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ALTEX 2015The whole blood pyrogen test was first described in this journal exactly twenty years ago. It employs the cytokine response of blood monocytes for the detection of...
The whole blood pyrogen test was first described in this journal exactly twenty years ago. It employs the cytokine response of blood monocytes for the detection of microbiological contaminants with the potential to finally replace the still broadly used rabbit pyrogen test. The article reviews its development process, the current status of the test as well as the challenges and missed opportunities. The article highlights the enormous efforts of many people to get the test to where it is today. But it also shows the incredible missed opportunities for implementation and thus sparing about 400,000 rabbits still used for this purpose per year worldwide; in the EU, since the official acceptance of the test, the number of animals used for pyrogen testing did not fall but increased by about 10,000 to 170,000. The test is the first solution enabling adequate pyrogen testing of cell therapies, including blood transfusions, and medical devices, but has not been implemented for either application by authorities. As the test can quantitatively assess human-relevant airborne pyrogens, the contribution of pyrogens to chronic obstructive lung diseases and childhood asthma can for the first time be defined and home and workplace safety improved in the future.
Topics: Animal Testing Alternatives; Animals; Drug Contamination; History, 20th Century; History, 21st Century; Humans; Lipopolysaccharides; Monocytes; Pyrogens; Rabbits
PubMed: 25863033
DOI: 10.14573/altex.1503241 -
The Journal of Physiology Jul 19751. Several possible mechanisms of the antipyretic action of indomethacin administered cat. 2. Indomethacin did not decrease bacterial endotoxin-induced release of...
1. Several possible mechanisms of the antipyretic action of indomethacin administered cat. 2. Indomethacin did not decrease bacterial endotoxin-induced release of endogenous pyrogen in vivo. 3. Indomethacin (5-40 mug/kg) inhibited the pyrogenic effect of peripherally or centrally administered leucocytic progen. A dose of 10 mug/kg caused a parallel shift to the right of the log dose-response curve for I.V. leucocytic pyrogen and reduced the potency of the pyrogen at least 50%. 4. Incubation of leucocytic pyrogen with indomethacin did not alter its pyrogenic potency. 5. Indomethacin exerted only a slight non-dose-related hypothermic effect in afebrile animals. 6. Indomethacin (up to 1 mg/kg) did not diminish the hyperthermic response to intraventricular administration of prostaglandin E1. 7. This pattern of activity indicates that indomethacin acts centrally to inhibit an effect of leucocytic pyrogen.
Topics: Animals; Body Temperature; Brain; Cats; Cerebral Ventricles; Dose-Response Relationship, Drug; Fever; Hypothermia; Indomethacin; Injections, Intravenous; Prostaglandins; Pyrogens
PubMed: 1151840
DOI: 10.1113/jphysiol.1975.sp010992 -
The Journal of Physiology Jan 19761. Cholinergic synapses in the hypothalamus may transmit information in those thermoregulatory pathways which function to raise body temperature. The effect of atropine,...
Effects of atropine, injected into a lateral cerebral ventricle of the rabbit, on fevers due to intravenous leucocyte pyrogen and hypothalamic and intraventricular injections of prostaglandin E1.
1. Cholinergic synapses in the hypothalamus may transmit information in those thermoregulatory pathways which function to raise body temperature. The effect of atropine, administered intracranially, on the febrile response to intravenous leucocyte pyrogen or intracranial prostaglandin E1 was therefore examined in conscious rabbits. 2. In rabbits exposed to a thermoneutral environment, micro-injections of PGE1, into the anterior hypothalamus, intraventricular injections of PGE1, and intravenous injection so leucocyte pyrogen all caused fever accompanied by vasoconstriction in the ears and reduced respiratory rate. Intraventricular injection of 200 mug atropine during such fevers attenuated their development. This was due to the activation of heat loss mechanisms through vasodilatation in the ears and an increase in the frequency of respiration. This suggests a similarity in the pattern of neuronal activity evoked by PGE1 and leucocyte pyrogen, at least at the site(s) where atropine directly or indirectly exerted its effect and in the efferent pathways from this site. 3. In rabbits exposed to a cold environment, intraventricular injection of PGE1 caused fever through the activation of shivering accompanied by increased O2 consumption. Intraventricular injection of atropine during the development of fever caused an inhibition of shivering accompanied by increased O2 consumption. Intraventricular injection of atropine during the development of fever caused an inhibition of shievering and a decrease in O2 consumption so that temperature ceased to rise and returned to normal. 4. During fever, reversal by atropine of the increased heat conservation of rabbits in a neutral environment, and of their increased heat production in a cold environment adds further support to the concept that cholinergic synapses provide an important link in central temperature-rasising pathways.
