Overview
The research goal of this study is to obtain CD34+ hematopoietic stem cells (HSC) from peripheral blood and/or bone marrow, and Mononuclear Cells (lymphocytes and monocytes), and granulocytes (grans) from peripheral blood that will be used in the laboratory and/or in the clinic to develop new cell therapies for patients with inherited or acquired disorders of immunity or blood cells. Development of novel cellular therapies requires access to HSC, Mononuclear Cells and/or granulocytes as the essential starting materials for the pre-clinical laboratory development of gene therapies and other engineered cell products. HSC or blood cells from healthy adult volunteers serve both as necessary experimental controls and also as surrogates for patient cells for clinical scale-up development. HSC or blood cells from patients serve both as the necessary experimental substrate for novel gene therapy and cellular engineering development for specific disorders and as pre-clinical scale up of cellular therapies. Collection of cells from adult patients collected in the NIH Department of Transfusion Medicine (DTM) under conditions conforming to accepted blood banking clinical practice may also be used directly in or cryopreserved for future use in other NIH protocols that have all required regulatory approvals allowing such use. In summary, the research goal of this protocol is the collection of HSC or blood cells that may be used for both laboratory research and/or for clinical treatment in other approved protocols.
Description
The research goal of this study is to obtain CD34+ hematopoietic stem cells (HSC) from peripheral blood and/or bone marrow, and Mononuclear Cells (lymphocytes and monocytes), and granulocytes (grans) from peripheral blood that will be used in the laboratory and/or in the clinic to develop new cell therapies for patients with inherited or acquired disorders of immunity or blood cells. Development of novel cellular therapies requires access to HSC, Mononuclear Cells and/or granulocytes as the essential starting materials for the pre-clinical laboratory development of gene therapies and other engineered cell products. HSC or blood cells from healthy adult volunteers serve both as necessary experimental controls and also as surrogates for patient cells for clinical scale-up development. HSC or blood cells from patients serve both as the necessary experimental substrate for novel gene therapy and cellular engineering development for specific disorders and as pre-clinical scale up of cellular therapies. Collection of cells from adult patients collected in the NIH Department of Transfusion Medicine (DTM) under conditions conforming to accepted blood banking clinical practice may also be used directly in or cryopreserved for future use in other NIH protocols that have all required regulatory approvals allowing such use. In summary, the research goal of this protocol is -the collection of HSC or blood cells that may be used for both laboratory research and/or for clinical treatment in other approved protocols.
Participants include: 1. Adult patients with any primary immune deficiency (PID) or other blood disorder where collection is both for clinical use in another approved treatment protocol to benefit the patient and for laboratory research; 2. Adult patients with any primary immune deficiency (PID) or blood disorder where collection is for laboratory research use only. 3. Healthy adult volunteers where the collection is for laboratory research use only.
The majority of subjects will have HSC collected from peripheral blood by apheresis. Daily subcutaneous injections of G-CSF (granulocyte colony stimulating factor/filgrastim) for 5 to 6 days is a standard of care method used to mobilize HSC to the peripheral blood prior to apheresis, and will be used for most subjects. Plerixafor (Mozobil) is approved as standard of care for use in combination with G-CSF to mobilize HSC.
Some adult patients will have a clinical scale aspiration collection of bone marrow to obtain HSC for clinical use in another approved treatment protocol. Some adult participants may have a small sample needle aspiration collection of bone marrow obtained for laboratory research purposes only.
Mononuclear cells and/or Granulocytes (gran) will be collected from peripheral blood by apheresis following no stimulation, using G-CSF alone, or a using a combined single dose of G-CSF (480mcg) and Dexamethasone (8mg) prior to collection as is the standard of care pre-treatment used in the NIH DTM for collection of granulocyte transfusions from healthy donors.
As noted, HSC, mononuclear cells, and gran collection from patients with PID or other blood disorders may be used for laboratory research or may be designated for future clinical treatment of the patient under separate treatment protocol. HSC, mononuclear cells, and gran collection from healthy volunteers will be designated entirely for laboratory research.
HSC will be used for the following clinical purposes, where clinical treatments would occur under separate IRB approved protocols: 1. Autologous HSC from patients may be genetically modified and infused into the patient for treatment of an infection or the underlying disease (Gene therapy); 2. Autologous HSC from patients may serve as back up (rescue product) for patients undergoing matched unrelated donor transplantation or haploidential related or unrelated donor transplantation.
Mononuclear Cells (lymphocytes and monocytes) and granulocytes will be used for the following clinical purposes, 1. Autologous lymphocytes may be genetically modified and infused into the patient for treatment of an infection or the underlying disease (Gene therapy); 2. Autologous lymphocytes, monocytes and granulocytes (neutrophils) may be transfected with mRNAs to transiently express a therapeutic protein for treatment of an infection or the underlying disease (Gene therapy).
