A Phase II Trial to Evaluate the Ability of a Dendritic Cell Vaccine to Immunize Melanoma or Epithelial Cancer Patients Against Defined Mutated Neoantigens Expressed by the Autologous Cancer
Phase: PHASE2
Conditions: Melanoma, Gastrointestinal Cancer, Breast Cancer, Ovarian Cancer, Pancreatic Cancer
Sponsor: National Cancer Institute (NCI)

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
1
Abbreviated Title: Personalized DC Vaccine
CC Protocol Number: 17-C-0177 B
IBC Number: RD-17-IV-07
NCT Number: NCT03300843
Version Date: March 26, 2018
PROTOCOL TITLE
A Phase II Trial to Evaluate the Ability of a Dendritic Cell Vaccine to Immunize Melanoma or 
Epithelial Cancer Patients Against Defined Mutated Neoantigens Expressed by the Autologous 
Cancer
NIH Principal Investigator:
Steven A. Rosenberg, M.D., Ph.D.
Chief of Surgery, Surgery Branch, NCI
Building 10, CRC, Room 3-3940
9000 Rockville Pike, Bethesda, MD 20892
Phone: 240-760-6218; Email: Steven.Rosenberg@nih.gov
Investigational Agents:
Drug Name:
Peptide loaded dendritic cell vaccine
IND Number:
17564
Sponsor:
Center for Cancer Research
Manufacturer:
Surgery Branch Cell Production Facility

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
2
PRÉCIS
Background:
x
Therapeutic vaccination against cancer has proven very challenging with little clinical 
benefit.
x
Vaccines against non-viral tumors have mainly targeted differentiation antigens, cancer 
testis antigens, and overexpressed antigens. However negative selection in the thymus 
against these normal nonmutated antigens severely limits the ability to generate high 
avidity anti-cancer T cells. Such depletion can impair their antitumor activity and limit
tumor elimination.
x
The National Cancer Institute Surgery Branch (NCI SB) has developed a pipeline for the 
identification of immunogenic T cell epitopes derived from neoantigens.
x
In recent studies, we identified the neoantigens recognized by TIL that mediated 
regression in patients with metastatic cancer. Using whole exome sequencing of a 
resected metastatic nodule followed by high throughput immunologic screening, we were 
able to demonstrate that tumor regressions were associated with the recognition by the 
administered TIL of unique somatic mutations that occurred in the cancer.
x
We, therefore, aim to use this pipeline to identify immunogenic neoantigens from 
epithelial cancer patients and to use these defined epitopes for a personalized therapeutic 
dendritic cell (DC) vaccine.
Objectives:
x
Primary objective:
o
To determine the clinical response rate in patients with metastatic melanoma or 
epithelial cancer who receive this DC vaccine
Eligibility:
x
$JH\HDUVDQG\HDUV
x
ECOG 0 – 2
x
Evaluable metastatic melanoma or epithelial cancer refractory to standard treatment
x
Metastatic melanoma or epithelial cancer lesion(s) that is resectable for TIL or in selected 
cases, available PBMC
Design:
x
Patients with metastatic melanoma or epithelial cancer will undergo surgical resection of 
tumor followed by exome and RNA sequencing to identify expressed mutations
(conducted under the NCI SB companion protocol 03-C-0277).
x
Patients will undergo apheresis and DC will be cryopreserved for vaccine preparation.
x
Immunogenic neoantigens will be identified from TIL and PBMC by high throughput
immunologic screening using long peptides and tandem minigenes covering all mutated 
epitopes.
x
Patient will be vaccinated with autologous mature dendritic cells loaded with long 
peptides and minimal epitopes from defined neoantigens or highly expressed mutations in 
tumor suppressor or driver genes.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
3
x
DC will be administered intravenously and subcutaneously for four cycles at biweekly
intervals.
x
Blood samples will be taken every two weeks, and patients will be monitored for the 
quantity and quality of circulating neoantigen-specific T cells.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
4
TABLE OF CONTENTS
PRÉCIS.......................................................................................................................................... 2
TABLE OF CONTENTS ............................................................................................................. 4 
1
INTRODUCTION............................................................................................................. 8 
 
Study Objectives ............................................................................................................. 8 
1.1.1 
Primary Objective....................................................................................................... 8 
1.1.2
Secondary Objectives.................................................................................................. 8
 
Background and Rationale.............................................................................................. 8 
1.2.1 
Cancer Vaccines.......................................................................................................... 8 
1.2.2 
Identification of Cancer-Specific Immunogenic Epitopes Derived from Somatic 
Mutations ................................................................................................................................ 9 
1.2.3 
Preclinical Studies for the Generation of a Dendritic Cell Vaccine ........................... 9 
1.2.4 
Rationale for Selecting Defined, Tumor Suppressor and Driver Neoantigens to use 
in this Trial............................................................................................................................ 11 
1.2.5 
Relevant Preclinical and Clinical Experience to Support the Proposed Route and 
Dose of the DC Vaccine ....................................................................................................... 12 
2
ELIGIBILITY ASSESSMENT AND ENROLLMENT.............................................. 12 
 
Eligibility Criteria......................................................................................................... 12 
2.1.1 
Inclusion Criteria ...................................................................................................... 12 
2.1.2 
Exclusion Criteria ..................................................................................................... 13 
 
Screening Evaluation .................................................................................................... 14 
2.2.1 
Within 3 Months Prior to Enrollment....................................................................... 14 
2.2.2 
Within 4 Weeks Prior to Enrollment ........................................................................ 14 
2.2.3 
Within 7 Days Prior to Enrollment........................................................................... 14 
 
Registration Procedures ................................................................................................ 15 
2.3.1 
Prior to Registration for this Protocol....................................................................... 15 
2.3.2 
Registration Procedure.............................................................................................. 15 
 
Treatment Assignment Procedures ............................................................................... 15 
2.4.1 
Cohorts...................................................................................................................... 15 
2.4.2 
Arms.......................................................................................................................... 15 
2.4.3 
Randomization and Arm Assignment....................................................................... 15 
3
STUDY IMPLEMENTATION...................................................................................... 16 
 
Study Design................................................................................................................. 16 
3.1.1 
Pre-Treatment Phase: DC Preparation...................................................................... 16 
3.1.2 
Treatment Phase........................................................................................................ 17 
3.1.3 
Protocol Stopping Rules ........................................................................................... 17 
 
Drug Administration..................................................................................................... 17 
 
On-Study Evaluation..................................................................................................... 18 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
5
3.3.1 
Within 7 Days Prior to the 1st Vaccination (Day 0).................................................. 18 
3.3.2
Within 2 Days Prior to the 1st Vaccination (Day 0).................................................. 18
3.3.3 
Within 2 Days Prior to Each Subsequent Vaccination (Days 14, 28, and 42 (± 5 
days)) 18
3.3.4 
After Each Vaccination............................................................................................. 19 
 
Post-Treatment Evaluation (Follow-Up) ...................................................................... 19 
3.4.1
1st Follow-up Evaluation (Day 56 ± 5 days)............................................................. 19
3.4.2 
Subsequent Follow-up Evaluations........................................................................... 19 
 
Study Calendar.............................................................................................................. 21 
 
Criteria for Removal from Protocol Therapy and Off-Study Criteria .......................... 23 
3.6.1 
Criteria for Removal from Protocol Therapy............................................................ 23 
3.6.2
Off-Study Criteria..................................................................................................... 23
3.6.3 
Off Protocol Therapy and Off-Study Procedure....................................................... 23 
4
CONCOMITANT MEDICATIONS/MEASURES...................................................... 23 
5
BIOSPECIMEN COLLECTION.................................................................................. 24 
 
Samples Sent to Figg Lab ............................................................................................. 24 
 
Samples Sent to Surgery Branch Cell Production Facility........................................... 24 
 
Prior to Each Vaccination............................................................................................. 24 
 
At Each Scheduled Follow-up Evaluation.................................................................... 24 
 
Immunological Testing................................................................................................. 24 
 
Sample Storage, Tracking and Disposition For Surgery Branch Cell Production 
Facility ...................................................................................................................................... 25 
 
Sample Storage, Tracking and Disposition for Dr. Figg’s Lab .................................... 25 
5.7.1 
Sample Data Collection ............................................................................................ 25 
5.7.2 
Sample Storage and Destruction............................................................................... 26 
6
DATA COLLECTION AND EVALUATION ............................................................. 26 
 
Data Collection ............................................................................................................. 26 
6.1.1 
Exclusions to Routine Adverse Event Recording..................................................... 27 
 
Data Sharing Plans........................................................................................................ 27 
6.2.1 
Human Data Sharing Plan......................................................................................... 27 
6.2.2 
Genomic Data Sharing Plan...................................................................................... 27 
 
Response Criteria.......................................................................................................... 27 
6.3.1 
Definitions................................................................................................................. 27 
6.3.2 
Disease Parameters ................................................................................................... 28 
6.3.3 
Methods for Evaluation of Measurable Disease....................................................... 29 
6.3.4 
Response Criteria...................................................................................................... 30 
6.3.5 
Duration of Response................................................................................................ 31 
 
Toxicity Criteria............................................................................................................ 31 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
6
7
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN 32
 
Definitions..................................................................................................................... 32 
7.1.1 
Adverse Event........................................................................................................... 32 
7.1.2 
Suspected Adverse Reaction..................................................................................... 32 
7.1.3 
Unexpected Adverse Reaction.................................................................................. 32 
7.1.4
Serious....................................................................................................................... 32
7.1.5 
Serious Adverse Event.............................................................................................. 32 
7.1.6 
Disability................................................................................................................... 33 
7.1.7 
Life-Threatening Adverse Drug Experience............................................................. 33 
7.1.8 
Protocol Deviation (NIH Definition)........................................................................ 33 
7.1.9
Non-Compliance (NIH Definition)........................................................................... 33
7.1.10 
Unanticipated Problem.......................................................................................... 33 
 
NCI-IRB Reporting and Clinical Director (CD) Reporting.......................................... 33 
7.2.1 
NCI-IRB and NCI CD Expedited Reporting of Unanticipated Problems and Deaths
33
7.2.2 
NCI-IRB Requirements for PI Reporting at Continuing Review............................. 33 
7.2.3 
NCI-IRB Reporting of IND Safety Reports.............................................................. 34 
 
IND Sponsor Reporting Criteria................................................................................... 34 
7.3.1 
Reporting Pregnancy................................................................................................. 34 
 
Institutional Biosafety Committee (IBC) Reporting Criteria........................................ 35 
7.4.1 
Serious Adverse Event Reports to IBC..................................................................... 35 
7.4.2 
Annual Reports to IBC.............................................................................................. 35 
 
Data and Safety Monitoring Plan.................................................................................. 36 
7.5.1 
Principal Investigator/Research Team ...................................................................... 36 
7.5.2 
Sponsor Monitoring Plan.......................................................................................... 36 
8
STATISTICAL CONSIDERATIONS .......................................................................... 37 
 
Statistical Hypothesis.................................................................................................... 37 
8.1.1 
Primary Efficacy Endpoint ....................................................................................... 37 
8.1.2 
Secondary Efficacy Endpoint ................................................................................... 37 
 
Sample Size Determination........................................................................................... 37 
 
Population for Analyses................................................................................................ 38 
 
Statistical Analyses ....................................................................................................... 38 
8.4.1 
General Approach ..................................................................................................... 38 
8.4.2 
Analysis of the Primary Endpoints ........................................................................... 38 
8.4.3 
Analysis of the Secondary Endpoint(s)..................................................................... 38 
8.4.4 
Safety Analyses......................................................................................................... 38 
8.4.5 
Baseline Descriptive Statistics.................................................................................. 38 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
7
8.4.6 
Planned Interim Analyses ......................................................................................... 38 
8.4.7
Sub-Group Analyses ................................................................................................. 38
8.4.8 
Tabulation of individual Participant Data................................................................. 38 
8.4.9 
Exploratory Analyses................................................................................................ 38 
9
COLLABORATIVE AGREEMENTS.......................................................................... 38 
10
HUMAN SUBJECTS PROTECTIONS........................................................................ 39 
Rationale for Patient Selection...................................................................................... 39
 
Participation of Children............................................................................................... 39 
 
Participation of Subjects Unable to Give Consent........................................................ 39 
 
Evaluation of Benefits and Risks/Discomforts............................................................. 39 
 
Risk/Benefit Analysis ................................................................................................... 40 
Consent Process and Documentation............................................................................ 40
10.6.1 
Informed Consent of Non-English Speaking Subjects ......................................... 40 
11
PHARMACEUTICAL INFORMATION..................................................................... 40 
 
Cell Preparation ............................................................................................................ 40 
 
Subcutaneous Injection and Intravenous Infusion Preparation..................................... 41 
12
REFERENCES................................................................................................................ 43 
13
FIGURES AND TABLES .............................................................................................. 44 
 
Figure 1: A Process for the Identification of Immunogenic Somatic Mutations.......... 44 
 
Figure 2: TLR Mix Induces the Secretion of IL-12p70, TNFD and Multiple 
Chemokines .............................................................................................................................. 45 
 
Figure 3: TLR Mix Alone is Sufficient to Induce Potent DC Maturation.................... 46 
 
Figure 4: Long Peptides, Minimal Epitopes and TMG mRNA used to Evaluate Antigen 
Presentation for CD4 and CD8 Cells........................................................................................ 47 
 
Figure 5: Long Peptides and Minimal Epitopes are Superior to TMG mRNA in 
Presenting Antigens to CD4 and CD8 Cells............................................................................. 48 
 
Figure 6: Peptide Loaded, TLR Stimulated DC’s Efficiently Present Both CD4 and 
CD8 Epitopes............................................................................................................................ 49 
 
Figure 7: Overview of the Method used to Evaluate the Activation of Neoantigen 
Specific T Cells......................................................................................................................... 50 
 
