In memory of Helen Laburn and Claus Jessen.

It is with great sadness that we report the passing of our dear colleagues: Professor Helen Laburn and Professor Claus Jessen. We will always remember them.

Mechanisms of fever production and lysis: lessons from experimental LPS fever.

Fever is a cardinal symptom of infectious or inflammatory insults, but it can also arise from noninfectious causes. The fever-inducing agent that has been used most frequently in experimental studies designed to characterize the physiological, immunological and neuroendocrine processes and to identify the neuronal circuits that underlie the manifestation of the febrile response is lipopolysaccharide (LPS). Our knowledge of the mechanisms of fever production and lysis is largely based on this model. Fever is usually initiated in the periphery of the challenged host by the immediate activation of the innate immune system by LPS, specifically of the complement (C) cascade and Toll-like receptors. The first results in the immediate generation of the C component C5a and the subsequent rapid production of prostaglandin E2 (PGE2). The second, occurring after some delay, induces the further production of PGE2 by induction of its synthesizing enzymes and transcription and translation of proinflammatory cytokines. The Kupffer cells (Kc) of the liver seem to be essential for these initial processes. The subsequent transfer of the pyrogenic message from the periphery to the brain is achieved by neuronal and humoral mechanisms. These pathways subserve the genesis of early (neuronal signals) and late (humoral signals) phases of the characteristically biphasic febrile response to LPS. During the course of fever, counterinflammatory factors, "endogenous antipyretics," are elaborated peripherally and centrally to limit fever in strength and duration. The multiple interacting pro- and antipyretic signals and their mechanistic effects that underlie endotoxic fever are the subjects of this review.

Age-dependent changes in temperature regulation - a mini review.

It is now well recognized that the body temperature of older men and women is lower than that of younger people and that their tolerance of thermal extremes is more limited. The regulation of body temperature does not depend on a single organ, but rather involves almost all the systems of the body, i.e. systems not exclusively dedicated to thermoregulatory functions such as the cardiovascular and respiratory systems. Since these deteriorate naturally with advancing age, the decrement in their functions resonates throughout all the bodily processes, including those that control body temperature. To the extent that the age-related changes in some of these, e.g. in the musculoskeletal system, can be slowed, or even prevented, by certain measures, e.g. fitness training, so can the decrements in thermoregulatory functions. Some deficits, however, are unavoidable, e.g. structural skin changes and metabolic alterations. These impact directly on the ability of the elderly to maintain thermal homeostasis, particularly when challenged by ambient thermal extremes. Since the maintenance of a relatively stable, optimal core temperature is one of the body's most important activities, its very survival can be threatened by these disorders. The present article describes the principal, age-associated changes in physiological functions that could affect the ability of seniors to maintain their body temperature when exposed to hot or cold environments.

Acetaminophen: antipyretic or hypothermic in mice? In either case, PGHS-1b (COX-3) is irrelevant.

Acetaminophen (AC) reduces the core temperatures (T(c)) of febrile and non-febrile mice alike. Evidence has been adduced that the selectively AC-sensitive PGHS isoform, PGHS-1b (COX-3), mediates these effects. PGHS-1b, however, has no catalytic potency in mice. To resolve this contradiction, AC was injected intravenously (i.v.) into conscious PGHS-1 gene-sufficient (wild-type (WT)) and -deficient (PGHS-1(-/-)) mice 60 min before or after pyrogen-free saline (PFS) or E. coli LPS (10 microg/kg) i.v. T(c) was monitored continuously; brain and plasma PGE(2) levels were determined hourly. AC at <160 mg/kg did not affect T(c) when given before PFS or LPS; at 160 mg/kg, it caused a approximately 2.5 degrees C T(c) fall in 60 min. LPS given after AC (all doses) induced a approximately 1 degrees C fever, not different from that in AC-untreated mice. But this rise was insufficient to overcome the hypothermia of the 160 mg/kg-treated mice; their T(c) culminated 1 degrees C below baseline. LPS given before AC similarly elevated T(c) approximately 1 degrees C. This rise was reduced to baseline in 30 min by 80 mg AC/kg; T(c) rebounded to its febrile level over the next 30 min. At 160 mg/kg, AC reduced T(c) to 4 degrees C below baseline in 60 min, where it remained until the end of the experiment. WT and PGHS-1(-/-) mice responded similarly to all the treatments. The basal brain and plasma PGE(2) levels of PFS mice and the elevated plasma levels of LPS mice were unchanged by AC at 160 mg/kg; but the latter's brain levels were reduced at 1h, then recovered. Thus, AC could exert an anti-PGHS-2 effect when this enzyme is upregulated in the brain of febrile mice. The hypothermia it induces in non-febrile mice, therefore, is due to another mechanism. PGHS-1b is not involved in either case.

