Phagocyte Luminescent Sensor

Abstract

The article describes basics of approach to build a device dedicated to sensitive assessment of phagocyte activity in biological fluids. The unique place of phagocytosis in the innate immune system, the mechanisms, localization and factors of oxidative phagocyte activity as well as the techniques used to detect respiratory burst and localization are briefly reviewed. Sensor design combines simplicity and artifact ‐absence in whole blood system, simultaneous record and analysis of extra‐cellular and intra‐cellular luminescence, sensitivity and efficiency of signal capturing of optical fiber–photomultiplier detection system and the convenience of disposable sample holder.

Dynamic Component Chemiluminescent Sensor for Assessing Circulating Polymorphonuclear Leukocyte Activity of Peritoneal Dialysis Patients

“…Peripheral whole blood (1:100 (v/v) final dilution) was used to avoid the appearance of artifacts due to the isolation of PMNs and to preserve conditions that are close to the in vivo cellular environment . It was collected from patients in heparinized tubes (20 U/mL).…”

“…Component analysis of CL kinetics was used to quantitate at a given time the status of the nonadaptive immune system contained in the collected blood sample. Incubation of PMNs with the fMLP chemoattractant assumed a change in PMN functionality termed priming factor . Moreover, blood storage has been described as modulating the phagocyte respiratory burst, as measured by CL, and was used as a kind of priming factor as well. ,, Thus, fMLP priming and blood storage (i.e., aging of blood) were chosen as factors of differential component assessment, reflecting well-controlled shifts in the phagocyte functional status.…”

“…During phagocytosis of microbial intruders, phagocytes increase their consumption of molecular oxygen. Activation of PMNs, induced by stimuli, results in the production of various reactive oxygen metabolites, including superoxide anion, hydrogen peroxide, hypochlorous acid, and hydroxyl radical, a process collectively termed “respiratory burst”. – This process is accompanied by light emission (chemiluminescence, CL) in the presence of luminol. CL, therefore, is a sensitive measure of the oxidative potential of phagocytes, which correlates well with antimicrobial activity, and can be detected as luminol-dependent CL (LCL). , Despite sensitivity and convenience of this approach, few clinical applications have been so far implemented. – …”

“…To gain as much information as possible from the CL kinetics pattern (related to the production of reactive oxygen species (ROS)), the extracellular, as well as intracellular components (phagocytosis- or nonphagocytosis-related), of the phagocyte-emitted whole-blood luminol-dependent CL were assessed simultaneously. ,, We used these components, and a previously described methodology, to derive CL kinetic parameters that allow quantitative comparison of the dynamic functional state of circulating blood PMNs . It was shown that to obtain even more kinetic information ,, from the PMNs, one can prime them artificially in order to obtain a dynamic picture of their circulating functional state. To do this we performed a laboratory-based ex vivo controlled shift in the phagocyte functional state by using two different priming systems (formylmethionyl-leucyl-phenylalanine (fMLP) chemoattractant priming and aging of blood) and comparing their dynamic change when measured in these two different scenarios.…”

“…10,19,20 We used these components, and a previously described methodology, 21 to derive CL kinetic parameters that allow quantitative comparison of the dynamic functional state of circulating blood PMNs. 10 It was shown that to obtain even more kinetic information 10,20,22 from the PMNs, one can prime them artificially in order to obtain a dynamic picture of their circulating functional state. To do this we performed a laboratory-based ex vivo controlled shift in the phagocyte functional state by using two different priming systems (formylmethionyl-leucyl-phenylalanine (fMLP) chemoattractant priming and aging of blood) and comparing their dynamic change when measured in these two different scenarios.…”