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10.3 participation of subjects unable

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 **10.6**), all subjects will be offered the opportunity to fill 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 (ACAT) for evaluation as needed for the following: an independent assessment of whether an individual has the capacity to provide consent; assistance in identifying and assessing an appropriate surrogate when indicated; and/or an assessment of the capacity to appoint a surrogate. For those subjects that become incapacitated and do not have pre-determined substitute decision maker, the procedures described in MAS Policy 87-4 and NIH HRPP SOP 14E 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.
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10.4 evaluation of benefits and

The discomforts of a 10-day CIV of rhIL-15 are expected to be the same pain and discomfort from venipuncture for blood drawing. The primary risk to patients participating in this research study is from expected or unforeseen toxicity or biological effects of recombinant human IL-15 as an investigational agent. Patients will be carefully monitored for any adverse events as described in Section 3.5 and appropriate care provided.  
The intent of the study is to evaluate the toxicity of rhIL-15 in this Phase I trial and to find a range of doses where it has an effect on humans (biological activity). Recently there has been new evidence to suggest that patients with solid tumors may respond to immunotherapeutics or that infiltration of their tumor deposits by activated effector T-cells is associated with an improved survival. These finding lend additional weight to the notion that the administration of rhIL-15 might be associated with a partial or complete response of the patient's tumor more commonly associated with the immune-based treatment of metastatic melanoma and metastatic renal cell cancer.
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10.5 strategies and procedures for

Patient accrual will be facilitated by the large number of clinical investigators who refer patients to the NCI and CCR. Contact letters or emails will be sent to the regional University referral center's Medical Oncologists informing them of this trial. In addition, the information will be provided on the NCI Clinical Trials and ClinicalTrials.gov web pages.
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10.6 risks/benefits analysis

The success of this effort cannot be predicted at this time. Because all patients in this protocol have incurable metastatic cancer and limited life expectancies, the potential benefit is thought to outweigh the potential risks. Given the prospect of direct benefit, this same risks/benefits analysis applies also to adults who may become unable to consent during participation.
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10.7 consent process and documentation

The investigational nature and the research objectives of this trial, the procedures and treatments involved and their attendant risks and discomforts, potential benefits, time commitments and potential alternative therapies will be carefully explained to the patients during the informed consent process. A signed informed consent document will be obtained prior to entry onto the study by the Principal Investigator or an Associate Investigator. The original signed consent goes to Medical Records and a copy will be placed in the research record. If the patient agrees to have the optional biopsy for research, then the patient will consent at the time of the procedure. If the patient refuses the optional biopsy at that time, the refusal will be documented in the medical record and in the research record. 10.7.1 Telephone reconsent Reconsent on this study may be obtained via telephone according to the following procedure: the informed consent document will be sent to the subject. An explanation of the change (s) in the study will be provided over the telephone after the subject has had the opportunity to read the consent form. The subject will sign and date the informed consent. A witness to the subject's signature will sign and date the consent.  
The original informed consent document will be sent back to the consenting investigator who will sign and date the consent form with the date the consent was obtained via telephone. A fully executed copy will be returned via mail for the subject's records. The informed consent process will be documented on a progress note by the consenting investigator and a copy of the informed consent document and note will be kept in the subject's research record. 10.7.2 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. 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 and will notify the IRB at the time of continuing review of the frequency of the use of the Short Form.
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11 pharmaceutical information 11.1 recombinant

Chemical Name: Recombinant human interleukin-15 (rhIL-15) Other Names: Interleukin-T (IL-T)—no longer recognized. Classification: Recombinant human cytokine.  
Molecular Structure: Member of the four α-helix bundle family of human cytokines consisting of 115 amino acids manufactured in *E. coli* by recombinant technology. Nonglycosylated.  
Description: Sterile, clear, colorless, particulate-free liquid; endotoxin free.  