The onset of fever: new insights into its mechanism.

The classical view of fever production is that it is modulated in the ventromedial preoptic area (VMPO) in response to signaling by pyrogenic cytokines elaborated in the periphery by mononuclear phagocytes and the consequent induction of cyclooxygenase (COX)-2-dependent prostaglandin (PG)E(2) in the VMPO. This mechanism has, however, been questioned, in particular because the appearance of circulating cytokines lags the onset of the febrile response to intravenously (iv) injected bacterial endotoxic lipopolysaccharide (LPS), an exogenous pyrogen. Moreover, COX-2, in this case, is itself an inducible enzyme, the de novo synthesis of which similarly lags significantly the onset of fever. Issues also exist regarding the accessibility of the POA to blood-borne cytokines. New data adduced over the past 10 years indicate that the peripheral febrigenic message is conveyed to the VMPO via a neural rather than a humoral route, specifically by the vagus to the nucleus tractus solitarius (NST), and that the peripheral trigger is PGE(2), not cytokines; vagal afferents express PGE(2) receptors (EP(3)). Thus, the initiation of the febrile responses to both iv and intraperitoneal (ip) LPS is temporally correlated with the appearance of LPS in the liver's Kupffer cells (Kc), its arrival immediately activating the complement (C) cascade and the consequent production of the anaphylatoxin C5a; the latter is the direct stimulus for PGE(2) production, catalyzed non-differentially by constitutive COX-1 and -2. From the NST, the signal proceeds to the VMPO via the ventral noradrenergic bundle, causing the intrapreoptic release of norepinephrine (NE) which then evokes two distinct core temperature (T(c)) rises, viz., one alpha(1)-adrenoceptor (AR)-mediated, rapid in onset, and PGE(2)-independent, and the other alpha(2)-AR-mediated, delayed, and COX-2/PGE(2)-dependent, i.e., the prototypic febrile pattern induced by iv LPS. The release of NE is itself modulated by nitric oxide contemporaneously released in the VMPO.

Preoptic nitric oxide attenuates endotoxic fever in guinea pigs by inhibiting the POA release of norepinephrine.

Lipopolysaccharide (LPS) administration induces hypothalamic nitric oxide (NO); NO is antipyretic in the preoptic area (POA), but its mechanism of action is uncertain. LPS also stimulates the release of preoptic norepinephrine (NE), which mediates fever onset. Because NE upregulates NO synthases and NO induces cyclooxygenase (COX)-2-dependent PGE(2), we investigated whether NO mediates the production of this central fever mediator. Conscious guinea pigs with intra-POA microdialysis probes received LPS intravenously (2 mug/kg) and, thereafter, an NO donor (SIN-1) or scavenger (carboxy-PTIO) intra-POA (20 mug/mul each, 2 mul/min, 6 h). Core temperature (T(c)) was monitored constantly; dialysate NE and PGE(2) were analyzed in 30-min collections. To verify the reported involvement of alpha(2)-adrenoceptors (AR) in PGE(2) production, clonidine (alpha(2)-AR agonist, 2 mug/mul) was microdialyzed with and without SIN-1 or carboxy-PTIO. To assess the possible involvement of oxidative NE and/or NO products in the demonstrated initially COX-2-independent POA PGE(2) increase, (+)-catechin (an antioxidant, 3 mug/mul) was microdialyzed, and POA PGE(2), and T(c) were determined. SIN-1 and carboxy-PTIO reduced and enhanced, respectively, the rises in NE, PGE(2), and T(c) produced by intravenous LPS. Similarly, they prevented and increased, respectively, the delayed elevations of PGE(2) and T(c) induced by intra-POA clonidine. (+)-Catechin prevented the LPS-induced elevation of PGE(2), but not of T(c). We conclude that the antipyretic activity of NO derives from its inhibitory modulation of the LPS-induced release of POA NE. These data also implicate free radicals in POA PGE(2) production and raise questions about its role as a central LPS fever mediator.

