The usefulness of flow cytometric analysis of cytokines in peripheral blood and bone marrow plasma

Abstract

Recently attention has been paid to the role of cytokines in clinical pathology, since they can mediate a wide variety of biological effects. For these reasons multiplex methods have been developed to simultaneously measure numerous cytokines in individual small volume specimens. The aim of the study was to assess the usefulness of flow cytometric analysis of cytokines in peripheral blood and bone marrow plasma with Cytometric Bead Array (CBA) kits. The study involved 59 children. Tests were performed in peripheral blood and bone marrow plasma. Human Inflammatory Cytokine Kit (IL-8, -1β, -6, -10, TNF, -12p70) and Human Th₁/Th₂/Th₁₇ Cytokine Kit (IL-2, -4, -6, -10, TNF, INF-γ, -17A) (BD Bioscience) were used. Samples were analyzed on a Cytomics FC500 flow cytometer (Beckman Coulter). In patients diagnosed for hemophagocytic lymphohistiocytosis (HLH) (n=10) and for acute lymphoblastic leukemia (ALL) (n=12) Human Inflammatory Cytokine Kit was used. In almost all samples individual cytokines were detected in a wide range of concentrations (0.47 - 653.74 pg/ml). In samples from patients suffering from allergy (n=12) and in healthy children (n=25) Human Th₁/Th₂/Th₁₇ Cytokine Kit was used. Detection of individual cytokines was much lower: concentration range 0.09-30.17 pg/ml. Based on our analysis the CBA test is suitable for analysis of several cytokines in small volumes of samples. A simple flow cytometer can be used for this test. The CBA test is more suitable for samples with expected increased levels of cytokines. When the levels of cytokines are low, the sensitivity of the CBA test can be too low.