The purpose of this study is to determine whether the administration of highly effective "killer" cells (cytotoxic T cells), along with Interleukin-2 (IL-2) and Recombinant Human Granulocyte Colony Stimulating Factor (GM-CSF) immediately following Autologous Peripheral Blood Stem Cell Transplantation (APBSCT) will enhance anti-tumor immune reconstitution and improve outcome of Multiple Myeloma patients. The overall hypothesis of this proposal is that immediately following APBSCT the immune reconstitution is optimal to administer "killer" cells, combined with the administration of IL-2 and GM-CSF.
Study Type
INTERVENTIONAL
Allocation
NA
Purpose
TREATMENT
Masking
NONE
Enrollment
23
This trial will test if the combination of infusing ex vivo expanded cytotoxic effector cells with IL-2 and GM-CSF post-transplant will accelerate immune reconstitution, resulting in an effector cell-versus-myeloma effect and, possibly, improved clinical outcomes.
Dartmouth-Hitchcock Medical Center
Lebanon, New Hampshire, United States
Number of Participants With Adverse Events in All Subjects
To establish the safety (toxicity) of myeloma patients treated with high dose melphalan, autologous peripheral blood stem cell transplantation (APBSCT) \& adoptive transfer of cytotoxic effector cells with Interleukin-2 (IL-2) and Recombinant Human Granulocyte Colony Stimulating Factor (GM-CSF).
Time frame: From initiation of treatment on protocol until Day 100
Count of Participants With Increased CD3+CD8+, CD8+ and CD56+ Concentrations Between Day 15 Post-Transplant and Days 21 to 28 Post-transplant
To demonstrate that the effector cell infusions result in a clinical effect, phenotypic analyses of blood samples using flow cytometry will be performed prior to, and after each infusion, focusing on the CD8 + populations (CD3+CD8+, CD8+CD56+). As a complement to flow cytometry, the following assays will be used to identify cell subset precursor frequencies, subset proliferation, and cytokine production: * Dye Dilution Proliferation Assay (DDPA) 36 - Evaluation of CD8+ T Cell Precursors * ELISPOT-Quantifying cytokine producing T cells:
Time frame: Day 15 post-transplant and between days 21 to 28 post-transplant
Time to Recovery of Absolute Neutrophil Count
To establish the time to engraftment of myeloma patients treated with high dose melphalan, APBSCT\& adoptive transfer of cytotoxic effector cells with IL-2 and GM-CSF.
Time frame: From initiation of treatment on protocol until Day 100
Time to Recovery of Platelet Count
To establish the time to engraftment of myeloma patients treated with high dose melphalan, APBSCT\& adoptive transfer of cytotoxic effector cells with IL-2 and GM-CSF.
Time frame: From initiation of treatment on protocol until Day 100
Assessment of Disease Response to Treatment
To establish the disease response of myeloma patients treated with high dose melphalan, APBSCT\& adoptive transfer of cytotoxic effector cells with IL-2 and GM-CSF.
Time frame: From initiation of treatment on protocol until Day 100
Number of Participants With Increased Expression of DAP10 and NKG2D on the CD8 Cell Population
* Isolate CD3+CD8+ and CD8+CD56+ from patients' blood following transplant. * Identify NKG2D and DAP10 expression. * Determine the mechanism of tumor cell killing and the relationship to NKG2D or DAP 10 expression. After isolating CD8+ cells from patient's blood samples using the AutoMACS, we will evaluate the expression of NKG2D and DAP10 on all CD8+ cells (CD3+CD8+ and CD8+CD56+cells) pre-transplant (Baseline) and following the third and fourth cellular infusions using phenotypic analysis. We postulate the increased expression of both DAP10 and NKG2D on the CD8 population immediately following APSCT and effector cell infusions when compared to baseline.
Time frame: Pre-transplant and following the third and fourth cellular infusions
Determine the Methods of Tumor Cell Killing of the in Vivo CD8+ Cells: Cytotoxicity Assays, Blocking Experiments, Analysis of T-cell Receptor (TCR)
We will isolate the CD8+ populations (CD3+CD8+, CD8+CD56+) using the Auto MACS (Miltenyi) and then identify the mechanisms of tumor cell killing by the CD8+ cells obtained pre-transplant (Baseline) and following the third and fourth cellular infusions. We will examine mechanisms of tumor cell killing through NKG2D receptor, major histocompatibility complex (MHC) Class I molecules or through the T cell receptor. We postulate the CD8+ cells obtained from patient's blood will kill tumor cells both via MHC Class I and through the NKG2D receptor.
Time frame: Pre-transplant and following the third and fourth cellular infusions.
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