Preoptic norepinephrine mediates the febrile response of guinea pigs to lipopolysaccharide.

Norepinephrine (NE) microdialyzed in the preoptic area (POA) raises core temperature (T(c)) via 1) alpha(1)-adrenoceptors (AR), quickly and independently of POA PGE(2), and 2) alpha(2)-AR, after a delay and PGE(2) dependently. Since systemic lipopolysaccharide (LPS) activates the central noradrenergic system, we investigated whether preoptic NE mediates LPS fever. We injected LPS (2 microg/kg iv) in guinea pigs prepared with intra-POA microdialysis probes and determined POA cerebrospinal (CSF) NE levels. We similarly microdialyzed prazosin (alpha(1) blocker, 1 microg/microl), yohimbine (alpha(2) blocker, 1 microg/microl), SC-560 [cyclooxygenase (COX)-1 blocker, 5 microg/microl], acetaminophen (presumptive COX-1v blocker, 5 microg/microl), or MK-0663 (COX-2 blocker, 0.5 microg/microl) in other animals before intravenous LPS and measured CSF PGE(2). All of the agents were perfused at 2 microg/min for 6 h. T(c) was monitored constantly. POA NE peaked within 30 min after LPS and then returned to baseline over the next 90 min. T(c) increased within 12 min to a first peak at approximately 60 min and to a second at approximately 150 min and then declined over the following 2.5 h. POA PGE(2) followed a concurrent course. Prazosin pretreatment eliminated the first T(c) rise but not the second; PGE(2) rose normally. Yohimbine pretreatment did not affect the first T(c) rise, which continued unchanged for 6 h; the second rise, however, was absent, and PGE(2) levels did not increase. SC-560 and acetaminophen did not alter the LPS-induced PGE(2) and T(c) rises; MK-0663 prevented both the late PGE(2) and T(c) rises. These results confirm that POA NE is pivotal in the development of LPS fever.

Endotoxic fever: new concepts of its regulation suggest new approaches to its management.

Endotoxic fever is regulated by endogenous factors that provide pro- and anti-pyretic signals at different points along the febrigenic pathway, from the periphery to the brain. Current evidence indicates that the febrile response to invading Gram-negative bacteria and their products is initiated upon their arrival in the liver via the circulation and their uptake by Kupffer cells (Kc). These pathogens activate the complement cascade on contact, hence generating complement component 5a. It, in turn, very rapidly stimulates Kc to release prostaglandin (PG)E2. Pyrogenic cytokines (TNF-alpha, etc.) are produced later and are no longer considered to be the immediate triggers of fever. The Kc-generated PGE2 either (1) may be transported by the bloodstream to the ventromedial preoptic-anterior hypothalamus (POA, the locus of the temperature-regulating center), presumptively diffusing into it and acting on thermoregulatory neurons; PGE2 is thus taken to be the final, central fever mediator. Or (2) it may activate hepatic vagal afferents projecting to the medulla oblongata, thence to the POA via the ventral noradrenergic bundle. Norepinephrine consequently secreted stimulates alpha1-adrenoceptors on thermoregulatory neurons, rapidly evoking an initial rise in core temperature (Tc) not associated with any change in POA PGE2; this neural, PGE2-independent signaling pathway is quicker than the blood-borne route. Elevated POA PGE2 and a secondary Tc rise occur later, consequent to alpha2 stimulation. Endogenous counter-regulatory factors are also elaborated peripherally and centrally at different points during the course of the febrile response; they are, therefore, anti-pyretic. These multiple interacting pathways are the subject of this review.

Kupffer cell-generated PGE2 triggers the febrile response of guinea pigs to intravenously injected LPS.

