[Novel methods of cytokine detection: real-time PCR, ELISPOT, and intracellular cytokine staining].
Cytokines are small hormone-like proteins that play important roles in immune system control. Cytokines regulate the proliferation and differentiation of cells and hematopoiesis and act as mediators in the inflammatory reaction. Changes in cytokine levels are found in many diseases, such as sepsis, bowel inflammatory disease, autoimmune diseases, as well as graft-versus-host disease. Cytokines levels can be detected using in vivo, in vitro, and ex vivo techniques. The level of cytokine produced can be measured by immunoenzymatic test (ELISA) in supernatant after cell culture with the addition of stimulant and in plasma by techniques that measure the level of cytokine secretion in cells (e.g. immunohistochemical staining, ELISPOT, and intracellular cytokine staining), and by molecular biological methods (RPA, real-time PCR, in situ hybridization,and Northern blot). Detection of cytokine mRNA in tissues is useful in the direct determination of heterogenic populations of cytokine-producing cells. Nowadays the most frequently used methods for measuring cytokine level are ELISPOT, intracellular cytokine staining with fl ow cytometry detection, and real-time PCR. These methods have an important clinical role in vaccine efficacy, in viral, bacterial, and verminous diagnostics, and in determining the efficacy of cancer treatment.
Turlej
Eliza
E
Laboratorium i Zakład Immunologii Klinicznej, Instytut Immunologii i Terapii Doświadczalnej PAN im. Ludwika Hirszfelda we Wrocławiu, 53-114 Wrocław. turlej@iitd.pan.wroc.pl
D016207:Cytokines / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013194:Staining and Labeling
D016207:Cytokines