11.1.1 Source Agent Ordering and Agent Accountability NCI-supplied agents may be requested by the Principal Investigator (or their authorized designee) at each participating institution. Pharmaceutical Management Branch (PMB) policy requires that agent be shipped directly to the institution where the patient is to be treated. PMB does not permit the transfer of agents between institutions (unless prior approval from PMB is obtained). The CTEP-assigned protocol number must be used for ordering all CTEP-supplied investigational agents. The responsible investigator at each participating institution must be registered with CTEP, DCTD through an annual submission of FDA Form 1572 (Statement of Investigator), Curriculum Vitae, Supplemental Investigator Data Form (IDF), and Financial Disclosure Form (FDF). If there are several participating investigators at one institution, CTEPsupplied investigational agents for the study should be ordered under the name of one lead investigator at that institution. In general, sites may order initial agent supplies when a subject is being screened for enrollment onto the study. Active CTEP-registered investigators and investigator-designated shipping designees and ordering designees can submit agent requests through the PMB Online Agent Order Processing (OAOP) application. Access to OAOP requires the establishment of a CTEP Identity and Access Management (IAM) account and the maintenance of an "active" account status and a "current" password. For questions about drug orders, transfers, returns, or accountability, call or email PMB any time. Refer to the PMB's website for specific policies and guidelines related to agent
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management.

Agent Inventory Records - The investigator, or a responsible party designated by the investigator, must maintain a careful record of the receipt, dispensing and final disposition of all agents received from the PMB using the appropriate NCI Investigational Agent (Drug)
Accountability Record (DARF) available on the CTEP forms page. Store and maintain separate NCI Investigational Agent Accountability Records for each agent, strength, formulation and ordering investigator on this protocol.
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useful links and contacts

- CTEP Forms, Templates, Documents: http://ctep.cancer.gov/forms/
- NCI CTEP Investigator Registration: PMBRegPend@ctep.nci.nih.gov
- PMB policies and guidelines: http://ctep.cancer.gov/branches/pmb/agent_management.htm
- PMB Online Agent Order Processing (OAOP) application: https://eappsctep.nci.nih.gov/OAOP/pages/login.jspx
- CTEP Identity and Access Management (IAM) account: https://eapps-ctep.nci.nih.gov/iam/
- CTEP Associate Registration and IAM account help: ctepreghelp@ctep.nci.nih.gov
- PMB email: PMBAfterHours@mail.nih.gov  
- PMB phone and hours of service: (240) 276-6575 Monday through Friday between 8:30 am and 4:30 pm (ET)
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11.1.2 toxicity

From the experience in the ongoing rhIL-15 IVB trial, side effects of fatigue, fever, rigors or chills, capillary leak syndrome, fluid retention and edema are likely. Although there has not been any evidence of meaningful pulmonary dysfunction or changes in renal function, the preclinical NHP data and experience with other similar biological compounds indicates that these toxicities could potentially occur in patients treated with the CIV schedule. Gastrointestinal toxicities in the IVB IL-15 patients have included nausea, vomiting and in one case explosive diarrhea exclusively in the patient treated with the 3 mcg/kg/day dose, and data from the NHP toxicology study also showed diarrhea and dehydration—these could also be more common in patients treated with the CIV regimen. Although there has not been significant granulocytopenia in the rhIL-15 patients treated to date and lymphopenia has been transient, there is still some potential for infectious issues if the granulocytopenia associated with CIV treatment of macaques occurs in the patients enrolled in this protocol.
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comprehensive adverse events and potential

The Comprehensive Adverse Event and Potential Risks list (CAEPR) provides a single list of reported and/or potential adverse events (AE) associated with an agent using a uniform presentation of events by body system.  
