Functional states of polymorphonuclear leukocytes determined by chemiluminescent kinetic analysis

Abstract

During the respiratory burst, upon stimulation with both soluble and particulate matter, polymorphonuclear leukocytes (PMN) generate reactive oxygen species (ROS) and emit chemiluminescence (CL) as a result of metabolic activation. The measurement of CL has been demonstrated to be a useful tool for in vitro assessment of the opsonophagocytic function of PMN. Using component analysis of CL kinetics, we characterized the functional state of PMN by three parameters of the respiratory burst: capacity, effectiveness and velocity (CEV space). The possibility of delimiting eight different functional states of PMN is discussed. The CL kinetics shown by blood PMN in different functional states was analysed, and revealed six out of eight functional states. We conclude that CEV-estimated functional states of PMN are relative, depending on both PMN readiness to generate ROS and conditions of the CL test.

Differentiation between Viral and Bacterial Acute Infections Using Chemiluminescent Signatures of Circulating Phagocytes

“…To conclude, the two infections exhibit completely different types of chemiluminescent signatures, each one described by its own characteristics, proving that the oxidative potential of the innate immunity can indeed serve as a prognostic or diagnostic marker. Moreover, we show herein that the infectious states do show an effective type of phagocytic behavior, as supported by the behavior of peritonitis and pulmonary abscess cases …”

“…The CL kinetics of bacterial infection are associated with high capacity, lower effectiveness than in controls, and high velocity in the standard system (Figures 1A and 2B), corresponding to an "effective" (C H E H V H ), very close to the "fighting" (C H E L V H ) functional state of the phagocytes. This state corresponds to the activated state of the immune system in case of infection (as it was already shown before 14,34 ): the phagocytes have high capacity to generate ROS, which occurs predominantly inside the cells. This oxidative potential of cells is achieved by processes directly associated with phagocytosis.…”

“…Figure presents the main parameters (capacity, effectiveness, and velocity) that are used in the estimation of functionality of the immune system and Figure represents the CL curves in case of different infections. The data was analyzed and compared to healthy, noninfected controls demonstrating the earlier described “resting” pattern (C L E H V L = Capacity Low Effectiveness High Velocity Low ). , The resting pattern is characterized with a low capacity to generate ROS, while phagocytosis of the prey is performed with high effectiveness, expressing the healthy state of the cells; and low velocity, as the cells are not activated and not ready for the defense and killing. In comparison to controls, it can be observed that the two infected groups have higher capacity and velocity in the standard system, revealed by the presence of infection, leading to an elevated cellular CL response (Figure A).…”

“…Moreover, we show herein that the infectious states do show an effective type of phagocytic behavior, as supported by the behavior of peritonitis 14 and pulmonary abscess cases. 34 Phagocyte CL Information Can Differentiate between Clinical Groups. The kinetic parameters previously described were calculated in all clinical cases, and the data set was checked to evaluate to what extent the proposed methodology can serve as a relevant tool to describe clinically primed and challenged phagocyte diversity.…”

Chemiluminescence of non‐differentiated THP‐1 promonocytes: developing an assay for screening anti‐inflammatory milk proteins and peptides

“…We have observed quite similar accelerated neutrophil CL reaction kinetics in patients suffering from various respiratory infections (unpublished). Shorter CL peak times were also obtained by priming ‘resting’ neutrophils in vitro with E. coli culture supernatant (41) or with N‐formylated methionine–leucine–phenylalanine (42) prior to stimulation with polystyrene particles or OZ, respectively. To our knowledge, the effect of priming on CL kinetics of nd‐THP‐1 cells has not been described in the literature before.…”