Figure 8: DCs Matured with TLR Mix are Capable of Stimulating Naïve, Antigen-
Experienced and Terminally Differentiated Effector Cells ...................................................... 51 
 
Figure 9: Process for Epitope Selection and Synthesis................................................. 52 
 Table 1: Defined Mutated Antigens Recognized by TIL Identified at the Surgery 
Branch53
14
APPENDICES................................................................................................................. 54 
 
Appendix 1: Cell Infusion Instructions......................................................................... 54 
 
Appendix 2: Certificate of Analysis for Immature Dendritic Cells.............................. 57 
 
Appendix 3: Certificate of Analysis for Mature Peptide Pulsed Dendritic Cells ......... 58 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
8
1
INTRODUCTION
STUDY OBJECTIVES
1.1.1
Primary Objective
x
To determine the clinical response rate in patients with metastatic melanoma or epithelial 
cancer who receive this DC vaccine.
1.1.2
Secondary Objectives
x
To determine whether administration of a DC vaccine presenting defined mutated 
neoantigens can increase the quantity and quality of circulating antigen-specific T cells.
x
To determine the safety of the vaccine.
BACKGROUND AND RATIONALE
1.2.1
Cancer Vaccines 
Protective vaccination against infectious diseases has proven to be one of the most effective 
health measures. Therapeutic vaccination against established diseases such as persistent 
infections and cancer has proven much more challenging. Cancer vaccines are designed to target 
antigens that can elicit an immune response that selectively attacks cancer cells. Ideally these 
antigens should be exclusively presented on cancer cells. Until recently, vaccines against non-
viral tumors mainly targeted differentiation antigens, cancer testis antigens, and overexpressed 
antigens. However, central immunological tolerance has limited the generation of high avidity 
reactivity against these normal antigens1,2. Thus, T cells prompted using these vaccines 
underwent depletion of high avidity clones directed against such antigens. This depletion causes 
the loss of T cells bearing high-affinity TCRs for their cognate antigens which have the superior
cytotoxic capacity, longer persistence in the tumor microenvironment, and decreased 
susceptibility to immune suppression3. Taken together, such depletion can lead to impaired 
clinical efficiency following vaccine administration. In recent years our group and others have 
extensively studied the importance of neoantigens as targets for immunotherapy. It is now clear 
that neoantigen specific T cells are present in most cancers. Neoantigens derived from somatic 
mutations offer a specific and highly immunogenic target for vaccination. Today, with the rapid 
development of technologies for DNA sequencing, the identification of those mutations becomes 
widely feasible. Evidence for the power of neoantigen vaccines in pre-clinical and clinical 
settings was recently published. The first proof of concept of neoantigen vaccines was shown in 
the B16 mice melanoma model4. By prophylactic vaccination with two mutated epitopes, the 
researchers achieved complete tumor protection in 40% of the mice. In therapeutic settings, they 
observed significant tumor growth inhibition induced by mutation-specific peptide vaccination. 
Recently, the same group published evidence for the potential of neoantigen vaccines as tested in 
three independent murine models5. In this paper, the researchers showed that a considerable 
fraction of the non-synonymous mutations in the cancer were immunogenic, and the majority of 
them were recognized by CD4+ T cells. Vaccination with these CD4+ epitopes induced potent 
tumor control and rejection of established tumors. Similar results were published in another 
independent mutation based cancer model. In this work the authors developed a pipeline to 
identify immunogenic mutant peptides using whole-exome and transcriptome sequencing 
together with mass spectrometry. Again, the researchers were able to show that vaccination with 
mutation specific peptides induced therapeutically active T cell responses. Results of the current 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
9
pre-clinical data support the evaluation of neoantigen vaccines as an effective cancer treatment. 
Although studied extensively in pre-clinical models, only one limited human trial has been 
published. In this trial, a group from Washington University in St. Louis reported on a first-in-
man clinical trial evaluating neoantigen vaccine in patients with metastatic melanoma6. The 
researchers vaccinated three patients with mature dendritic cells presenting seven predicted 
mutation-specific HLA-A*02:01 restricted epitopes. They showed that vaccination with high 
affinity, patient specific mutated epitopes augmented T cell immunity directed against naturally 
occurring dominant neoantigens and broadened the response by revealing subdominant 
neoantigens; but no clinical responses were observed. However, this did show that vaccination 
against mutated epitopes was safe as none of the three patients experienced any autoimmune 
adverse events. 
1.2.2
Identification of Cancer-Specific Immunogenic Epitopes Derived from Somatic 
Mutations
The NCI SB has developed a process to identify immunogenic T cell epitopes derived from 
neoantigens (Figure 1). The identification of these antigens is done in four main stages: 1) 
Whole exome sequencing and RNA-seq analysis of the tumor and a matched normal apheresis
sample to identify highly expressed somatic mutations; 2) Construction of tandem minigenes 
(TMGs) and synthesis of long peptides covering these mutations; 3) expression of the constructs 
in autologous antigen presenting cells; 4) In vitro co-culture assay to identify T cells recognizing 
mutated epitopes from TIL and PBMC7. In earlier studies done by the NCI SB (published and 
unpublished data), 25 patients with melanoma were screened in the above manner, 64 antigenic 
somatic mutations were identified with no overlapping between patient tumors8,9. Recently, a 
study to identify antigenic mutations from patients with epithelial cancers including those of the 
GI tract, genitourinary tract and breast, identified 57 non-overlapping somatic mutations. 
In this study, two patients were identified who developed T-cell response against the mutated 
KRAS oncogene10. Work published by the NCI SB in Nature Medicine proved for the first time 
that neoantigen specific T cells can also be isolated from patients’ PBMC7. By using this 
process, neoantigen specific lymphocytes were identified in the peripheral blood of three of four 
patients with melanoma. 
Despite their low frequency in the circulation, these CD8+PD-1+ cell populations had 
lymphocytes that targeted unique patient-specific neoantigens. This method provides a novel 
noninvasive approach to identify antigenic neoantigens. 
Thus far, more than 190 immunogenic epitopes have been identified (Table 1) from multiple 
cancer types including melanoma, ovarian, colorectal, lung and breast cancers7,8,10-13. This 
process, which can be applied to any cancer type and can be completed in several weeks, ensures 
that the selected antigens are not just expressed in the tumor but can also prompt a significant T 
cell-mediated immune response.
1.2.3
Preclinical Studies for the Generation of a Dendritic Cell Vaccine
Development of a personalized neoantigen vaccine puts forward several hurdles. The vaccination 
must address those issues that are unique to the cancer model. The number of epitopes is 
variable.
x
The vaccine should optimally include both CD4 and CD8 epitopes.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
10
x
The vaccine may induce a cell-mediated Th1 response.
x
The vaccine manufacturing should avoid expensive and time-consuming processes. 
To address these issues, ex vivo generated DC will be used. Manufacturing of DC from patient 
PBMC is a well-established and straightforward method. DC vaccines are proven to induce T 
cell responses against multiple malignancies and infectious diseases. Numerous clinical trials 
incorporating and using different administration routes have demonstrated the safety and 
tolerance of DC vaccines14. In addition, the use of DCs both as antigen presenting cells in the 
initial high throughput screening for recognition of neoantigens and in the delivery of these 
antigens in the vaccine product, should ensure that all antigens will be properly processed and
presented to T cells upon vaccination.
DCs Matured with PolyI:C, R848, and IFNJUpregulate Co-Stimulatory Molecules and 
Secrete Pro-Inflammatory Cytokines
DCs are one of the most effective antigen-presenting cells to induce T cell immunity. Even 
though immature DCs (iDCs) can uptake, process, and present antigens, they fail to secrete 
proinflammatory cytokines, and therefore have been shown to be tolerogenic, or at best weakly 
immunogenic15. Multiple maturation cocktails have been used in the clinic to produce fully 
mature DCs. Until recently, researchers used mainly immature DCs or cells activated by a 
cytokine cocktail (Il-1E, IL-6, TNFD and PGE2). Lately, several reports citing the use of TLR 
ligands such as PolyI:C and R848 as DCs stimulators have been published6. To evaluate which 
maturation method generated the highest number of DCs secreting IL-12p70/TNFDday six 
monocyte-derived DCs were incubated with LPS, CD40L expressing 3T3 cells, TLR mix 
(PolyI:C, R848, and IFNJ) and cytokine cocktail (Il-1E, IL-6, TNFD and PGE2). Sixteen hours 
post incubation the level of costimulatory molecules was measured by flow cytometry and the 
secretion of multiple cytokines and chemokines was measured by multiplex assay (Figure 2). 
From Figure 2 it can be clearly seen that the TLR mix induced the secretion of IL-12p70, TNFD,
IP-12, MCP-1 and MIP-1E. The TLR cocktail is capable of maturing DCs to secrete TH1 
polarizing cytokines and multiple chemokines. Recently, a group from Washington University in 
St. Louis (WUSTL) used a combination of K562-CD40L cells with PolyI:C, R848, and IFNJto 
mature monocyte-derived DCs. To evaluate if the addition of K562-CD40L cells to the TLR mix 
could induce superior DCs maturation, clinical grade K562-CD40L cells (K602C14) were 
produced and their contribution to DC stimulationwas evaluated. The original cells used by the 
WUSTL group (K463H) (Figure 3) were used as a positive control. This indicates that TLR mix 
alone is enough to produce fully mature DCs, and thus no additional stimulation is needed. 
DCs Loaded with Long Peptides and Minimal Epitopes Stimulate both CD4 and CD8 T 
Cells 
To evaluate the best method of loading DCs with antigenic determinants, mRNA electroporation 
of TMGs was compared to loading with long peptides and minimal epitopes. 
A screening system to evaluate the presentation of three CD8 and three CD4 epitopes in one 
assay was developed. A series of TMG constructs harboring different signal peptides and 
targeting signals and encoding both long peptides and minimal epitopes covering all antigenic 
determinants (Figure 4) were produced. 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
11
To test antigen presentation, DCs were transfected with the TMG constructs or, loaded with long 
peptide or minimal epitopes. DCs were co-cultured with PBMC transduced with TCRs
recognizing the corresponding antigens and antigen recognition was evaluated by flow cytometry 
for 41BB expression (Figure 5). The results clearly showed that while long peptides and 
minimal epitopes were recognized by CD4 and CD8 cells, TMGs elicited only CD8 responses 
likely indicating that CD4 epitopes cannot be processed through cytosolic proteins thus 
abrogating the use of TMGs as an efficient platform for antigen presentation. To test if mature 
DCs could efficiently present both CD4 and CD8 epitopes DCs were loaded with long peptides 
and minimal epitopes and stimulated for 16 hours using the TLR mix (Figure 6). As shown the 
mature DCs efficiently presented both minimal and long epitopes for CD4 and CD8 epitopes.
Thus, mature DCs loaded with long peptides or minimal epitopes covering both CD8 and CD4 
epitopes will be used in this trial.
Mature DCs can Stimulate Both Memory, Effector, and Naïve Neoantigen Specific T 
Cells
An efficient vaccine should be able to augment neoantigen specific T cells and to expand the 
breadth of the anti-tumor immune response. To test if DCs matured with TLR mix are capable of 
stimulating naïve, antigen-experienced and terminally differentiated effector cells, an in vitro 
stimulation experiment was conducted. Apheresis samples from a patient with colorectal cancer 
were thawed and then incubated in an appropriately sized tissue culture flask. Adherent 
monocytes were differentiated into DCs, loaded with neoantigen peptide pools and matured 
using a TLR mix. Non-adherent cells were collected, and memory, naïve and terminally 
differentiated effector memory cells (TEMRA) were sorted and in vitro stimulated with mature 
peptide-pulsed autologous DCs (Figure 7). Ten days after the first stimulation T cells were 
restimulated with DCs loaded with all peptide pools, and sorted based on 41BB and OX40 
expression to enrich for neoantigen-specific cells. Cells were then tested for neoantigen specific
recognition using single peptides derived from the most reactive peptide pools. By applying this 
method, neoantigen specific T cells from naïve, antigen-experienced and TEMRA cells (Figure 8)
could be identified. These results show that peptide-loaded, TLR mix stimulated DCs are capable 
of in vitro activation of T cells derived from memory or naïve precursors.
1.2.4
Rationale for Selecting Defined, Tumor Suppressor and Driver Neoantigens to use in this 
Trial
Historically, preclinical and clinical neoantigen vaccines used epitopes predicted in silico. 
Although in silico prediction can select for proper antigens, in many cases there is no evidence 
that those antigens are processed and presented by the tumor or antigen presenting cells in the 
microenvironment. As described above, patient TIL and PBMC will be screened to identify 
defined mutated antigens recognized by T cells guaranteeing that those antigens are processed 
and presented for T cells in the tumor or its draining lymph nodes. 
There will be one exception for using defined neoantigens in this trial. There is growing evidence 
to suggest that targeting tumor driver and suppressor genes can elicit superior clinical response. 
Mutations in these genes are highly common and have been found in many tumors. When 
mutations are identified in KRAS, NRAS, p53, EGFR, IDH1, PIK3CA or any other potential 
driver mutations, peptides covering those epitopes will be synthesized and used in the vaccine 
composition. The method/process for epitope selection and synthesis is shown in Figure 9.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
12
Augmenting T cell responses against defined neoantigens, should increase the likelihood of a 
significant clinical response. 
1.2.5
Relevant Preclinical and Clinical Experience to Support the Proposed Route and Dose of 
the DC Vaccine
As shown by the NCI SB, neoantigen specific cells can be found both in the tumor and in the 
blood of cancer patients7,10. To boost responses against defined neoantigens, we are using a 
systemic vaccine dose and schedule covering both the blood and peripheral tissues. For that 
purpose, we decided to use two routes of administration: Intravenous (IV) to stimulate T cells 
that reside in the lungs, spleen and the bone marrow of the patients and subcutaneously (SQ) to 
stimulate T cells residing in peripheral lymph nodes. This concept was also shown before in a 
mouse model16. Recently, a group from Washington University in St. Louis (WUSTL) reported 
on a first-in-man clinical trial evaluating neoantigen vaccine in patients with metastatic 
melanoma6. The researchers vaccinated three patients IV with mature dendritic cells presenting 
seven predicted mutation-specific HLA-A*02:01 restricted epitopes. They showed that 
vaccination with patient specific mutated epitopes augmented T cell immunity directed against 
naturally occurring dominant neoantigens and broadened the response by revealing subdominant 
neoantigens. The initial priming dose they used was 1.5x107 DC per peptide (9 peptides in total, 
1.35X108 DC total), and all the subsequent doses were 5.0x106 DC per peptide (4.5X107 DC 
total). In our trial, we plan to load DC with a peptide pool containing all defined epitopes 
recognized by TIL. Therefore, we chose a dose level similar to the doses used during the 
WUSTL trial for the IV administration. In a different phase I/II clinical trial of sequential SQ and 
IV delivery of a dendritic cell vaccine for refractory multiple myeloma17, patients were 
vaccinated with increasing doses of 5 × -, 10× -, 50×106 cells and 10× -, 50×106 cells 
respectively. The treatment was well tolerated, and no severe adverse events occurred. Therefore, 
we decided to vaccinate with DC doses which are not higher than 50×106 for the SQ and the IV 
route of administration. We are planning to give the maximal dose possible according to our 
manufacturing yields to allow efficient activation of circulating and tissue resident neoantigen 
specific cells.
2
ELIGIBILITY ASSESSMENT AND ENROLLMENT
ELIGIBILITY CRITERIA
2.1.1
Inclusion Criteria
a.
Metastatic melanoma or epithelial cancer with at least one lesion that is resectable or in
selected cases, available PBMCs.
b. Measurable and evaluable metastatic disease per RECIST 1.1 criteria.
c.
Confirmation of the diagnosis of metastatic cancer by the Laboratory of Pathology of 
NCI.
d. All patients must be refractory to approved standard systemic therapy.
e.
Patients with 3 or fewer brain metastases that are less than 1 cm in diameter and 
asymptomatic are eligible. Lesions that have been treated with stereotactic radiosurgery 
must be clinically stable for one month after treatment for the patient to be eligible. 
Patients with surgically resected brain metastases are eligible. 
f.
$JH \HDUVDQG\HDUV

