Clinical trial NCT00001529: Improved Methods of Cell Selection for Bone Marrow Transplant Alternatives
Official title: Use of Granulocyte Colony Stimulating Factor (G-CSF) Mobilized Leukapheresis Collections From Healthy Volunteers to Develop Improved Methods of Stem Cell and Lymphocyte Selection for Allogeneic Transplantation
Study type: OBSERVATIONAL
Overall status: RECRUITING
Conditions: Graft-Versus-Host Disease
Graft-versus-leukemia
Donor Apheresis
Keywords: G-CSF
Donor Apheresis
Graft Versus Host Disease
graft versus leukemia
Natural History
Normal Volunteer
Lead sponsor: National Institutes of Health Clinical Center (CC)
Sponsor class: NIH
Collaborators: name: New York University and New York Genome Center
First posted: 1999-11-04
Enrollment: 500
Sex: ALL
Minimum age (years): 18
Maximum age (years): 60
Standard ages: ADULT
Accepts healthy volunteers: yes
Observational model: CASE_ONLY
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Healthy Volunteers

Brief summary: Bone marrow transplants (BMT) are one form of treatment for disorders of the blood, including leukemia. However, because the procedure is often associated with potentially life-threatening reactions, it is usually reserved for patients with serious illnesses under the age of 60 years old.

One serious reaction complicating bone marrow transplants is referred to as graft-versus-host disease (GVHD). GVHD is a potentially fatal incompatibility reaction. The reaction is caused by antigens found on the cells of the patient that are not present on the cells of the donor. The antigens are recognized by transplanted white blood cells (lymphocytes). These lymphocytes begin attacking the recipient s cells and tissues and may lead to death.

In order to avoid GVHD, researchers have developed a technique using peripheral blood instead of bone marrow that allows transplantation of stem cells and removal of lymphocytes. Stem cells are the cells responsible for returning blood cell production to normal. Lymphocytes are the white blood cells that can cause GVHD.

The technique requires two steps. In the first step blood cells are collected from donors who have received doses of a growth factor. The growth factor (granulocyte colony stimulating factor) is designed to increase the production of donor stem cells.

In the second step white blood cell lymphocytes are removed from the collected blood, leaving only the stem cells.

The main goal of this study is to develop and improve the method of processing cells that are collected after stimulation with growth factor (G-CSF), by removing the white blood cell lymphocytes which can cause graft-versus-host disease (GVHD) while keeping the stem cells necessary for healthy blood cell building. In addition, researchers are interested in studying whether giving G-CSF has an effect on lymphocyte function, which may influence the immune reactions occurring in bone marrow transplantation.

Detailed description: The NHLBI Stem Cell Transplantation program is exploring ways to make allogeneic transplantation safer and more widely applicable. Prior NHLBI transplant protocols have evaluated the strategy of using T cell depleted marrow transplants followed by delayed lymphocyte add-back to control or prevent GVHD while conserving useful donor immune function against residual leukemia and infectious agents. Over the past ten years, a number of increasingly efficient methods have been used to deplete T cells but retain stem cells, and we have shown the safety and utility of the delayed T cell add-back approach. We have also found a positive relationship between the administration of higher CD34+ cell doses and outcome. Investigation of highly purified grafts with the add-back of specific T cell populations is ongoing, and the ability to test new purification approaches and devices on clinical-scale PBSC products is critical to the continued development of new transplantation approaches in our program. This requires testing the approaches on G-CSF mobilized PBSCs collected by apheresis from healthy donors, since this is the cell source that will be used in all clinical allogeneic transplantation protocols in our program.

Therefore, the primary intent of this protocol is to provide a mechanism for mobilizing, collecting, storing, and analyzing G-CSF mobilized apheresis samples from healthy volunteers. Cells will be used to develop a method of processing the cells that are collected after stimulation with G-CSF, by removing the lymphocytes, which can mediate GVHD while retaining the stem cells which are necessary for hematopoietic reconstitution. At the same time we will study whether G-CSF administration has an effect on lymphocyte, function which may influence the immune reactions occurring in allogeneic bone marrow transplantation. Furthermore the CD34+ cells collected will be a valuable resource for experimental studies of lymphocyte-stem cell interactions in our laboratory.

