CD25hi depletion, and the unmanipulated product will be 
given to the patient, per institutional standards of care. The patient will not be assessable for the 
study objectives, and will be replaced by another patient.
If the total product collected contains 1.5-fold excess of CD3+ cells/kg per planned infusion 
dose, then the product will be CD25hi depleted with the CliniMACS CD25 device according to 
manufacturer instructions. After depletion, the volume of processed product infused will be 
adjusted to the targeted CD3+ cell dose per the dose level schema, while maintaining 0.5% 
CD4+CD25hi cells. There will be no add back of the depleted fraction enriched for CD4+CD25hi 
cells.
If the composition of the product after processing cannot meet both targets above, then the 
volume infused will be adjusted to best target their CD3+ dose per dose-level cohort. The patient 
will be unevaluable for DLT assessment.

DF/HCC Protocol #: 19-142
 Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
15
Table 1: Product Release Criteria
Test Performed
Test Method
Release Criteria
Viable CD3+ cell dose*
Flow Cytometry
Dose Level +2: 3 x 107/kg
Dose Level +1: 3 x 107/kg
Dose Level   0: 3 x 107/kg
Dose Level  -1: 1 x 107/kg
CD4+ CD25hi cells
Flow cytometry
≤0.5% of viable lymphocytes
Viability
Trypan blue
≥70%
Endotoxin
Charles River Endosafe®
-PTS™ System
< 5.0 EU/Kg/hr
Gram stain
BWH Microbiology
laboratory
No organisms
* Acceptable range: 0.8-1 x 107 viable CD3+ cells/kg for DL -1; 2.5-3 x 107 viable CD3+ cells/kg for DL 0, +1,
+2
Donor Lymphocyte Infusion: The donor lymphocyte product will not be administered until 
it has passed the release criteria for infection control monitoring, as defined by the DFCI 
CMGTL and the FDA (Table 1). If release criteria (except CD3+ cell dose and CD4+CD25hi 
cell fraction) are not met, the DLI product will NOT be infused, and the patient will be 
unevaluable for DLT assessment and will be replaced.
Once release criteria have been met, the DLI product will be infused into the recipient 
through a central or peripheral venous catheter, per the cell infusion SOP. The donor 
lymphocytes will be infused over 5-15 minutes. Acetaminophen 650mg PO and 
diphenhydramine 25mg IV or PO will be given as premedication prior to the infusion. 
Patients will be monitored for infusional reactions for 1 hour after the end of the infusion. 
Excess product, if any, will be cryopreserved per standard operating procedure for future use 
if necessary.
CD25/Treg-depleted DLI Dose Schema: Patients will receive a defined dose of CD25hi 
Treg depleted DLI. 5 patients will be enrolled, initially at dose level 0, and subsequent 
cohorts will be dose adjusted per the CD3+ dose escalation/de-escalation schema detailed 
below.
Table 2: CD25/Treg-depleted DLI Dose Escalation/De-escalation Schema Per Cohort
Cohort
Viable CD3+ Dose* 
(#cells/kg**)
Dose Level +2
3 x 107
Dose Level +1
3 x 107
Dose Level 0
3 x 107
Dose Level -1
1 x 107
** Recipient’s body weight in Kg
** Acceptable range: 0.8-1 x 107 viable CD3+ cells/kg for DL -1; 2.5-3 x 107 viableD3+ cells/kg for DL 0, +1, 
and +2

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
16
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
5.1.4 Ipilimumab administration.
Ipilimumab will be administered intravenously over 90 ± 10 minutes after the DLI product 
observation period is completed, and dosing will continue in 3-week intervals for 4 cycles, unless 
unacceptable toxicity or symptomatic disease progression occurs (see Appendices E-F for 
disease progression criteria). MTD will be assessed at day 43 by DLT criteria as defined in 
Section 5.4. At their day 92 evaluation, patients without evidence of progressive disease or 
unacceptable toxicity can receive maintenance Ipilimumab per their assigned dose level on a q 
3 monthly basis starting from week 24, for 4 doses. 
5.1.5 Dose-escalation/De-escalation schema: Four dose-level combinations of CD25/Treg- 
depleted DLI plus Ipilimumab will be evaluated to determine MTD.
Table 4: CD25/Treg-depleted DLI plus Ipilimumab Dose Escalation/De-escalation Schema Per 
Cohort
Cohort
Viable CD3+
Dose 
(#cells/kg*)
Ipilimumab Dose
Dose Level +2
3 x 107
10 mg/kg
Dose Level +1
3 x 107
3 mg/kg
Dose Level 0
3 x 107
1 mg/kg
Dose Level -1
1 x 107
1 mg/kg
** Recipient’s body weight in Kg
** Acceptable range: 0.8-1 x 107 viable CD3+ cells/kg for DL -1; 2.5-3 x 107 viableD3+ cells/kg for DL 0, +1, 
and +2
To ensure at least 3 patients will be evaluable within a group, a cohort of 5 eligible patients will 
enter at each dose level. If there are fewer than 3 evaluable patients in a cohort, additional patients 
will be added at that dose-level. Patients may be enrolled in the next cohort when 3 or more 
evaluable patients from the previous cohort have completed their day 43 assessment. Patients 
will be considered unevaluable for the determination of MTD if they require removal from the 
study due to alternative treatments for progressive disease (except hydroxyurea, with PI 
approval) or other reasons not related to an adverse event within 6 weeks after initial dosing. 
Additional patients may be enrolled at a specific dose cohort to substitute for patients within the 
cohort who are considered unevaluable for MTD. There will be at least one-week interval 
between consecutive enrollments at the same dose level during phase 1A.
Dose escalation or de-escalation is as follows: If ≤1 DLTs are observed within a cohort of up to 
5 evaluable patients at dose level 0, then a dose escalation will take place. If ≤1 DLTs are
observed at dose-level -1 or dose-level +2, this dose will be the MTD. If ≥2 DLTs are observed 
in a cohort of up to 5 evaluable patients, then the MTD is considered exceeded. If this is dose 
level 0, dose de-escalation will take place, and up to 5 patients will be enrolled at dose level -1. 
If ≥2 DLTs are observed at dose level +2, then dose level+1 will be the MTD. If ≥2 DLTs are 
observed at dose level -1, accrual will stop.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
17
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
5.1.7 Further Therapy for Patients with Progressive Disease: Treatment with hydroxyurea to 
temporarily control counts, with PI approval, will be acceptable on study. Patients who require 
additional therapy due to progressive disease will come off study to be treated at the discretion 
of the treating physician. Patients who receive off-study therapy (e.g. chemotherapy) will be 
followed for 1 year following DLI infusion for survival outcomes. Phase 1a patients who are 
removed from study without evidence of toxicity prior to their week 6 assessment (e.g. for 
progressive disease) will be considered unevaluable for MTD assessment, and will be replaced 
as necessary. There will be at least one-week interval between consecutive enrollments at 
the same dose level during phase 1A.
5.1.7 Phase 1b: In phase Ib, an additional 10 patients will be enrolled at the MTD to better 
define toxicity and describe preliminary efficacy at this dose level. No patients will be replaced 
in this phase.
5.2
Pre-Treatment Criteria
5.2.1
Cycle 1, Day 1
In order to start on therapy, subjects must meet all of the inclusion criteria outlined in Section 
3.1, and must not have any of the exclusion criteria outlined in Section 3.2.
5.2.2
Other Cycles
To proceed with subsequent cycles, subjects must not have:

grade 2 or higher acute GVHD (see Appendix C for GVHD grading) requiring 
systemic immunosuppression

extensive (or severe) chronic GVHD requiring systemic immunosuppression

ongoing non-hematologic toxicity of grade 2 or higher unrelated to underlying 
malignancy or GVHD
Subjects will be allowed to proceed to subsequent cycles if they have ongoing:

immune-related adverse events not defined as GVHD (e.g. diarrhea, pruritus, 
endocrinopathy, and rash) that have responded to corticosteroids and improve to 
grade 1 or less
5.3
Agent Administration
Treatment will be administered on an outpatient basis. Reported adverse events and 
potential risks are described in Section 6. Dose modifications are not allowed in this
study per Section 6.1.3. No investigational or commercial agents or therapies other 
than those described below may be administered with the intent to treat the patient's 
malignancy.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
18
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
5.3.1
Ipilimumab
Unit: Ipilimumab 5 mg/mL, in 10-mL or 40-mLvials 
Route: Intravenous infusion
Appearance: Clear, colorless solution
Preparation, Handling, and Storage of Study Drug:
Please refer to Section 8.1 for detailed information in this regard.
Drug administration and dosage schedule:
Induction Phase:
Ipilimumab is administered by IV infusion every 3 weeks for a total of four doses (Cycles 
1-4). Infusions should be given over 90 +/- 10 minutes (not bolus or IV push).
Maintenance Phase:
Ipilimumab is administered by IV infusion every 12 weeks (3 months), beginning 
at Cycle 5, 24 weeks after the first dose of Ipilimumab, for up to 4 doses. Patients 
who show clinical benefit may be eligible to continue to receive Ipilimumab every 
12 weeks beyond Cycle 8 with prior approval from the Study PI.
Dose delays are allowed as per the dosing criteria described later in this Section.
Dose Levels:
There will be three Ipilimumab dose levels in this study in separate 
cohorts:
Dose levels -1, 0: 1mg/kg
Dose level +1: 3 mg/kg
Dose level +2: 10 mg/kg
5.4
Definition of Dose-Limiting Toxicity (DLT)
Toxicities are to be assessed according to the NCI Common Toxicity Criteria for Adverse 
Events (CTCAE), Version 5.0 (Appendix D). DLT observation period is 42 days (6 weeks) 
after cell infusion plus Ipilimumab on day 1. 
Definition of DLTs:
1.
Development of solid organ/non-hematologic toxicity (exclusive of those attributable to 

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
19
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
GVHD or Immune-related AEs) of persistent CTCAE grade 3 (i.e. without 
improvement to Grade 1 or less within 3 weeks) or grade 4 or higher (any duration) 
within 6 weeks of cell infusion and considered possibly, probably, or definitely related 
to cells and/or Ipilimumab. Non-clinically significant metabolic adverse events 
(abnormalities in serum sodium, 
potassium, chloride, 
magnesium, 
calcium, 
phosphorous, blood urea nitrogen,
and glucose that last less than 48 hours that respond to appropriate medical therapy) will 
not be considered DLTs regardless of CTCAE grade. Infections are expected in the 
context of allotransplantation and relapse, and will not considered DLTs.
2.
Severe cytopenias (ANC < 500/µl and/or Plts < 20,000/µl) not related to the underlying 
disease or infections (e.g CMV) or other etiologies (e.g. emptying of relapsed bone 
marrow), occurring within 6 weeks after cell infusion, that persist despite 1 week of 
growth factor support (e.g., G-CSF or GM-CSF). Patients would be evaluated with a BM 
biopsy to assess for underlying disease vs. infection (e.g., CMV) vs. loss of relapsed host 
hematopoiesis. If cytopenias are persistent, not explained by these or other transplant- 
or disease-associated causes, and possibly, probably or definitely related to cells and/or 
Ipilimumab, then the pancytopenia will be deemed a DLT.
3.
Acute GVHD: grade 3 or greater acute GVHD that does not improve to grade 2 or lower within 
14 days of treatment.
4.
Grade 3 or higher non-neurologic immune-related adverse events (e.g. diarrhea, pruritus, 
rash, endocrinopathies) that respond to corticosteroids and improve to grade 1 or less 
within 3 weeks will NOT count as DLTs. Biopsy of affected organs will be encouraged 
to help determine whether the immune-related condition is due to direct effects of 
Ipilimumab versus GVHD
5.
CTCAE grade 3 and 4 infusion reactions related to the study DLI and/or the initial 
infusion of Ipilimumab.
6.
Any grade 5 toxicity not due to underlying malignancy.
Management and dose modifications associated with the above adverse events are outlined 
in Section 6 (Expected Toxicities and Dosing Delays).
Dose escalation will proceed within each cohort according to the scheme outlined in
Section 5.1.5.
5.5
General Concomitant Medication and Supportive Care Guidelines
Antiviral, antifungal and antibacterial prophylaxis and monitoring should follow 
institutional practice as per the BMT-SOP. These typically include: daily acyclovir (or 
equivalent) for HSV prophylaxis, bactrim (or equivalent) for PCP prophylaxis, IV 
gammaglobulin for hypo-gammaglobulinemia, and azole (with mold activity) use for 
fungal prophylaxis as well as monitoring of beta-glucan and galactomannan levels. 
Growth factor support (e.g. GCSF, erythropoietin) and transfusion support is allowed, 
and can be used at the discretion of the investigators.
5.6
Duration of Therapy
Duration of therapy will depend on individual response, evidence of disease 
progression, and tolerance. In the absence of treatment delays due to adverse events,

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
20
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
treatment (including induction followed by maintenance therapy) may continue for 60 
weeks or until one of the following criteria applies:

Disease progression (as defined in Appendices E-F). As T-cell infiltration of the tumor 
site (e.g. granulocytic sarcoma, skin) can look like progressive disease, please discuss
the results of clinical or radiographic assessments (e.g. leukemia cutis site 
inflammation; granulocytic sarcoma scans) showing progressive disease with the study 
PI before removing a patient from the study for disease progression. Patients with 
radiographic progression who are deriving clinical benefit will be allowed to remain on 
treatment until the next schedule efficacy assessment.

