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Preclinical studies on deoxycoformycin and deoxyadenosine as pharmacologic T cell purging tools
W Sheridan1, D S Gordon, A J Fullen
1Department of Medicine, Emory University School of Medicine, Atlanta, GA 30322.
Abstract:
Deoxycoformycin (dCf) is a potent toxin to T lymphocytes in human peripheral blood and bone marrow. In the presence of deoxyadenosine (dAdo), dCf inhibits T cell function as measured by DNA synthesis induced by stimuli in the rank order of mixed lymphocyte culture greater than murine monoclonal antibody OKT3 greater than phytohemagglutinin. Approximately two logs of human bone marrow T cells were removed by 24 h of incubation with dCf and dAdo at doses that preserved colony-forming ability of the treated marrow. A semi-closed system of isolating mononuclear cells in large volumes using density gradient centrifugation was developed. Using this system, it was found that dCf and dAdo could remove about two logs of T lymphocytes after 24 h of incubation when the cells were incubated in large volumes at a relatively high concentration (10(7) cells/ml). This system appears well suited for purging T lymphocytes from human bone marrow before clinical transplantation.
Insights
Deoxycoformycin (dCf) and deoxyadenosine (dAdo) effectively remove T lymphocytes from bone marrow. This method preserves marrow stem cell function, showing promise for T cell purging in transplantation.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- T lymphocytes are crucial for immune responses but can cause graft-versus-host disease after transplantation.
- Purging T cells from donor bone marrow is a strategy to prevent this complication.
Purpose of the Study:
- To evaluate the efficacy of deoxycoformycin (dCf) in combination with deoxyadenosine (dAdo) for T lymphocyte depletion.
- To assess the feasibility of using this method for purging T cells from human bone marrow for transplantation.
Main Methods:
- Developed a semi-closed system for isolating mononuclear cells using density gradient centrifugation.
- Incubated human bone marrow T cells with dCf and dAdo in large volumes at high cell concentrations (10^7 cells/ml).
- Assessed T cell depletion by measuring DNA synthesis and cell counts after 24 hours.
Main Results:
- dCf and dAdo significantly inhibited T cell function, as measured by DNA synthesis.
- Approximately two logs of T lymphocytes were removed within 24 hours.
- The treatment preserved the colony-forming ability of the remaining bone marrow cells.
Conclusions:
- The combination of dCf and dAdo effectively purges T lymphocytes from human bone marrow.
- The developed method is suitable for clinical application in bone marrow transplantation to reduce T cell contamination.