Version 1.2, February 01, 20161  
| Specific Protocol                                                                                                                                                                                                                                   |                                    |
|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------|
| Exceptions to Expedited  Reporting (SPEER)                                                                                                                                                                                                          |                                    |
| Adverse Events with Possible                                                                                                                                                                                                                        |                                    |
| Relationship to Recombinant Human IL-15 (CTCAE 4.0 Term)                                                                                                                                                                                            |                                    |
| BLOOD AND LYMPHATIC SYSTEM DISORDERS Anemia                                                                                                                                                                                                         | Anemia (Gr 2)                      |
| Bone marrow hypocellular CARDIAC DISORDERS Sinus tachycardia                                                                                                                                                                                        | Sinus tachycardia (Gr 2)           |
| GASTROINTESTINAL DISORDERS Abdominal pain Diarrhea Nausea                                                                                                                                                                                           | Nausea (Gr 2)                      |
| Vomiting                                                                                                                                                                                                                                            | Vomiting (Gr 2)                    |
| GENERAL DISORDERS AND ADMINISTRATION SITE CONDITIONS Chills                                                                                                                                                                                         | Chills (Gr 2)                      |
| Edema limbs Fatigue                                                                                                                                                                                                                                 | Fatigue (Gr 2)                     |
| Fever                                                                                                                                                                                                                                               | Fever (Gr 2)                       |
| Injection site reaction INFECTIONS AND INFESTATIONS Sepsis INVESTIGATIONS Alanine aminotransferase increased Aspartate aminotransferase increased Blood bilirubin increased Creatinine increased Lymphocyte count decreased                         | Lymphocyte count decreased  (Gr 2) |
| Lymphocyte count increased Neutrophil count decreased Platelet count decreased White blood cell decreased METABOLISM AND NUTRITION DISORDERS Hypoalbuminemia Hypophosphatemia                                                                       | Hypophosphatemia (Gr 2)            |
| MUSCULOSKELETAL AND CONNECTIVE TISSUE DISORDERS Generalized muscle weakness NERVOUS SYSTEM DISORDERS Dizziness Headache RESPIRATORY, THORACIC AND MEDIASTINAL DISORDERS Dyspnea SKIN AND SUBCUTANEOUS TISSUE DISORDERS Dry skin Erythema multiforme | Erythema multiforme (Gr 2)         |
| Skin and subcutaneous tissue disorders - Other (rash) VASCULAR DISORDERS Capillary leak syndrome Hypertension                                                                                                                                       | Hypertension (Gr 2)                |
| Hypotension                                                                                                                                                                                                                                         | Hypotension (Gr 2)                 |  
1This table will be updated as the toxicity profile of the agent is revised.  
Adverse events reported on Recombinant Human IL-15 trials, but for which there is insufficient evidence to suggest that there was a reasonable possibility that Recombinant Human IL-15 caused the adverse event:
Blood and lymphatic system disorders - Febrile neutropenia Cardiac disorders - Atrial fibrillation; Chest pain - cardiac; Palpitations; Pericardial effusion; Pericardial tamponade; Sinus bradycardia; Ventricular tachycardia Gastrointestinal disorders - Ascites; Constipation; Duodenal hemorrhage; Gastritis; Gastrointestinal disorders - Other (increased appetite); Ileus; Mucositis oral; Pancreatitis General disorders and administration site conditions - Edema face; Infusion related reaction; Infusion site extravasation; Multi-organ failure; Pain Immune system disorders - Autoimmune disorder Infections and infestations - Tooth infection; Upper respiratory infection; Urinary tract infection Injury, poisoning and procedural complications - Bruising Investigations - Alkaline phosphatase increased; Cardiac troponin I increased; Electrocardiogram QT corrected interval prolonged; GGT increased; INR increased; Lipase increased; Serum amylase increased; Weight gain; Weight loss Metabolism and nutrition disorders - Anorexia; Dehydration; Hyperkalemia; Hypocalcemia; Hypokalemia Musculoskeletal and connective tissue disorders - Back pain; Bone pain; Muscle weakness upper limb; Myalgia; Pain in extremity Neoplasms benign, malignant and unspecified (incl cysts and polyps) - Tumor pain Nervous system disorders - Dysgeusia; Peripheral sensory neuropathy; Presyncope; Vasovagal reaction Psychiatric disorders - Anxiety; Psychosis Reproductive system and breast disorders - Genital edema Respiratory, thoracic and mediastinal disorders - Adult respiratory distress syndrome; Bronchopulmonary hemorrhage; Cough; Hypoxia; Laryngeal inflammation; Pleural effusion; Pneumonitis; Pulmonary edema; Wheezing Skin and subcutaneous tissue disorders - Erythroderma; Palmar-plantar erythrodysesthesia syndrome; Pruritus; Rash acneiform; Skin and subcutaneous tissue disorders - Other (skin plaques)
Vascular disorders - Hot flashes; Visceral arterial ischemia Note: Recombinant Human IL-15 in combination with other agents could cause an exacerbation of any adverse event currently known to be caused by the other agent, or the combination may result in events never previously associated with either agent.  
11.1.3 Formulation Sterile rhIL-15 drug product is vialed at a concentration of 0.52 mg/mL (0.16 mg/0.3 ml) solution of 25 mM sodium phosphate, dibasic and 500 mM sodium chloride (pH 7.0 ±0.4) in a 5 mL borosilicate glass vial. rhIL-15 drug product for intravenous administration should be withdrawn from the vial and prepared as described in Section **11.1.5**.  