HSC, lymphocytes, monocytes and granulocytes will be used for laboratory research studies that include: Delineating the pathophysiology of inherited immune deficiencies; Delineating the physiology of and improving engraftment of hematopoietic stem cells; Determining how hematopoietic stem cells may be maintained in ex vivo culture without losing pluripotent potential; Delineating the molecular mechanisms responsible for lineage specific differentiation; Developing efficient methods for gene transfer into hematopoietic stem cells for corrective gene therapy; Developing methods for restoration of function in defective peripheral blood monocytes and/or granulocytes; Further characterization of peripheral blood monocytes and/or granulocytes from patients with PID.
Eligibility
- ELIGIBILITY CRITERIA:
Patients (Patients with a genetically defined PID or other blood disorder or clinical
history consistent with PID or other blood disorder)
1. Patients will have a genetically defined PID or have a clinical history of recurrent
infections or other problems suggestive of PID or other blood disorder, must be 18-70
years of age,
2. Some patients may have active bacterial or fungal infection at the time of study
entry.
3. Preserved renal function (creatinine less than or equal to 2.5 mg/dL; less than or
equal to 3+ proteinuria); preserved hepatic function (bilirubin less than or equal to
2.0 mg/dl); preserved hematologic function (WBC greater than or equal
to1000/mm^3;granulocytes greater than or equal to 500/mm^3; platelets greater than or
equal to 100,000; hematocrit greater than or equal to 25). Of note, patients with PID
often have associated chronic thrombocytopenia. Patients with stable chronic
thrombocytopenia will be eligible for collection, at the investigator s discretion,
with the caveat that patients with platelet count <40,000 the day prior to collection
will be transfused with platelets on the morning of collection. Platelets may also be
given to these patients following the collection if medically indicated..
4. Patients of childbearing potential may be entered if using effective contraception and
having a negative serum or urine pregnancy test within one week of beginning G-CSF
administration.
5. Patients may remain on their regimen of prophylactic treatments as deemed necessary by
the investigator.
6. Willingness to allow blood cell samples to be stored
7. Willingness to allow blood and/or bone marrow samples to be modified to iPS cells
Healthy Adult Volunteers
1. Healthy adults aged 18-65 without active current infection or history of recurrent
infection,
2. Weighs at least 50kg.
3. Normal renal function (creatinine less than or equal to 1.5 mg/dL; less than or equal
to 1+ proteinuria); normal hepatic function (bilirubin less than or equal to 1.5
mg/dL); normal hematologic function (WBC greater than or equal to 2500/mm^3;
granulocytes greater than or equal to 1200/mm^3; platelets greater than or equal to
120,000; hematocrit greater than or equal to 38).
4. Normal female volunteers of childbearing potential may be entered if using effective
contraception and having a negative serum or urine pregnancy test within one week of
beginning GCSF administration.
5. Willingness to allow blood cell samples to be stored
6. Willingness to allow blood and/or bone marrow samples to be modified to iPS cells
For PBMCs and grans collections, adult subjects with known genetic mutations may
participate as healthy volunteers for research purposes as long as the other criteria
listed above are fulfilled.
EXCLUSIONS:
Patients
1. Patients who are hemodynamically unstable (systolic or diastolic blood pressure fall
of 20 mm Hg from the stable patient s baseline measurement) or requiring mechanical
respiratory assistance are excluded.
2. Female patients who are pregnant or lactating as determined by history and/or positive
pregnancy test are excluded.
3. Must be negative by routine blood donor eligibility testing criteria including tests
for syphilis (RPR) and TTV Donor Transplant Panel testing (list is modified
periodically, but may include hepatitis B and C, HIV and HTLV, T. cruzi). This does
not apply to leukapheresis patients, as these tests are not required by DTM.
1. XSCID patients do not make antibodies and false positives may occur because they
receive periodic infusions of pooled donations of IVIG. We have observed positive
anti-HBc testing in these patients. If this occurs, more specific DNA or antigen
testing will be done and must be negative.
2. Patients with CGD and other patients with autoimmunity as part of their PID
phenotype may have false positive antibody tests and if this occurs more specific
DNA or antigen testing will be done and must be negative.
3. Autologous HSC Transplant patients - may be positive for Hepatitis B and C if the
investigator deems it necessary to be collected and used as a safety back-up
Healthy Volunteers
1. Active bacterial, fungal or viral infection as evidenced by history, physical exam
(temperature >38 degress C), or WBC >9000 are excluded.
2. Females who are pregnant or lactating as determined by history and/or pregnancy test
are excluded.
3. Must be negative by routine blood donor eligibility testing criteria, including tests
for syphilis (RPR) and TTV Recipient Transplant Panel (list is modified periodically,
but may include hepatitis B and C, HIV and HTLV, T. cruzi) This does not apply to
leukaphersis patients, as these tests are not required by DTM policy.
4. Someone without peripheral venous access in arm veins adequate for apheresis (healthy
volunteers only).
5. If in the opinion of the investigator participation in this study places the healthy
adult volunteer at undue risk.
Patients being considered for clinical scale bone marrow harvesting
1. Who are unable to lie prone during the bone marrow harvesting procedure.
2. Who are unable to tolerate general anesthesia during the bone marrow harvesting
procedure.