Abstract
The evaluation of glucose metabolic activity in immune cells is becoming an increasingly standard task in immunological research. In this study, we described a sensitive, inexpensive, and non-radioactive assay for the direct and rapid measurement of the metabolic activity of CD4+ T cells in culture. A portable, custom-built Cell Culture Metabolite Biosensor device was designed to measure the levels of acidification (a proxy for glycolysis) in cell-free CD4+ T cell culture media. In this assay, ex vivo activated CD4+ T cells were incubated in culture medium and mini electrodes were placed inside the cell free culture filtrates in 96-well plates. Using this technique, the inhibitors of glycolysis were shown to suppress acidification of the cell culture media, a response similar to that observed using a gold standard lactate assay kit. Our findings show that this innovative biosensor technology has potential for applications in metabolic research, where acquisition of sufficient cellular material for ex vivo analyses presents a substantial challenge.
Highlights
Whilst considerable research has focused on the role of antigenic signals, co-stimulation, and cytokines in guiding T cell responses, at a fundamental level, it is cellular metabolism that regulates T cell function and differentiation, and that influences the final outcome of the adaptive immune response [1]
The growth, function, and survival of an activated lymphocyte depend on a pronounced increase in the glucose metabolism that is directly regulated and that has a profound impact on T cell survival and function
It is currently established that T cell metabolic reprogramming from oxidative to glycolytic metabolism affects the functions of T cells, and it impacts T cell physiology and pathological processes [2,3,4,5,6]
Summary
Crowe 1,2,3 , Spyridon Kintzios 4 , Grigoris Kaltsas 5 and Clovis S. Infectious Diseases Department, The Alfred hospital, Melbourne, VIC 3004, Australia. Laboratory of Cell Technology, School of Food Science, Biotechnology and Development, Agricultural
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