Vials are for single use only. 11.1.4 Stability and Storage Long-term at ≤ -70oC.  
Storage of rhIL-15 diluted in 0.1% HSA in infusion bags is limited to a maximum of 4 hours at room temperature or 24 hours at 2-8°C prior to initiation of the 24- hour infusion. Drug stability studies are ongoing and will be continued throughout the clinical trial. Long-term drug stability information will be periodically updated and communicated to the Clinical Center Pharmacy Department. 11.1.5 Preparation and Administration Procedures For all dose levels, the dose of rhIL-15 will be diluted in the appropriate volume of 0.1% human serum albumin (HSA) in 5% dextrose in water, USP (D5W) to reach a final rhIL-15 concentration of 1 mcg/mL. The rhIL-15 infusion will be administered to the patient by continuous intravenous infusion (CIV) over 24 hours using a portable ambulatory pump on the inpatient unit for either a total of 240 or 120 hours depending on the treatment cohort. Treatment with rhIL-15 will begin within 4 hours of preparation of the infusion bag and the infusion must be completed within 24 hours from the time drug administration begins.Otherwise a new infusion bag must be prepared to complete administration of the remaining dose. The infusion solutions will be mixed in an MMI bag. The rhIL-15 infusion solution is stable at a concentration of 1.0 mcg/mL with 0.1 HSA for 4 hours at controlled room temperature (15°C–30°C) prior to initiation of the 24-hour infusion or 24 hours at 2-8ºC prior to initiation of the 24-hour infusion. This stability information was previously documented by the Biopharmaceutical Development Program (BDP) of Leidos Biomedical Research, Inc., the drug manufacturer. 11.1.6 Drug Incompatibilities None known.
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57. Waldmann TA, Lugli E, Roederer M, Perera LP, Smedley JV, Macallister RP, et al.  
Safety (toxicity), pharmacokinetics, immunogenicity, and impact on elements of the normal immune system of recombinant human IL-15 in rhesus macaques. Blood. 2011;117(18):4787-95. 58. Conlon KCL, E.; Rosenberg, S.A.; Fojo, A.T.; Morris, J.C.; Fleisher, T.A.; Welles, H.C.; Dubois, S.P.; Perera, L.P.; Stewart, D.M.; Goldman, C.K.; Bryant, B.; Decker, J.M.; Chen, J.; Shih, J.; Worthy, T.A.; Figg, W.D.; Peer, C.J.; Sneller, M.C.; Lane, H.C.; Yovandich, J.L.; Creekmore, S.P.; Roederer, M.; Waldmann, T.A. First-in-human phase I trial of intravenous infusions of rhIL-15 to patients with refractory metastatic cancer was associated with activation and redistribution of NK and CD8 T-cells, and inflammatory cytokine secretion. In press J Clin Oncol. 2014. 59. Dillman RO, Church C, Oldham RK, West WH, Schwartzberg L, Birch R. Inpatient Continuous-Infusion Interleukin-2 in 788 Patients with Cancer - the National Biotherapy StudyGroup Experience. Cancer. 1993;71(7):2358-70.  
60. Thompson JA, Lee DJ, Lindgren CG, Benz LA, Collins C, Levitt D, et al. Influence of Dose and Duration of Infusion of Interleukin-2 on Toxicity and Immunomodulation. J Clin Oncol. 1988;6(4):669-78. 61. Negrier S, Perol D, Ravaud A, Bay JO, Oudard S, Chabaud S, et al. Randomized study of intravenous versus subcutaneous interleukin-2, and IFN alpha in patients with good prognosis metastatic renal cancer. Clin Cancer Res. 2008;14(18):5907-12.  
62. Yu AL, Gilman AL, Ozkaynak MF, London WB, Kreissman SG, Chen HX, et al. AntiGD2 Antibody with GM-CSF, Interleukin-2, and Isotretinoin for Neuroblastoma. New Engl J
Med. 2010;363(14):1324-34. 63. Conlon KC, Lugli E, Rosenberg SA, Morris JC, Fleisher T, Welles H, et al. Results from the first-in-human phase I trials of recombinant human Interleukin 15 (rhIL-15) administered as a daily 30 minute intravenous infusion (IVB) for 12 consecutive days or as continuous intravenous infusion (CIV) for 240 hours in patients with refractory metastatic cancers. Cancer Res. 2014;74(19). 64. Morishima C, McNeel DG, Patel MR, Kohrt HE, Waldmann TA, Thompson JA, et al.  