Topics: Animals; Atropine; Body Temperature Regulation; Efferent Pathways; Hypothalamus; Injections, Intraventricular; Male; Prostaglandins E; Pyrogens; Rabbits; Receptors, Cholinergic; Respiration
PubMed: 1255503
DOI: 10.1113/jphysiol.1976.sp011255 -
The Journal of Clinical Investigation Feb 1980Fever not explained by infection may occur in patients with malignant lymphoma presumably caused by a release of endogenous pyrogen. Although pyrogen has been found in...
Fever not explained by infection may occur in patients with malignant lymphoma presumably caused by a release of endogenous pyrogen. Although pyrogen has been found in some tumors with a mixed cell population, production of endogenous pyrogen by the neoplastic cells has not been demonstrated. This report documents the apparently spontaneous synthesis and release of such pyrogen by two human tumor cell lines derived from patients with Hodgkin's disease and histiocytic lymphoma. The endogenous pyrogen from the two cell lines was similar and closely resembled that produced by normal human monocytes in antigenic properties as well as heat and pronase sensitivity. The Hodgkin's disease and histiocytic lymphoma cell lines do not require specific stimulation for the production of endogenous pyrogen suggesting that the mechanism of pyrogen release by neoplastic macrophage-related cells differs from that of normal phagocytic cells. The tumor-associated fever in some patients with malignant lymphoma may be caused by a release of endogenous pyrogen by proliferating neoplastic cells.
Topics: Cell Line; Fever; Hodgkin Disease; Humans; Lymphoma, Large B-Cell, Diffuse; Macrophages; Pyrogens
PubMed: 6985918
DOI: 10.1172/JCI109695 -
The Journal of Experimental Medicine Jun 19611. The mechanism of release of a pyrogen from leucocytes has been studied in cells obtained from sterile rabbit peritoneal exudates and from rabbit blood. Attempts were...
1. The mechanism of release of a pyrogen from leucocytes has been studied in cells obtained from sterile rabbit peritoneal exudates and from rabbit blood. Attempts were made to induce human leucocytes-from blood-to release a pyrogen. 2. Rabbit leucocytes, kept below 4 degrees C., were not pyrogenic and did not release any pyrogen when disintegrated. Incubating such cells, in various media, at 37 degrees C. led to the formation of a pyrogen which was heat-labile. The maximum yield was attained after 1(1/2) hours' incubation. 3. The formation of rabbit leucocytic pyrogen was prevented by freezing and thawing the leucocytes, by heating them to 56 degrees C. for half an hour before incubation, and by ageing them in the cold. 4. Nitrofurazone (5-nitro-2-furaldehyde semicarbazone) prevents the formation of leucocytic pyrogen when given by mouth to the cell-donor animals, or when added to leucocytes in intro. 5. Leucocytes from rabbit blood formed leucocytic pyrogen, on incubation in saline, and this formation was also inhibited by nitrofurazone. 6. No leucocytic pyrogen was released from human leucocytes subjected to mechanical, osmotic, or thermal damage, and it was not formed when the cells were incubated in saline. 7. The source of rabbit leucocytic pyrogen, the action of nitrofurazone on leucocytes, and the supposed role of leucocytic pyrogen in fever are discussed.
Topics: Animals; Fever; Furaldehyde; Humans; Interleukin-1; Leukocytes; Pyrogens; Rabbits
PubMed: 13699218
DOI: 10.1084/jem.113.6.1127 -
British Journal of Experimental... Feb 1971SJL mice kept in a 23° environment and injected intravenously with endotoxin developed a marked hypothermia compared with animals given pyrogen-free saline (PFS). In...
SJL mice kept in a 23° environment and injected intravenously with endotoxin developed a marked hypothermia compared with animals given pyrogen-free saline (PFS). In contrast, giving endotoxin to SJL mice which had been “pre-conditioned” for 4 hr at 36° caused relative hyperthermia. Both responses were best observed at 1½ hr after injection. An endotoxin dose of 0·02 μg. was at the threshold of detectability in mice pre-conditioned at 36°, while with 23° animals, the threshold dose was 0·2 μg. Dose-response curves, with an index of precision (λ) of about 0·73, were obtained for mice in both environments, endotoxin doses of about 20 μg. being in the plateau regions of maximum responses. Mouse strains SWR/J and CMRL behaved similarly to SJL, but the temperature responses at both 23° and 36° were smaller. BALB/cJ and AKR/J mice showed a hyperthermic response to endotoxin at 36° but no hypothermia at 23°, while ST/bJ mice showed the converse pattern of hypothermia after endotoxin at 23° but no hyperthermia at 36°. Thus the strain of mouse is an important variable. We suggest that a hypothermia test in SJL mice may provide a simple and convenient bioassay procedure for endotoxin. Although its sensitivity is much less than the rabbit pyrogenicity test, it may be useful for the quantitative measurement of endotoxin activity of such preparations as typhoid, pertussis and cholera vaccines which are rich in endotoxin.