Intercurrent illness that prevents further administration of treatment,

Unacceptable adverse event(s), requiring temporary or permanent stopping of 
Ipilimumab defined in Section 6.1.3

Participant decides to withdraw from the study, or

General or specific changes in the participant's condition render the participant

unacceptable for further treatment in the opinion of the treating investigator
5.7
Criteria for Taking a Participant Off Protocol Therapy
Participants will be removed from study when any of the criteria listed in Section 5.7 
apply. The reason for study removal and the date the participant was removed must be 
documented in the study-specific case report form (CRF). Alternative care options will 
be discussed with the participant.
Patients will be removed from the study for any of the following reasons:

Death

Adverse event(s) that, in the judgment of the investigator, may cause severe or 
permanent harm or which rule out continuation of study drug

General or specific changes in the patient’s condition render the patient
unacceptable for further treatment in the judgment of the investigator

Suspected pregnancy

Lost to follow-up

Patient withdraws consent

Investigator removes the patient from study

Dana Farber Cancer Institute decides to close the study
The reason for taking a participant off study, and the date the participant was removed, 
must be documented in the case report form (CRF). In addition, the study team will 
ensure Off Treatment/Off Study information is updated in OnCore in accordance with 
DF/HCC policy REGIST-101.
In the event of unusual or life-threatening complications, participating investigators must 
immediately notify the Principal Investigator.
John Koreth MBBS DPhil, 
Ph: 617-632-3470
Pager: 41341

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
21
DF/HCC Protocol #: 19-142
Protocol Version Date: 6/26/23
5.8
Duration of Follow Up
Participants removed from treatment for unacceptable AEs will be tracked for 30 days 
from last study treatment to document AE outcome (resolution, improvement, 
persistence or progression). For all patients, data on survival, disease status 
(remission/progression/relapse), and alternative therapies (if any) will be tracked for 1 
year after completion of last study treatment. 
Participants will also be asked to allow for long-term follow up so that survival, relapse 
and late toxicities, should they occur beyond 1 year, may also be identified. Long-term 
follow up will be at DFCI if participants live locally or with their local oncology 
providers if they live remotely. For participants living remotely, phone calls to their local 
oncology providers may be made on a 6-month basis.
5.9
End of Study Definition.
The end of study is defined as any one of the following (whichever occurs first):

The date of the 1-year follow-up

The date of death

Lost to follow-up

Patient withdraws consent
6. DOSING DELAYS/DOSE MODIFICATIONS
Dose delays and modifications during the study therapy will be made using the following 
recommendations. Toxicity assessments will be done using the CTEP Version 5.0 of the 
NCI Common Terminology Criteria for Adverse Events (CTCAE) which is identified and 
located on the CTEP website at: 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm.
If possible, symptoms should be managed symptomatically. In the case of toxicity, 
appropriate medical treatment should be used (including anti-emetics, anti-diarrheals, etc.).
All CTCAE grade 3 and higher events experienced by participants will be collected from the 
time of the first dose of study treatment, through the study and until the final study visit. 
Participants continuing to experience toxicity at the off study visit may be contacted for 
additional assessments until the toxicity has resolved or is deemed irreversible.
6.1
Anticipated Toxicities
A list of the adverse events and potential risks associated with the agents administered 
in this study appear below and will determine whether dose delays and modifications 
will be made or whether the event requires expedited reporting in addition to routine 
reporting.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
22
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
6.1.1 Adverse Event List for CD25/Treg-depleted DLI
Unselected DLI for treatment of malignant disease relapse has been associated with 
Teff-mediated GVHD flare, and with myelosuppression primarily in cases of 
significant leukemia/lymphoma involving the bone marrow (owing to replacement of 
host hematopoiesis). However, infusion of the donor CD25/Treg-depleted cells was 
found to be safe in our phase I clinical trial. All patients will be closely monitored for 
any adverse effects including GVHD and marrow suppression.

GVHD
The severity of acute GVHD is measured according to the extent of 
involvement of each of the three organs: skin, liver, and GI tract. The current 
grading system for acute GVHD ranges from grade 0 (no GVHD) to grade IV 
(severe, life threatening), and is shown in Appendix 1. Details on the 
management of acute GVHD will be discussed in Section 6.1.7

Pancytopenia
Myelosuppression can be seen after DLI and may be pronounced in patients 
with active disease in the bone marrow at the time of DLI. The management of 
pancytopenia is outlined in Section 6.1.8
6.1.2 Adverse Event List for Ipilimumab
Ipilimumab can result in severe and fatal immune-mediated adverse events probably 
due to T-cell activation and proliferation. A listing is provided in Section 7.2.1.2. These 
can include (but are not limited to) autoimmune hemolytic anemia, acquired anti-factor 
VIII immune response, autoimmune aseptic meningitis, autoimmune hepatitis, 
autoimmune thyroiditis, hepatic failure, pure red cell aplasia, pancreatitis, ulcerative 
and hemorrhagic colitis, endocrine disorders (e.g., autoimmune thyroiditis, 
hyperthyroidism, hypothyroidism, autoimmune hypophysitis/hypopituitarism, and 
adrenal insufficiency), ocular manifestations (e.g., uveitis, iritis, conjunctivitis, 
blepharitis, and episcleritis), sarcoid granuloma, myasthenia gravis, polymyositis, and 
Guillain-Barre syndrome. The majority of these reactions manifested early during 
treatment; however, a minority occurred weeks to months after discontinuation of 
Ipilimumab especially with the initiation of additional treatments.
6.1.3 Ipilimumab Dose Modifications/delays
Intra-patient dose modifications of Ipilimumab are not allowed in this study 
as lower doses have not been found to be less likely to lead to resolution of 
immune-related AEs that were seen at higher doses.
Decisions to delay an Ipilimumab dose will be made based on specified safety 
criteria. Treatment with Ipilimumab will be delayed or discontinued if the subject 
experiences at least one adverse event, specified below, considered by the

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
23
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
investigator to meet the criteria as outlined. The investigator should contact the 
Protocol Chair for any adverse event that will prompt a delayed dose or 
discontinuation of Ipilimumab. The following criteria will be used to determine 
dose delays, restarting doses, or discontinuing Ipilimumab:
It is necessary to delay study drug dosing for the following related adverse 
event(s):
• Any ≥ Grade 2 non-skin related adverse event (including irAEs), except for 
asymptomatic laboratory abnormalities
• Any ≥ Grade 3 laboratory abnormality
• Any adverse event, laboratory abnormality or intercurrent illness that, in the 
judgment of the investigator, presents a substantial clinical risk to the subject with 
continued dosing
• Any ≥ Grade 3 skin related adverse event regardless of causality
Restart Ipilimumab dosing if/when the adverse event(s) resolve(s) to ≤ Grade 
1 severity or return to baseline within 3 weeks of initiating the dose delay (no 
more than 6 weeks since the most recent prior dose administration).
The maximum amount of time that induction Ipilimumab can be delayed due to 
adverse events before patients must be taken off treatment is 3 weeks, (i.e. 6 
weeks between doses, respectively). Patients on maintenance Ipilimumab dosing 
must be taken off treatment if dosing is delayed for more than 4 weeks due to 
toxicity.
In the event of a dose delay:
• Certain study assessments, including bone marrow biopsy, T-cell chimerism, 
tumor measurement (CT or PET/CT), and/or correlative study samples associated 
with day 1 of the delayed cycle may be assessed on the original planned cycle day 
1 (first day of dose delay) only (do not need to be re-assessed on actual cycle day 
1, the day of cells and Ipilimumab infusion)
• In the event of a dose delay of the first dose (cycle 1, day 1) baseline bone 
marrow biopsy, radiographic assessment, T-cell chimerism do not need to be 
reassessed, even if those tests fall outside the required baseline time frames.
• If Ipilimumab dosing must be delayed for > 3 weeks (more than 6 weeks since 
the prior dose administration) for any of the above reasons, patients must come 
off treatment, except after approval by the study chair for individual cases with 
ongoing clinical benefit.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
24
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
6.1.4 Ipilimumab Discontinuation
Criteria that Require Permanent Discontinuation of Ipilimumab:
Subjects MUST discontinue study treatment for any of the following reasons 
that are not related to a related adverse event:
•
Withdrawal of informed consent (subject’s decision to withdraw for any reason)
•
Pregnancy
•
Termination of the study
•
Loss of ability to freely provide consent through imprisonment or involuntary 
incarceration for treatment of either a psychiatric or physical (e.g., infectious 
disease) illness.
Subjects must permanently discontinue Ipilimumab if they experience any adverse 
event, laboratory abnormality or intercurrent illness that, in the judgment of the 
investigator, presents a substantial clinical risk to the subject with continued dosing.
The following neurological adverse events require permanent discontinuation 
of Ipilimumab and defines unacceptable neurotoxicity:
•
Any neurologic motor toxicity ≥ Grade 3 regardless of causality
•
Any ≥ Grade 3 treatment-related sensory neurologic toxicity
Ipilimumab need NOT be discontinued in the following situations:
•
Potentially reversible inflammation (< Grade 4), attributable to a local antitumor 
reaction and a potential therapeutic response
•
Subjects with the following conditions where in the investigator’s opinion
continuing study drug administration is justified
•
Ocular toxicity that has responded to topical therapy
•
Endocrinopathies where clinical symptoms are controlled with appropriate 
hormone replacement therapy.
Note: Ipilimumab may not be restarted while the subject is being treated with 
systemic corticosteroids except for subjects on stable doses of hormone replacement 
therapy such as hydrocortisone for adrenal insufficiency.
6.1.5 Concomitant Treatments
Prohibited and/or Restricted Treatments
Subjects in this study may not have anti-tumor therapy (except hydroxyurea, with PI 
approval) for up to 4 weeks before and after dosing with Ipilimumab. Subjects must 
have not had immune suppression within 4 weeks of starting Ipilimumab. Patients 
started on systemic anti-neoplastic chemotherapy or immune suppression while on 
treatment will be taken off the study. The administration of live vaccinations while 
on treatment is prohibited. Short courses of corticosteroids for disease symptom 
control may be permitted after discussion with the Study Chair.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
25
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
6.1.6 Study Procedures by Visit and Treatment Cycle
Results of all safety laboratory tests (that is, all chemistry and all hematology 
results) must be obtained and reviewed before day 1 Ipilimumab administration, as 
applicable. All laboratory results must be within the established range before 
Ipilimumab is administered. All induction period baseline laboratory samples must 
be collected within a window of up to 7 days before administration of Ipilimumab.
6.1.7 Suggested Guidelines for the Management of GVHD
Biopsy of affected organs (e.g. liver, gut) should be considered to distinguish 
GVHD vs. immune-related toxicity when clinically appropriate (e.g. prior to 
systemic immune suppressive therapy). Clinically significant acute GVHD 
(Grade ≥ 2) may be treated with systemic steroids equivalent to 1-2 mg/kg/d of 
prednisone equivalent given daily until the disease is under control, then tapered 
10-20% per week thereafter at the treating physician’s discretion. Cyclosporine, 
tacrolimus, or other medications may be added as adjunctive therapy at the 
discretion of the treating SCT physician.
6.1.8 Suggested Guidelines for the Management of Pancytopenia
Pancytopenia is a common and expected condition for patients after chemotherapy 
and conventional DLI. As such, we will institute the following plan of action to 
minimize the duration of pancytopenia to reduce the risk of infection:
If any patient has persistent severe cytopenia post cell infusion with ANC <500/µl 
and/or Plts <20,000/µl persisting for >2 weeks even after 1 week of growth factor 
support, they would be evaluated with a BM biopsy to assess for diseaseprogression 
vs. GVHD vs. infection (e.g., CMV) vs. loss of chimerism.
Patients with pancytopenia will be also evaluated for infections (e.g. CMV) or graft 
loss/disease progression which can also cause pancytopenia in post HCT setting. If 
the pancytopenia is deemed on bone marrow biopsy to be secondary to disease 
progression and graft loss (e.g. increased blasts, loss of donor chimerism), disease 
response (emptying of a diseased marrow, and fall of host chimerism) or if an 
alternate cause is found (e.g. CMV infection), it will not be considered a DLT.
A hematopoietic cell infusion (CD34 selected ‘stem cell boost’) from the original 
donor can be considered in cases where there is no evidence of persistent disease and 
persistent donor chimerism.
6.1.9 Immune-Related Adverse Events (irAEs) Reactions and Immune- mediated 
Adverse Reactions: Definition, Monitoring, and Treatment
For the purposes of this study, an immune-related adverse reaction is defined as an 
adverse reaction of unknown etiology associated with drug exposure and consistent 
with an immune phenomenon. Efforts should be made to rule out neoplastic,