CITN11-02 interim trial results: subcutaneous administration of recombinant human IL-15 (rhIL15) is associated with expansion of peripheral blood CD56+ NK cells and CD8+ T cells. Journal for ImmunoTherapy of Cancer. 2015;3(2):P203. 65. Waldmann TaF. T unpublished data. 66. Eisenbeis CF, Lesinski GB, Anghelina M, Parihar R, Valentino D, Liu J, et al. Phase I study of the sequential combination of interleukin-12 and interferon alfa-2b in advanced cancer: Evidence for modulation of interferon signaling pathways by interleukin-12. J Clin Oncol. 2005;23(34):8835-44.  
67. Muller WA. Mechanisms of Leukocyte Transendothelial Migration. Annu Rev PatholMech. 2011;6:323-44.  
68. Klempner MS, Noring R, Mier JW, Atkins MB. An Acquired Chemotactic Defect in Neutrophils from Patients Receiving Interleukin-2 Immunotherapy. New Engl J Med. 1990;322(14):959-65. 69. Rosenberg SA, Dudley ME. Adoptive cell therapy for the treatment of patients with metastatic melanoma. Curr Opin Immunol. 2009;21(2):233-40. 70. Wang E, Miller LD, Ohnmacht GA, Mocellin S, Perez-Diaz A, Petersen D, et al.  
Prospective molecular profiling of melanoma metastases suggests classifiers of immune responsiveness. Cancer Res. 2002;62(13):3581-6. 71. Rustin GJS, Quinn M, Thigpen T, du Bois A, Pujade-Lauraine E, Jakobsen A, et al. Re: New guidelines to evaluate the response to treatment in solid tumors (ovarian cancer). J Natl Cancer I. 2004;96(6):487-8. 72. Bubley GJ, Carducci M, Dahut W, Dawson N, Daliani D, Eisenberger M, et al. Eligibility and response guidelines for phase II clinical trials in androgen-independent prostate cancer: Recommendations from the prostate-specific antigen working group. J Clin Oncol. 1999;17(11):3461-7. 73. Scher HI, Halabi S, Tannock I, Morris M, Sternberg CN, Carducci MA, et al. Design and end points of clinical trials for patients with progressive prostate cancer and castrate levels of testosterone: Recommendations of the prostate cancer clinical trials working group. J Clin Oncol. 2008;26(7):1148-59.  
74. Vergote I, Rustin GJS, Eisenhauer EA, Kristensen GB, Pujade-Lauraine E, Parmar MKB, et al. Re: New guidelines to evaluate the response to treatment in solid tumors [ovarian cancer]. J Natl Cancer I. 2000;92(18):1534-5. 75. Eisenhauer EA, Therasse P, Bogaerts J, Schwartz LH, Sargent D, Ford R, et al. New response evaluation criteria in solid tumours: Revised RECIST guideline (version 1.1). Eur J Cancer. 2009;45(2):228-47.
