Assessment of donor T-cell function in cellular blood components by the CD69 induction assay: effects of storage, gamma radiation, and photochemical treatment.
BACKGROUND
Functional donor T-lymphocytes in blood components may cause a variety of transfusion complications. A flow cytometric assay based on the measurement of induced CD69 expression may be an alternative to cell proliferation methods in determining the functional status of these cells in blood components.


METHODS
Seven units of whole blood, RBCs, and platelet concentrates (PCs) were stored under blood bank conditions. Half of 3 PCs each were gamma-radiated or treated with UVA+psoralen; the other half served as controls. Samples were analyzed for phorbolester-induced expression of CD69 as an indicator of cell responsiveness and for exclusion of propidium iodide as a measure of cell membrane integrity and viability.


RESULTS
CD69 inducibility and propidium iodide exclusion decreased exponentially (half-life, 3. 3 and 8.1 days, respectively) during cold blood storage. Irradiation and UVA+psoralen treatment of PCs immediately reduced CD69 inducibility to 21 percent (controls, 82%; p = 0.004) and 12 percent (controls, 95%; p = 0.0008), respectively. The proportion of cells capable of propidium iodide exclusion was similar in treated samples and controls, but it declined faster in the treated samples during subsequent storage.


CONCLUSIONS
Flow cytometric measurement of CD69 induction can be adapted to provide quantitative assessment of T-cell function in blood components. Results obtained by the CD69 assay are in general agreement with those previously reported by use of proliferation methods; the assay may be useful for special applications in transfusion medicine.
Clinical Laboratories, San Francisco General Hospital, CA 94110, USA. Fiebig@itsa.ucsf.edu
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Fiebig
D F
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Hirschkorn
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Maino
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Grass
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Lin
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Busch
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D001681:Biological Assay; D001771:Blood Banks; D001782:Blood Donors; D001793:Blood Preservation; D005363:Ficusin / Q000494:pharmacology; D005720:Gamma Rays; D006801:Humans; D037181:Lectins, C-Type; D017319:Photosensitizing Agents / Q000494:pharmacology; D013601:T-Lymphocytes / Q000502:physiology*; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D005363:Ficusin; D037181:Lectins, C-Type; D017319:Photosensitizing Agents; D000099192:CD69 Antigens