Eligibility criteria: * INCLUSION CRITERIA:

Healthy individual aged between 18 and 60 years.

No active infection or history of recurrent infection.

Normal renal function: creatinine less than 1.5 mg/dL, proteinuria less than 1+.

Normal liver function: bilirubin less than 1.5 mg/dL, transaminase less than 1.5- fold upper limit of normal

Normal blood count: WBC 3,000-10,000/microliter, ANC \>1,500/microliter, platelets \>150,000/microliter, hemoglobin \>12.5g/dL.

Normal cardiovascular function, no history of chest pain, myocardial infarction, peripheral vascular disease, transient ischemic attack, or stroke.

Healthy female subjects of childbearing age should have a negative serum pregnancy test within one week of beginning G-CSF administration.

Female subjects should not be lactating.

Subject must be eligible for normal blood donation. He or she must be tested negative for syphilis (RPR), hepatitis B and C (HBsAg, Anti-HBc, Anti-HCV), HIV, HTLV-1, West Nile virus, T. Cruzi and Babesia test.

Subject must be able to comprehend the investigational nature of the study and provide informed consent to participate in the protocol.

Antecubital veins must be adequate for peripheral access during apheresis. Potential participants must be screened by an apheresis nurse to check venous access before protocol entry.

EXCLUSION CRITERIA:

Active viral, bacterial, fungal or parasite infection.

Female with positive pregnancy test or lactating.

History of autoimmune disease such as rheumatoid arthritis, systemic lupus erythematosus.

History of cancer excluding squamous carcinoma of the skin.

History of any hematologic disorders.

History of cardiovascular disease or related symptoms such as chest pain, shortness of breath, history of cerebrovascular disease.

Any positive serum screening test as listed in eligibility.

Allergy to G-CSF or bacterial E coli products.

Administration of NSAID within 10 days of starting protocol.

History of G-CSF administration and leukapheresis within past 3 months.

Primary outcomes:
measure: Provide a source of primitive hematopoietic cells from mobilized blood for laboratory studies including optimization of culture and expansion, preservation techniques, gene transfer, analysis of cell surface antigens, &amp; analysis of migra...
timeFrame: End of Study
Secondary outcomes:
measure: Use the cells to develop a reliable technique for T cell depletion of peripheral blood stem cell transplants, which conserves sufficient CD34 cells for safe engraftment while minimizing the risk of GVHD.
measure: Study the effect of G-CSF on lymphocyte subsets and helper cytotoxic function.
Other outcomes:
Locations:
city: Bethesda
country: United States
facility: National Institutes of Health Clinical Center
state: Maryland
Study officials:
affiliation: National Heart, Lung, and Blood Institute (NHLBI)
name: Andre Larochelle, M.D.
role: PRINCIPAL_INVESTIGATOR
References:
citation: Goldman J. Peripheral blood stem cells for allografting. Blood. 1995 Mar 15;85(6):1413-5. No abstract available.
citation: Grigg AP, Roberts AW, Raunow H, Houghton S, Layton JE, Boyd AW, McGrath KM, Maher D. Optimizing dose and scheduling of filgrastim (granulocyte colony-stimulating factor) for mobilization and collection of peripheral blood progenitor cells in normal volunteers. Blood. 1995 Dec 15;86(12):4437-45.
citation: Cottler-Fox M, Cipolone K, Yu M, Berenson R, O'Shaughnessy J, Dunbar C. Positive selection of CD34+ hematopoietic cells using an immunoaffinity column results in T cell-depletion equivalent to elutriation. Exp Hematol. 1995 Apr;23(4):320-2.