untitled h1
13 appendices appendix a performance

| ECOG Performance Status Scale                                                                                                                                                             | Karnofsky Performance Scale                                       |                                                              |             |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------|--------------------------------------------------------------|-------------|
| Grade                                                                                                                                                                                     | Descriptions                                                      | Percent                                                      | Description |
| 100                                                                                                                                                                                       | Normal, no complaints, no  evidence of disease.                   |                                                              |             |
| Normal activity. Fully active, able                                                                                                                                                       |                                                                   |                                                              |             |
| 0                                                                                                                                                                                         | to carry on all pre-disease  performance without restriction.     | Able to carry on normal activity;                            |             |
| 90                                                                                                                                                                                        | minor signs or symptoms of  disease. Normal activity with effort; |                                                              |             |
| 80                                                                                                                                                                                        | some signs or symptoms of  disease.                               |                                                              |             |
| Symptoms, but ambulatory. Restricted in physically strenuous  activity, but ambulatory and able  to carry out work of a light or  sedentary nature (e.g., light  housework, office work). |                                                                   |                                                              |             |
| 1                                                                                                                                                                                         | Cares for self, unable to carry on                                |                                                              |             |
| 70                                                                                                                                                                                        | normal activity or to do active  work.                            |                                                              |             |
| In bed <50% of the time. Ambulatory and capable of all  self-care, but unable to carry out  any work activities. Up and about  more than 50% of waking hours.                             | Requires occasional assistance,                                   |                                                              |             |
| 60                                                                                                                                                                                        | but is able to care for most of  his/her needs.                   |                                                              |             |
| 2                                                                                                                                                                                         | 50                                                                | Requires considerable assistance  and frequent medical care. |             |
| 40                                                                                                                                                                                        | Disabled, requires special care  and assistance.                  |                                                              |             |
| In bed >50% of the time. Capable  of only limited self-care, confined  to bed or chair more than 50% of  waking hours.                                                                    |                                                                   |                                                              |             |
| 3                                                                                                                                                                                         | Severely disabled,                                                |                                                              |             |
| 30                                                                                                                                                                                        | hospitalization indicated. Death  not imminent.                   |                                                              |             |
| 100% bedridden. Completely  disabled. Cannot carry on any  self-care. Totally confined to bed  or chair.                                                                                  | 20                                                                | Very sick, hospitalization  indicated. Death not imminent.   |             |
| 4                                                                                                                                                                                         | 10                                                                | Moribund, fatal processes  progressing rapidly.              |             |
| 5                                                                                                                                                                                         | Dead.                                                             | 0                                                            | Dead.       |
untitled h1
appendix b immune-based studies 1.

- Flow cytometry analysis for peripheral blood lymphocyte subset enumeration will be performed using standard techniques in the College of American pathologists
(CAP) certified flow cytometry laboratory within the Clinical Pathology Department, NIH Clinical Center.  
- Special flow cytometry also performed by Dr. Enrico Lugli with Dr. Mario Roederer in the Vaccine Research Center, Building 40, Room 5044.  
2. Assay of rhIL-15 by R&D Systems Quantikine ELISA Kit (Catalog \# D1500) to define concentration of IL-15 in pharmacokinetic assay.  
3. Assay for antibodies to infused IL-15 (see APPENDIX C).  
APPENDIX C: ASSAY FOR ANTIBODIES TO RH**IL-15**  
  
  
- Plates are coated with human IL-15 for 3 hours at 37°C, washed, blocked with 3% FBS
and washed again.  
- A standard curve for assay quantitation and quality control is constructed using serial dilutions of a commercial affinity purified goat anti-human IL-15 that is diluted in heatinactivated normal human serum. The standard curve samples are incubated for 2 hours at 37°C and washed.  
- Biotin conjugated IL-15 is added to each well, incubated 2 hours at 37°C, and the plates are washed.  
- Alkaline phosphatase–conjugated streptavidin is added to each well for 2 hours at 37°C
and then washed.  
- The assay is developed with the addition of diethanolamine buffer with p-Nitrophenyl Phosphatase for 1 hour at 37°C and then immediately read at 405 nm.  
- To detect antibodies to human IL-15 in test samples, serum from the test subject will be assayed in duplicate at dilutions of 1/3 and 1/9 concomitantly with the standard curve samples as above, and the resultant OD obtained used to quantitate the level of antibody present.