Because the onset of fever induced by intravenously (i.v.) injected bacterial endotoxic lipopolysaccharides (LPS) precedes the appearance in the bloodstream of pyrogenic cytokines, the presumptive peripheral triggers of the febrile response, we have postulated previously that, in their stead, PGE2 could be the peripheral fever trigger because it appears in blood coincidentally with the initial body core temperature (Tc) rise. To test this hypothesis, we injected Salmonella enteritidis LPS (2 microg/kg body wt i.v.) into conscious guinea pigs and measured their plasma levels of LPS, PGE2, TNF-alpha, IL-1beta, and IL-6 before and 15, 30, 60, 90, and 120 min after LPS administration; Tc was monitored continuously. The animals were untreated or Kupffer cell (KC) depleted; the essential involvement of KCs in LPS fever was shown previously. LPS very promptly (<10 min) induced a rise of Tc that was temporally correlated with the elevation of plasma PGE2. KC depletion prevented the Tc and plasma PGE2 rises and slowed the clearance of LPS from the blood. TNF-alpha was not detectable in plasma until 30 min and in IL-1beta and IL-6 until 60 min after LPS injection. KC depletion did not alter the times of appearance or magnitudes of rises of these cytokines, except TNF-alpha, the maximal level of which was increased approximately twofold in the KC-depleted animals. In a follow-up experiment, PGE2 antiserum administered i.v. 10 min before LPS significantly attenuated the febrile response to LPS. Together, these results support the view that, in guinea pigs, PGE2 rather than pyrogenic cytokines is generated by KCs in immediate response to i.v. LPS and triggers the febrile response.

Cytokines, PGE2 and endotoxic fever: a re-assessment.

The innate immune system serves as the first line of host defense against the deleterious effects of invading infectious pathogens. Fever is the hallmark among the defense mechanisms evoked by the entry into the body of such pathogens. The conventional view of the steps that lead to fever production is that they begin with the biosynthesis of pyrogenic cytokines by mononuclear phagocytes stimulated by the pathogens, their release into the circulation and transport to the thermoregulatory center in the preoptic area (POA) of the anterior hypothalamus, and their induction there of cyclooxygenase (COX)-2-dependent prostaglandin (PG)E(2), the putative final mediator of the febrile response. But data accumulated over the past 5 years have gradually challenged this classical concept, due mostly to the temporal incompatibility of the newer findings with this concatenation of events. Thus, the former studies generally overlooked that the production of cytokines and the transduction of their pyrogenic signals into fever-mediating PGE(2) proceed at relatively slow rates, significantly slower certainly than the onset latency of fever produced by the i.v. injection of bacterial endotoxic lipopolysaccharides (LPS). Here, we review the conflicts between the earlier and the more recent findings and summarize new data that reconcile many of the contradictions. A unified model based on these data explicating the generation and maintenance of the febrile response is presented. It postulates that the steps in the production of LPS fever occur in the following sequence: the immediate activation by LPS of the complement (C) cascade, the stimulation by the anaphylatoxic C component C5a of Kupffer cells, their consequent, virtually instantaneous release of PGE(2), its excitation of hepatic vagal afferents, their transmission of the induced signals to the POA via the ventral noradrenergic bundle, and the activation by the thus, locally released norepinephrine (NE) of neural alpha(1)- and glial alpha(2)-adrenoceptors. The activation of the first causes an immediate, PGE(2)-independent rise in core temperature (T(c)) [the early phase of fever; an antioxidant-sensitive PGE(2) rise, however, accompanies this first phase], and of the second a delayed, PGE(2)-dependent T(c) rise [the late phase of fever]. Meanwhile-generated pyrogenic cytokines and their consequent upregulation of blood-brain barrier cells COX-2 also contribute to the latter rise. The consecutive steps that initiate the febrile response to LPS would now appear, therefore, to occur in an order different than conceived originally.

Putative antihyperpyretic factor induced by LPS in spleen of guinea pigs.