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
13
g. Clinical performance status of ECOG 0, 1, 2.
h. Patients of both genders must be willing to practice birth control from the time of 
enrollment on this study and for four months after treatment.
i.
Serology: 
x
Seronegative for HIV antibody. (The experimental treatment being evaluated in this 
protocol depends on an intact immune system. Patients who are HIV seropositive can 
have decreased immune-competence and thus are less responsive to the experimental 
treatment and more susceptible to its toxicities.) 
x
Seronegative for hepatitis B antigen, and seronegative for hepatitis C antibody. If 
hepatitis C antibody test is positive, then the patient must be tested for the presence of 
antigen by RT-PCR and be HCV RNA negative. 
j.
Hematology
x
Absolute neutrophil count > 1000/mm3 without the support of filgrastim
x
:%&PP3
x
3ODWHOHWFRXQWPP3
x
Hemoglobin > 8.0 g/dL. Subjects may be transfused to reach this cut-off. 
x
CD4 count > 200/uL
k. Chemistry:
x
Serum ALT/AST < 5.0 x ULN
x
Serum cUHDWLQLQHPJGL
x
7RWDOELOLUXELQ2.0 mg/dL, except in patients with Gilbert’s Syndrome, who must 
have a total bilirubin  3.0 mg/dL.
l.
More than four weeks must have elapsed since any prior systemic therapy at the time the 
patient receives the vaccine, and patients’ toxicities must have recovered to a grade 1 or 
less (except for toxicities such as alopecia or vitiligo).
Note: Patients may have undergone minor surgical procedures within the past 3 weeks, as 
long as all toxicities have recovered to grade 1 or less. 
m. Ability of subject to understand and the willingness to sign a written informed consent 
document.
n. Subjects must be co-enrolled on protocol 03-C-0277.
2.1.2
Exclusion Criteria
a.
Women of child-bearing potential who are pregnant or breastfeeding because of the 
potentially dangerous effects of the treatment on the fetus or infant.
b. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency 
Disease).
c.
Concurrent opportunistic infections. (The experimental treatment being evaluated in this 
protocol depends on an intact immune system. Patients who have decreased immune 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
14
competence may be less responsive to the experimental treatment and more susceptible to 
its toxicities).
d. Active systemic infections requiring anti-infective treatment, coagulation disorders or any 
other active or uncompensated major medical illnesses.
e.
Patients who are receiving any other investigational agents.
SCREENING EVALUATION
Note: Testing for screening evaluation is conducted under the NCI SB companion protocol, 99-
C-0128 (Evaluation for NCI Surgery Branch Clinical Research Protocols).
Note: The date of enrollment is the date the subject signs the written informed consent 
document. The patient will be consented and enrolled on this protocol once the patient has 
adequate leukapheresis material banked by the Surgery Branch Cell Production Facility (SB 
CPF) and prior to administration of the first vaccine.
2.2.1
Within 3 Months Prior to Enrollment
x
HIV antibody titer, HBsAg determination, and anti HCV 
x
Confirmation of diagnosis of metastatic melanoma or epithelial cancer by the Laboratory 
of Pathology of NCI. (Note: Testing is permitted to be conducted at any time prior to 
enrollment.)
x
Anti CMV antibody titer, HSV serology, and EBV panel. (Note: Patients who are known 
to be positive do not need to be retested.)
2.2.2
Within 4 Weeks Prior to Enrollment
x
Complete history and physical examination. (Note: Patient history may be obtained
within 8 weeks prior to enrollment.)
x
Baseline imaging to determine the status of disease. This may include CT, MRI, PET, 
and/or photography.
2.2.3
Within 7 Days Prior to Enrollment
x
Physical examination, including ECOG of 0, 1, or 2
x
Vital signs
x
CBC w/differential
x
Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN),
Hepatic Panel (alkaline phosphatase, AST/GOT, ALT/GPT, total bilirubin, direct 
bilirubin), Mineral Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, 
Creatinine Kinase, Lactate Dehydrogenase, Total Protein
x
PT/PTT
x
ȕ-HCG pregnancy test (serum or urine) on all females of child-bearing potential
x
Urinalysis, with culture if indicated 
x
Chest x-ray
x
EKG

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
15
REGISTRATION PROCEDURES
2.3.1
Prior to Registration for this Protocol
Patients will initially sign the consent for and enroll on the NCI SB companion protocol, 03-C-
0277 (Cell Harvest and Preparation for Surgery Branch Adoptive Cell Therapy Protocols), prior 
to tumor harvest and leukapheresis for generation of the cell product. Patients will sign the 
consent for and enroll on this treatment protocol once the patient has adequate leukapheresis
material banked by the SB CPF and prior to administration of the first vaccine.
Immature and mature dendritic cells must pass the FACS and sterility or gram stain assay as 
noted in the Certificates of Analysis (CoAs) (Appendix 2 and Appendix 3). If cells do not meet 
CoA requirements then the patient will be taken off-study (Section 3.6.2). Once it is confirmed 
that immature and mature dendritic cells have met the CoA requirements (Appendix 2 and 
Appendix 3), the vaccines will be administered to the patient.
2.3.2
Registration Procedure
Authorized staff must register an eligible candidate with NCI Central Registration Office (CRO) 
within 24 hours of signing consent. A registration Eligibility Checklist from the web site 
(http://home.ccr.cancer.gov/intra/eligibility/welcome.htm) must be completed and sent via 
encrypted email to: NCI Central Registration Office at ncicentralregistration-l@mail.nih.gov.
After confirmation of eligibility at Central Registration Office, CRO staff will call pharmacy to 
advise them of the acceptance of the patient on the protocol prior to the release of any 
investigational agents. Verification of Registration will be forwarded electronically via email to 
the research team. A recorder is available during non-working hours. 
TREATMENT ASSIGNMENT PROCEDURES
2.4.1
Cohorts
Subjects will be enrolled in one of two cohorts:
Number
Name
Description
1
Melanoma
Patients with metastatic melanoma
2
Epithelial 
Patients with epithelial cancer
2.4.2
Arms
Number
Name
Description
1
Experimental 
Therapy
Peptide loaded dendritic cell vaccine on days 
0, 14, 28, and 42
2.4.3
Randomization and Arm Assignment
This is a non-randomized study. All subjects will be directly assigned based on cohort as 
follows:
x
Subjects in Cohorts 1-2 will be directly assigned to Arm 1.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
16
3
STUDY IMPLEMENTATION
STUDY DESIGN
3.1.1
Pre-Treatment Phase: DC Preparation
Tumor Harvest
 or possible 
apheresis
Initial Growth of 
Cells
 Sequencing
 Identification 
of mutations
 Manufacture  
peptides
Cryo preserve cells
Patient Apheresis
 Manufacture 
Dendritic Cells 
 Cryo-preserve 
DCs
 Thaw DCs
 Pulse with 
identified 
peptides
 Mature DCs
 Prepare final 
product
Administer
Vaccines 1 
(day 0)
 Thaw cells
 Test reactivity 
of peptides
 Identify 
peptides
Order GLP Peptides
6-8  weeks
5-6 weeks
Receive GLP 
Peptides
 Thaw DCs
 Pulse with 
identified 
peptides
 Mature DCs
 Prepare final 
product
Administer
Vaccines  2
(~day 14)
 Thaw DCs
 Pulse with 
identified 
peptides
 Mature DCs
 Prepare final 
product
Administer
Vaccines 3
(~day 28)
 Thaw DCs
 Pulse with 
identified 
peptides
 Mature DCs
 Prepare final 
product
Administer
Vaccines 4
(~day 42)
3 days
Under 03-C-0277, patients with evaluable metastatic melanoma or epithelial cancers will 
undergo resection of the tumor and apheresis. Whole exome and RNA sequencing will be 
performed to identify all cancer mutations using our standard procedures. TIL will be grown and 
expanded for this trial according to standard operating procedures submitted in the IND. After a 
sufficient yield of TIL (5x107 cells) is expanded and PBMCs are available, immunogenic 
neoantigens will be identified by high throughput immunologic screening using long peptides 
and tandem minigenes covering all mutated epitopes to identify the exact mutations recognized 
by autologous T cells. For vaccine preparation, patients will undergo apheresis and DCs will be 
produced and cryopreserved for vaccine preparation. Peptides (15-25mers with the mutation at 
positions 13) or minimal epitopes (9-10mers) will be synthesized and pulsed on autologous 
antigen presenting cells (DC). The antigen pulsed DC will then be matured using a TLR mix.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
17
Patients will receive peptide-loaded mature dendritic cells intravenously and subcutaneously at 
biweekly intervals for 4 cycles. Immunologic tests will be performed as described in Section 5.5.
3.1.2
Treatment Phase
The protocol will consist of two cohorts: cohort 1, patients with melanoma and cohort 2, patients 
with epithelial cancer.
The study will utilize an optimal design where initially 21 evaluable patients will be enrolled to 
each cohort. If 0 or 1 of the 21 patients in a given cohort experiences a clinical response, then no 
further patients will be enrolled; if 2 or more of the first 21 evaluable patients enrolled have a 
clinical response, then accrual will continue until a total of 41 evaluable patients have been 
enrolled to that cohort.
Patients will receive no other experimental agents while on this protocol. All patients will 
receive one course of treatment (4 vaccination cycles). The start date of the course will be the 
start date of the first vaccination; the end date will be the day of the first post-treatment
evaluation (week 8 if the patient receives all 4 cycles).
Enrollment will be staggered within each cohort with three weeks between start dates for the first 
three patients to allow for a 3-week interval for safety assessment before the next enrolled patient 
is scheduled to receive their first dose of the DC vaccine.
3.1.3
Protocol Stopping Rules 
New subject enrollment to the protocol will be temporarily halted if any of the following 
conditions are met, and discussions will be had with the FDA or NIH IRB regarding protocol 
revisions if applicable:
x
If one or more treatment-related death occurs due to vaccination, we will promptly 
discuss this with the NCI IRB and the FDA. 
x
Two or more patients develop a grade 3 or greater toxicity at any point in the study not 
attributable to disease progression (or circumstances unrelated to the study).
o
The following grade 3 reactions commonly associated with immunization will not be 
included in this protocol stopping rule:

Pruritus/itching: intense or widespread and interfering with ADL lasting <72 
hours 

Fatigue: severe fatigue interfering with ADL lasting < 72 hours 

Fever: > 40.0°C for < 24 hours 

Local lymphadenopathy lasting < 1 week
x
If one of the first three patients (or 2 of the first 6 patients, or 3 of the first 9 patients, or 4 
of the first 12 patients) develop grade 3 autoimmunity, that cannot be resolved to less 
than or equal to a grade 2 autoimmune toxicity within 10 days, or any grade 4 or greater 
autoimmune toxicity.
DRUG ADMINISTRATION
Patients will receive peptide pulsed dendritic cell vaccines on days 0, 14 (± 5 days), 28 (± 5
days), and 42 (± 5 days). The vaccine will be administered as both an intravenous (IV) infusion