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
26
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
infectious, metabolic, toxin or other etiologic causes prior to labeling an event an 
irAEs. Serologic, immunologic, and histologic (biopsy) data should be used to 
support the diagnosis of an immune-related toxicity. Suspected immune-related 
adverse reactions must be documented on an AE or SAE form. Another term for an 
irAE is an immune-mediate adverse reaction, as it is termed in the Ipilimumab US 
Prescribing Information. Both terms may be used in this protocol document.
Patients should be informed of and carefully monitored for evidence of clinically 
significant systemic immune-mediated adverse reactions (e.g., systemic lupus 
erythematosus-like diseases) or organ-specific immune-mediated adverse reaction 
(e.g., rash, colitis, uveitis, hepatitis or thyroid disease). If an immune-mediated 
adverse reaction is noted, appropriate work-up (including biopsy if possible) should 
be performed, and steroid therapy may be considered if clinically necessary.
It is unknown if systemic corticosteroid therapy has an attenuating effect on 
Ipilimumab activity. However, clinical anti-tumor responses have been maintained 
in patients treated with corticosteroids and discontinued from Ipilimumab. If 
utilized, corticosteroid therapy should be individualized for each patient.
Recommended guidelines for specific immune-mediated adverse reactions are 
included in Section 6.1.13 below, in Appendix G, and the Ipilimumab package 
insert. These recommendations should be utilized as clinically appropriate for the 
treatment of individual patients. Please contact the study PI directly for any 
questions.
6.1.10
Other Guidance
The following guidance is provided for the management of Ipilimumab treatment 
related events. These recommendations, treatment algorithms in Appendix G, and 
further information in the IB, should be considered in the context of appropriate 
medical treatment for each patient. Subjects with ongoing adverse effects due to 
Ipilimumab at the conclusion of the study will be provided additional medical care 
as needed by the investigators.
6.1.11
Treatment of infusion reactions associated with Ipilimumab
Since Ipilimumab contains only human protein sequences, it is less likely that any 
allergic reaction will be seen in patients. However, it is possible that infusion of 
Ipilimumab will induce a cytokine release syndrome that could be evidenced by 
fever, chills, rigors, rash, pruritus, hypotension, hypertension, bronchospasm, or 
other symptoms. Such reactions were however not seen in our prior phase 1 trial for 
relapse after HCT. No prophylactic pre-medication should be given unless indicated 
by previous experience in an individual patient. Reactions should be treated based 
upon the following recommendations.
For mild symptoms (e.g., localized cutaneous reactions such as mild pruritus, 
flushing, rash):

Decrease the rate of infusion until recovery from symptoms, remain at bedside

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
27
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
and monitor patient.

Complete the Ipilimumab infusion at the initial planned rate.

Diphenhydramine 50 mg IV may be administered at the discretion of the 
treating physician and patients may receive additional doses with close 
monitoring

Premedication with diphenhydramine may be given at the discretion of the 
investigator for subsequent doses of Ipilimumab
For moderate symptoms (any symptom not listed above [mild symptoms] or below 
[severe symptoms] such as generalized pruritus, flushing, rash, dyspnea, 
hypotension with systolic BP >80 mmHg):
Interrupt Ipilimumab.
Administer diphenhydramine 50 mg IV.
Monitor patient closely until resolution of symptoms.
Corticosteroids may abrogate any beneficial immunologic effect, but may be 
administered after approval by Study PI
Resume Ipilimumab infusion after recovery of symptoms
At the discretion of the treating physician, Ipilimumab infusion may be resumed 
at one half the initial infusion rate, then increased incrementally to the initial 
infusion rate.
If symptoms develop after resumption of the infusion, the infusion should be 
discontinued and no additional Ipilimumab should be administered that day
The next dose of Ipilimumab will be administered at its next scheduled time and 
may be given with pre-medication (diphenhydramine and acetaminophen) and 
careful monitoring, following the same treatment guidelines outlined above.
At the discretion of the treating physician, additional oral or IV antihistamine 
may be administered prior to dosing with Ipilimumab
For severe symptoms (e.g., any reaction such as bronchospasm, generalized 
urticaria, systolic blood pressure <80 mm Hg, or angioedema):
Immediately discontinue infusion of Ipilimumab, and disconnect infusion tubing 
from the subject.
Consider bronchodilators, epinephrine 1 mg IV or subcutaneously, and/or 
diphenhydramine 50 mg IV, with solumedrol 100 mg IV, as needed
Patients should be monitored until the investigator is comfortable that the 
symptoms will not recur
No further Ipilimumab will be administered without approval from study PI
In case of late-occurring hypersensitivity symptoms (e.g., appearance within one 
week after treatment of a localized or generalized pruritus), symptomatic treatment 
may be given (e.g., oral antihistamine, or corticosteroids).
6.1.12
Treatment of Ipilimumab-Related Isolated Drug Fever

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
28
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
In the event of isolated drug fever, the investigator must use clinical judgment to 
determine if the fever is related to the Ipilimumab or to an infectious etiology. If a 
patient experiences isolated drug fever, for the next dose, pre-treatment with 
acetaminophen or non-steroidal anti-inflammatory agent (investigator discretion) 
should be instituted and a repeated antipyretic dose at approximately 6 and 12 hours 
after Ipilimumab infusion, should be administered. The infusion rate will remain 
unchanged for future doses. If a patient experiences recurrent isolated drug fever 
following premedication and post dosing with an appropriate antipyretic, the 
infusion rate for subsequent dosing should be decreased to 50% of the previous rate. 
If fever recurs following infusion rate change, the investigator should assess the 
patient’s level of discomfort with the event and use clinical judgment to determine 
if the patient should receive further Ipilimumab.
6.1.13
Monitoring and Management of Immune-mediated Adverse Reactions
We anticipate that Ipilimumab immune-mediate adverse reactions and GVHD will 
be difficult to distinguish clinically. No increase in GVHD was seen in the low 
dose, single-administration study of Ipilimumab in patients with hematologic 
malignancies post alloSCT. Therefore, for the present study, will need to use the 
experience with Ipilimumab in studies of solid tumor patients to guide us in 
distinguishing immune-related AEs from GVHD. Below is an overview of the 
approach to immune-related AEs utilized on prior Ipilimumab studies, which we 
will follow for the present study. In the present study, we will also encourage 
biopsy of the affected organ so that more can be learned about how to distinguish 
immunerelated AEs from GVHD pathologically.
Immune-mediated Enterocolitis
The clinical presentation of GI immune-related AEs included diarrhea, increase in 
the frequency of bowel movements, abdominal pain, or hematochezia, with or 
without fever. However inflammation may occur in any part of the GI tract 
including esophagitis and gastritis. Fatalities due to GI perforation have been 
reported in clinical trials of Ipilimumab. Patients should be carefully monitored for 
GI symptoms that may be indicative of immune-related colitis, diarrhea, or GI 
perforation. Diarrhea or colitis occurring after initiation of Ipilimumab therapy 
should be evaluated to exclude infectious or alternate etiologies.
In symptomatic patients, rule out infectious etiologies and consider endoscopic 
evaluation with biopsy to establish etiology and for persistent or severe symptoms. 
Withhold Ipilimumab dosing for any patients with enterocolitis pending 
evaluation; administer anti-diarrheal treatment and, if persistent evaluate with 
colonoscopy and biopsy (to distinguish GVHD vs. immune-mediated 
inflammation) and initiate systemic corticosteroids at a dose of 0.5 mg/kg/day 
prednisone or equivalent.
Permanently discontinue Ipilimumab in patients with severe enterocolitis and 
evaluate with colonoscopy and biopsy (to distinguish GVHD vs. immune- 
mediated inflammation), and initiate systemic corticosteroids at a dose of 1 to 2

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
29
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
mg/kg/day of prednisone or equivalent. Upon improvement to grade 1 or less, 
initiate corticosteroid taper and continue to taper over at least one month. In 
clinical trials, rapid corticosteroid tapering has resulted in recurrence or 
worsening symptoms of enterocolitis in some patients.
Immune-mediated Hepatitis and Pancreatitis
Hepatic immune-related AEs were mostly clinically silent and manifested as 
transaminase or bilirubin laboratory abnormalities. Fatal hepatic failure has been 
reported in clinical trials of Ipilimumab. Serum transaminase and bilirubin and 
lipase levels must be evaluated before each dose of Ipilimumab as early laboratory 
changes may be indicative of emerging immune-related hepatitis/ pancreatitis and 
elevations in liver function tests (LFTs) may develop in the absence of clinical 
symptoms. Increase in LFT or total bilirubin should be evaluated to exclude other 
causes of hepatic injury, including infections, disease progression, or other 
medications, and monitored until resolution. Liver biopsies from patients who had 
immune-related hepatotoxicity showed evidence of acute inflammation 
(neutrophils, lymphocytes, and macrophages), and biopsy should be considered in 
patients with grade 2 and higher hepatotoxicity (to distinguish GVHD vs.
immune-related hepatotoxicity).
Monitor liver function tests (hepatic transaminase and bilirubin levels, lipase) and 
assess patients for signs and symptoms of hepatotoxicity/ pancreatitis before each 
dose of Ipilimumab. Withhold Ipilimumab in patients with grade 2 hepatotoxicity 
and consider liver biopsy (to distinguish GVHD vs. immune-related 
hepatotoxicity).
Permanently discontinue Ipilimumab in patients with grade 3–5 hepatotoxicity/ 
pancreatitis, consider liver biopsy (to distinguish GVHD vs. immune-related 
hepatotoxicity), and administer systemic corticosteroids at a dose of 1 to 2 
mg/kg/day of prednisone or equivalent. When liver function tests show sustained 
improvement or return to baseline, initiate corticosteroid tapering and continue to 
taper over 1 month.
Immune-mediated Dermatitis
Skin immune-related AEs presented mostly frequently as a rash and/or pruritus. 
Some subjects reported vitiligo associated with Ipilimumab administration. Fatal 
toxic epidermal necrolysis has been reported in clinical trials of Ipilimumab.
Monitor patients for signs and symptoms of dermatitis such as rash and pruritus. 
Unless an alternate etiology has been identified, signs or symptoms of dermatitis 
should be considered immune-mediated.
For mild to moderate dermatitis, such as grade 2 localized rash and pruritus, treat 
symptomatically. For persistent grade 2, grade 3, or greater, topical steroids may be 
administered. Administer topical or systemic corticosteroids as indicated if there is 
no improvement of symptoms within 1 week.
Withhold Ipilimumab dosing in patients with moderate to severe signs and 
symptoms.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
30
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
Permanently discontinue Ipilimumab in patients with Stevens-Johnson syndrome, 
toxic epidermal necrolysis, or rash complicated by full thickness dermal ulceration,
or necrotic, bullous, or hemorrhagic manifestations. Administer systemic 
corticosteroids at a dose of 1 to 2 mg/kg/day of prednisone or equivalent. When 
dermatitis is controlled, corticosteroid tapering should occur over a period of at 
least 1 month.
Immune-related Neurological Events
Fatal Guillain-Barré syndrome has been reported in clinical trials of Ipilimumab. 
Patients may present with muscle weakness and myasthenia gravis, cranial nerve 
palsy (nVII Bells palsy), and aseptic meningitis, encephalopathy.
Unexplained motor neuropathy, muscle weakness, or sensory neuropathy lasting 
more than 4 days should be evaluated and non-inflammatory causes such as disease 
progression, infections, metabolic syndromes, nerve entrapment, and medications 
should be excluded as causes.
Withhold Ipilimumab dosing in patients with any evidence of neuropathy pending 
evaluation.
Permanently discontinue Ipilimumab in patients with severe neuropathy (interfering 
with daily activities) such as Guillain-Barré-like syndromes. Consider initiation of 
systemic corticosteroids at a dose of 1 to 2 mg/kg/day prednisone or equivalent for 
severe neuropathies.
Immune-mediated Endocrinopathies
Ipilimumab can cause inflammation of endocrine organs including thyroid 
(Hashimoto’s thryroiditis with positive antibodies) and adrenal glands, 
hypophysitis, hypopituitarism, and resulting thyroid and adrenal insufficiency, low 
ADH, prolactin, FSH, LH. Hyperthyroid with Graves’ disease and positive antibody 
has been reported.
Patients may present with subtle and nonspecific symptoms. The most common 
clinical presentation includes headache and fatigue. Symptoms may also include 
visual field defects, behavioral changes, and electrolyte disturbances including 
hyponatremia and hypotension. Adrenal crisis as a cause of the patient’s symptoms 
should be excluded. Most of the subjects symptomatically improved with hormone 
replacement therapy. Long-term hormone replacement therapy with HC and 
levothyroxine will typically be required for subjects developing 
hypophysitis/hypopituitarism after treatment with Ipilimumab. Some patients have 
regained partial function following steroid treatment.
Unless an alternate etiology has been identified, signs or symptoms of 
endocrinopathies should be considered immune-mediated and drug withheld 
pending evaluation. Patients may demonstrate both central (hypophysitis) and 
peripheral adrenal and thyroid insufficiency. Evaluation of hypophysitis should 
include pituitary MRI.

