A single-photon imaging system for the simultaneous quantitation of luminescent emissions from multiple samples

Maly, Friedrich E.; Urwyler, Adrian; Rolli, Hanspeter; Dahinden, Clemens A.; Weck, A.L. de

doi:10.1016/0003-2697(88)90344-2

Cited by 42 publications

(15 citation statements)

References 10 publications

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“…Herein, the lucigenin-dependent CL was measured using a single-photon imaging system with a two-dimensional photon counting system (MTP reader, Hamamatsu Photonics, Herrsching, Germany) as described elsewhere [22,231 .With this system 96 wells can be measured and analyzed simultaneously. In brief, eosinophils were suspended to a density of 5 X lo4 cells/ml in HBSS + BSA, respectively, containing 200 mM lucigenin (Sigma).…”

Section: Lucigenin-dependent Chemiluminescence (Cl)mentioning

confidence: 99%

C3a activates reactive oxygen radical species production and intracellular calcium transients in human eosinophils

et al. 1994

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Whereas C5a is a well-established potent activator of eosinophils, the functional role of C3a in the activation of eosinophils is, so far, poorly understood. Here, the activation of human eosinophils stimulated with C3a was analyzed and compared to C5a activation. Flow-cytometrical measurements revealed that stimulation of eosinophils by C3a resulted in a transient elevation of the intracellular calcium concentration ([Ca2+]i) in a dose-dependent manner. In addition, the production of reactive oxygen radical species (ROS) of eosinophils after C3a and C5a stimulation was measured by lucigenin-dependent chemiluminescence and quantified by superoxide dismutase-inhibitable reduction of ferricytochrome C. Half maximal and maximal ROS production in response to C3a was observed at 50 ng/ml and 1000 ng/ml, respectively, whereas C3a-desArg was inactive. To ensure that C3a stimulation was not caused by contamination with C5a, monoclonal antibodies were used to demonstrate the specificity of C3a. The effect of C3a was completely abolished in the presence of monovalent antigen-binding fragments of a functionally blocking anti-C3a monoclonal antibody. In addition, blockade of the C5a receptor by the monoclonal anti-C5a receptor antibody S5/1 totally inhibited the C5a-evoked ROS production, whereas the C3a response in the presence of S5/1 was unaffected. Finally, desensitization experiments revealed a homologous desensitization of C3a after restimulation with C3a. In contrast, no cross-desensitization was observed upon stimulation with C5a. Furthermore, the C3a- and C5a-induced production of ROS of eosinophils was totally inhibited by pertussis toxin, indicating the involvement of guanine nucleotide-binding proteins (Gi-proteins). In summary, these results demonstrate that C3a is a potent activator for eosinophils initiating transient [Ca2+]i changes and production of reactive oxygen species. C3a therefore may play a part in the pathophysiology of diseases with eosinophil and complement activation.

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Section: Lucigenin-dependent Chemiluminescence (Cl)mentioning

confidence: 99%

C3a activates reactive oxygen radical species production and intracellular calcium transients in human eosinophils

et al. 1994

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“…Cells were stimulated with 20 ng/ml of phorbol myristyl acetate in the presence of 100 /*M lucigenin (JV,N-dimethyl-acridinium nitrate; both from Sigma, Miinchen, FRG). The 60 min integral of CL was then measured using a computer-aided microtitre plate luminometer as described in more detail elsewhere (Maly et al, 1988). It was thus possible to complete the evaluation of all serum samples in triplicates on MNC of one donor within <3 h.…”

Section: Methodsmentioning

confidence: 99%

Enhancement of mononuclear cell oxidative burst by early post-transfer sera from human patients after unsuccessful embryo transfer

Maly

Sterzik²,

Henderson³

et al. 1990

Human Reproduction

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Early pregnancy sera were earlier shown to modulate T lymphocyte rosetting with sheep erythrocytes. We set out to investigate whether early pregnancy sera also modulate the state of activation of another immunologically relevant cell type, the monocyte. As an index of monocyte activation, we measured phorbol ester-triggered oxidative burst activity by a highly sensitive chemiluminescence method. Contrary to our expectations, incubation of mononuclear cells with sera taken early after embryo transfer from patients with a successfully developing pregnancy had no effect. Unexpectedly, such sera from patients from a control group of patients in whom no pregnancy developed after embryo transfer caused enhancement of mononuclear cell chemiluminescence. Stimulatory activity of these sera appeared between days 4 and 6 and was maximal between days 7 and 9 post embryo transfer. Whether this phenomenon is causally related to, or represents a consequence of the failure of embryo transfer, can currently not be decided.

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“…The culture medium was DMEM (200 m l/well) containing 10% heat-inactivated FCS, and antibiotics. Then 5 m l of the chemiluminescent probe (L-012, 5 mg/ml) was added to the cell suspension [11] and the reaction mixture was incubated for another 2 min in a dark room at 37 ЊC [12]. The reactions were initiated by the addition of 5 m l of ligands.…”

Section: Inflammation Researchmentioning

confidence: 99%

Hyaluronic acid inhibits interleukin-1-induced superoxide anion in bovine chondrocytes

Fukuda

Takayama

Ueno

et al. 1997

Inflammation Research

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A single-photon imaging system for the simultaneous quantitation of luminescent emissions from multiple samples

Cited by 42 publications

References 10 publications

C3a activates reactive oxygen radical species production and intracellular calcium transients in human eosinophils

C3a activates reactive oxygen radical species production and intracellular calcium transients in human eosinophils

Enhancement of mononuclear cell oxidative burst by early post-transfer sera from human patients after unsuccessful embryo transfer

Hyaluronic acid inhibits interleukin-1-induced superoxide anion in bovine chondrocytes

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