Abbreviated Title: Ph I Study of CIV rhIL-15 Version Date: 06/13/2018  
| Work sheet for 12-C-0113 Cycle 1                                     |                                                               |          |                  |                |      |         |           |
|----------------------------------------------------------------------|---------------------------------------------------------------|----------|------------------|----------------|------|---------|-----------|
| Start Date of Treatment: __/__/20                                    | | Dose Level: _                                               |          |                  |                |      |         |           |
| Patient Name:                                                        | mcg/kg/day                                                    |          |                  |                |      |         |           |
| Start Time of Treatment:                                             |                                                               |          |                  |                |      |         |           |
| All samples should be kept at Room Temperature in Clean Utility Room |                                                               |          |                  |                |      |         |           |
| Page Figg Lab (pager 102-11964) for Sample pick up                   | Page LyMB research nurse (pager 102-11029) for sample pick up |          |                  |                |      |         |           |
| Specimen Information                                                 | Ideal                                                         | Actual   | Pick up          | Comments       |      |         |           |
| Study                                                                | Sample name                                                   | Tube     | Sample           | Sample         | Time | (Missed | Signature |
| Day                                                                  | Time                                                          | Drawn    | Specimens, etc.) |                |      |         |           |
| type                                                                 |                                                               |          |                  |                |      |         |           |
| IL-15 antibodies                                                     | 1, 1                                                          | 4 ml SST | ELISA            |                |      |         |           |
| IL-15 Level PreRx                                                    | PK                                                            |          |                  |                |      |         |           |
| 8 ml SST                                                             |                                                               |          |                  |                |      |         |           |
| 1                                                                    |                                                               |          |                  |                |      |         |           |
| Cytokines MesoScale Pre Rx                                           | ELISA                                                         |          |                  |                |      |         |           |
| IL-15 level 10 min                                                   | 1                                                             | 4 ml SST | PK               |                |      |         |           |
| 1                                                                    | IL-15 level 1hr                                               | 1        | 4 ml SST         | PK             | Figg |         |           |
| IL-15 level 2hr                                                      | .1                                                            | 4 ml SST | PK               |                |      |         |           |
| IL-15 level 4hr                                                      | pk                                                            |          |                  |                |      |         |           |
| 8 ml SST                                                             |                                                               |          |                  |                |      |         |           |
| Cytokines MesoScale 4 hr.                                            | a                                                             | ELISA    |                  |                |      |         |           |
| IL-15 level 8hr                                                      | PK                                                            |          |                  |                |      |         |           |
| .                                                                    | 8 ml SST                                                      |          |                  |                |      |         |           |
| Cytokines MesoScale 8 hr.                                            | ELISA                                                         |          |                  |                |      |         |           |
| IL-15 level 12hr                                                     |                                                               | 4 ml SST | PK               |                |      |         |           |
| IL-15 level 24hr                                                     | PK                                                            |          |                  |                |      |         |           |
| Cytokines MesoScale  24 hr.                                          | 1                                                             | 8 ml SST | ELISA            |                |      |         |           |
| 2                                                                    | Roederer FACS day 2                                           | 2        | 10 ml Lavender   | Immune subsets | LyMB |         |           |
| 3                                                                    | IL-15 level 48hr                                              | 4mlSST   | Figg             |                |      |         |           |
| .BK                                                                  |                                                               |          |                  |                |      |         |           |
| IL-15 level day 7                                                    | Draw with AM Labs                                             | Figg     |                  |                |      |         |           |
| 4 ml SSTE                                                            | PK                                                            | 1        |                  |                |      |         |           |  
Version Date: 06/13/2018  
| Start Date of Treatment:    /_/20_                                                      | Dose Level:                                                   | .mcg/kg/day       |                |                   |                   |           |
|-----------------------------------------------------------------------------------------|---------------------------------------------------------------|-------------------|----------------|-------------------|-------------------|-----------|
| Patient Name:                                                                           | Start Time of Treatment:                                      |                   |                |                   |                   |           |
| All samples should be kept at Room Temperature in Clean Utility Room Page until Pick up |                                                               |                   |                |                   |                   |           |
| Page Figg Lab (pager 102-11964) for Sample pick up                                      | Page LyMB research nurse (pager 102-11029) for sample pick up |                   |                |                   |                   |           |
| Specimen Information                                                                    | Ideal                                                         | Actual            | Comments       |                   |                   |           |
| Study                                                                                   | Pick up                                                       |                   |                |                   |                   |           |
| Sample                                                                                  | Time                                                          | (Missed           |                |                   |                   |           |
| Sample name                                                                             | Tube                                                          | Sample            | Time           | Drawn             | Specimens, etc.)  | Signature |
| Type                                                                                    |                                                               |                   |                |                   |                   |           |
| IL-15 level day 8                                                                       | PK                                                            | Figg              |                |                   |                   |           |
| Draw with AM Labs                                                                       |                                                               |                   |                |                   |                   |           |