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
18
and a subcutaneous (SQ) injection. The total dose will be divided equally between IV and SQ 
containing 1.0x107 to 8.0x107 cells per cycle depending upon the manufacturing yield.
The patient’s dendritic cell products will be delivered to the Patient Care Unit by a staff member 
from the NCI SB. Prior to infusion, the cell product identity labels on the syringe and cell 
infusion bag will be double-checked by two authorized staff (MD or RN), an identification of the 
product and documentation of administration are entered in the patient’s chart, as is done for 
blood banking protocols.
x
Cells for SQ injection will be administered first, followed by cells for IV infusion.
x
Cells for SQ injection will be administered in a total of 1.0 mL (0.5x107 to 4.0x107 cells)
into the deep subcutaneous tissue of a proximal extremity. Injections will preferentially 
be administered in the anterior thigh. Injections will not be given into extremities that 
have had radical lymph node dissections. The same extremity will be used for each cycle 
of the peptide loaded dendritic cell vaccine administration.
x
Cells for IV administration (0.5x107 to 4.0x107 cells) will be infused over 20 to 30 
minutes, or as clinically determined by an investigator for patient safety, via non-filtered 
tubing, gently agitating the bag during infusion to prevent cell clumping. See Appendix 1
for cell infusion instructions. 
x
Vital signs will be monitored hourly (±15 minutes) for two hours. The post-
administration monitoring period may be adjusted by the PI.
x
All patients will receive their peptide pulsed dendritic cell vaccines in the NIH Clinical 
Center. The first three patients will receive their Day 0 SQ injection and IV infusion on
the In-Patient Care Unit, and then will receive their subsequent SQ injections and IV 
infusions (days 14, 28, and 42) as outpatient procedures in the 3SE-S Day Hospital. All 
patients enrolled after the first three patients will receive their full course of SQ injections 
and IV infusions (days 0, 14, 28, and 42) as outpatient procedures in the 3SE-S Day
Hospital. This schedule may be adjusted by the PI as clinically appropriate. 
ON-STUDY EVALUATION
3.3.1
Within 7 Days Prior to the 1st Vaccination (Day 0)
x
Repeat all screening procedures from Section 2.2.3 if not completed within 7 days prior 
to the 1st vaccination (Day 0). 
3.3.2
Within 2 Days Prior to the 1st Vaccination (Day 0)
x
Lymphocyte phenotyping (TBNK)
x
Review of baseline symptoms
x
Research samples as described in Section 5
3.3.3
Within 2 Days Prior to Each Subsequent Vaccination (Days 14, 28, and 42 (± 5 days))
x
Physical examination, including ECOG
x
Vital signs 
x
CBC w/differential

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
19
x
Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN), 
Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct bilirubin), Mineral 
Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, Creatinine Kinase, Lactate 
Dehydrogenase, Total Protein
x
ȕ-HCG pregnancy test (serum or urine) on all females of child-bearing potential
x
Toxicity assessment, including a review of systems
x
Research samples as described in Section 5
3.3.4
After Each Vaccination
x
Vital signs will be monitored hourly (± 15 minutes) for two hours. The post-
administration monitoring period may be adjusted by the PI.
POST-TREATMENT EVALUATION (FOLLOW-UP)
3.4.1
1st Follow-up Evaluation (Day 56 ± 5 days)
All patients will return to the NIH Clinical Center for their first follow-up evaluation on day 56 
(±5 days), which is approximately 2 weeks from the 4th vaccination.
At the first follow-up evaluation, patients will undergo:
x
Physical examination, including ECOG
x
Vital signs
x
CBC w/differential 
x
Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN), 
Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct bilirubin), Mineral 
Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, Creatinine Kinase, Lactate 
Dehydrogenase, Total Protein
x
Lymphocyte phenotyping (TBNK)
x
ȕ-HCG pregnancy test (serum or urine) on all females of child-bearing potential
x
Toxicity assessment, including a review of systems
x
Research samples as described in Section 5
x
Imaging studies as performed at baseline to determine tumor response. If clinically 
indicated, other scans or x-rays may be performed, e.g., brain MRI, bone scan.
x
A 5-liter leukapheresis may be performed. If the patient is unable to undergo 
leukapheresis, approximately 96 mL of blood may be obtained. This will be conducted 
under 03-C-0277.
3.4.2
Subsequent Follow-up Evaluations
The second follow-up evaluation will take place 1 month (± 5 days) after the first follow-up
evaluation (Day 56). Subsequent follow-up evaluations will take place every 1-2 months for the 
first year, and then annually or as clinically appropriate as determined by the PI. As noted in 
Section 3.4.1, additional leukapheresis may be performed only at the first follow-up evaluation.
It will not be performed at any other post-treatment evaluations.
Note: Patients may be seen more frequently as clinically indicated.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
20
At each scheduled follow-up evaluation, patients will undergo:
x
Physical examination, including ECOG
x
Vital signs
x
CBC w/differential
x
Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN), 
Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct bilirubin), Mineral 
Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, Creatinine Kinase, Lactate 
Dehydrogenase, Total Protein
x
Lymphocyte phenotyping (TBNK)
x
ȕ-HCG pregnancy test (serum or urine) on all females of child-bearing potential
(complete at all follow-up evaluations within 4 months after the last vaccine)
x
Toxicity assessment, including a review of systems
x
Research samples as described in Section 5
x
Imaging studies as performed at baseline to determine tumor response. If clinically 
indicated, other scans or x-rays may be performed, e.g., brain MRI, bone scan.
Note: Patients who are unable or unwilling to return for follow-up evaluations may be followed
via phone or e-mail contact. Patients may be asked to send laboratory, imaging and physician 
exam reports performed by their treating physician.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
21
STUDY CALENDAR
Assessments
Screening/Baseline
(prior to enrollment on treatment protocol)
On-Study/Course of Treatment 
(1 vaccine = 1 cycle)
Post-Treatment Follow-up
Within 3 
Months
Within 4 
Weeks
Within 7 
Days
Day 01
Day 14 
(±5 d)
Day 28 
(±5 d)
Day 42
(±5 d)
Day 56 
(±5 d)
Subsequent 
Follow-up Visits2
Informed consent3
Confirmation of diagnosis by NCI Lab 
of Pathology4
X
Medical history5
X
Physical exam
X
X
X
X
X
X
X
X
Performance score (ECOG)6
X
X
X
X
X
X
X
Vital signs7
X
X
X
X
X
X
X
ȕ-HCG pregnancy test8
X
X
X
X
X
X
X
Urinalysis9
X
X
EKG
X
X
Toxicity assessment
X
X
X
X
X
Laboratory Procedures
Viral testing10
X
1 Assessments (except TBNK and research samples) may be done within 7 days prior to the first vaccination on Day 0. Lymphocyte phenotyping (TBNK) and 
research samples must be drawn within 2 days prior to the first vaccination on Day 0.
2 The second follow-up evaluation will take place 1 month (± 5 days) after the first follow-up evaluation (Day 56). Subsequent follow-up evaluations will take place 
every 1-2 months for the first year, and then annually or as clinically appropriate as determined by the PI. Patients may be seen more frequently as clinically indicated.
Patients who are unable or unwilling to return for follow-up evaluations may be followed via phone or email contact. Patients may be asked to send laboratory, 
imaging and physician exam reports performed by their treating physician.
3 The date of enrollment is the date the subject signs the written informed consent document. The patient will be consented and enrolled on this protocol once the 
patient has adequate leukapheresis material banked by the SB CPF and prior to administration of the first vaccine.
4 Confirmation of diagnosis of metastatic melanoma or epithelial cancer. Testing is permitted to be conducted at any time prior to enrollment.
5 Patient history may be obtained within 8 weeks prior to enrollment.
6 ECOG of 0, 1, or 2.
7 After each vaccination, vital signs will be monitored hourly (± 15 minutes) for two hours. The post-administration monitoring period may be adjusted by the PI.
8 Serum or urine; on all females of child-bearing potential. Complete at all follow-up evaluations within 4 months after the last vaccine.
9 With culture if indicated.
10 HIV antibody titer, HBsAg determination, anti HCV, anti CMV antibody titer, HSV serology, and EBV panel. Patients who are known to be positive for CMV, 
HSV and/or EBV do not need to be retested.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
22
Assessments
Screening/Baseline
(prior to enrollment on treatment protocol)
On-Study/Course of Treatment 
(1 vaccine = 1 cycle)
Post-Treatment Follow-up
Within 3 
Months
Within 4 
Weeks
Within 7 
Days
Day 01
Day 14 
(±5 d)
Day 28 
(±5 d)
Day 42
(±5 d)
Day 56 
(±5 d)
Subsequent 
Follow-up Visits2
CBC w/differential
X
X
X
X
X
X
X
Chemistry panel11
X
X
X
X
X
X
X
PT/PTT
X
X
Lymphocyte phenotyping (TBNK)
X
X
X
Additional leukapheresis12
X
Correlatives13
CPT tubes (8) (SB CPF)
X
X
X
X
X
X
SST tube (1) (Figg Lab)
X
X
X
X
X
X
Imaging
CT, MRI, PET, and/or photography14
X
X
X
Chest x-ray
X
X
Treatment/Intervention
Peptide loaded dendritic cell vaccine15
X
X
X
X
11 Chemistry panel: Sodium (Na), Potassium (K), Chloride (Cl), Total CO2 (bicarbonate), Creatinine, Glucose, Urea nitrogen (BUN), Albumin, Calcium total, 
Magnesium total (Mg), Inorganic Phosphorus, Alkaline Phosphatase, ALT/GPT, AST/GOT, Total Bilirubin, Direct Bilirubin, LD, Total Protein, Total CK, Uric Acid
12 A 5 liter leukapheresis may be performed. If the patient is unable to undergo leukapheresis, approximately 96 ml of blood may be obtained. This will be conducted 
under 03-C-0277, and will only be performed at the first follow-up evaluation. 
13 Research samples, as described in Section 5, prior to each vaccination and at each scheduled follow-up evaluation. Includes evaluation of specific lysis and 
cytokine release, metabolomic and bioenergetic studies (using Seahorse), intracellular FACS of cytokine production, ELISA-spot assays, and lymphocyte subset 
analysis may be used to evaluate the immunological correlates of treatment.
14 As performed at baseline to determine tumor response. If clinically indicated, other scans or x-rays may be performed, e.g., brain MRI, bone scan.
15 Administered subcutaneously and intravenously.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
23
CRITERIA FOR REMOVAL FROM PROTOCOL THERAPY AND OFF-STUDY CRITERIA
Prior to removal from study, effort must be made to have all subjects complete a safety visit 
approximately 30 days following the first follow-up evaluation on Day 56.
3.6.1
Criteria for Removal from Protocol Therapy
Patients will be taken off treatment for the following:
x
Completion of treatment
x
Grade 3 or 4 toxicity due to the vaccine, excluding those known events such as local 
injection site reactions, skin rash, pruritus, fatigue, fever and local adenopathy.
x
Progression of disease
x
Participant requests to be withdrawn from active therapy
x
Investigator discretion
x
Positive pregnancy test
3.6.2
Off-Study Criteria
Patients will be taken off study for the following:
x
Cells do not meet CoA requirements
x
Completed study follow-up period
x
Patient requests to be withdrawn from the study
x
There is significant noncompliance
x
Lost to follow-up
x
Investigator discretion
x
Progression of disease
x
Death
All patients will be co-enrolled on the NCI SB companion protocol, 09-C-0161 (Follow up
Protocol for Subjects Previously Enrolled in NCI Surgery Branch Studies). Patients who are 
taken off-study for progressive disease or study closure on the treatment protocol may be 
followed on protocol 09-C-0161.
Once a patient is taken off study, no further data can be collected.
3.6.3
Off Protocol Therapy and Off-Study Procedure
Authorized staff must notify the Central Registration Office (CRO) when a subject is taken off 
protocol therapy and when a subject is taken off-study. A Participant Status Updates Form from 
the web site (http://home.ccr.cancer.gov/intra/eligibility/welcome.htm) main page must be 
completed and sent via encrypted email to: NCI Central Registration Office at
ncicentralregistration-l@mail.nih.gov.
4
CONCOMITANT MEDICATIONS/MEASURES
Over the counter medications, such as ibuprofen, may be used to provide relief of local injection 
site reactions.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
24
5
BIOSPECIMEN COLLECTION
Blood and tissue are tracked at the patient level and can be linked to all protocols on which the 
patient has been enrolled. Samples will be used to support the specific objectives listed in the 
treatment protocol(s), e.g., immunologic monitoring, cytokine levels, persistence. Patients who 
agree to have specimens and data stored to support long-term research efforts within the NCI SB
will be consented and enrolled on 03-C-0277.
The amount of blood that may be drawn from adult patients for research purposes shall not 
exceed 10.5 mL/kg or 550 mL, whichever is smaller, over any eight-week period.
Samples will be ordered in CRIS and tracked through Clinical Trial Data Management system.
Should a CRIS screen not be available, the CRIS downtime procedures will be followed.
Samples will not be sent outside NIH without IRB notification and an executed MTA.
SAMPLES SENT TO FIGG LAB
x
Venous blood samples will be collected in either a 4 mL or an 8 mL SST tube to be 
processed for serum and stored for future research. Record the date and exact time of 
draw on the tube. Blood tubes may be kept in the refrigerator until pick up.
x
For sample pick up, page 102-11964.
x
For immediate help, call 240-760-6180 (main blood processing core number) or, if no 
answer, 240-760-6190 (main clinical pharmacology lab number).
x
For questions regarding sample processing, contact Julie Barnes by e-mail or at 240-760-
6044.
x
The samples will be processed, barcoded, and stored in Dr. Figg’s lab until requested by 
the investigator.
SAMPLES SENT TO SURGERY BRANCH CELL PRODUCTION FACILITY
x
Venous blood samples will be collected in 8 mL CPT tubes to be processed and stored for 
future research. Record the date and exact time of draw on the tube. Blood tubes are kept 
at room temperature until pick up.
x
Samples will be picked up by the research nurse or designee and transported to the SB 
CPF within 24 hours of blood draw.
x
The samples will be processed, barcoded, and stored in the SB CPF.
PRIOR TO EACH VACCINATION
x
8 CPT tubes (8 mL each): SB CPF
x
1 SST tube (8 mL): Figg Lab
AT EACH SCHEDULED FOLLOW-UP EVALUATION
x
8 CPT tubes (8 mL each): SB CPF
x
1 SST tube (8 mL): Figg Lab
IMMUNOLOGICAL TESTING
x
A variety of tests including evaluation of specific lysis and cytokine release, metabolomic 
and bioenergetic studies (using Seahorse), intracellular FACS of cytokine production, 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
25
ELISA-spot assays, and lymphocyte subset analysis may be used to evaluate the 
immunological correlates of treatment. In general, differences of 2-3 fold in these assays 
over the baseline measurement are indicative of true biologic differences. 
Note: The collection and analysis of research labs will be monitored by the TIL lab and not by 
the Center for Cancer Research’s data contractor.
SAMPLE STORAGE, TRACKING AND DISPOSITION FOR SURGERY BRANCH CELL
PRODUCTION FACILITY
Blood and tissue collected during the course of this study will follow the Cell Tracking and 
Labeling System established by the SB CPF. The Cell Tracking and Labeling System is designed 
to unambiguously ensure that patient/data verification is consistent. The patients’ cell samples 
(blood or tissue) are tracked by distinct identification labels that include a unique patient 
identifier and date of specimen collection. Cryopreserved blood and tissue samples also bear the 
date the sample was frozen. All cryopreserved samples are tracked for freezer location and 
storage criteria. All samples are stored in monitored freezers/refrigerators in 3NW NCI SB
laboratories at specified temperatures with alarm systems in place. Serum samples will be sent to 
the Blood Processing Core (BPC) for storage. Samples will be barcoded and stored on site or 
offsite at NCI Frederick Central Repository Services in Frederick, MD. All collected samples 
(blood or tissue) are entered into a central computer database with identification and storage 
location, and this database is backed up every night.
If, at any time, a patient withdraws from the study and does not wish for their existing samples to 
be utilized, the individual must provide a written request. Following receipt of this request, the 
samples will be destroyed and reported as such to the IRB. Any samples lost (in transit or by a 
researcher) or destroyed due to unknown sample integrity (i.e. broken freezer allows for 
extensive sample thawing, etc.) will be reported as such to the IRB.
Blood and tissue collected during the course of this study will be stored, tracked and disposed of 
as specified in protocol 03-C-0277.
SAMPLE STORAGE, TRACKING AND DISPOSITION FOR DR. FIGG’S LAB
5.7.1
Sample Data Collection
All samples sent to the BPC will be barcoded, with data entered and stored in the LABrador (aka 
LabSamples) utilized by the BPC, and data will be updated to the NCI SB central computer 
database weekly. This is a secure program, with access to LABrador limited to defined Figg lab 
personnel, who are issued individual user accounts. Installation of LABrador is limited to 
computers specified by Dr. Figg. These computers all have a password restricted login screen. 
All Figg lab personnel with access to patient information annually complete the NIH online 
Protection of Human Subjects course. 
LABrador creates a unique barcode ID for every sample and sample box, which cannot be traced 
back to patients without LABrador access. The data recorded for each sample includes the 
patient ID, name, trial name/protocol number, time drawn, cycle time point, dose, material type, 
as well as box and freezer location. Patient demographics associated with the clinical center 
patient number are provided in the system. For each sample, there are notes associated with the 
processing method (delay in sample processing, storage conditions on the ward, etc.).

