Radioimmunoassay of Prostaglandins and Other Cyclooxygenase Products of Arachidonate Metabolism

Peskar, Bernhard A.; Simmet, Th.; Peskar, B. M.

doi:10.1007/978-3-642-71809-0_18

Cited by 5 publications

(4 citation statements)

References 162 publications

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“…After 3 days of culturing MTT was added, and reduced MTT was measured after 4 h as described above. Radioimmunoassay of 6-keto-PGF,.. Flot (6-keto PGFia) was determined radioimmunologically as described previously (18). RESULTS MDHM-mediated production of inflammatory mediators by peritoneal macrophages.…”

Section: Methodsmentioning

confidence: 99%

MDHM, a macrophage-stimulatory product of Mycoplasma fermentans, leads to in vitro interleukin-1 (IL-1), IL-6, tumor necrosis factor, and prostaglandin production and is pyrogenic in rabbits

Mühlradt

Schade

1991

Infect Immun

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Mycoplasmafermentans-derived high-molecular-weight material (MDHM) was originally discovered because of its capacity to generate, through the induction of monokine synthesis, cytolytic T lymphocytes in concanavalin A-stimulated thymocyte cultures. This study shows that MDHM-activated macrophages not only released interleukin-6 (IL-6) but also exhibited increased synthesis of cell-associated IL-1 as well as liberation of tumor necrosis factor and prostaglandin. We determined 6-keto prostaglandin Fla since it is the stable metabolite of the bioactive prostacyclin. MDHM appeared to be as potent as lipopolysaccharide in inducing the synthesis of these mediators. Priming with gamma interferon further increased MDHM-mediated IL-6 release. Since monokines can be pyrogenic, we tested the effects of an intravenous injection of MDHM on rectal temperatures and leukocyte counts in rabbits. At 1 h after a bolus injection of MDHM, leukocyte counts dropped to about 35% of the initial values, reflecting a decrease in both lymphocytes and granulocytes. At 4 to 6 h after injection, granulocyte counts began to increase again, whereas lymphocyte counts remained low. No leukocytosis was noted during this time. The lack of leukocytosis can be explained by the failure of MDHM-stimulated macrophages to release IL-1. The property of MDHM to cause IL-6 release from macrophages and the IL-6 growth dependency of the 7TD1 hybridoma cell line were made use of in a coculture assay system to quantitate the activity of MDHM. With this method and macrophages from C3HtHeJ lipopolysaccharide-nonresponder mice, MDHM activity was found to be equally distributed in the mycoplasma growth medium and the sedimented mycoplasmas after sonication.

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Section: Methodsmentioning

confidence: 99%

MDHM, a macrophage-stimulatory product of Mycoplasma fermentans, leads to in vitro interleukin-1 (IL-1), IL-6, tumor necrosis factor, and prostaglandin production and is pyrogenic in rabbits

Mühlradt

Schade

1991

Infect Immun

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“…Iscove's medium was obtained from Gibco (Karlsruhe, FRG). A monoclonal antibody directed against LTC 4 was kindly donated by Dr. M. Reinke (Universit~it Erlangen, FRG) [8], and an antiserum against 6-keto-PGFl, was raised in rabbits as described previously [9]. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was from Sigma (St. Louis, MO).…”

Section: Reagentsmentioning

confidence: 99%

“…PGI 2 (in the form of the stable metabolite 6-keto-PGF~) and LTC 4 were determined radioimmunologically as described previously [8,9]. Aliquots of the supernates were mixed with [3H]-labelled 6-keto-PGFl~ or LTC4, respectively and a rabbit antiserum (6-keto-PGFl~) or a monoclonal antibody (LTC4).…”

Section: Quantification Of Pgi 2 and Ltc 4 In The Supernates Of Macromentioning

confidence: 99%

Suppression of endotoxin mitogenicity of spleen cells by lipoxygenase inhibitors and its reversal by 13-hydroxyoctadecadienoic acid

Elekes¹,

Jakobs²,

Schade³

1993

FEMS Immunology &Amp; Medical Microbiology

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Abstract:The effects of several inhibitors of lipoxygenases were investigated in murine spleen cell cultures activated with endotoxin (lipopolysaccharide) It was found that these inhibitors interfere with the proliferative response of the cultures. Indomethacin, a specific cyclooxygenase inhibitor, had no such effect. Endotoxin induced the synthesis of tumour necrosis factor a in spleen cells which was prevented by treatment with a lipoxygenase inhibitor. The inhibition of the mitogenic effect of endotoxin could be reversed by addition of 13-hydroxyoctadecadienoic acid. This was not the case with leukotriene B 4 and C 4 or 15-hydroxyeicosatetraenoic acid. In contrast, these substances had inhibitory effects on the mitogenicity of spleen cells. It is suggested that 13-hydroxyoctadecadienoic acid is involved in the development of the mitogenic reaction, possibly on the level of tumour necrosis factor a production of macrophages present in the cultures.

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“…6-Kcto-prostaglandin F," (6-keto-PGF,") was determined in the supernatants by a radioimmunoassay as described earlier [23]. Platelet-activating factor (PAF) was measured by a bioassay as previously described [24],…”

Section: Measurement Of Mediatorsmentioning

confidence: 99%

Release of Mediators from Human Gastric Mucosa and Blood in Adverse Reactions to Benzoate

Schaubschläger¹,

Becker²,

Schade³

et al. 1991

Int Arch Allergy Immunol

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A study was carried out on 29 patients to investigate the amount of histamine liberation and release of platelet-activating factor and 6-keto-prostaglandin F_1α from gastric mucosa and whole blood or mononuclear cells by sodium benzoate. The patients suffered from asthma (10), atopic dermatitis (7) and chronic urticaria (4). 8 patients with unrelated, non-immunologic diseases served as controls. In the oral provocation test (OPT) 3 patients experienced a recurrence of their original disease, whilst 1 asthmatic patient reacted with abdominal disorder. The release of histamine and prostaglandin from mucosa was significantly increased by sodium benzoate in comparison to the spontaneous release observed in patients. The mucosa of the control persons did not react to sodium benzoate. Furthermore, there was a significant difference in prostaglandin release between patients with positive OPT and the control persons. No difference could be found between patients with negative OPT and those with positive OPT. Additionally, in the mediator release from whole blood or mononuclear cells there was no obvious difference apparent. These results suggest a possible involvement of prostacyclin and histamine in adverse reactions to benzoate. Due to the sensitivity of the method, a mediator release from mucosa can already be demonstrated in a preclinical state of the pseudoallergic reaction in the absence of clinical symptoms.

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Radioimmunoassay of Prostaglandins and Other Cyclooxygenase Products of Arachidonate Metabolism

Cited by 5 publications

References 162 publications

MDHM, a macrophage-stimulatory product of Mycoplasma fermentans, leads to in vitro interleukin-1 (IL-1), IL-6, tumor necrosis factor, and prostaglandin production and is pyrogenic in rabbits

MDHM, a macrophage-stimulatory product of Mycoplasma fermentans, leads to in vitro interleukin-1 (IL-1), IL-6, tumor necrosis factor, and prostaglandin production and is pyrogenic in rabbits

Suppression of endotoxin mitogenicity of spleen cells by lipoxygenase inhibitors and its reversal by 13-hydroxyoctadecadienoic acid

Release of Mediators from Human Gastric Mucosa and Blood in Adverse Reactions to Benzoate

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