| 1                                                                                       | 8 ml SST                                                      | ELISA             |                |                   |                   |           |
| 8                                                                                       | Cytokines MesoScale day 8                                     | Draw with AM Labs |                |                   |                   |           |
| Roederer FACS day 8                                                                     | 2                                                             | 10 ml Lavender    | Immune subsets | LyMB              |                   |           |
| 9                                                                                       | IL-15 level day 9                                             | 4ml SST           | PK             | Draw with AM Labs |                   |           |
| 1                                                                                       |                                                               |                   |                |                   |                   |           |
| 10                                                                                      | IL-15 level day 10                                            | 1                 | 4 ml SST       | PK                | Draw with AM Labs |           |
| IL-15 Level End of Rx                                                                   | 1                                                             | 4 ml SST          | PK             |                   |                   |           |
| 1                                                                                       | 4 ml SST                                                      | PK                |                |                   |                   |           |
| IL-15 Level 10 min post Rx                                                              |                                                               |                   |                |                   |                   |           |
| IL-15 Level 30 min post Rx                                                              | 1                                                             | 4 ml SST          | PK             | Figg              |                   |           |
| 11                                                                                      | IL-15 Level 1 hr.  post Rx                                    | 1                 | 4 ml SST       | PK                |                   |           |
| IL-15 Level 2 hr.  post Rx                                                              | 1                                                             | 4 ml SST          | PK             |                   |                   |           |
| IL-15 Level 4 hr.  post Rx                                                              | 1                                                             | 4 ml SST          | PK             |                   |                   |           |
| IL-15 Level 24 hr.  post Rx                                                             | PK                                                            |                   |                |                   |                   |           |
| 1                                                                                       | 8 ml SST                                                      |                   |                |                   |                   |           |
| Cytokines MesoScale day 12                                                              | ELISA                                                         |                   |                |                   |                   |           |
| 12                                                                                      | Roederer FACS day 12                                          | 2                 | 10 ml Lavender | Immune subsets    | Draw with AM Labs | LyMB      |
| Draw with AM Labs                                                                       | Figg                                                          |                   |                |                   |                   |           |
| IL-15 antibodies                                                                        | 1                                                             | 4 ml SST          | ELISA          |                   |                   |           |  
Abbreviated Title: Ph I Study of CIV rhIL-15 Version Date: 06/13/2018 APPENDIX E:  CIV RHIL-15 BIOSPECIMEN WORKSHEET FOR 12-C-0113 CYCLE 2 AND BEYOND  
| Work sheet for 12-C-0113 Cycle 2                                                        | Dose Level:                                                   | _mcg/kg/day    |                |                   |                   |           |
|-----------------------------------------------------------------------------------------|---------------------------------------------------------------|----------------|----------------|-------------------|-------------------|-----------|
| Start Date of Treatment:                                                                | _/__/20_                                                      |                |                |                   |                   |           |
| Patient Name:                                                                           | Start Time of Treatment:                                      |                |                |                   |                   |           |
| All samples should be kept at Room Temperature in Clean Utility Room Page until Pick up |                                                               |                |                |                   |                   |           |
| Page Figg Lab (pager 102-11964) for Sample pick up                                      | Page LyMB research nurse (pager 102-11029) for sample pick up |                |                |                   |                   |           |
| Comments                                                                                |                                                               |                |                |                   |                   |           |
| Specimen Information                                                                    | Ideal                                                         | Actual         |                |                   |                   |           |
| Study                                                                                   | (Missed                                                       |                |                |                   |                   |           |
| Sample name                                                                             | Tube                                                          | Sample         | Sample         | Time              | Pick up           | Signature |
| day                                                                                     | Specimens,                                                    |                |                |                   |                   |           |
| type                                                                                    | Drawn                                                         |                |                |                   |                   |           |
| Time                                                                                    | etc)                                                          |                |                |                   |                   |           |
| Figg                                                                                    |                                                               |                |                |                   |                   |           |
| 1                                                                                       | IL-15 antibodies                                              | 1              | ELISA          | Draw with AM Labs |                   |           |
| 4 ml SST                                                                                |                                                               |                |                |                   |                   |           |
| 2                                                                                       | Roederer FACS day 2                                           | 10 ml Lavender | Immune subsets | Draw with AM Labs |                   |           |
| 2                                                                                       |                                                               |                |                |                   |                   |           |
| 8                                                                                       | LyMB                                                          |                |                |                   |                   |           |
| Roederer FACS day 8                                                                     | 2                                                             | 10 ml Lavender | Immune subsets | Draw with AM Labs |                   |           |
| Roederer FACS day 12                                                                    | 2                                                             | 10 ml Lavender | Immune subsets | Draw with AM Labs |                   |           |
| 12                                                                                      | IL-15 antibodies                                              | 1              | 4 ml SST       | ELISA             | Draw with AM Labs | Figg      |