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
26
5.7.2
Sample Storage and Destruction
Barcoded samples are stored in barcoded boxes in a locked freezer at either -20 or -80°C
according to stability requirements. These freezers are located onsite in the BPC and offsite at 
NCI Frederick Central Repository Services in Frederick, MD. Visitors to the laboratory are 
required to be accompanied by laboratory staff at all times. 
Access to stored clinical samples is restricted. Samples will be stored until requested by a 
researcher named on the protocol. All requests are monitored and tracked in LABrador. All 
researchers are required to sign a form stating that the samples are only to be used for research 
purposes associated with this trial (as per the IRB approved protocol) and that any unused 
samples must be returned to the BPC. It is the responsibility of the Principal Investigator to 
ensure that the samples requested are being used in a manner consistent with IRB approval.
Following completion of this study, samples will remain in storage as detailed above. Access to 
these samples will only be granted following IRB approval of an additional protocol, granting 
the rights to use the material. 
If, at any time, a patient withdraws from the study and does not wish for their existing samples to 
be utilized, the individual must provide a written request. Following receipt of this request, the 
samples will be destroyed and reported as such to the IRB. Any samples lost (in transit or by a 
researcher) or destroyed due to unknown sample integrity (i.e. broken freezer allows for 
extensive sample thawing, etc.) will be reported as such to the IRB.
Sample barcodes are linked to patient demographics and limited clinical information. This 
information will only be provided to investigators listed on this protocol, via registered use of the 
LABrador. It is critical that the sample remains linked to patient information such as race, age, 
dates of diagnosis and death, and histological information about the tumor, in order to correlate 
genotype with these variables.
6
DATA COLLECTION AND EVALUATION
DATA COLLECTION
The PI will be responsible for overseeing entry of data into an in-house password protected 
electronic system (C3D), and ensuring data accuracy, consistency and timeliness. The PI,
associate investigators/research nurses and/or a contracted data manager will assist with the data 
management efforts. All data obtained during the conduct of the protocol will be kept in secure 
network drives or in approved alternative sites that comply with NIH security standards. Primary 
and final analyzed data will have identifiers so that research data can be attributed to an 
individual human subject participant. Data will be entered into the NCI CCR C3D database. 
All adverse events (AEs), including clinically significant abnormal findings on laboratory 
evaluations, regardless of severity, will be followed until return to baseline or stabilization of 
event. Patients will be followed for adverse events from the time the patient receives the 
investigational agent/intervention to the time of the second follow-up evaluation, or until off-
study, whichever comes first. 
An abnormal laboratory value will be recorded in the database as an AE only if the laboratory 
abnormality is characterized by any of the following:
x
Results in discontinuation from the study

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
27
x
Is associated with clinical signs or symptoms 
x
Requires treatment or any other therapeutic intervention
x
Is associated with death or another serious adverse event, including hospitalization. 
x
Is judged by the Investigator to be of significant clinical impact
x
If any abnormal laboratory result is considered clinically significant, the investigator will 
provide details about the action taken with respect to the test drug and about the patient’s 
outcome.
All AEs must be recorded on the AE case report form.
End of study procedures: Data will be stored according to HHS and FDA regulations, and NIH 
Intramural Records Retention Schedule as applicable. 
Loss or destruction of data: Should we become aware that a major breach in our plan to protect 
subject confidentiality and trial data has occurred, the IRB will be notified.
6.1.1
Exclusions to Routine Adverse Event Recording
There are no exclusions to routine adverse event reporting/recording for this protocol.
DATA SHARING PLANS
6.2.1
Human Data Sharing Plan
De-identified human data generated for use in future and ongoing research will be shared 
through a NIH-funded or approved repository (ClinicalTrials.gov) and BTRIS. At the completion 
of data analysis, data will be submitted to ClinicalTrials.gov either before publication or at the 
time of publication or shortly thereafter. Data may also be used to support long-term research 
efforts within the NCI SB and de-identified data may also be shared with collaborators as 
specified in protocol 03-C-0277.
6.2.2
Genomic Data Sharing Plan
The NIH Genomic Data Sharing Policy does not apply to this study.
RESPONSE CRITERIA
For the purposes of this study, patients should be re-evaluated for response approximately two
weeks following the last vaccination, then one month later x1, and then every 1-2 months for the 
first year, and then annually or as clinically indicated. In addition to a baseline scan, 
confirmatory scans should also be obtained at least 4 weeks following initial documentation of 
objective response.
Response and progression will be evaluated in this study using the new international criteria 
proposed by the revised Response Evaluation Criteria in Solid Tumors (RECIST) guideline 
(version 1.1)18. Changes in the largest diameter (unidimensional measurement) of the tumor 
lesions and the shortest diameter in the case of malignant lymph nodes are used in the RECIST
criteria.
6.3.1
Definitions
Evaluable for toxicity: All patients will be evaluable for toxicity from the time of their first 
treatment with peptide loaded dendritic cell vaccine.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
28
Evaluable for objective response: Only those patients who have measurable disease present at 
baseline, have received at least one cycle of therapy, and have had their disease re-evaluated will 
be considered evaluable for response. These patients will have their response classified 
according to the definitions stated below. (Note: Patients who exhibit objective disease 
progression prior to the end of cycle 1 will also be considered evaluable.)
Evaluable Non-Target Disease Response: Patients who have lesions present at baseline that are 
evaluable but do not meet the definitions of measurable disease, have received at least one cycle 
of therapy, and have had their disease re-evaluated will be considered evaluable for non-target 
disease. The response assessment is based on the presence, absence, or unequivocal progression 
of the lesions. 
6.3.2
Disease Parameters
Measurable disease: Measurable lesions are defined as those that can be accurately measured in 
at least one dimension (longest diameter to be recorded) as:
x
By chest x-ray:  20 mm;
x
By CT scan: 
o
Scan slice thickness 5 mm or under:  10 mm
o
Scan slice thickness > 5 mm: double the slice thickness
x
With calipers on clinical exam:  10 mm.
All tumor measurements must be recorded in millimeters (or decimal fractions of centimeters).
Note: Tumor lesions that are situated in a previously irradiated area might or might not be 
considered measurable. If the investigator thinks it appropriate to include them, the conditions 
under which such lesions should be considered must be defined in the protocol.
Malignant lymph nodes: To be considered pathologically enlarged and measurable, a lymph 
node must be >15 mm in short axis when assessed by CT scan (CT scan slice thickness 
recommended to be no greater than 5 mm). At baseline and in follow-up, only the short axis will 
be measured and followed.
Non-measurable disease: All other lesions (or sites of disease), including small lesions (longest 
GLDPHWHUPPRUSDWKRORJLFDOO\PSKQRGHVZLWKWRPPVKRUWD[LVDUHFRQVLGHUHG
non-measurable disease. Bone lesions, leptomeningeal disease, ascites, pleural/pericardial 
effusions, lymphangitis cutis/pulmonitis, inflammatory breast disease, and abdominal masses 
(not followed by CT or MRI), are considered as non-measurable.
Target lesions: All measurable lesions up to a maximum of 2 lesions per organ and 5 lesions in 
total, representative of all involved organs, should be identified as target lesions and recorded 
and measured at baseline. Target lesions should be selected on the basis of their size (lesions 
with the longest diameter), be representative of all involved organs, but in addition should be 
those that lend themselves to reproducible repeated measurements. It may be the case that, on 
occasion, the largest lesion does not lend itself to reproducible measurement in which 
circumstance the next largest lesion which can be measured reproducibly should be selected. A 
sum of the diameters (longest for non-nodal lesions, short axis for nodal lesions) for all target 
lesions will be calculated and reported as the baseline sum diameters. If lymph nodes are to be 
included in the sum, then only the short axis is added into the sum. The baseline sum diameters 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
29
will be used as reference to further characterize any objective tumor regression in the measurable 
dimension of the disease.
Non-target lesions: All other lesions (or sites of disease) including any measurable lesions over 
and above the 5 target lesions should be identified as non-target lesions and should also be 
recorded at baseline. Measurements of these lesions are not required, but the presence, absence, 
or in rare cases unequivocal progression of each should be noted throughout follow-up.
6.3.3
Methods for Evaluation of Measurable Disease
All measurements should be taken and recorded in metric notation using a ruler or calipers. All 
baseline evaluations should be performed as closely as possible to the beginning of treatment and 
never more than 4 weeks before the beginning of the treatment.
The same method of assessment and the same technique should be used to characterize each 
identified and reported lesion at baseline and during follow-up. Imaging-based evaluation is 
preferred to evaluation by clinical examination unless the lesion(s) being followed cannot be 
imaged but are assessable by clinical exam.
Clinical lesions: Clinical lesions will only be considered measurable when they are superficial 
(e.g., skin nodules and palpable lymph nodes) and t10 mm diameter as assessed using calipers 
(e.g., skin nodules). In the case of skin lesions, documentation by color photography, including a 
ruler to estimate the size of the lesion, is recommended. 
Conventional CT and MRI: This guideline has defined measurability of lesions on CT scan based 
on the assumption that CT slice thickness is 5 mm or less. If CT scans have slice thickness 
greater than 5 mm, the minimum size for a measurable lesion should be twice the slice thickness.
MRI is also acceptable in certain situations (e.g. for body scans). Ideally, the same type of 
scanner should be used and the image acquisition protocol should be followed as closely as 
possible to prior scans. Body scans should be performed with breath-hold scanning techniques, if 
possible.
FDG-PET: While FDG-PET response assessments need additional study, it is sometimes 
reasonable to incorporate the use of FDG-PET scanning to complement CT scanning in 
assessment of progression (particularly possible 'new' disease). New lesions on the basis of FDG-
PET imaging can be identified according to the following algorithm: 
x
Negative FDG-PET at baseline, with a positive FDG-PET at follow-up is a sign of PD 
based on a new lesion.
x
No FDG-PET at baseline and a positive FDG-PET at follow-up: If the positive FDG-PET 
at follow-up corresponds to a new site of disease confirmed by CT, this is PD. If the 
positive FDG-PET at follow-up is not confirmed as a new site of disease on CT, 
additional follow-up CT scans are needed to determine if there is truly progression 
occurring at that site (if so, the date of PD will be the date of the initial abnormal FDG-
PET scan). If the positive FDG-PET at follow-up corresponds to a pre-existing site of 
disease on CT that is not progressing on the basis of the anatomic images, this is not PD.
x
FDG-PET may be used to upgrade a response to a CR in a manner similar to a biopsy in 
cases where a residual radiographic abnormality is thought to represent fibrosis or 
scarring. The use of FDG-PET in this circumstance should be prospectively described in 
the protocol and supported by disease-specific medical literature for the indication.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
30
However, it must be acknowledged that both approaches may lead to false positive CR 
due to limitations of FDG-PET and biopsy resolution/sensitivity.
Note: A ‘positive’ FDG-PET scan lesion means one which is FDG avid with an uptake greater 
than twice that of the surrounding tissue on the attenuation corrected image.
6.3.4
Response Criteria
Evaluation of Target Lesions
Complete Response (CR): Disappearance of all target lesions. Any pathological lymph nodes 
(whether target or non-target) must have reduction in short axis to <10 mm.
Partial Response (PR): At least a 30% decrease in the sum of the diameters of target lesions, 
taking as reference the baseline sum of diameters.
Progressive Disease (PD): At least a 20% increase in the sum of the diameters of target lesions, 
taking as reference the smallest sum on study (this includes the baseline sum if that is the 
smallest on study). In addition to the relative increase of 20%, the sum must also demonstrate an 
absolute increase of at least 5 mm. (Note: the appearance of one or more new lesions is also 
considered progressions).
Stable Disease (SD): Neither sufficient shrinkage to qualify for PR nor sufficient increase to 
qualify for PD, taking as reference the smallest sum of diameters while on study.
Evaluation of Non-Target Lesions
Complete Response (CR): Disappearance of all non-target lesions and normalization of tumor 
marker level. All lymph nodes must be non-pathological in size (<10 mm short axis).
Note: If tumor markers are initially above the upper normal limit, they must normalize for a 
patient to be considered in complete clinical response.
Non-CR/Non-PD: Persistence of one or more non-target lesion(s) and/or maintenance of tumor 
marker level above the normal limits.
Progressive Disease (PD): Appearance of one or more new lesions and/or unequivocal 
progression of existing non-target lesions. Unequivocal progression should not normally trump 
target lesion status. It must be representative of overall disease status change, not a single lesion 
increase.
Although a clear progression of “non-target” lesions only is exceptional, the opinion of the 
treating physician should prevail in such circumstances, and the progression status should be 
confirmed at a later time by the review panel (or PI).
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 patient's best response assignment will 
depend on the achievement of both measurement and confirmation criteria.
For Patients with Measurable Disease (i.e., Target Disease)