We reported previously that the onset of LPS-induced fever, irrespective of its route of administration, is temporally correlated with the appearance of LPS in the liver and that splenectomy significantly increases both the febrile response to LPS and the uptake of LPS by Kupffer cells (KC). To further evaluate the role of the spleen in LPS fever production, we ligated the splenic vein and, 7 and 30 days later, monitored the core temperature changes over 6 h after intraperitoneal (ip) injection of LPS (2 microg/kg). Both the febrile response and the uptake of LPS by KC were significantly augmented. Like splenectomy, splenic vein ligation (SVL) increased the febrile response and LPS uptake by KC until the collateral circulation developed, suggesting that the spleen may normally contribute an inhibitory factor that limits KC uptake of LPS and thus affects the febrile response. Subsequently, to verify the presence of this factor, we prepared splenic extracts from guinea pigs pretreated with LPS (8 microg/kg ip) or pyrogen-free saline, homogenized and ultrafiltered them, and injected them intravenously into splenectomized (Splex) guinea pigs pretreated with LPS (8 microg/kg ip). The results confirmed our presumption that the splenic extract from LPS-treated guinea pigs inhibits the exaggerated febrile response and the LPS uptake by the liver of Splex guinea pigs, indicating the presence of a putative splenic inhibitory factor, confirming the participation of the spleen in LPS-induced fever, and suggesting the existence of a novel antihyperpyretic mechanism. Preliminary data indicate that this factor is a lipid.

LPS-activated complement, not LPS per se, triggers the early release of PGE2 by Kupffer cells.

The intravenous injection of LPS rapidly evokes fever. We have hypothesized that its onset is mediated by prostaglandin (PG)E(2) quickly released by Kupffer cells (Kc). LPS, however, does not stimulate PGE(2) production by Kc as rapidly as it induces fever; but complement (C) activated by LPS could be the exciting agent. To test this hypothesis, we injected LPS (2 or 8 microg/kg) or cobra venom factor (CVF, an immediate activator of the C cascade that depletes its substrate, ultimately causing hypocomplementemia; 25 U/animal) into the portal vein of anesthetized guinea pigs and measured the appearance of PGE(2), TNF-alpha, IL-1beta, and IL-6 in the inferior vena cava (IVC) over the following 60 min. LPS (at both doses) and CVF induced similar rises in PGE(2) within the first 5 min after treatment; the rises in PGE(2) due to CVF returned to control in 15 min, whereas PGE(2) rises due to LPS increased further, then stabilized. LPS given 3 h after CVF to the same animals also elevated PGE(2), but after a 30- to 45-min delay. CVF per se did not alter basal PGE(2) and cytokine levels and their responses to LPS. These in vivo effects were substantiated by the in vitro responses of primary Kc from guinea pigs to C (0.116 U/ml) and LPS (200 ng/ml). These results indicate that LPS-activated C rather than LPS itself triggers the early release of PGE(2) by Kc.

Lipopolysaccharide challenge causes exaggerated fever and increased hepatic lipopolysaccharide uptake in vinblastine-induced leukopenic guinea pigs.

OBJECTIVE: To better understand the pathophysiology of the fever often manifested by immunocompromised patients undergoing chemotherapy that become neutropenic and suffer a bacterial infection. DESIGN: Prospective animal study. SETTING: A physiology laboratory in a medical school setting. MEASUREMENTS AND MAIN RESULTS: We induced leukopenia in guinea pigs with vinblastine (0.7 mg/kg, intravenously, 4 days before) and measured the animals' febrile response to 2 microg of lipopolysaccharide/kg and the uptake of 75 microg of fluorescein isothiocyanate-labeled lipopolysaccharide/kg by Kupffer cells. The leukopenic animals exhibited significantly higher fevers and greater hepatic fluorescein isothiocyanate-lipopolysaccharide uptake than their controls. CONCLUSION: Lipopolysaccharide-challenged, vinblastine-induced leukopenic guinea pigs exhibit hyperpyrexia and significantly elevated uptake of lipopolysaccharide by Kupffer cells, the major source of pyrogenic mediators. This could explain "febrile neutropenia."

Fever onset is linked to the appearance of lipopolysaccharide in the liver.