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
31
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
Monitor thyroid function tests and clinical chemistries at the start of treatment and 
recheck if signs/symptoms of thyroid dysfunction arise. Withhold Ipilimumab 
dosing in symptomatic patients. Initiate systemic corticosteroids at a dose of 1 to 2 
mg/kg/day of prednisone or equivalent, and initiate appropriate hormone 
replacement therapy.
Other Immune-mediated Adverse Reactions, Including Ocular Manifestations 
Ocular inflammation, manifested as grade 2 or grade 3 episcleritis or uveitis, was 
associated with concomitant diarrhea in a few subjects (<1%) and occasionally 
occurred in the absence of clinically apparent GI symptoms. Other presumed 
immune-related AEs reported include, but were not limited to, arthritis/arthralgias, 
pneumonitis, pancreatitis, immune-related (aseptic) meningitis, immune-related 
nephritis, pure red cell aplasia, noninfective myocarditis, polymyositis, and 
myasthenia gravis, of which were individually reported for <1% of subjects.
Initiate systemic corticosteroids at a dose of 1 to 2 mg/kg/day prednisone or 
equivalent for severe immune-mediated adverse reactions.
Administer corticosteroid eye drops to patients who develop uveitis, iritis, or 
episcleritis. Permanently discontinue Ipilimumab for immune-mediated ocular 
disease that is unresponsive to local immunosuppressive therapy.
Immune-related Cardiac Toxicities

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
32
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
Overall, immune-related AEs commonly started within 3 to 10 weeks from first 
dose, were successfully managed in most instances by omitting doses, discontinuing 
dosing, and/or through administering symptomatic or immunosuppressive therapy, 
including corticosteroids, as detailed above. Immune-related AEs generally 
resolved within days to weeks in the majority of subjects.
7. ADVERSE EVENTS: LIST AND REPORTING REQUIREMENTS
7.1
Definitions
7.1.1
Adverse Event

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
33
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
An adverse event (AE) is any undesirable sign, symptom or medical condition or 
experience that develops or worsens in severity after starting the first dose of 
study treatment or any procedure specified in the protocol, even if the event is not 
considered to be related to the study.
Abnormal laboratory values or diagnostic test results constitute adverse events 
only if they induce clinical signs or symptoms or require treatment or further 
diagnostic tests.
7.1.2
Expectedness
Adverse events can be 'Expected' or 'Unexpected.'
7.1.2.1 Expected Adverse Event
Expected adverse events are those that have been previously identified as resulting 
from administration of the agent. For the purposes of this study, an adverse event is 
considered expected when it appears in the current adverse event list, the 
Investigator’s Brochure, the package insert or is included in the informed consent 
document as a potential risk.
7.1.2.2 Unexpected Adverse Event
For the purposes of this study, an adverse event is considered unexpected when it 
varies in nature, intensity or frequency from information provided in the current 
adverse event list, the Investigator’s Brochure, the package insert or when it is not 
included in the informed consent document as a potential risk.
7.1.2.3
Additional Protocol-Specific Expedited Adverse Event Reporting 
Exclusions
For this protocol only, the AEs/grades listed below do not require expedited reporting. 
However, they still must be reported through the routine reporting mechanism (see Section 
7.4):
Adverse Event

Infection: CTCAE gr 3 or less

Immune-related events (e.g. diarrhea, pruritus, rash, endocrinopathies) that respond to 
corticosteroids and improve to grade 1 or less with steroids: CTCAE gr 3 or less

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
34
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
7.2
Expected Toxicities
7.2.1
Adverse Events Lists
7.2.1.1
Adverse Event List for CD25/Treg-depleted Cells.
Unselected donor lymphocyte infusion for treatment of malignant disease relapse has been 
associated with Teff-mediated GVHD flare, and with myelosuppression with depletion of host 
hematopoiesis primarily in cases of significant disease relapse involving the bone marrow. 
However, infusion of the CD25/Treg-depleted cells was found to be safe in our phase 1 clinical 
trial. These patients will be closely monitored for any adverse effects including GVHD and 
marrow suppression. Biopsy of affected organs (e.g. gut, liver) is recommended in appropriate 
patients to distinguish GVHD from immune-related inflammation, as per section 6.1.13.
7.2.1.2
Adverse Event List(s) for Ipilimumab
A list of reported and/or potential adverse events (AE) associated with Ipilimumab is 
provided in the investigators brochure (IB), and is briefly summarized below. In 
addition to the comprehensive list, a subset is identified with bold and italicized text. 
This subset of AEs (indicated with *) is a list of events that are protocol-specific 
exceptions to expedited reporting (except as noted below).
NOTE: Report expedited AEs on the subset ONLY IF they exceed the grade noted in 
parentheses next to the AE. If this is part of a combination protocol using multiple 
investigational agents and has an AE also listed on different agent/therapy, use the 
lower of the grades to determine if expedited reporting is required.
The following side effects have been reported in clinical trials in patients receiving 3 
mg/kg Ipilimumab:
Very common (may affect more than 1 in 10 people)
anorexia
diarrhea* (gr 3), vomiting, or nausea* (gr 3) 
skin rash* (gr 3), pruritis* (gr 3)
fatigue* (gr 3), reaction at site of injection, fever* (gr 2)
Common (may affect up to 1 in 10 people)
tumor pain
hypothyroidism, hypopituitarism 
dehydration
confusion
neuropathy (causing pain, weakness and cramps), dizziness, headache, 
blurred vision, pain in the eye
low blood pressure, temporary redness of the face and neck, feeling of intense heat 
with sweating and rapid heart beat

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
35
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
shortness of breath, cough
bleeding in the stomach or intestine, inflammation of the intestines (colitis)* (gr 3), 
constipation, heartburn, stomach pain
abnormal function of the liver
inflammation of the inner surface lining of a particular organ
inflammation and redness of the skin, skin color change in patches (vitiligo), hives, 
hair loss or thinning, excessive sweating at night, dry skin
pain in muscles and joints, muscle spasms 
shivering, lack of energy, swelling, pain 
flu-like illness
weight loss
Uncommon (may affect up to 1 in 100 people)
sepsis, septic shock, meningitis, gastritis, colitis, urinary tract infection, infection of 
the respiratory tract
paraneoplastic syndromes such as hypercalcemia, hypercholesterolemia and 
hypoglycemia
allergic reaction
adrenal insufficiency, hyperthyroidism, defect of the glands producing sex hormones 
decreased function of the adrenal glands caused by an underactive hypothalamus 
tumor lysis syndrome
changes in mental health, depression, lowered sex drive
Guillain-Barré syndrome, fainting, inflammation of the nerves within the brain, 
hydrocephalus, ataxia, shaking, brief involuntary muscle contraction, difficulty in 
speaking
inflammation of the eye which causes redness or pain, bleeding in the eye, iritis, 
reduced vision, foreign body sensation in the eyes, swollen runny eyes, swelling of the 
eye, inflammation of the eyelids
irregular or abnormal heart beat
inflammation of the blood vessels, disease of the blood vessels, restriction in the 
blood supply to the extremities, low blood pressure when standing up
extreme difficulty in breathing, pleural effusion, pneumonitis, hay fever
bowel perforation, inflammation of the membrane of the stomach wall and small 
intestine, pancreatitis, peptic ulcer, esophagitis, intestinal obstruction
liver failure, inflammation of the liver, enlarged liver, jaundice 
toxic epidermal necrolysis
myositis, arthralgia
inflammation of the thyroid gland, the kidney, or the central nervous system 
multi organ inflammation
muscle weakness
kidney function failure, kidney disease 
absence of menstrual periods
multi organ dysfunction, reaction related to infusion of the medicine 
change in hair color
Rare (may affect up to 1 in 1,000 people)

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
36
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
inflammatory disease of blood vessels (most commonly cranial arteries) 
inflammation of the anus and the rectal wall (marked by bloody stools and a frequent
urge to defecate) 
psoriasis
erythema multiforme
a type of severe skin reaction characterized by rash accompanied by one or more of 
the following features: fever, swelling of the face or lymph glands, increase of 
eosinophils, effects on liver, kidneys or lungs (DRESS).
In addition, the following uncommon (may affect up to 1 in 100 people) side effects 
have been reported in patients who received doses other than 3mg/kg of Ipilimumab in 
clinical trials:
meningism: neck stiffness, intolerance of bright light and headache, flu-like 
discomfort
inflammation of the heart muscle, weakness of the heart muscle, fluid around the 
heart
inflammation of the liver or the pancreas, nodules of inflammatory cells in various 
organs of your body
infection within the abdomen 
erythema nodosum 
overactive pituitary gland
decreased function of the parathyroid gland 
inflammation of the eye, eye muscle inflammation 
decreased hearing
poor blood circulation which makes toes and fingers numb or pale
damage to the tissues of the hands and feet resulting in redness, swelling and blisters 
Ipilimumab may cause changes in the results of laboratory tests. These include:
a variation in the number of red blood cells, white blood cells or platelets 
an abnormal variation of hormones and liver enzyme levels in the blood 
abnormal liver function test
abnormal levels of calcium, sodium, phosphate or potassium in the blood 
hematuria, proteinuria
an abnormally high alkalinity of the blood and other body tissues 
kidneys unable to remove acids from blood normally
presence of autoantibodies
Please refer to the Ipilimumab IB for a comprehensive list of adverse events.
7.3
Adverse Event Characteristics

CTCAE term (AE description) and grade: The descriptions and grading 
scales found in the revised NCI Common Terminology Criteria for Adverse 
Events (CTCAE) version 5.0 will be utilized for AE reporting. All appropriate

CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
37
DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
treatment areas should have access to a copy of the CTCAE version 5.0. A 
copy of the CTCAE version 5.0 can be downloaded from the CTEP web site 
http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm.

For expedited reporting purposes only:
-
AEs for the agent(s) that are listed above should be reported only if the 
adverse event varies in nature, intensity or frequency from the expected 
toxicity information which is provided.
-
Other AEs for the protocol that do not require expedited reporting are 
outlined in the next Section (Expedited Adverse Event Reporting) under the 
sub-heading of Protocol-Specific Expedited Adverse Event Reporting 
Exclusions.

Attribution of the AE:
-
Definite – The AE is clearly related to the study treatment.
-
Probable – The AE is likely related to the study treatment.
-
Possible – The AE may be related to the study treatment.
-
Unlikely – The AE is doubtfully related to the study treatment.
-
Unrelated – The AE is clearly NOT related to the study treatment.
7.4
Adverse Event Reporting
7.4.1
In the event of an unanticipated problem or life-threatening complications treating 
investigators must immediately notify the Overall PI. Additionally, BMS must be 
notified within 24 hours.
BMS Contact:
SAE Email Address: Worldwide.Safety@BMS.com
SAE Facsimile Number: +1 609-818-3804
7.4.2
Investigators must report to the Overall PI any adverse event (AE) that occurs after the 
initial dose of study treatment, during treatment, or within 90 days of the last dose of 
treatment on the local institutional SAE form. Per BMS guidelines, Ipilimumab follow 
up for AE/SAEs is 90 days from the last dose of treatment.
7.4.3
DF/HCC Adverse Event Reporting Guidelines
Investigative sites within DF/HCC will report AEs directly to the DFCI Office for 
Human Research Studies (OHRS) per the DFCI IRB reporting policy.
7.4.4
Protocol-Specific Adverse Event Reporting Exclusions
For this protocol only, the AEs/grades listed below do not require expedited reporting 
to the Overall PI or the DFCI IRB. However, they still must be reported through the 
routine reporting mechanism (i.e. case report form).