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
31
Target 
Lesions
Non-Target 
Lesions
New 
Lesions
Overall 
Response
Best Overall Response when 
Confirmation is Required*
CR
CR
No
CR
>4 wks. Confirmation**
CR
Non-CR/Non-PD
No
PR
>4 wks. Confirmation**
CR
Not evaluated
No
PR
PR
Non-CR/Non-
PD/not evaluated
No
PR
SD
Non-CR/Non-
PD/not evaluated
No
SD
Documented at least once >4
wks. from baseline**
PD
Any
Yes or No
PD
no prior SD, PR or CR
Any
PD***
Yes or No
PD
Any
Any
Yes
PD
*
See RECIST v1.1 manuscript for further details on what is evidence of a new lesion.
**
Only for non-randomized trials with response as primary endpoint.
***
In exceptional circumstances, unequivocal progression in non-target lesions may be accepted as disease 
progression.
Note:
Patients with a global deterioration of health status requiring discontinuation of treatment without objective 
evidence of disease progression at that time should be reported as “symptomatic deterioration.” Every effort should be made 
to document the objective progression even after discontinuation of treatment.
6.3.5
Duration of Response
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 
recurrent or progressive disease is objectively documented (taking as reference for progressive 
disease the smallest measurements recorded since the treatment started).
The duration of overall CR is measured from the time measurement criteria are first met for CR 
until the first date that progressive disease is objectively documented.
Duration of stable disease: Stable disease is measured from the start of the treatment until the 
criteria for progression are met, taking as reference the smallest measurements recorded since the 
treatment started, including the baseline measurements.
TOXICITY CRITERIA
The following adverse event management guidelines are intended to ensure the safety of each 
patient while on the study. 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 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).

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
32
7
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN
DEFINITIONS
7.1.1
Adverse Event
An adverse event is defined as any untoward medical occurrence in a human subject, including 
any abnormal sign (for example, abnormal physical exam or laboratory finding), symptom, or 
disease, temporally associated with the subject’s participation in research, whether or not 
considered related to the subject’s participation in the research.
7.1.2
Suspected Adverse Reaction
Suspected adverse reaction means any adverse event for which there is a reasonable possibility 
that the drug caused the adverse event. For the purposes of IND safety reporting, ‘reasonable 
possibility’ means there is evidence to suggest a causal relationship between the drug and the 
adverse event. A suspected adverse reaction implies a lesser degree of certainty about causality 
than adverse reaction, which means any adverse event caused by a drug.
7.1.3
Unexpected Adverse Reaction
An adverse event or suspected adverse reaction is considered “unexpected” if it is not listed in 
the investigator brochure or is not listed at the specificity or severity that has been observed; or, 
if an investigator brochure is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current application. 
"Unexpected” also refers to adverse events or suspected adverse reactions that are mentioned in 
the investigator brochure as occurring with a class of drugs or as anticipated from the 
pharmacological properties of the drug, but are not specifically mentioned as occurring with the 
particular drug under investigation.
7.1.4
Serious 
An Unanticipated Problem or Protocol Deviation is serious if it meets the definition of a Serious 
Adverse Event or if it compromises the safety, welfare or rights of subjects or others.
7.1.5
Serious Adverse Event
An adverse event or suspected adverse reaction is considered serious if in the view of the 
investigator or the sponsor, it results in any of the following:
x
Death,
x
A life-threatening adverse drug experience
x
Inpatient hospitalization or prolongation of existing hospitalization
x
Persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions
x
A congenital anomaly/birth defect.
x
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse drug experience when, based upon
appropriate medical judgment, they may jeopardize the patient or subject and may require 
medical or surgical intervention to prevent one of the outcomes listed in this definition.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
33
7.1.6
Disability
A substantial disruption of a person’s ability to conduct normal life functions.
7.1.7
Life-Threatening Adverse Drug Experience
Any adverse event or suspected adverse reaction that places the patient or subject, in the view of 
the investigator or sponsor, at immediate risk of death from the reaction as it occurred, i.e., it
does not include a reaction that had it occurred in a more severe form, might have caused death.
7.1.8
Protocol Deviation (NIH Definition)
Any change, divergence, or departure from the IRB-approved research protocol.
7.1.9
Non-Compliance (NIH Definition)
The failure to comply with applicable NIH Human Research Protections Program (HRPP) 
policies, IRB requirements, or regulatory requirements for the protection of human research 
subjects.
7.1.10 Unanticipated Problem
Any incident, experience, or outcome that:
x
Is unexpected in terms of nature, severity, or frequency in relation to:
(a) the research risks that are described in the IRB-approved research protocol and 
informed consent document; Investigator’s Brochure or other study documents, and
(b) the characteristics of the subject population being studied; AND
x
Is related or possibly related to participation in the research; AND
x
Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized.
NCI-IRB REPORTING AND CLINICAL DIRECTOR (CD) REPORTING
7.2.1
NCI-IRB and NCI CD Expedited Reporting of Unanticipated Problems and Deaths
The Protocol PI will report in the NIH Problem Form to the NCI-IRB and NCI Clinical Director:
x
All deaths, except deaths due to progressive disease 
x
All Protocol Deviations
x
All Unanticipated Problems
x
All non-compliance
Reports must be received within 7 days of PI awareness via iRIS.
7.2.2
NCI-IRB Requirements for PI Reporting at Continuing Review
The protocol PI will report to the NCI-IRB: 
1. A summary of all protocol deviations in a tabular format to include the date the deviation 
occurred, a brief description of the deviation and any corrective action.
2. A summary of any instances of non-compliance
3. A tabular summary of the following adverse events:

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
34
x
All Grade 2 unexpected events that are possibly, probably or definitely related to the 
research; 
x
All Grade 3 and 4 events that are possibly, probably or definitely related to the 
research; 
x
All Grade 5 events regardless of attribution; 
x
All serious events regardless of attribution. 
Note: Grade 1 events are not required to be reported. 
7.2.3
NCI-IRB Reporting of IND Safety Reports
Only IND Safety Reports that meet the definition of an unanticipated problem will need to be 
reported to the NCI IRB.
IND SPONSOR REPORTING CRITERIA 
From the time the subject receives the investigational agent/intervention to the time of the second 
follow-up evaluation, the investigator must immediately report to the sponsor, using the 
mandatory MedWatch form 3500a or equivalent, any serious adverse event, whether or not 
considered drug related, including those listed in the protocol or investigator brochure and must 
include an assessment of whether there is a reasonable possibility that the drug caused the event.
For serious adverse events that occur after the second follow-up evaluation, only those events
that have an attribution of at least possibly related to the agent/intervention will be reported.
Required timing for reporting per the above guidelines:
x
Deaths (except death due to progressive disease) must be reported via email within 24 
hours. A complete report must be submitted within one business day. 
x
Other serious adverse events as well as deaths due to progressive disease must be 
reported within one business day
Events will be submitted to the Center for Cancer Research (CCR) at: CCRsafety@mail.nih.gov
and to the CCR PI and study coordinator.
7.3.1
Reporting Pregnancy
Maternal Exposure
If a patient becomes pregnant during the course of the study, the study treatment should be 
discontinued immediately and the pregnancy reported to the Sponsor. The potential risk of 
exposure of the fetus to the investigational agent(s) or chemotherapy agents(s) should be 
documented in box B5 of the MedWatch form “Describe Event or Problem”.
Pregnancy itself is not regarded as an SAE. However, as patients who become pregnant on study 
risk intrauterine exposure of the fetus to agents which may be teratogenic, the CCR is requesting 
that pregnancy should be reported in an expedited manner as Grade 3 “Pregnancy, puerperium 
and perinatal conditions - Other (pregnancy)” under the Pregnancy, puerperium and perinatal 
conditions SOC. 
Congenital abnormalities or birth defects and spontaneous miscarriages should be reported and 
handled as SAEs. Elective abortions without complications should not be handled as AEs. The 
outcome of all pregnancies (spontaneous miscarriage, elective termination, ectopic pregnancy, 
normal birth, or congenital abnormality) should be followed up and documented.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
35
If any pregnancy occurs in the course of the study, then the investigator should inform the 
Sponsor within one day, i.e., immediately, but no later than 24 hours of when he or she 
becomes aware of it.
The designated Sponsor representative will work with the investigator to ensure that all relevant 
information is provided to the Sponsor within 1 to 5 calendar days for SAEs and within 30 days 
for all other pregnancies.
The same timelines apply when outcome information is available.
Paternal Exposure
Male patients should refrain from fathering a child or donating sperm during the study and for 
120 days after the last vaccination.
Pregnancy of the patient’s partner is not considered to be an AE. However, the outcome of all 
pregnancies (spontaneous miscarriage, elective termination, ectopic pregnancy, normal birth, or 
congenital abnormality) occurring from the date of the first dose until 120 days after the last dose 
should, if possible, be followed up and documented.
INSTITUTIONAL BIOSAFETY COMMITTEE (IBC) REPORTING CRITERIA
7.4.1
Serious Adverse Event Reports to IBC
Serious adverse events are not required to be reported to the NIH IBC, as the research is not 
subject to the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid 
Molecules (NIH Guidelines). Therefore, the reporting requirements as described in Appendix M 
do not apply for this study.
7.4.2
Annual Reports to IBC
Within 60 days after the one-year anniversary of the date on which the IBC approved the initial 
protocol, and after each subsequent anniversary until the trial is completed, the Principal 
Investigator (or delegate) shall submit the information described below. Alternatively, the IRB 
continuing review report can be sent to the IBC in lieu of a separate report. Please include the 
IBC protocol number on the report.
Clinical Trial Information 
A brief summary of the status of the trial in progress or completed during the previous year. The 
summary is required to include the following information: 
x
the title and purpose of the trial 
x
clinical site
x
the Principal Investigator
x
clinical protocol identifiers; 
x
participant population (such as disease indication and general age group, e.g., adult or 
pediatric); 
x
the total number of participants planned for inclusion in the trial; the number entered into 
the trial to date whose participation in the trial was completed; and the number who 
dropped out of the trial with a brief description of the reasons 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
36
x
the status of the trial, e.g., open to accrual of subjects, closed but data collection ongoing, 
or fully completed, 
x
if the trial has been completed, a brief description of any study results. 
Progress Report and Data Analysis 
Information obtained during the previous year's clinical and non-clinical investigations, 
including: 
x
a narrative or tabular summary showing the most frequent and most serious adverse 
experiences by body system
x
a summary of all serious adverse events submitted during the past year
x
a summary of serious adverse events that were expected or considered to have causes not 
associated with the use of the gene transfer product such as disease progression or 
concurrent medications
x
if any deaths have occurred, the number of participants who died during participation in 
the investigation and causes of death
x
a brief description of any information obtained that is pertinent to an understanding of the 
gene transfer product’s actions, including, for example, information about dose-response, 
information from controlled trials, and information about bioavailability. 
DATA AND SAFETY MONITORING PLAN
7.5.1
Principal Investigator/Research Team 
The clinical research team will meet on a regular basis when patients are being actively treated 
on the trial to discuss each patient. Decisions about dose level enrollment and dose escalation if 
applicable will be made based on the toxicity data from prior patients. 
All data will be collected in a timely manner and reviewed by the Principal Investigator. Adverse 
events will be reported as required above. Any safety concerns, new information that might 
affect either the ethical and or scientific conduct of the trial, or protocol deviations will be 
immediately reported to the IRB using iRIS.
The Principal Investigator will review adverse event and response data on each patient to ensure
safety and data accuracy. The Principal Investigator will personally conduct or supervise the 
investigation and provide appropriate delegation of responsibilities to other members of the 
research staff. 
7.5.2
Sponsor Monitoring Plan
As a sponsor for clinical trials, FDA regulations require the CCR to maintain a monitoring 
program. The CCR’s program allows for confirmation of: study data, specifically data that could 
affect the interpretation of primary study endpoints; adherence to the protocol, regulations, and 
SOPs; and human subject’s protection. This is done through independent verification of study 
data with source documentation focusing on:
x
Informed consent process
x
Eligibility confirmation
x
Drug administration and accountability
x
Adverse events monitoring

