Abstract:
:In several clinical contexts, the measurement of ATP concentration in T lymphocytes has been proposed as a biomarker of immune status, predictive of secondary infections. However, the use of such biomarker in lymphopenic patients requires some adaptations in the ATP dosage protocol. We used blood from healthy volunteers to determine the optimal experimental settings. We investigated technical aspects such as the type of anticoagulant for blood sampling, the effect of freeze and thaw cycles, the reagent and sample mixing sequence, and the optimal dilution buffer. We also shortened the incubation time to 8h, and even showed that a 30min incubation may be sufficient. To evaluate the ATP rise upon lymphocyte activation, the optimal dose of stimulant was defined to be 4μg/mL of phytohaemagglutinin. Lastly, we determined that the number of T cells needed for this measurement was as low as 50,000, which is compatible with the existing lymphopenia in clinical settings. This optimized protocol appears ready to be assessed in lymphopenic patients to further investigate the interconnection between T lymphocyte metabolism and impaired phenotype and functions.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Girardot T,Mouillaux J,Idealisoa E,Poujol F,Rouget C,Rimmelé T,Monneret G,Textoris J,Venet Fdoi
10.1016/j.jim.2016.10.001subject
Has Abstractpub_date
2016-12-01 00:00:00pages
59-66eissn
0022-1759issn
1872-7905pii
S0022-1759(16)30259-9journal_volume
439pub_type
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