Functional stability of ACD anticoagulated blood samples stored for 72 hours at room temperature.

Abstract

Abstract Characterization of the immune system in remote locations is challenging, requiring technical expertise and specific analytical equipment. Immune functional assessments require live cells, and the window of viability following blood donation is limited. To test the ability of ACD blood collection tubes to expand this window of viability, cellular function was analyzed over 4 consecutive days, beginning on the date of blood collection. T-cell activation, characterized by CD69 and CD25 expression, was measured following a 24-hour stimulation with soluble anti-CD3 and anti-CD28 antibodies (CD3/CD28) or with staphylococcus enterotoxins A and B (SEA+SEB). Mitogen stimulated cytokine profiles were determined following a 48-hour stimulation with CD3/CD28, phytohemagglutinin and ionomycin (PMA+I), or lipopolysaccharide (LPS). T-cell activation following stimulation with SEA+SEB was not altered by the processing delay; however, following stimulation with CD3/CD28, T-cell activation increased over the 72 hour period. The processing delay did not affect cytokine production following CD3/CD28 stimulation. Following stimulation with LPS and PMA+I, results were varied. While some cytokine responses were unaltered following the processing delay, others were impaired. These findings confirm the general viability of whole blood stored at room temperature (68 to 72°F) for 48 to 72 hours in ACD tubes and suggest many phenotypic and functional assays are feasible on blood samples aged as described. Investigators using this protocol should confirm their specific culture technique and measured output. This protocol may assist studies of subjects in remote locations with no immediate access to laboratory instrumentation.