Topics: Animals; Biological Assay; Body Temperature; Endotoxins; Escherichia coli; Fever; Hot Temperature; Hypothermia; Mice; Species Specificity
PubMed: 4926536
DOI: No ID Found -
Infection and Immunity Dec 1986Synthetic lipid A analogs, beta(1-6)glucosamine disaccharide 1,4'-bisphosphates, which possesses four tetradecanoyl groups at the 2- and 2'-amino, and 3- and 3'-hydroxyl...
Synthetic lipid A analogs, beta(1-6)glucosamine disaccharide 1,4'-bisphosphates, which possesses four tetradecanoyl groups at the 2- and 2'-amino, and 3- and 3'-hydroxyl groups (LA-17-PP), and each two of the (R)-3-hydroxytetradecanoyl and tetradecanoyl groups at the 2- and 2'-amino and 3- and 3'-hydroxyl groups, respectively (LA-18-PP), were far less endotoxic than synthetic (506, LA-15-PP) and bacterial Escherichia coli type lipid A's; neither compound showed any detectable lethal toxicity in chicken embryos or preparatory activity for the local Shwartzman reaction in rabbits. Also both compounds were only weakly pyrogenic and comparably less lethally toxic in galactosamine-loaded mice than the reference synthetic and bacterial lipid A's and a synthetic counterpart to biosynthetic lipid A precursor Ia (406, LA-14-PP). Nevertheless, LA-17-PP and LA-18-PP exhibited definite in vivo immunoadjuvant activity in mice, and the ability to induce a possible tumor necrosis factor and alpha/beta interferon in Mycobacterium bovis BCG and Propionibacterium acnes-primed mice, respectively, although these activities were weaker than those of the reference lipid A's. 4'-Monophosphate analogs of the above two test compounds exhibited neither endotoxic nor beneficial activities, but they showed remarkable in vitro bioactivities comparable to those of the corresponding bisphosphate compounds; the ability to activate the human complement system and the clotting enzyme cascade of horseshoe crab amoebocyte lysate, stimulatory effects on guinea pig and murine peritoneal macrophages, and murine splenocytes.
Topics: Adjuvants, Immunologic; Animals; Bacterial Toxins; Complement Activation; Endotoxins; Glycoproteins; Guinea Pigs; Interferon Type I; Limulus Test; Lipid A; Macrophage Activation; Mice; Mice, Inbred Strains; Pyrogens; Shwartzman Phenomenon; Structure-Activity Relationship; Tumor Necrosis Factor-alpha
PubMed: 3781622
DOI: 10.1128/iai.54.3.673-682.1986 -
Journal of Bacteriology May 1967The efficiency of ionizing radiation in detoxifying the lethal determinant(s) of the lipopolysaccharide (LPS) of Salmonella typhimurium, S. enteritidis, and Escherichia...
The efficiency of ionizing radiation in detoxifying the lethal determinant(s) of the lipopolysaccharide (LPS) of Salmonella typhimurium, S. enteritidis, and Escherichia coli in aqueous solution and associated with heat-killed S. typhimurium cells in suspension decreased with doses above 1 Mrad. The 50% end point of inactivation was more than 7.0 Mrad for heat-killed salmonellae and 4.8, 4.5, and 1.0 Mrad for the LPS of S. typhimurium, S. enteritidis, and E. coli, respectively. After exposure to 20 Mrad, S. typhimurium LPS retained a small portion of its lethal properties although the ld(50) was much greater than 9.5 mg per 20-g mouse. However, at -184 C, no inactivation of the lethal determinant(s) occurred after exposure to as much as 20 Mrad. This demonstrated the significance of the indirect effect and the mobility and formation of free radicals. At 22 C, the optical density at 400 mmu increased and the pH decreased with increasing radiation dose, but no qualitative changes were observed in the infrared spectrum. No change was observed in the pyrogenicity of S. typhimurium LPS; a slight decrease in antigenicity was revealed when 6 days, but not when 1 day, elapsed between vaccination and challenge in the mouse protection test. The results were interpreted as evidence of the existence of two or more lethal and antigenic determinants. The differential effect of radiation on these properties and on the pyrogenic component(s) probably are indicative of separate functional sites for lethal, antigenic, and pyrogenic activities.