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
38
Events not considered to be serious adverse events are hospitalizations for:

routine treatment or monitoring of the studied indication, not associated with 
any deterioration in condition, or for elective procedures

elective or pre-planned treatment for a pre-existing condition that did not 
worsen

emergency outpatient treatment for an event not fulfilling the serious criteria 
outlined above and not resulting in inpatient admission

respite care
7.5
Reporting to the Food and Drug Administration (FDA)
The Overall PI, as study sponsor, will be responsible for all communications with the 
FDA. The Overall PI will report to the FDA, regardless of the site of occurrence, any 
serious adverse event that meets the FDA’s criteria for expedited reporting following 
the reporting requirements and timelines set by the FDA.
7.6
Reporting to Hospital Risk Management
Participating investigators will report to their local Risk Management office any participant 
safety reports, sentinel events or unanticipated problems that require reporting per institutional 
policy.
7.7
Routine Adverse Event Reporting
All Adverse Events must be reported in routine study data submissions to the Overall PI on the 
toxicity case report forms. AEs reported through expedited processes (e.g., reported to the 
IRB, FDA, etc.) must also be reported in routine study data submissions.
8. PHARMACEUTICAL INFORMATION
A list of the adverse events and potential risks associated with the investigational or other 
agents administered in this study can be found in Section 6.
8.1
Ipilimumab

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
39
Bristol-Myers Squibb Inc. (BMS) supplies distributes Ipilimumab at no cost for 
this study.
8.1.1.1
Description
Chemical Name or Amino Acid Sequence: 4 polypeptide chains, 2 identical 
heavy chains with 447 amino acids and 2 identical light chains consisting of 215 
amino acids.
Other Names: Anti-CTLA-4 monoclonal antibody, MDX-010
Classification: Human monoclonal antibody
M.W.: 147,991 Daltons
Mode of Action: Ipilimumab is specific for the CTLA-4 antigen expressed on a 
subset of activated T-cells. CTLA-4 interaction with the B7 molecule, one of its 
ligands expressed on professional antigen presenting cells, can down-regulate T-cell 
response. Ipilimumab is thought to act by blocking the interaction of CTLA-4 with 
the B7 ligand, resulting in a blockade of the inhibitory effect of T-cell activation.
The CTLA-4/B7 creates the interaction.
Description: Ipilimumab is a fully human immunoglobulin (IgG1
) with two
manufacturing processes – ongoing trials have been using substances manufactured 
using Process B. New clinical trials will be using Ipilimumab that is manufactured 
by Process C. The Process C has been developed using a higher producing sub- 
clone of the current Master Cell Bank, and modified cell culture and purification 
steps.
8.1.2
Form
Ipilimumab will be provided in open-label containers. Container labels will contain 
the batch number, contents, storage conditions, and Investigational New Drug 
caution.
Ipilimumab is available as a 200 mg /40 mL or 50 mg /10 mL (5 mg/mL) single-use 
vial. The vial is a clear, colorless, sterile, isotonic aqueous solution (pH of 7) that 
may contain particles. Each vial is a Type I flint glass vial with gray butyl stoppers 
and sealed with aluminum seals.
Process C
Component
50 mg/ viala
50 mg/ viala
Ipilimumab
53.5 mg
213 mg
Sodium Chloride, USP
62.6 mg
249 mg
TRIS-hydrochloride
33.7 mg
134.3 mg
Diethylenetriamine pentacetic acid
0.42 mg
1.67 mg
Mannitol, USP
107 mg
426 mg
Polysorbate 80 (plant-derived)
1.18 mg
4.69 mg
Sodium Hydroxide
QS to pH 7
Hydrochloric acid
QS to pH 7
Water for Injection
QS: 10.7 mL
QS: 42.6 mL

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
40
Nitrogenc
Processing agent
aIncludes 0.7 overfill; bIncludes 2.6 mL overfill.
cNitrogen is used to transfer the bulk solution through the pre-filled and sterilizing 
filters into the aseptic area.
8.1.3
Storage and Stability
Storage: Store intact vials refrigerated at (2-8 C), protected from light. Do not 
freeze.
Stability: Shelf-life surveillance of the intact vials is ongoing. Solution as described 
above is stable up to 24 hours refrigerated at (2-8 C) or at room temperature/ room 
light.
CAUTION: Ipilimumab does not contain antibacterial preservatives. Use prepared 
IV solution immediately. Discard partially used vials.
8.1.4
Compatibility
Ipilimumab will be administered individually on this study, and thus issues of 
compatibility with other agents are not anticipated.
8.1.5
Handling
The investigational product must be stored in a secure area according to local 
regulations. The investigator is responsible for ensuring that it is dispensed only to 
study subjects. The investigational product must be dispensed only from official study 
sites by authorized personnel according to local regulations. The investigator must 
ensure that the investigational product is stored in accordance with the environmental 
conditions (temperature, light, and humidity) determined by the sponsor. If concerns 
regarding the quality or appearance of the investigational product arise, the 
investigational product must not be dispensed and BMS must be contacted 
immediately, via:
Brittney McLaughlin
Project Manager, Syneos Health
Providing services to Bristol-Myers Squibb 
3401 Princeton Pike
Lawrenceville, NJ 08648
syneoshealth.com
Ipilimumab (BMS-734016) Injection (5 mg/ml), must be stored refrigerated (2ºC - 
8ºC) with protection from light. In preparation of infusion, Ipilimumab may be 
stored in IV infusion bags (PVC, non-PVC/non-DEHP) or glass infusion containers 
at room temperature or refrigerated (2ºC - 8ºC) for up to 24 hours. Drug must be

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
41
completely delivered to the subject within 24 hours of preparation. This includes any 
time in transit plus the total time for the infusion.
As with all injectable drugs, care should be taken when handling and preparing 
Ipilimumab. Whenever possible, it should be prepared in a laminar flow hood or 
safety cabinet using standard precautions for the safe handling of intravenous agents, 
applying aseptic technique. Gloves are required. If Ipilimumab concentrate or 
solution comes in contact with skin or mucosa, immediately and thoroughly wash 
with soap and water. After final drug reconciliation, unused Ipilimumab solution 
should be disposed of at the site following procedures for the disposal of anticancer 
drugs.
8.1.6
Availability
Ipilimumab is an investigational agent supplied by BMS.
8.1.7
Preparation and Administration
The following instructions are suggested guidelines for preparation of Ipilimumab 
which may be prepared per institutional standards and package insert. The supplies 
needed for Ipilimumab preparation and administration include calibrated syringes and 
infusion containers. Ipilimumab is to be administered as an IV infusion over 90 +/-10 
minutes through an IV line containing a sterile, nonpyrogenic, low-protein-binding in-
line filter per institutional standard practice. Flush IV line with NS or D5W after each 
dose per institutional standard practice. The total dose for each subject’s first 
Ipilimumab infusion (Cycle 1, Day 1) must be calculated using the most recent subject 
weight (obtained within 3 days of the dosing visit, and prior to the infusion). The same 
total dose may be given in the subsequent infusions if the most recent subject weight 
(obtained within 3 days of dosing visit, and prior to the infusion) is within 5% of the 
subject weight prior to the first infusion. If the most recent subject weight (obtained 
within 3 days of the dosing visit, and prior to the infusion) at a subsequent dosing visit 
is not within 5% of the subject weight prior to the first infusion, the total dose must be 
recalculated based on the most recent subject weight for that dosing visit.
1) As Ipilimumab is stored at refrigerated temperatures (2 - 8ºC), allow the appropriate 
number of vials of Ipilimumab to stand at room temperature for approximately 5 
minutes.
2) Aseptically withdraw the required volume of Ipilimumab solution into a syringe. 
Insert the needle at an angle into the Ipilimumab vial by placing the needle bevel 
side down against the glass, with the tip touching the neck of the vial. The initial 
solution concentration is 5 mg/mL. [Note: A sufficient excess of Ipilimumab is 
incorporated into each vial to account for withdrawal losses].

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
42
3) Ensure that the Ipilimumab solution is clear, colorless, and essentially free from 
particulate matter on visual inspection. If multiple vials are needed for a subject, it 
is important to use a separate sterile syringe and needle for each vial to prevent 
problems such as dulling of needle tip, stopper coring, repeated friction of plunger 
against syringe barrel wall, and so on.
4) Preparation of ipilimumab will follow package insert and/or institutional standard 
practice.
5) Mix by GENTLY inverting several times. DO NOT shake.
6) Visually inspect the final solution. If the initial diluted solution or final dilution for 
infusion is not clear, or the contents appear to contain precipitate, the solution should be 
discarded.
7) Do not draw into each vial more than once. Any partial vials should be safely 
discarded and should not be stored for reuse.
8) Ipilimumab should be administered under the supervision of a physician experienced 
in the use of intravenous agents. Ipilimumab is administered as an intravenous infusion 
only.
9) Ipilimumab is administered over 90 +/- 10 minutes, and hydration is allowed but not 
required per protocol. Patients should be observed closely for infusion reactions during 
treatment, with vital signs checked at least every 30 minutes, including after an 
observation period of 30 minutes after completing the infusion. A detailed description 
of common infusion reactions and their management is outlined in Section 6.1.11
8.1.8
Ordering
Ipilimumab will be provided by BMS. Investigational supplies of study drug will be 
provided to DFCI research pharmacy as per standard procedures and agreements with 
BMS.
BMS protocol#: CA184-559
BMS vendor contact for drug orders is 
distribution.allentown@thermofisher.com
8.1.9
Accountability
The investigator, or a responsible party designated by the investigator, should maintain

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
43
a careful record of the inventory and disposition of the agent using a drug 
accountability form.
8.1.10 Destruction and Return
Expired ipilimumab supplies will be destroyed on site per SOP. At the end of the study, 
unused supplies of ipilimumab will be destroyed on site per SOP. Destruction will be 
documented in the computerized institutional Drug Accountability Record Form.
9. CORRELATIVE STUDIES
9.1
Laboratory Correlative Study Sample Collection
All laboratory tests are mandatory and will be processed at the Cell Manipulation Core 
Facility at DFCI. Please refer to the study operations manual for details regarding 
processing and shipping study specimens.
Samples will be obtained pre-dose at screening or Cycle 1 Day 1 (C1D1), per the schema 
in the required data table (Table 6).
9.1.1 Peripheral Blood Correlatives 
Each time point will require a PASQ3P (three - 10mL EDTA tubes) for correlative 
analysis. These research tubes will preferably be drawn at the same time as clinical labs 
drawn at the below time points, though can be drawn at an alternative time for scheduling 
convenience. There are two exceptions:
a) Cycle 1 Day 1 (C1D1): Order a PASQ6P – six – 10mL EDTA tubes. Each 
lab involved requires a baseline sample. This needs to be collected prior to 
the patient receiving any study treatment. 
b) Cycle 3 Day 1, Cycle 4 Day 29, Cycle 5 Day 1, Cycle 6 Day 1, Cycle 8 
Day 1, Off Rx: For every time point when a BM aspirate is collected, 
instead of collecting a PASQ3P (three-10mL EDTA tubes) order a PASQ4P 
(four-10 mL EDTA tubes). 
9.1.2 Bone Marrow Aspirate and Biopsy
Bone marrow aspirate and biopsy samples will be obtained per the schema in the required 
data table (Tables 6, 7). Two-10 mL research bone marrow aspirates and one bone 
marrow core biopsy should be collected for correlatives at each time point a biopsy is 
noted in the study calendar below. 

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
44
9.1.3 Skin/Extra-Medullary Disease Biopsies
Skin/other sites of extra-medullary disease will also be biopsied per the required data 
table schema, if deemed safe and clinically feasible. Tissue should be sent in a sterile, 
leak proof container that contains DMEM at room temperature to the TIGL. Sample 
should be delivered within 4 hours and be at least 3 mm in diameter.
Archival tissue from patients on study may be utilized for correlatives studies as 
necessary.
9.2
Planned Correlative Studies
The goals of the correlative analyses include:
To assess the immunologic impact of infusions of CD25/Treg-depleted DLI plus 
Ipilimumab. These include immunophenotypic analysis of T-cell subsets, B and NK 
cells; phenotypic and functional readouts of effector T (Teff) and regulatory T (Treg) cell 
activation 
status; 
changes 
in 
plasma 
cytokines 
and 
chemokines; 
tumor 
microenvironment; and tumor mutation profile. We also seek to correlate clinical 
response with changes in circulating Teff vs. Treg cell counts and activation; the 
persistence of adoptively transferred Teff cells; changes in cytokine and chemokine 
levels; tumor infiltration by activated CD8+ T cells; and tumor mutation profile, blast 
phenotype and tumor microenvironment. In an exploratory analysis, we will perform 
tumor neoantigen discovery to identify novel T cell targets and correlate tumor 
mutational load with response.
The specific studies include:

Assessment of donor cell chimerism (molecular testing and cell based assays).

Immune cell frequencies and numbers (flowcytometry, mass cytometry, immune 
histochemistry).

CD4 Teff and Treg, CD8 T, and NK cell frequency, number, phenotype, proliferation 
(flow or mass cytometry).

Teff cell function assays and phenotypic readouts in vivo and in vitro.

Teff TCR sequencing

Teff, Treg, CD8 T, NK cell gene expression / transcriptome profiling.

Next generation sequencing of measurable residual disease.

AML/MDS/MPN immune-evasion assessments (flow or mass cytometry, genome 
sequencing of baseline and relapse samples, including non-malignant tissue).

Serum cytokine, chemokine assessments
Samples will be collected at time points mentioned above (Tables 6, 7).