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
37
x
Response assessment
The monitoring program also extends to multi-site research when the CCR is the coordinating 
center.
This trial will be monitored by personnel employed by a CCR contractor. Monitors are qualified 
by training and experience to monitor the progress of clinical trials. Personnel monitoring this 
study will not be affiliated in any way with the trial conduct.
8
STATISTICAL CONSIDERATIONS
STATISTICAL HYPOTHESIS
8.1.1
Primary Efficacy Endpoint
The primary objectives of this trial is to determine the clinical response rate in patients 
with metastatic melanoma or epithelial cancers who receive this dendritic cell vaccine.
The objective response rate (responses/evaluable patients) is the measurable endpoint, and 
it will be of interest to determine if it exceeds 10% and is consistent with 25%.
8.1.2
Secondary Efficacy Endpoint
The secondary objectives are to determine the safety of the vaccine and to determine 
whether treatment administration can increase the quantity and quality of circulating 
antigen-specific T cells.
SAMPLE SIZE DETERMINATION
Assays to assess quality and quantity of the vaccine induced cells will include Eliza and Elispot 
assays assessing reactivity to the mutated peptide compared to the non-mutated peptide. Patients 
will be enrolled into 2 individual cohorts; melanoma and common epithelial cancers. For each of 
the 2 cohorts, the study will be conducted using a phase II optimal design19 (Simon R, Controlled 
Clinical Trials 10:1-10, 1989). The objective will be to determine if the vaccine is able to be 
associated with a clinical response rate that can rule out 5% (p0=0.05) in favor of a modest 20% 
PR + CR rate (p1=0.20).
In patients in each of the two cohorts, the following design will be used. For each cohort, with 
alpha=0.05 (5% probability of accepting a poor therapy) and beta=0.10 (10% probability of 
rejecting a good therapy,), initially 21 evaluable patients will be enrolled. If 0 or 1 of the 21 
patients experiences a clinical response, then no further patients will be enrolled. If 2 or more of 
the first 21 evaluable patients enrolled have a clinical response, then accrual will continue until a 
total of 41 evaluable patients have been enrolled. As it may take several weeks to determine if a 
patient has experienced a clinical response, a temporary pause of up to 6 months in the accrual to
the trial may be necessary to ensure that enrollment to the second stage is warranted. If 2 to 4 of 
the 41 have a clinical response, then this will be considered inadequate for further investigation. 
If 5 or more of 41 patients have a clinical response, then this will indicate that this strategy 
provides a new approach that may be worthy of further consideration. Under the null hypothesis 
(5% response rate), the probability of early termination is 72%.
A total of up to 86 patients may be required (allowing up to 2 inevaluable patients per cohort).
Provided that about 1-2 patients per month will be able to be enrolled onto this trial, 
approximately 6 years may be needed to accrue the maximum number of required patients.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
38
However, as adequate responses to proceed to the second stage of accrual may not occur, the trial 
may end up accruing as few as 21 patients in each cohort.
POPULATION FOR ANALYSES
Modified intention to treat analysis dataset. Patients who receive adequate treatment (defined as 
the administration of peptide pulsed antigen presenting cells) will be considered evaluable and 
included in the statistical analyses
STATISTICAL ANALYSES
8.4.1
General Approach
Descriptive statistical analyses will be performed without any formal hypothesis testing.
8.4.2
Analysis of the Primary Endpoints
For each cohort, the clinical response rate (PR+CR/number of evaluable subjects) will be 
determined and reported along with the corresponding 95% two-sided confidence interval.
8.4.3
Analysis of the Secondary Endpoint(s)
To determine whether treatment administration can increase the quantity and quality of 
circulating antigen-specific T cells, by measuring the reactivity of circulating cells against the 
immunizing peptides in Elispot and 41BB upregulation assays.
8.4.4
Safety Analyses
Safety will be monitored by identifying the type and severity of any adverse events. The fraction 
of patients who experience a DLT will be identified, with information reported about the number 
and grade of each type of DLT identified.
8.4.5
Baseline Descriptive Statistics
Not applicable.
8.4.6
Planned Interim Analyses
As noted in Section 8.2, results after the first stage of a Simon optimal two-stage design in each 
cohort will be examined to determine if the minimum number of responses has been obtained.
8.4.7
Sub-Group Analyses
There are no planned analyses based on demographic characteristics since the power to do 
subgroup analyses would be limited and make interpretation difficult.
8.4.8
Tabulation of individual Participant Data
None planned.
8.4.9
Exploratory Analyses
None planned.
9
COLLABORATIVE AGREEMENTS
None.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
39
10 HUMAN SUBJECTS PROTECTIONS
RATIONALE FOR PATIENT SELECTION
Subjects from both genders and all racial/ethnic groups are eligible for this study if they meet the 
eligibility criteria. To date, there is no information that suggests that differences in drug 
metabolism or disease response would be expected in one group compared to another. Efforts 
will be made to extend accrual to a representative population, but in this preliminary study, a 
balance must be struck between patient safety considerations and limitations on the number of 
individuals exposed to potentially toxic and/or ineffective treatments on the one hand and the 
need to explore gender and ethnic aspects of clinical research on the other hand. If differences in 
outcome that correlate to gender or to ethnic identity are noted, accrual may be expanded or a 
follow-up study may be written to investigate those differences more fully.
We will limit enrollment to patients 70 years of age or less because based on our admittedly
limited experience with prior ACT clinical trials, younger patients tolerate and recover from 
these toxicities better than elderly patients.
PARTICIPATION OF CHILDREN
Since the efficacy of this experimental procedure is unknown, it does not seem reasonable to 
expose children to this risk without further evidence of benefit. Should results of this study 
indicate efficacy in treating metastatic cancer, which is not responsive to other standard forms of 
therapy, future research can be conducted in the pediatric population to evaluate potential benefit 
in that patient population.
PARTICIPATION OF SUBJECTS UNABLE TO GIVE CONSENT
Adults unable to give consent are excluded from enrolling in the protocol. However, re-consent 
may be necessary and there is a possibility, though unlikely, that subjects could become 
decisionally impaired. For this reason and because there is a prospect of direct benefit from 
research participation (Section 10.5DOOVXEMHFWVDJHZLOOEHRIIHUHGWKHRSSRUWXQLW\WRILOO
in their wishes for research and care, and assign a substitute decision maker on the “NIH 
Advance Directive for Health Care and Medical Research Participation” form so that another 
person can make decisions about their medical care in the event that they become incapacitated 
or cognitively impaired during the course of the study. Note: The PI or AI will contact the NIH 
Ability to Consent Assessment Team for evaluation. For those subjects that become 
incapacitated and do not have pre-determined substitute decision maker, the procedures 
described in MAS Policy 87-4 for appointing a surrogate decision maker for adult subjects who 
are (a) decisionally impaired, and (b) who do not have a legal guardian or durable power of 
attorney, will be followed.
EVALUATION OF BENEFITS AND RISKS/DISCOMFORTS
The experimental treatment has a chance to provide clinical benefit though this is unknown. The 
NCI SB has extensive experience with ACT following treatment with high-dose aldesleukin, 
however this experimental treatment does not include the standard NCI SB preparative regimen 
or high-dose aldesleukin. Although we have seen responses to prior NCI SB ACT treatments, we 
do not know if this change in our process will improve patient outcome. The risks associated 
with ACT are substantial, including, a delay in treatment due to the need to harvest and grow the 
cells, a surgical procedure (possible major) to obtain tumor for the cell product, the possibility 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
40
that a cell product cannot be generated, infection due to the surgical procedure, and death. The 
risks in this treatment are detailed in Section 11. The risks associated with taking blood draw 
samples are pain, bruising, or infection.
RISK/BENEFIT ANALYSIS
Because all patients in this protocol have metastatic or recurrent/refractory locally advanced 
cancer and limited life expectancies the potential benefit is thought to outweigh the potential 
risks.
CONSENT PROCESS AND DOCUMENTATION
If the patient meets the thorough screening for eligibility, the patient, with family members or 
friends at the request of the patient, will be presented with a detailed description of the protocol 
treatment. The specific requirements, objectives, and potential advantages and disadvantages will 
be presented. The informed consent document is given to the patient, who is requested to review 
it and to ask questions prior to agreeing to participate in the treatment portion of this protocol.
The patient is reassured that participation on trial is entirely voluntary and that he/she can 
withdraw or decide against treatment at any time without adverse consequences. The Principal 
Investigator, associate investigator, or clinical fellow is responsible for obtaining written consent 
from the patient.
10.6.1 Informed Consent of Non-English Speaking Subjects
If there is an unexpected enrollment of a research participant for whom there is no translated 
extant IRB approved consent document, the Principal Investigator and/or those authorized to 
obtain informed consent will use the Short Form Oral Consent Process as described in MAS 
Policy M77-2, OHSRP SOP 12, 45 CFR 46.117 (b) (2), and 21 CFR 50.27 (b) (2). The summary 
that will be used is the English version of the extant IRB approved consent document. Signed 
copies of both the English version of the consent and the translated short form will be given to 
the subject or their legally authorized representative and the signed original will be filed in the 
medical record.
Unless the PI is fluent in the prospective subject’s language, an interpreter will be present to 
facilitate the conversation (using either the long translated form or the short form). Preferably 
someone who is independent of the subject (i.e., not a family member) will assist in presenting 
information and obtaining consent. Whenever possible, interpreters will be provided copies of 
the relevant consent documents well before the consent conversation with the subject (24 to 48 
hours if possible). 
We request prospective IRB approval of the use of the short form process for non-English 
speaking subjects and will notify the IRB at the time of continuing review of the frequency of the 
use of the Short Form.
11
PHARMACEUTICAL INFORMATION
CELL PREPARATION
The procedure for growing and expanding the autologous DC and the Certificate of Analysis are 
similar to those approved by the Food and Drug Administration and used in other NCI SB
clinical studies. This product will be provided for investigational use only under a Sponsor IND. 
The Certificate of Analysis is in Appendix 2 and Appendix 3, and the Standard Operating 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
41
Procedures for the growth of dendritic cells is in the IND and in the NCI SB BB Master File 
13782. DC will be administered at a dose of between 1x107 to 8x107 dendritic cells.
SUBCUTANEOUS INJECTION AND INTRAVENOUS INFUSION PREPARATION
Classification: Peptide Loaded Dendritic Cell Vaccine
Mode of Action: Autologous monocyte derived DC pulsed with peptide neoantigens are 
recognized in vivo by tumor-specific cytotoxic T-lymphocytes, leading to a major 
histocompatibility complex-restricted cytotoxic response against tumor cells bearing these 
neoantigens. 
Product Description: The vaccine is derived from autologous DCs produced ex vivo from 
monocytes isolated from an apheresis product. DCs are produced using clinical grade cytokines 
(IL-4 and GM-CSF). Neoantigens will be identified from a sequence comparison of a patient’s 
tumor and normal DNA sample. An algorithm will predict mutated sequences with a high 
probability of binding to a patient’s specific HLA repertoire. Neoantigens will be prepared as 
peptides, either as 25mers, with the mutated amino acid residue situated at position 13, or as 
minimal epitopes. The patient’s tumor infiltrating lymphocytes, cultured from a biopsy sample or 
PBMC, will be screened for their ability to recognize these predicted neoantigens pulsed onto 
DCs. Neoantigens recognized by the patient’s TIL or PBMC will be used to formulate the 
vaccine. The vaccine will be formulated for each administration as follows. The identified 
peptides will be pulsed onto thawed, previously cryopreserved DCs overnight. The following 
day, the DCs pulsed with peptides will be matured with Poly I:C, R848 and IFN-J (TLR mix). 
The DCs will be washed and 1-8x107 will be loaded into syringes for IV and SQ administration.
How Supplied: Neoantigen Peptides: The peptides will be manufactured by Peptides and 
Elephants (PE) GmBH, Potsdame, Germany. The peptides are supplied as a sterile, GLP grade, 
lyophilized powder (TFA salt) in vials. Interferon-J is purchased from Miltenyi as a lyophilized 
powder. Poly I:C will be purchased from a qualified supplier as a lyophilized powder. R848 will 
be purchased from a qualified supplier as a lyophilized powder. 
Preparation: Peptides for pulsing DCs will be reconstituted in sterile water for DC pulsing to a 
final concentration of 1 mg/mL. Poly I:C is reconstituted in sterile water at a concentration of 
1mg/ml. R848 is reconstituted in sterile water for injection at a concentration of 1mg/mL.
Interferon-J is reconstituted in sterile water for injection at a concentration of 4x106 units per mL.
Storage: Peptides for Pulsing: Lyophilized vials will be stored at -20°C. Reconstituted peptide 
solutions will be stored at -20°C. DCs will be cryopreserved in Cryostor10 and maintained in 
vapor phase liquid nitrogen (-140 to -180°C) until used. Poly I:C and R848 are stored at -20°C as 
lyophilized powders and as reconstituted solutions. Interferon-J is stored as lyophilized powder 
at -20°C and as a reconstituted solution at 4°C.
Stability: The peptides, Poly I:C, R848 and DC are stable at the recommended storage 
temperatures for at least one year. Interferon-J will be made up fresh for each maturation 
process. The long-term stability of these vaccine components is under evaluation through an on-
going stability testing program. 
Route of Administration:
SQ Injection: A total of 1 mL (0.5 to 4x107) DCs will be administered as an injection into the 
deep SQ tissue of a proximal extremity. Injections will preferentially be administered in the 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
42
anterior thigh. Injections will not be given into extremities that have had radical lymph node 
dissections. The same extremity will be used for each cycle of the peptide loaded dendritic cell 
vaccine administration.
IV Infusion: A total of (0.5 to 4x107) cells for IV administration will be infused over 20-30
minutes via non-filtered tubing, gently agitating the bag during infusion to prevent cell clumping.
Potential Adverse Events and Risks: The peptide antigens pulsed on to the DCs will be unique 
to each patient and may mimic portions of natural proteins found naturally throughout the body, 
there is a chance for the development of autoimmune reactions to cells containing these proteins. 
Special Handling: The peptides are NOT a cytotoxic or infectious agent and require no special 
handling. The DCs are a blood derived product. All personnel should treat blood derived 
products as potentially infectious material. Universal precautions and infection control should be 
practiced at all times.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
43
12 REFERENCES
1.
Klein L, Hinterberger M, Wirnsberger G, et al: Antigen presentation in the 
thymus for positive selection and central tolerance induction. Nat Rev Immunol 9:833-44, 2009
2.
Abramson J, Giraud M, Benoist C, et al: Aire's partners in the molecular control 
of immunological tolerance. Cell 140:123-35, 2010
3.
Bos R, Marquardt KL, Cheung J, et al: Functional differences between low- and 
high-affinity CD8(+) T cells in the tumor environment. Oncoimmunology 1:1239-1247, 2012
4.
Castle JC, Kreiter S, Diekmann J, et al: Exploiting the mutanome for tumor 
vaccination. Cancer Res 72:1081-91, 2012
5.
Kreiter S, Vormehr M, van de Roemer N, et al: Mutant MHC class II epitopes 
drive therapeutic immune responses to cancer. Nature 520:692-6, 2015
6.
Carreno BM, Magrini V, Becker-Hapak M, et al: Cancer immunotherapy. A 
dendritic cell vaccine increases the breadth and diversity of melanoma neoantigen-specific T 
cells. Science 348:803-8, 2015
7.
Gros A, Parkhurst MR, Tran E, et al: Prospective identification of neoantigen-
specific lymphocytes in the peripheral blood of melanoma patients. Nat Med, 2016
8.
Lu YC, Yao X, Crystal JS, et al: Efficient identification of mutated cancer 
antigens recognized by T cells associated with durable tumor regressions. Clin Cancer Res 
20:3401-10, 2014
9.
Robbins PF, Lu YC, El-Gamil M, et al: Mining exomic sequencing data to 
identify mutated antigens recognized by adoptively transferred tumor-reactive T cells. Nat. Med 
19:747-752, 2013
10.
Tran E, Ahmadzadeh M, Lu YC, et al: Immunogenicity of somatic mutations in 
human gastrointestinal cancers. Science 350:1387-1390, 2015
11.
Prickett TD, Crystal JS, Cohen CJ, et al: Durable Complete Response from 
Metastatic Melanoma after Transfer of Autologous T Cells Recognizing 10 Mutated Tumor 
Antigens. Cancer Immunol Res 4:669-78, 2016
12.
Tran E, Turcotte S, Gros A, et al: Cancer immunotherapy based on mutation-
specific CD4+ T cells in a patient with epithelial cancer. Science 344:641-645, 2014
13.
Cohen CJ, Gartner JJ, Horovitz-Fried M, et al: Isolation of neoantigen-specific T 
cells from tumor and peripheral lymphocytes. J Clin Invest 125:3981-91, 2015
14.
Ueno H, Schmitt N, Klechevsky E, et al: Harnessing human dendritic cell subsets 
for medicine. Immunol Rev 234:199-212, 2010
15.
Steinman RM: Decisions about dendritic cells: past, present, and future. Annu 
Rev Immunol 30:1-22, 2012
16.
Mullins DW, Sheasley SL, Ream RM, et al: Route of immunization with peptide-
pulsed dendritic cells controls the distribution of memory and effector T cells in lymphoid tissues 
and determines the pattern of regional tumor control. J Exp Med 198:1023-34, 2003
17.
Curti A, Tosi P, Comoli P, et al: Phase I/II clinical trial of sequential 
subcutaneous and intravenous delivery of dendritic cell vaccination for refractory multiple 
myeloma using patient-specific tumour idiotype protein or idiotype (VDJ)-derived class I-
restricted peptides. Br J Haematol 139:415-24, 2007
18.
Eisenhauer EA, Therasse P, Bogaerts J, et al: New response evaluation criteria in 
solid tumours: revised RECIST guideline (version 1.1). Eur. J. Cancer 45:228-247, 2009
19.
Simon R: Optimal two-stage designs for phase II clinical trials. Control Clin. 
Trials 10:1-10, 1989

