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Flow cytometric determination of leukocytes and lymphocyte subsets in thrombocytapheresis products
1Institut für Transfusionsmedizin und Transplantationsimmunologie, Universität Münster, Germany.
To determine the nature of the leukocyte contamination of platelet concentrates we developed a method to identify the immunophenotype of the leukocytes in platelet concentrates. Flow cytometry is ideally suited for this type of rare event detection. Using LDS-751, Forward Scatter and Side Scatter and a modified lysis protocol we were able to discriminate leukocytes from platelets and erythrocytes leaving 2 fluorescent channels free for immunophenotypic analyses. With this protocol we analyzed the leukocyte content and lymphocyte subset distribution pattern of peripheral blood and of the resulting thrombocytapheresis product of 40 blood donors at our institute. We found that the leukocyte content of the platelet concentrate existed almost purely (median 97%, 95-99%) of lymphocytes and monocytes (median 2%, 0-5%), granulocytes were < 1%. Within the lymphocyte population in thrombocytapheresis products the distribution of CD3, CD19, CD4, CD8, CD57, CD5, HLA-DR and gamma/delta T cells did not differ significantly from the corresponding distribution in the peripheral blood of the donors. We conclude that this flow cytometric method can be used as quality control to monitor leukocyte content and lymphocyte distribution of platelet concentrates.
To determine the nature of the leukocyte contamination of platelet concentrates we developed a method to identify the immunophenotype of the leukocytes in platelet concentrates. Flow cytometry is ideally suited for this type of rare event detection. Using LDS-751, Forward Scatter and Side Scatter and a modified lysis protocol we were able to discriminate leukocytes from platelets and erythrocytes leaving 2 fluorescent channels free for immunophenotypic analyses. With this protocol we analyzed the leukocyte content and lymphocyte subset distribution pattern of peripheral blood and of the resulting thrombocytapheresis product of 40 blood donors at our institute. We found that the leukocyte content of the platelet concentrate existed almost purely (median 97%, 95-99%) of lymphocytes and monocytes (median 2%, 0-5%), granulocytes were < 1%. Within the lymphocyte population in thrombocytapheresis products the distribution of CD3, CD19, CD4, CD8, CD57, CD5, HLA-DR and gamma/delta T cells did not differ significantly from the corresponding distribution in the peripheral blood of the donors. We conclude that this flow cytometric method can be used as quality control to monitor leukocyte content and lymphocyte distribution of platelet concentrates.