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
45
10. STUDY CALENDAR
Baseline disease assessments (e.g. BM, Scans), cardiac/pulmonary evaluations, and 
Labs/IDM must be done ≤4 weeks prior to the start of therapy. Baseline other labs (e.g. 
CBC, chemistries) should be repeated within 1 week prior to start of therapy. In the event 
that the participant’s condition is deteriorating, laboratory evaluations should be repeated 
within 48 hours prior to initiation of the next cycle of therapy.
There will be at least one-week interval between consecutive enrollments at the same dose 
level during phase 1A. All assessments must be performed prior to administration of any study 
medication. All study assessments should be performed within ± 3 days of the protocol- 
specified date during induction and ± 7 days during maintenance, unless otherwise noted.
Ipilimumab should be administered within ± 3 days of the protocol-specified date during 
induction (Table 6) and ± 7 days during maintenance (Table 7), unless otherwise noted. Note 
that in the calendar below, entry to maintenance Ipilimumab during weeks 24-60 are only for 
patients with stable disease or better at time of Cycle 4 day 29 (D92) response assessment 
time point.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
46
Table 6: Induction Phase
Screening
C1 
D1 
(D1)
C1 
D8 
(D8)
C1 
D15 
(D15)
C2 
D1 
(D22)
C2 
D8 
(D29)
C3 
D1 
(D43) 
(DLT
Endpoint)
C4 
D1 
(D64)
C4 
D29 
(D92)
(Response 
Endpoint)
CD25/Treg depleted 
DLI
X
Ipilimumab*
X
X
X
X
Informed Consent
X
EKG1
X
Cardiac Evaluation1,2
X
PFT
X
X
-HCG Test3
X
Complete History
X
Interval History
X
X
X
X
X
X
X
X
Physical exam (Ht, 
Wt, VS4, PS)
X4
X
X
X
X
X
X
X
X
CBC and differential
X
X
X
X
X
X
X
X
X
Serum Chemistry5
X
X
X
X
X
X
X
X
X
TSH
X
X
Chimerism testing
X
X
X
PB Correlative 
studies6
X
X11
X
X
X
X
X12
X
X12
BM aspirate and 
core7
X
X
X
aGvHD assessment8
X
X
X
X
X
X
X
X
X
cGVHD assessment9
X
X
X
Viral Panel10
X
Table 7: Maintenance Phase (For patients with SD/PR/CR at D92 Response Assessment)
C4 
D57
C5 
D1
(Wk 24)
C5 
D29
C5 
D57
C6 
D1
(Wk 36)
C6 
D29
C6 
D57
C7 
D1
(Wk 48)
C7 
D29
C7 
D57
C8D1 
(Wk 
60)
and 
beyond
Off 
Rx**
Ipilimumab
X
X
X
X
PFT
X
X
X
X
X
Complete History
X
Interval History
X
X
X
X
X
X
X
X
X
X
X
Physical exam4 (Ht, 
Wt, VS, PS)
X
X
X
X
X
X
X
X
X
X
X
X
CBC and differential
X
X
X
X
X
X
X
X
X
X
X
X
Serum Chemistry5
X
X
X
X
X
X
X
X
X
X
X
X
TSH
X
X
Chimerism testing
X
X
PB Correlative studies6
X
X12
X12
X
X12
X12
BM aspirate and core7
X
X
X
X
aGvHD assessment8
X
X
X
X
X
X
X
X
X
X
X
cGVHD assessment9
X
X
X
X
X

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
47
* – Patients will be dosed at start of week 1, 4, 7, and 10 (induction) and return for an end of induction response evaluation on 
start of week 14. Maintenance dosing will then occur at weeks 24, 36, 48 and 60. Patients who experience clinical benefit may 
continue to receive Ipilimumab q12 weeks till week 60 unless there is unacceptable toxicity, disease progression, or withdrawal 
of consent.
**– End of study off-treatment visit will be 3 months after last dose of maintenance Ipilimumab. Note: for IND/IDE trials, follow up 
visits or other contact are required in order to identify SAEs during the 30 days following the end of study treatment
1 – A baseline EKG and cardiac ECHO (or MUGA) is required for all patients, and anytime thereafter as clinically indicated.
2 – For those with evidence of CHF, myocardial infarction, myocarditis or cardiomyopathy at any time during the study, a detailed 
cardiac evaluation including labs (CPK, Troponin), cardiology consult, with EKG and ECHO (or MUGA) at minimum is required, 
as clinically indicated. Please refer to Section 6.1.13 for guidance on cardiac toxicity management for Ipilimumab.
3 – For females of childbearing potential only
4 – 02 Saturation on room air required only at screening; Weight on day closest prior to infusion will be used for study dosing 
calculations, height only needs to be collected at screening.
5 – Includes: comprehensive metabolic panel, LFTs, magnesium, phosphorus, uric acid, LDH, lipase, 
eGFR
6 – See Section 9.1.1.
7 – See Section 9.1.2 and 9.1.3. If there is no bone marrow involvement at baseline and/or extra-medullary disease is 
documented, the other sites of disease relapse will be assessed as clinically appropriate (section 9.1.3 e.g. measurement of 
lesions of leukemia cutis including skin biopsy, PET/CT assessments for other extramedullary sites (e.g. granulocytic sarcoma)
8 – See Appendix C for aGVHD assessment
9 – Per NIH consensus criteria for cGVHD assessment 
10 – Viral Panel includes HIV, Hep B and C
11 – Exception under 9.1.1 a). Draw a PASQ6P
12 – Exception under 9.1.1 b). Draw a PASQ4P       
Supplemental Scheduling Table 
Induction Phase
Total Days
Maintenance Phase
Total Days
Cycle 1 Day 1 (Ipi)
1
Cycle 4 Day 57
120
Cycle 1 Day 8 
8
Cycle 5 Day 1 (Ipi)
169
Cycle 1 Day 15
15
Cycle 6 Day 1 (Ipi)
253
Cycle 2 Day 1 (Ipi)
22
Cycle 7 Day 1 (Ipi)
337
Cycle 2 Day 8 
29
Cycle 8 Day 1 (Ipi)
421
Cycle 3 Day 1 (Ipi)
43
Off Treatment 
Cycle 4 Day 1 (Ipi)
64
Cycle 4 Day 29
92
     
*Induction Phase +/- 3 days for appointment window
**Maintenance Phase +/- 7 days  for appointment window
***Patients who do not complete the entirety of the Induction and Maintenance (8 cycles) phases are not required to complete 
the Off-Treatment visit outlined in the Study Calendar. However, any corresponding data that can be gathered through Stand of 
Care appointments should be collected. Toxicities will be recorded until 3 months after the last dose of Ipilimumab.                        

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
48
11. MEASUREMENT OF EFFECT / CLINICAL RESPONSE
Evaluable for toxicity- Participants who receive cell infusion at the specified dose plus 
Ipilimumab will be evaluated for toxicity. Evaluable patients are those who experience 
a DLT before day 43, or are assessable for DLT at day 43. Phase 1a patients who are 
unevaluable for DLT may be replaced.
Evaluable for response- Only those participants who have measurable disease present at 
baseline, have received cell infusion at the specified dose and least two cycles of 
Ipilimumab therapy, and have had their disease re-evaluated will be considered evaluable 
for response. These participants will have their response classified according to the 
disease-specific definitions stated in Appendices E-F. Patients unevaluable for response 
will not be replaced.
Response Criteria will be disease-specific, and are provided in detail in the following 
appendices:
Appendix E: Acute Myelogenous Leukemia
Appendix F: Myelodysplastic Syndrome / Myeloproliferative Neoplasm
Evaluation of Best Overall Response The best overall response is the best response 
recorded from the start of the treatment until disease progression/recurrence (taking as 
reference for progressive disease the smallest measurements recorded since the treatment 
started). The participant's best response assignment will depend on the achievement of 
both measurement and confirmation criteria
Duration of overall response: The duration of overall response is measured from the time 
measurement criteria are met for CR or PR (whichever is first recorded) until the first date 
that recurrence or PD is objectively documented, taking as reference for PD the smallest 
measurements recorded since the treatment started.
Duration of overall complete response: The duration of overall CR is measured from the 
time measurement criteria are first met for CR until the first date that recurrent disease is 
objectively documented.
Progression-Free Survival (PFS) is defined as the duration of time from start of treatment 
to time of objective disease progression or death whichever occurs first.
Overall Survival (OS) is defined as the duration of time from start of treatment to time of 
death.
12. DATA REPORTING / REGULATORY REQUIREMENTS
Adverse event lists, guidelines, and instructions for AE reporting can be found in Section 7.0 
(Adverse Events: List and Reporting Requirements).

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
49
12.1
Data Reporting
12.1.1 Method
The Office of Data Quality (ODQ) will collect, manage, and perform quality checks on 
the data for this study.
Note: If your study has been assigned to CDUS-Complete reporting, all adverse events 
(both routine and expedited) that have occurred on the study and meet the mandatory 
CDUS reporting guidelines must be reported via the monitoring method identified 
above. If your study has been assigned to CDUS-Abbreviated reporting, no adverse 
event reporting (routine or expedited) is required to be reported via CDUS.
12.1.2 Responsibility for Data Submission
Investigative sites within DF/HCC or DF/PCC are responsible for submitting data 
and/or data forms to the Office of Data Quality (ODQ) in accordance with DF/HCC 
policies.
12.2
Data Safety Monitoring
The DF/HCC Data and Safety Monitoring Committee (DSMC) will review and monitor 
toxicity and accrual data from this study. The committee is composed of medical oncologists, 
research nurses, pharmacists and biostatisticians with direct experience in cancer clinical 
research. Information that raises any questions about participant safety will be addressed with 
the Overall PI and study team.
The DSMC will review each protocol up to four times a year with the frequency determined by 
the outcome of previous reviews. Information to be provided to the committee may include:
up-to-date participant accrual; current dose level information; DLT information; all grade 2 or 
higher unexpected adverse events that have been reported; summary of all deaths occurring 
within 30 days of intervention for Phase I or II protocols; for gene therapy protocols, summary 
of all deaths while being treated and during active follow-up; any response information; audit 
results, and a summary provided by the study team. Other information (e.g. scans, laboratory 
values) will be provided upon request.
12.3
Collaborative Agreements Language
Not applicable
13. STATISTICAL CONSIDERATIONS

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
50
13.1
Study Design
This is an open-label, single center phase 1 study with the primary objective of 
establishing the safety and exploring the efficacy of infusing CD25/Treg-depleted cells 
plus Ipilimumab in AML, MDS and MPN patients who relapse after 8/8 HLA-matched 
donor hematopoietic cell transplant.
13.2
Primary endpoints
The primary endpoint of this study is to determine the maximum tolerated dose (MTD) of 
CD25/Tre-depleted DLI followed by Ipilimumab in AML, MDS and MPN patients relapsed 
after HLA-matched donor stem cell transplantation.
13.3
Secondary endpoints

ORR (To determine complete remission (CR/CRi) rate at 6 weeks (day 43) after 
the cell infusion plus Ipilimumab

To determine the rate of progression-free survival (PFS) and overall survival (OS) 
at ~3 months (day 92) post cell infusion plus Ipilimumab

To determine the rate of progression-free survival (PFS) and overall survival (OS) 
at ~ 1-year (60 weeks) post cell infusion plus Ipilimumab

To determine the incidence and severity of acute GVHD rates after cell infusion 
plus Ipilimumab

To determine the incidence and severity of chronic GVHD rates after cell infusion 
plus Ipilimumab
.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
51
13.4
Determining MTD
Four escalating/de-escalating doses of the combination therapy will be considered to determine 
the MTD: CD25/Treg depleted DLI 3x107 CD3+ cells/kg + Ipilimumab 10 mg/kg (Dose level
+2), CD25/Treg depleted DLI 3x107 CD3+ cells/kg + Ipilimumab 3 mg/kg (Dose level +1), 
CD25/Treg depleted DLI 3x107 CD3+ cells/kg + Ipilimumab 1 mg/kg (Dose level 0, starting 
level), and CD25/Treg depleted DLI 1x107 CD3+ cells/kg + Ipilimumab 1 mg/kg (Dose level - 
1). Considering the DLT observation period of 6 weeks and the accrual rate, the study will enroll 
5 eligible patients at each dose level to minimize delays in accrual. Patients will be unevaluable 
for MTD if they receive DLI product with cells not meeting the dose-level criteria (Section 
5.1.3), die of progressive disease without DLT, or have disease progression or other reason 
requiring removal from the study without DLT within the 6 week DLT observation period 
(Section 5.4).
Since the MTDs of CD25/Treg depleted DLI and Ipilimumab were previously defined in 
monotherapy studies. Dose level 0 will be the initial dose level for this combination therapy. 
To ensure at least 3 patients will be evaluable in each dose level, a cohort of 5 eligible patients 
will enter at each dose level. If there are fewer than 3 evaluable patients in a cohort, additional 
patients can be added at that dose level. At any dose level, if 2 or more patients experience 
DLT, MTD is considered exceeded. If this is dose level -1, then the study will be terminated. If 
this is dose level+1 or +2, the next lower level (dose level 0 or +1, respectively) will be 
declared to be the MTD. If this is dose level 0, dose de-escalation will take place (decision rule 
I). If no DLT is seen in the first 3, 4, or 5 evaluable patients at dose level 0 or +1, then dose 
escalation will take place. If no DLT is seen in the first 3, 4, or 5 evaluable patients at dose 
level -1 or +2, that dose level will be declared to be the MTD. If there are 3 evaluable patients 
in a cohort of 5 eligible patients and if 1 of the 3 evaluable patients experiences DLT then 2 
additional evaluable patients will be treated at the same dose level. If 1 or more of the 2 
additional patients experience(s) DLT then MTD is considered to have been exceeded, and the 
decision rule I will be applied. If there are 4 evaluable patients and if 1 of the 4 evaluable 
patients experiences DLT then 1 additional evaluable patient will be treated at the same dose 
level. If this additional patient experiences DLT then MTD is considered to have been exceeded, 
and the decision rule I will be applied. If 1 DLT is seen in a total of 5 evaluable patients treated 
at dose level 0, then dose escalation will take place. If 1 DLT is seen in a total of 5 evaluable 
patients treated at dose level -1 or +2, then this dose level will be declared to be the MTD. 
Table 8 shows the probability of escalation under various true DLT rates for the cohort. With 
this design, there is 92-93% probability of dose escalation if the true rate of DLT is 10% and 
74-76% probability if the true DLT rate is 20%. Of note, the probability of dose de- escalation 
is one minus the probability of dose escalation.
Table 8. Probability of dose escalation
True rate of DLT
Prob. of Escalation
for (3+2) evaluable 
cases
Prob. of Escalation
for (4+1) evaluable 
cases
Prob. of Escalation 
for 5 evaluable cases
10%
0.926
0.919
0.919
20%
0.758
0.737
0.737
30%
0.559
0.528
0.528
40%
0.372
0.337
0.337
50%
0.219
0.189
0.189