Topics: Animals; Endotoxins; Lipopolysaccharides; Male; Mice; Polysaccharides, Bacterial; Rabbits; Radiation Effects; Salmonella typhimurium; Spectrophotometry; Typhoid-Paratyphoid Vaccines
PubMed: 5337846
DOI: 10.1128/jb.93.5.1607-1614.1967 -
The Journal of Physiology Sep 19741. Experiments were carried out in unanaesthetized cats to find out if a prostaglandin is the mediator (a) for the long lasting fever which often follows injections of...
1. Experiments were carried out in unanaesthetized cats to find out if a prostaglandin is the mediator (a) for the long lasting fever which often follows injections of phsyiological salt solutions into the cerebral ventricles or into the cisterna magna, as well as their perfusions through the cerebral ventricles, and (b) for the sodium fever which occurs during a perfusion of the cerebral ventricles with calcium-free artificial c.s.f. A fever mediated by prostaglandin should be accompanied by an increase of prostaglandin activity in cisternal c.s.f., and be abolished or prevented by antipyretics like paracetamol or indomethacin which inhibit prostaglandin synthesis. Both criteria were applied.2. The fever which follows injections or perfusions of physiological salt solutions appears to be mediated by a prostaglandin of the E series, probably E(2) (PGE(2)) because it was accompanied by increased prostaglandin E-like activity in the c.s.f. and abolished by paracetamol and indomethacin. During the first few days after pre-treatment of the cats with intramuscular chloramphenicol the injections were rarely followed by fever.3. The fever which occurs during a perfusion with calcium-free artificial c.s.f. appears not to be mediated by prostaglandin, because it was not associated with increased prostaglandin activity in the cisternal effluent, and not prevented by paracetamol or indomethacin, although these antipyretics usually attenuated the fever.4. A perfusion of the cerebral ventricles with artificial c.s.f. containing calcium in an abnormally high concentration (6.25 mM) brought down fever produced by PGE(1), or PGE(2), or bacterial pyrogen.
Topics: Acetaminophen; Animals; Body Temperature; Calcium; Cats; Cerebral Ventricles; Cerebrospinal Fluid; Chloramphenicol; Cisterna Magna; Fever; Indomethacin; Injections; Perfusion; Prostaglandin Antagonists; Prostaglandins; Pyrogens; Shigella dysenteriae; Sodium Chloride
PubMed: 4215879
DOI: 10.1113/jphysiol.1974.sp010675 -
The Journal of Physiology Oct 19791. Intravenous injections of leucocytic pyrogen in doses of 15, 30 and 60 mul./kg caused febrile reactions in male rabbits that were related to age of the animal:...
1. Intravenous injections of leucocytic pyrogen in doses of 15, 30 and 60 mul./kg caused febrile reactions in male rabbits that were related to age of the animal: rabbits under 2 yr of age developed fevers that were related to dose of pyrogen, while rabbits 2-3 yr old showed large febrile responses which were not dose-related.2. Female rabbits of comparable ages generally showed smaller febrile reactions to I.V. leucocytic pyrogen, and still older females (3-5 yr) developed fever only after the largest dose.3. Dose-related febrile responses to 2.5, 5 and 10 mul. leucocytic pyrogen given intracerebroventricularly (I.C.V.) were greater in male rabbits 1-3 yr old than in females of comparable age. Female rabbits 3-5 yr old showed dose-related fevers that were smaller than those of younger animals of both sexes.4. There were no major differences in response to 125, 250 and 500 ng PGE(2), given I.C.V., between male and female rabbits under 2 yr of age. Females 2-3 yr of age had greater responses to PGE(2) than males of comparable age whilst the oldest females showed smaller responses.5. It is concluded that the febrile response of the rabbit to peripheral and central leucocytic pyrogen varies with both age and sex. Differences in sensitivity of central fever controls to endogenous pyrogen in animals of different ages and sexes may account for the different responses to peripheral pyrogen.
Topics: Aging; Animals; Body Temperature; Dose-Response Relationship, Drug; Female; Injections, Intravenous; Injections, Intraventricular; Male; Prostaglandins E; Pyrogens; Rabbits; Sex
PubMed: 521933
DOI: 10.1113/jphysiol.1979.sp012967