To assess the relative contributions of different phagocytes to the febrile response of guinea pigs to intravenous (i.v.) and intraperitoneal (i.p.) bacterial endotoxic lipopolysaccharide (LPS), we injected fluorescein isothiocyanate (FITC)-labeled LPS at doses of 37.5, 75, 150, 300 and 900 microg/kg, and measured its distribution and corresponding core temperature (T(c)) changes before and at 15, 30, 60, 90, and 120 min after injection. At all times, i.v. FITC-LPS appeared as granular fluorescent patches in circulating leukocytes and hepatic macrophages; its density was proportional to dose. At all doses, the density of i.v. FITC-LPS labeling decreased from its peak 15 min after injection at a rate commensurate with its dose. Intraperitoneal FITC-LPS was also present dose- and time-dependently in peritoneal macrophages, but it appeared later and accumulated more slowly except at the highest dose. Compared with i.v. FITC-LPS, its maximal appearance was always lower in density. No labeling was found at any time in brain and kidney following any dose of i.v. or i.p. FITC-LPS injection. The initiation of T(c) rises was best correlated with the presence of FITC-LPS in liver, irrespective of its route of injection. Pretreatment with gadolinium chloride 3 days before LPS injection attenuated the febrile response and reduced FITC-LPS labels in liver. These results suggest that the Kupffer cells may be central to the initiation of the febrile response of guinea pigs to i.v. and i.p. LPS.

Preoptic alpha 1- and alpha 2-noradrenergic agonists induce, respectively, PGE2-independent and PGE2-dependent hyperthermic responses in guinea pigs.

We have shown previously that norepinephrine (NE) microdialyzed into the preoptic area (POA) of conscious guinea pigs stimulates local PGE(2) release. To identify the cyclooxygenase (COX) isozyme that catalyzes the production of this PGE(2) and the adrenoceptor (AR) subtype that mediates this effect, we microdialyzed for 6 h NE, cirazoline (alpha(1)-AR agonist), and clonidine (alpha(2)-AR agonist) into the POA of conscious guinea pigs pretreated intrapreoptically (intra-POA) with SC-560 (COX-1 inhibitor) or nimesulide or MK-0663 (COX-2 inhibitors) and measured the animals' core temperature (T(c)) and intra-POA PGE(2) responses. Cirazoline induced T(c) rises promptly after the onset of its dialysis without altering PGE(2) levels. NE and clonidine caused early falls followed by late rises of T(c); intra-POA PGE(2) levels were closely correlated with this thermal course. COX-1 inhibition attenuated the clonidine-induced T(c) and PGE(2) falls but not the NE-elicited hyperthermia, but COX-2 inhibition suppressed both the clonidine- and NE-induced T(c) and PGE(2) rises. Coinfused cirazoline and clonidine reproduced the late T(c) rise of clonidine but not its early fall and also not the early rise produced by cirazoline; on the other hand, the PGE(2) responses were similar to those to NE. Prazosin (alpha(1)-AR antagonist) and yohimbine (alpha(2)-AR antagonist) blocked the effects of their respective agonists. These results indicate that alpha(1)- and alpha(2)-AR agonists microdialyzed into the POA of conscious guinea pigs evoke distinct T(c) responses: alpha(1)-AR activation produces quick, PGE(2)-independent T(c) rises, and alpha(2)-AR stimulation causes an early T(c) fall and a late, COX-2/PGE(2)-dependent T(c) rise.

Signaling the brain in systemic inflammation: the role of complement.

The complement (C) cascade is activated in almost immediate reaction to the appearance in the body of pathogenic microorganims and their products, e.g., bacterial endotoxic lipopolysaccharide (LPS), resulting in the generation of a series of potent bioactive fragments that have critical roles in the innate immune response of the afflicted host, including, potentially, the production of the fever that so characteristically marks bacterial infections. For instance, its derivatives C3a, C3b, iC3b, C5a, and C5b-9 independently induce the production by myeloid and non-myeloid cells of the cytokines interleukin (IL)-1(, IL-6 and tumor necrosis factor-(, and of prostaglandin (PG)E2, all putative mediators of fever. Therefore, any one of these C components could be involved, centrally or peripherally, in the induction of the febrile response to LPS. Indeed, we have shown that hypocomplementation by cobra venom factor (CVF) dose-dependently attenuates LPS-induced fever in guinea pigs and wild-type (WT) mice, and that C5 gene-ablated mice are unable to develop fever after LPS. In further studies, we found that a specific antagonist to the C5a receptor, C5aR1a, prevents the LPS-induced febrile rise of WT and C3 null mutant mice, implicating C5a as the responsible factor. Various lines of evidence from our laboratory suggest that the macrophages of the liver (Kupffer cells [Kc]) may be the specific target cells of C5a and that the product they release may be PGE2. PGE2, in turn, may be the substance that binds to vagal afferents in the liver that convey the pyrogenic message to the brain. Other studies by our group (not included in this review) have separately traced the neural pathway by which this message may be transmitted from the liver to the brain and processed there for action. The purpose of this article is to review the studies that have led us to conclude that C5a, Kc and Kc-generated PGE2 may be integrally involved in the pathogenesis of LPS fever. If further verified, these results will be important for better understanding how infectious stimuli may trigger the multivariate acute-phase responses generally, and fever particularly, that promptly spring into action to defend the continued well-being of the afflicted host.