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
52
60%
0.11
0.087
0.087
Once the MTD is established, 10 more patients will be treated at the MTD in phase 1b to provide 
a better estimate of the toxicity and to describe preliminary efficacy at this dose level.
To ensure the MTD determined in Phase 1a is safe for a larger Phase 2 study, we will monitor 
toxicity in this expansion cohort closely. That is, in a total of 10 evaluable patients in the 
expansion cohort, if ≤2 DLTs are observed, this MTD will be the recommended Phase II dose. 
Conversely, if 3 or more DLTs are observed, further accrual will be halted pending DSMC 
review. Depending on the result of this review, the DSMC may recommend permanent closure 
or testing of the next lower dose in the expansion cohort. With this design the probability of 
observing ≤2 DLTs in 10 is 0.07 if the true but unknown DLT rate is 10%, but 0.83 if the rate is 
40%. Patients will receive maintenance Ipilimumab per their assigned dose level.
Amendment, December 23, 2020
The study team reviews AEs at each dose level closely on an ongoing basis. As of December 
20, 2020, 10 patients have been treated on study: 3 at dose level 0, 6 at dose level 1, and 1 at 
dose level 2.  At dose level 2, the first patient treated had DLT (severe aGVHD that was fatal), 
deemed attributable to study therapy. Due to a concern that we would observe more than 1 
DLT in a cohort of 5 patients at this dose level if we continue the accrual, the protocol was 
amended to deem MTD exceeded at this dose level. 
Further accrual of the expansion cohort was due for dose level 1, where 1 of 6 patients had 
experienced DLT (GVHD) within the DLT observation period. Recently, however, 1 additional 
patient has experienced delayed SAE with histology suggesting a possible role for remote 
Ipilimumab effect. Additionally, 1 other patient had a late SAE, albeit outside of the DLT 
window, that was concerning for treatment toxicity. We therefore further amend the protocol to 
enroll 2 more patients at dose level 0 to a total of 5 evaluable patients. If 1 or none of these 2 
additional patients experiences DLT, dose level 0 will be declared to be the presumptive MTD 
and thereafter proceed to a phase 1b expansion cohort. If, however, we observe 2 DLTs at dose 
level 0, dose de-escalation will take place to dose level -1 and apply the decision rule above.
 
Amendment June 25, 2021
No DLTs were experienced during the DLT period in the final 2 patients enrolled at dose level 
0. Therefore, the phase 1b expansion cohort will proceed at dose level 0. 
13.5
Sample Size/Accrual Rate
The sample size will approximately range from 4-25 evaluable patients, depending on the 
number of dose levels tested. Based on the current practice, we anticipate that the accrual rate 
will be approximately 8-10 patients per year, thus requiring up to 38 months for the completion 
of the accrual. This projection does not count a mandatory waiting period prior to dose de- 
escalation. Follow-up after termination of accrual will be 1 year after the last patient ends 
therapy.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
53
13.6
Stratification Factors
None
13.7
Analysis plan
Due to limited sample size, the analysis will be primarily descriptive. AEs/SAEs will be 
summarized by patient, type and grade as defined by the CTCAE v4.0. Rates will be 
described as proportions with exact binomial confidence intervals. CR/CRi rate at day 43 
also will be described as a proportion with exact binomial confidence interval. Kaplan- 
Meier models will be used to estimate PFS and OS at day 100 and 1 year with exact 
pointwise confidence intervals. Acute and chronic GVHD will be described as cumulative 
incidence in the competing risks framework treating death without GVHD as a competing 
event. The association between clinical outcomes (response, PFS and OS and acute or 
chronic GVHD) and correlative endpoints (e.g., circulating Teff and Treg cells, number 
of AML/MDS/MPN blasts and other characteristics of the BM microenvironment pre- 
therapy and at post-treatment relapse) will be explored using univariable Cox, Fine and 
Gray regression analysis or exact Wilcoxon-Rank-Sum test, as appropriate. Analyses will 
be carried out in SAS 9.4 or R 3.3.2 or higher.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
54
14. PUBLICATION PLAN
The initial study results will be made public within 12 months of the end of data collection. 
The study report is planned for publication in a peer-reviewed journal, and the initial release 
may be an abstract that meets the requirements of the International Committee of Medical 
Journal Editors. A final report of study outcomes will be made public no later than 3 years 
after the end of data collection.
The study PI holds the primary responsibility for publication of study results. Permission of 
the study PI will be required before any unpublished study information can be used or passed 
on to a third party.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
55
REFERENCES
1.
Kekre N, Koreth J. Novel strategies to prevent relapse after allogeneic 
haematopoietic stem cell transplantation for acute myeloid leukaemia and myelodysplastic 
syndromes. Curr Opin Hematol 2015;22:116-22.
2.
Saber W, Opie S, Rizzo JD, Zhang MJ, Horowitz MM, Schriber J. Outcomes after 
matched unrelated donor versus identical sibling hematopoietic cell transplantation in adults 
with acute myelogenous leukemia. Blood 2012;119:3908-16.
3.
Fuchs EJ. Related haploidentical donors are a better choice than matched unrelated 
donors: Point. Blood Adv 2017;1:397-400.
4.
Schmid C, Labopin M, Nagler A, et al. Donor lymphocyte infusion in the treatment 
of first hematological relapse after allogeneic stem-cell transplantation in adults with acute 
myeloid leukemia: a retrospective risk factors analysis and comparison with other strategies by 
the EBMT Acute Leukemia Working Party. J Clin Oncol 2007;25:4938-45.
5.
Goldsmith SR, Slade M, DiPersio JF, et al. Donor-lymphocyte infusion following 
haploidentical hematopoietic cell transplantation with peripheral blood stem cell grafts and 
PTCy. Bone Marrow Transplant 2017;52:1623-8.
6.
Nishiyama Y, Saikawa Y, Nishiyama N. Interaction between the immune system 
and acute myeloid leukemia: A model incorporating promotion of regulatory T cell expansion 
by leukemic cells. Biosystems 2018;165:99-105.
7.
van den Brink M, Uhrberg M, Jahn L, DiPersio JF, Pulsipher MA. Selected 
biological issues affecting relapse after stem cell transplantation: role of T-cell impairment, NK 
cells and intrinsic tumor resistance. Bone Marrow Transplant 2018.
8.
Armand P. Immune checkpoint blockade in hematologic malignancies. Blood 
2015;125:3393-400.
9.
Nikiforow S, Kim HT, Daley H, et al. A phase I study of CD25/regulatory T-cell- 
depleted donor lymphocyte infusion for relapse after allogeneic stem cell transplantation. 
Haematologica 2016;101:1251-9.
10.
Davids MS, Kim HT, Bachireddy P, et al. Ipilimumab for Patients with Relapse 
after Allogeneic Transplantation. N Engl J Med 2016;375:143-53.
11.
Piccirillo CA, Shevach EM. Naturally-occurring CD4+CD25+ immunoregulatory T 
cells: central players in the arena of peripheral tolerance. Semin Immunol 2004;16:81-8.
12.
Fehervari Z, Sakaguchi S. Development and function of CD25+CD4+ regulatory T 
cells. Curr Opin Immunol 2004;16:203-8.
13.
Azuma T, Takahashi T, Kunisato A, Kitamura T, Hirai H. Human CD4+ CD25+ 
regulatory T cells suppress NKT cell functions. Cancer Res 2003;63:4516-20.
14.
Cederbom L, Hall H, Ivars F. CD4+CD25+ regulatory T cells down-regulate co- 
stimulatory molecules on antigen-presenting cells. Eur J Immunol 2000;30:1538-43.
15.
Maloy KJ, Salaun L, Cahill R, Dougan G, Saunders NJ, Powrie F. CD4+CD25+ 
T(R) cells suppress innate immune pathology through cytokine-dependent mechanisms. J Exp 
Med 2003;197:111-9.
16.
Serra P, Amrani A, Yamanouchi J, et al. CD40 ligation releases immature dendritic 
cells from the control of regulatory CD4+CD25+ T cells. Immunity 2003;19:877-89.
17.
Thornton AM, Shevach EM. CD4+CD25+ immunoregulatory T cells suppress 
polyclonal T cell activation in vitro by inhibiting interleukin 2 production. J Exp Med 
1998;188:287-96.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
56
18.
Janssens W, Carlier V, Wu B, VanderElst L, Jacquemin MG, Saint-Remy JM. 
CD4+CD25+ T cells lyse antigen-presenting B cells by Fas-Fas ligand interaction in an 
epitope-specific manner. J Immunol 2003;171:4604-12.
19.
Salomon B, Lenschow DJ, Rhee L, et al. B7/CD28 costimulation is essential for the 
homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes. 
Immunity 2000;12:431-40.
20.
Shevach EM, McHugh RS, Piccirillo CA, Thornton AM. Control of T-cell 
activation by CD4+ CD25+ suppressor T cells. Immunol Rev 2001;182:58-67.
21.
Ehrenstein MR, Evans JG, Singh A, et al. Compromised function of regulatory T 
cells in rheumatoid arthritis and reversal by anti-TNFalpha therapy. J Exp Med 2004;200:277- 
85.
22.
Viglietta V, Baecher-Allan C, Weiner HL, Hafler DA. Loss of functional 
suppression by CD4+CD25+ regulatory T cells in patients with multiple sclerosis. J Exp Med 
2004;199:971-9.
23.
Miyara M, Amoura Z, Parizot C, et al. Global natural regulatory T cell depletion in 
active systemic lupus erythematosus. J Immunol 2005;175:8392-400.
24.
Taylor PA, Lees CJ, Blazar BR. The infusion of ex vivo activated and expanded 
CD4(+)CD25(+) immune regulatory cells inhibits graft-versus-host disease lethality. Blood 
2002;99:3493-9.
25.
Hoffmann P, Ermann J, Edinger M, Fathman CG, Strober S. Donor-type 
CD4(+)CD25(+) regulatory T cells suppress lethal acute graft-versus-host disease after 
allogeneic bone marrow transplantation. J Exp Med 2002;196:389-99.
26.
Cohen JL, Trenado A, Vasey D, Klatzmann D, Salomon BL. CD4(+)CD25(+) 
immunoregulatory T Cells: new therapeutics for graft-versus-host disease. J Exp Med 
2002;196:401-6.
27.
Zorn E, Kim HT, Lee SJ, et al. Reduced frequency of FOXP3+ CD4+CD25+ 
regulatory T cells in patients with chronic graft-versus-host disease. Blood 2005;106:2903-11.
28.
Rieger K, Loddenkemper C, Maul J, et al. Mucosal FOXP3+ regulatory T cells are 
numerically deficient in acute and chronic GvHD. Blood 2006;107:1717-23.
29.
Rezvani K, Mielke S, Ahmadzadeh M, et al. High donor FOXP3-positive regulatory 
T-cell (Treg) content is associated with a low risk of GVHD following HLA-matched 
allogeneic SCT. Blood 2006;108:1291-7.
30.
Miura Y, Thoburn CJ, Bright EC, et al. Association of Foxp3 regulatory gene 
expression with graft-versus-host disease. Blood 2004;104:2187-93.
31.
Koreth J, Matsuoka K, Kim HT, et al. Interleukin-2 and regulatory T cells in graft- 
versus-host disease. N Engl J Med 2011;365:2055-66.
32.
Koreth J, Kim HT, Jones KT, et al. Efficacy, durability, and response predictors of 
low-dose interleukin-2 therapy for chronic graft vs. host disease. Blood 2016.
33.
Turk MJ, Guevara-Patino JA, Rizzuto GA, Engelhorn ME, Sakaguchi S, Houghton 
AN. Concomitant tumor immunity to a poorly immunogenic melanoma is prevented by 
regulatory T cells. J Exp Med 2004;200:771-82.
34.
Sugiyama D, Nishikawa H, Maeda Y, et al. Anti-CCR4 mAb selectively depletes 
effector-type FoxP3+CD4+ regulatory T cells, evoking antitumor immune responses in 
humans. Proc Natl Acad Sci U S A 2013;110:17945-50.
35.
Pastille E, Bardini K, Fleissner D, et al. Transient ablation of regulatory T cells 
improves antitumor immunity in colitis-associated colon cancer. Cancer Res 2014;74:4258-69.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
57
36.
Zhou Q, Bucher C, Munger ME, et al. Depletion of endogenous tumor-associated 
regulatory T cells improves the efficacy of adoptive cytotoxic T-cell immunotherapy in murine 
acute myeloid leukemia. Blood 2009;114:3793-802.
37.
Blazar BR, Carreno BM, Panoskaltsis-Mortari A, et al. Blockade of programmed 
death-1 engagement accelerates graft-versus-host disease lethality by an IFN-gamma- 
dependent mechanism. J Immunol 2003;171:1272-7.
38.
Blazar BR, Taylor PA, Panoskaltsis-Mortari A, Sharpe AH, Vallera DA. Opposing 
roles of CD28:B7 and CTLA-4:B7 pathways in regulating in vivo alloresponses in murine 
recipients of MHC disparate T cells. J Immunol 1999;162:6368-77.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
58
APPENDICES
APPENDIX A
PERFORMANCE STATUS CRITERIA
ECOG Performance Status Scale
Karnofsky Performance Scale
Grade
Descriptions
Percent
Description
100
Normal, no complaints, no evidence
of disease.
0
Normal activity. Fully active, able 
to carry on all pre-disease 
performance without restriction.
90
Able to carry on normal activity; 
minor signs or symptoms of disease.
80
Normal activity with effort; some 
signs or symptoms of disease.
1
Symptoms, but ambulatory. 
Restricted in physically strenuous 
activity, but ambulatory and able 
to carry out work of a light or 
sedentary nature (e.g., light 
housework, office work).
70
Cares for self, unable to carry on 
normal activity or to do active work.
60
Requires occasional assistance, but
is able to care for most of his/her 
needs.
2
In bed <50% of the time. 
Ambulatory and capable of all 
self-care, but unable to carry out 
any work activities. Up and about
more than 50% of waking hours.
50
Requires considerable assistance and 
frequent medical care.
40
Disabled, requires special care and 
assistance.
3
In bed >50% of the time. Capable 
of only limited self-care, confined 
to bed or chair more than 50% of 
waking hours.
30
Severely disabled, hospitalization
indicated. Death not imminent.
20
Very sick, hospitalization indicated.
Death not imminent.
4
100% bedridden. Completely 
disabled. Cannot carry on any
self-care. Totally confined to bed 
or chair.
10
Moribund, fatal processes 
progressing rapidly.
5
Dead.
0
Dead.