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
44
13 FIGURES AND TABLES
FIGURE 1: A PROCESS FOR THE IDENTIFICATION OF IMMUNOGENIC SOMATIC 
MUTATIONS

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
45
FIGURE 2: TLR MIX INDUCES THE SECRETION OF IL-12P70, TNFD AND MULTIPLE 
CHEMOKINES
Monocyte derived dendritic cells were stimulated with TLR mix (PolyI:C, R848, and IFNJ), 
3T3CD40L cells, LPS or a cytokine mix (Il-1E, IL-6, TNFD and PGE2), Non-treated cells (NT) 
were used as negative control. Sixteen hours post incubation the level of costimulatory molecules 
was evaluated by flow cytometry (A, B) and the secretion of multiple cytokines and chemokines 
was measured by multiplex assay (C, D).

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
46
FIGURE 3: TLR MIX ALONE IS SUFFICIENT TO INDUCE POTENT DC MATURATION
Monocyte derived dendritic cells were stimulated with TLR mix (PolyI:C, R848, and IFNg), 
K463H+TLR mix and K602C14 cells + TLRmix. As a negative control, we used DCs alone or 
DCs stimulated with K562 cells. Sixteen hours post incubation the level of costimulatory 
molecules was evaluated by flow cytometry (A) and the secretion of multiple cytokines was 
measured by multiplex assay (B).

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
47
FIGURE 4: LONG PEPTIDES, MINIMAL EPITOPES AND TMG MRNA USED TO EVALUATE 
ANTIGEN PRESENTATION FOR CD4 AND CD8 CELLS

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
48
FIGURE 5: LONG PEPTIDES AND MINIMAL EPITOPES ARE SUPERIOR TO TMG MRNA IN 
PRESENTING ANTIGENS TO CD4 AND CD8 CELLS
DCs were transfected with the TMG constructs or loaded with long peptide and minimal epitopes 
for 12 hours. DCs were co-cultured with PBMC transduced with TCR’s recognizing the 
corresponding antigens for 18 hours, and antigen recognition was evaluated for Class-I epitopes 
(A) and Class-II epitopes (B) by flow cytometry for 41BB expression. Irrelevant TMG and 
DMSO were used as negative control for TMG transfection and peptide loading, respectively.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
49
FIGURE 6: PEPTIDE LOADED, TLR STIMULATED DC’S EFFICIENTLY PRESENT BOTH 
CD4 AND CD8 EPITOPES
DCs were loaded with long peptides for 2, 4, and 10 hours and matured using a TLR mix. DCs 
were washed and co-cultured with PBMC transduced with TCRs recognizing the corresponding 
antigens for 18 hours (A); antigen recognition was evaluated by flow cytometry for 41BB 
expression (B).

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
50
FIGURE 7: OVERVIEW OF THE METHOD USED TO EVALUATE THE ACTIVATION OF 
NEOANTIGEN SPECIFIC T CELLS

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
51
FIGURE 8: DCS MATURED WITH TLR MIX ARE CAPABLE OF STIMULATING NAÏVE,
ANTIGEN-EXPERIENCED AND TERMINALLY DIFFERENTIATED EFFECTOR CELLS
Apheresis samples from a colorectal cancer patient were thawed and then incubated in an 
appropriately sized tissue culture flask. Adherent monocytes were differentiated into DCs, 
loaded with neoantigen peptide pools and matured using a TLR mix. Non-adherent cells were 
collected, and memory, naïve and terminally differentiated effector memory cells (TEMRA) 
were sorted, and in vitro stimulated with mature peptide-pulsed autologous DCs. Ten days after 
the first stimulation T cells were re-stimulated with DCs loaded with all peptide pools, and sorted 
based on 41BB and OX40 expression to enrich for neoantigen-specific cells. Sorted cells were 
stimulated by DCs loaded with all peptide pools and tested for upregulation of 41BB (data not 
shown). Memory (A), TEMRA (B), and naïve(C) T cells were then tested for neoantigen specific 
recognition by 41BB upregulation following co-culture with DCs loaded with single peptides 
derived from the most reactive peptide pools.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
52
FIGURE 9: PROCESS FOR EPITOPE SELECTION AND SYNTHESIS

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
53
TABLE 1: DEFINED MUTATED ANTIGENS RECOGNIZED BY TIL IDENTIFIED AT THE 
SURGERY BRANCH 

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
54
14 APPENDICES
APPENDIX 1: CELL INFUSION INSTRUCTIONS
Necessary equipment:
x
Primary IV tubing (2) (DO NOT USE AN INLINE FILTER FOR CELLS)
x
Secondary IV tubing (1)
x
NS (sodium chloride 0.9%)
x
IV infusion pump
x
Gloves
Steps:
Key Points:
1. Patient RN will be informed of the 
approximate time of cell arrival at the 
bedside.
2. Verify the physician orders:

to administer the cells

for the date of administration
3. Verify that the consent form for the 
protocol is signed.
4. Ensure that emergency and monitoring 
equipment are available in the patient’s 
room:

oxygen

suction

vital sign monitor with pulse oximeter
and thermometer
5. Provide patient education covering 
infusion procedure, potential 
complications and associated symptoms to 
report. 
6. Measure and record baseline vital signs, 
respiratory and circulatory assessments.
7. Verify the patency of the patient’s IV 
access.
Optimally, a central line or peripheral IV of 
20G or larger should be used.
8. Hang a primary line of NS at a kvo rate -
NEW bag and NEW tubing. 
This MUST be ready and infusing prior
to the cells being delivered to the unit.
Have a second bag of NS and tubing 
connected to the patient as an emergency 
line.
The primary line will be the dedicated NS line 
for infusing the cells. Under no circumstances 
are any other substances to be infused into the 
line. Do not use an inline filter for cells.
Cell death occurs quickly – the infusion must 
be initiated promptly.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
55
Do not infuse medication during the cell 
infusion. If emergency meds must be 
administered, use the emergency NS IV line. 
The second bag of NS will be the emergency 
IV solution and can be used for medication 
administration.
9. The patient RN will be notified 
approximately 10 minutes before the cells 
arrive on the unit. The cells will be hand 
delivered to the bedside.
It is important to be at the bedside awaiting 
the arrival of the cells for infusion; have 
baseline VS, assessment, and IV lines hooked 
up when the cells arrive. Cell death occurs 
quickly after the cells are removed from the 
laboratory. Initiate the infusion as quickly as 
possible.
10. Prior to spiking the cell bag, two RNs will 
perform the identification procedure. Both
RNs must sign the tag on the cell bag.
11. Infuse the cells by infusion pump over 20-
30 minutes or as clinically indicated.
a.
Piggyback the cells into the dedicated 
NS line; use the backflush technique 
to prime the line.
b. While the cells are infusing, gently
agitate the bag of cells every few 
minutes. When the cell bag is empty, 
backflush NS to rinse the bag and 
infuse this at the same rate as the cells; 
rinse bag until NS runs clear. 
Gently agitating the bag prevents the cells 
from clumping in the bag. 
12. Measure and record VS before and after 
the cell infusion, then hourly (± 15 
minutes) from the end of the infusion for 
two hours.
13. Documentation:
a.
After the cells have infused, remove 
the adhesive backed “cell therapy
product” tag from the cell bag and 
place it on a progress note. Sign and 
date the progress note then send it to 
medical records.
b. Document the cell infusion in CRIS 
using the Surgery Branch Cell Product 
Administration Flowsheet.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
56
c.
Two separate Surgery Branch Cell 
Product Administration Flowsheets 
should be completed: one for the IV 
and a second one for the SQ cell 
administration.

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
57
APPENDIX 2: CERTIFICATE OF ANALYSIS FOR IMMATURE DENDRITIC CELLS
Personalized Dendritic Vaccine Patient:
Date of preparation of final product:
Tests performed on final product:
Test
Method
Limits
Result
Tests Performed 
by
Initials/Date
Cell viability1
trypan blue 
exclusion
>50%
Total viable cell 
number1
visual microscopic 
count
>107
Identity2
FACS
> 60 % 
CD11C+CD86+
Microbiological 
studies 
aerobic culture1
no growth
anaerobic culture1
no growth
Fungal Culture1
no growth
gram stain1
no micro-organisms 
seen
mycoplasma test1
negative
1 Performed on the final product immediately prior to cryopreservation.
Prepared by:
_________________________________________
Date: ____________________
QC sign-off:
__________________________________________
Date: ____________________
Qualified laboratory or Clinical Supervisor
QA sign-off:
__________________________________________
Date: ____________________

Abbreviated Title: Personalized DC Vaccine
Version Date: March 26, 2018
58
APPENDIX 3: CERTIFICATE OF ANALYSIS FOR MATURE PEPTIDE PULSED DENDRITIC 
CELLS
Personalized Dendritic Vaccine Patient:
Date of preparation of final product:
Tests performed on final product:
Test
Method
Limits
Result
Tests Performed 
by
Initials/Date
Cell viability1
trypan blue 
exclusion
>50%
Total viable cell 
number1
visual 
microscopic 
count
between 107 and 
8 X 107
Identity1
FACS
> 60 % 
CD11C+CD86+
Microbiological 
studies 
aerobic culture2
no growth
anaerobic culture2
no growth
gram stain1
no micro-organisms 
seen
fungal culture
no growth
mycoplasma test2
negative
Endotoxin1
limulus assay
<5 E.U./kg
1 Performed on the final product prior to infusion. Results are available at the time of infusion.
2 Sample for test collected on the final product prior to infusion. Results will not be available before cells 
are infused into the patient.
Prepared by:
_________________________________________
Date: ____________________
QC sign-off:
__________________________________________
Date: ____________________
Qualified laboratory or Clinical Supervisor
QA sign-off:
__________________________________________
Date: ____________________