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
59
Appendix B: BODY SURFACE AREA AND CREATININE CLEARANCE
Body surface area (BSA) should be calculated using Dubois formula that yields the following 
results in meters squared (m2):
BSA = (W 0.425 x H 0.725) x 0.007184
where the weight is in kilograms and the height is in centimeters.
Creatinine clearance (CrCl) can be calculated using the Cockroft-Gault equation as follows:
CrCl (ml/min) =
(140 - age) (actual wt in kg) 
72  serum creatinine (mg/dl)
For females, use 85% of calculated CrCl value.
Note: In markedly obese participants, the Cockroft-Gault formula will tend to overestimate the 
creatinine clearance. (Adipose tissue tends to contribute little creatinine requiring renal 
clearance.)

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
60
Appendix C: ACUTE GVHD GRADING
Stage Skin (Active Erythema Only)
Liver 
(Bilirubin)
Upper GI
Lower GI (stool 
output/da
0
No active (erythematous) GVHD 
rash
<2 mg/dL
No or intermittent 
nausea, vomiting, 
or anorexia
Adult: <500 mL/day or 
<3 episodes/day
1
Maculopapular rash
<25% BSA
2-3 mg/dL
Persistent nausea, 
vomiting or 
anorexia
Adult: 500-999 
mL/day or 
episodes/day
2
Maculopapular rash 
25-50% BSA
3.1-
6 mg/dL
Adult: 1000-1500 
mL/day episodes/day
3
Maculopapular rash
>50% BSA
6.1-
15 mg/dL
Adult: >1500 mL/day 
or > episodes/day
4
Generalized erythroderma (>50% 
BSA) plus bullous formation and 
desquamation >5% BSA
>15 mg/dL
Severe abdominal 
pain w without ileus or 
grossly bl stool 
(regardless of stool 
volume).
Overall clinical grade (based on most severe target organ involvement):

Grade 0: No stage 1-4 of any organ.

Grade I: Stage 1-2 skin without liver, upper GI, or lower GI involvement.

Grade II: Stage 3 rash and/or stage 1 liver and/or stage 1 upper GI and/or stage 1 lower GI.

Grade III: Stage 2-3 liver and/or stage 2-3 lower GI, with stage 0-3 skin and/or stage 0-1 upper GI.

Grade IV: Stage 4 skin, liver, or lower GI involvement, with stage 0-1 upper GI.
Adapted from Harris AC, et al. BBMT 2016; 22:4

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
61
Appendix D: NCI COMMON TOXICITY CRITERIA
Please refer to the NIH website: 
https://ctep.cancer.gov/protocoldevelopment/electronic_applications/ctc.htm#ctc_40
Forms available upon request

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
62
Appendix E: AML RESPONSE CRITERIA
(Cheson, et al., 2003)
E.1. Antitumor Effect: AML
E.1.1 Morphological Complete Remission (CR):
-Normalization of the peripheral blood absolute neutrophil count > 1.0x109/L, platelets > than 
100x109/L no residual evidence of extramedullary disease and bone marrow aspirate with ≤ 
5% blasts, no blasts with Auer rods.
E.1.2 Morphological Complete Remission with incomplete blood count recovery (CRi):
-Same as CR but without normalization of the peripheral blood absolute neutrophil and platelet 
Count
E.1.3 Relapse from CR or CRi:
-Reappearance of leukemic blasts in the peripheral blood; or > 5% blasts in the bone marrow 
not attributable to another cause (e.g. recovery of normal cells following treatment –induced 
aplasia or use of growth factors) OR
-Appearance or reappearance of extramedullary disease.
-If there are no circulating blasts and no extramedullary disease and the bone marrow blast 
percentage is >5% but <20%, then a repeat bone marrow performed at least 7 days after the 
first marrow examination and documenting bone marrow blast percentage >5% is necessary 
to establish relapse.
E.1.4 Partial Remission (PR):
-Normalization of the peripheral blood absolute neutrophil count 1.0x109/L, platelets > than 
100x109/L, and at least a 50% decrease in the percentage of marrow aspirate blasts to 5-25% , 
or marrow blasts <5% with Auer rods.
E.1.5 Cytogenetic Responses
Complete cytogenetic response: An abnormal clone is detected in all metaphases prior to 
treatment and only normal metaphases are observed in all metaphases following treatment. 
Partial cytogenetic response: An abnormal clone is detected in all metaphases prior to 
treatment
and the post-treatment sample has 50% or fewer abnormal metaphases compared to the 
pretreatment value.
Cytogenetic non-response: An abnormal clone is detected in all metaphases prior to treatment 
and neither complete nor partial cytogenetic response is observed in post-treatment specimens 
Specimens with normal cytogenetic results before treatment will not be evaluated for 
cytogenetic
response. The method used to determine cytogenetic response will be standard metaphase 
cytogenetics.
E.1.6 Stable disease:
-Subjects who fail to achieve CR, Cri, or PR and who do not have criteria for PD will be 
defined as having the stable disease. If the subject dies prior to response assessment at the end 
of Cycle 1, then they will be classified as “indeterminate”.
E.1.7 Progressive disease (PD): may be defined as ONE of the following:
->50% increase in peripheral blood or bone marrow blasts from best assessment with 
minimum threshold of 20% blasts in the marrow or 1.0x109/L blasts in peripheral blood.
-Development of biopsy proven extramedullary leukemia (if the subject has extramedullary

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
63
disease at baseline, then PD will be defined by blood and marrow criteria or if new sites of 
extramedullary disease appear).
-If subject who present with an initial marrow blast percentage sufficiently high to preclude the 
ability to base disease progression on a >50% increase in marrow blast percentage, disease 
progression should be based on peripheral blood criteria or development of extramedullary 
leukemia as above.
E.1.8 Persistent Disease
Hematologic Persistence – Bone marrow or peripheral blasts >5% after d+30 following 
HCT

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
64
Appendix F: MDS/MPN RESPONSE CRITERIA
(Cheson, et al., 2006)
F.1 Antitumor Effect – Myelodysplastic Syndrome / Myeloproliferative Neoplasm
F.1.1. Complete Remission (CR)
Requires that all of the following be present:
Peripheral Blood Counts:
-Hemoglobin greater than 11 g/dL (untransfused, 4 weeks since last transfusion, subject not on 
erythropoietin).
-Neutrophils 1,000/mm3 or more (not on a myeloid growth factor)
-Platelets 100,000/mm3 or more (not on a thrombopoetic agent)
-Blasts, 0%
-No dysplasia
Bone Marrow Aspirate and Biopsy:
-Less than 5% myeloblasts
-Normal maturation of all cell lines
-No evidence for dysplasia
When erythroid precursors constitute less than 50% of bone marrow nucleated cells, the 
percentage of blasts is based on all nucleated cells; when there are 50% or more erythroid cells, 
the percentage blasts should be based on the nonerythroid cells.
F.1.2 Partial Remission (PR):
Requires that all of the criteria for complete remission be satisfied except blasts decreased by 
50% or more over pretreatment. Cellularity and morphology are not relevant. Persisting 
dysplasia is permitted.
Hematologic Improvement (HI)-major (must have at least one of the three listed below) 
must
persist for a minimum of 8 weeks to be considered a confirmed response.
HI-major Erythroid: For subjects with pretreatment hemoglobin less than 11 g/dL,
greater than 2 g/dL increase in hemoglobin; for RBC transfusion-dependent subjects 
transfusion independence.
HI-major Platelet: For subjects with a pretreatment platelet count less than
100,000/mm3, an absolute increase of 30,000/mm3 or more; for platelet transfusion 
dependent subjects, stabilization of platelet counts and platelet transfusion independence.
HI-major Neutrophil: For absolute neutrophil count (ANC) less than 1000/mm3 before
therapy, at least a 100% increase, or an absolute increase of more than 500/mm3, 
whichever is greater.
HI-major Trilineage: Trilineage response will require the demonstration of HI-major for 
each of the blood lineages (erythroid, platelet, neutrophil). Trilineage hematologic 
improvement will also apply to subjects who achieve adequate counts in all three lineages
even if one or more lineages was adequate prior to treatment. Thus, all subjects who normalize
all three lineages (within IWG definitions) for 8 weeks, but who do not meet the criteria for 
PR, will be considered HITrilineage.
F.1.3 Cytogenetic Responses

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
65
Complete cytogenetic response: An abnormal clone is detected in all metaphases prior to 
treatment and only normal metaphases are observed in all metaphases following
treatment. Partial cytogenetic response: An abnormal clone is detected in all metaphases
prior to treatment and the post-treatment sample has 50% or fewer abnormal metaphases 
compared to the pretreatment value.
Cytogenetic non-response: An abnormal clone is detected in all metaphases prior to
treatment and neither complete nor partial cytogenetic response is observed in posttreatment 
specimens
Specimens with normal cytogenetic results before treatment will not be evaluated for
cytogenetic response. The method used to determine cytogenetic response will be 
standard metaphase cytogenetics.
F.1.4 Progressive Disease
At least one of the following four criteria must be met:
1. An increase of ≥ 50% in blasts relative to baseline, to at least the level shown below for 
different baseline levels:
2. Baseline blasts must increase to
- < 5% > 5%
- ≥ 5 and < 10% > 10%
- ≥ 10 and < 20% > 20%
- ≥ 50% reduction from maximum remission/response levels in granulocytes or platelets.
- Reduction in Hb concentration by at least 2g/dL or development of transfusion dependence.
F.1.5 Relapse
Relapse after CR or PR: one or more of the following:
-Return to pretreatment bone marrow blast percentage,
-Decrement of 50% or greater from maximum remission/response levels in granulocytes or 
platelets,
-Reduction in hemoglobin concentration by at least 2 g/dL or transfusion dependence.
-Progression after Hematologic Improvement: One or more of the following:
-Decrement of 50% or greater decrement from maximum response levels in granulocytes 
or platelets,
-A reduction in hemoglobin concentration by at least 2 g/dL or transfusion dependence.
F.1.6 Persistent Disease
Hematologic Persistence – Cytopenias not attributable to other post-transplant causes 
accompanied by characteristic morphological changes > d +90 following alloSCT.
Cytogenetic Persistence – Persistence of clonal abnormality > d +90 following alloSCT

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
66
Appendix G: MANAGEMENT OF IMMUNE RELATED AEs DUE TO IPILIMUMAB
GI TOXICITY, HEPATOTOXICITY, ENDOCRINOPATHY, NEUROPATHY AND 
SKIN TOXICITY MANAGEMENT

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
67

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
68

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
69

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
70

DF/HCC Protocol # 19-142 
Protocol Version Date: 6/26/23
CONFIDENTIAL
This document is confidential. Do not disclose or use except as authorized.
71

Page 1