Intracerebroventricular interleukin-6, macrophage inflammatory protein-1 beta and IL-18: pyrogenic and PGE(2)-mediated?

This study was undertaken to determine whether cyclooxygenase (COX)-2, the critical enzyme in the production of febrigenic prostaglandin (PG) E(2), may be involved centrally in the fever induced in mice by homologous interleukin (IL)-6, macrophage inflammatory protein (MIP)-1 beta, and interleukin (IL)-18, a member of the pyrogenic IL-1 beta family. To this end, the core temperatures (Tc) of COX-1 and COX-2 gene-ablated mice and of their normal wild-type (WT) counterparts were recorded after intracerebroventricular (i.c.v.) challenge with recombinant murine (rm) IL-6 (10 ng/mouse), rmMIP-1 beta (20 pg/mouse), rmIL-18 (0.01-1 microgram/mouse), rmIL-1 beta (positive control; 0.1 microgram/mouse), or their vehicle (0.1% bovine serum albumin [BSA] in sterile phosphate-buffered saline [PBS]; 5 microl/mouse). rmIL-6 caused a approximately 1 degrees C T(c) rise in WT mice that peaked at approximately 120 min and gradually recovered over the next 3 h; COX-1(-/-) mice exhibited a relatively faster (peak at 45 min) and shorter (recovery at 150 min) febrile course, whereas COX-2(-/-) mice did not develop fever. rmMIP-1 beta induced a 1 degrees C fever (peak at 60 min) with a long time course (recovery incomplete at 300 min) in both WT and COX-2(-/-) mice; COX-1(-/-) mice displayed a quick-onset (peak at 40 min) and shorter (recovery at approximately 240 min) fever. rmIL-18 did not cause any thermal response at any dose whether administered intraperitoneally (i.p.) or i.c.v. in WT mice; COX gene-ablated mice, therefore, were not tested. These data indicate that COX-2-dependent PGE(2) is critical for the febrile response to IL-6, but not to MIP-1 beta. IL-18 i.p. or i.c.v. is not pyrogenic.

The spleen modulates the febrile response of guinea pigs to LPS.

The febrile responses of splenectomized (Splex) or sham-operated (Sham) guinea pigs challenged intravenously or intraperitoneally with lipopolysaccharide (LPS) 7 and 30 days after surgery were evaluated. FITC-LPS uptake by Kupffer cells (KC) was additionally assessed 15, 30, and 60 min after injection. LPS at 0.05 microg/kg iv did not evoke fever in Sham animals but caused a 1.2 degrees C core temperature (T(c)) rise in the Splex animals. LPS at 2 microg/kg iv induced a 1.8 degrees C greater T(c) rise of the Splex animals than of their controls. LPS at 2 and 8 microg/kg ip 7 days postsurgery induced 1.4 and 1.8 degrees C higher fevers, respectively, in the Splex than Sham animals. LPS at 2 and 8 microg/kg ip 30 days postsurgery also increased the febrile responses of the asplenic animals by 1.6 and 1.8 degrees C, respectively. FITC-LPS at 7 days was detected in the controls within KC 15 min after its administration; the label density was reduced at 30 min and almost 0 at 60 min. In the Splex group, in contrast, the labeling was significantly denser and remained unchanged through all three time points; this effect was still present 30 days after surgery. Similar results were obtained at 60 min after FITC-LPS intraperitoneal injection. Gadolinium chloride pretreatment (-3 days) of the Splex group significantly reduced both their febrile responses to LPS (8 microg/kg ip) and their KC uptake of FITC-LPS 7 days postsurgery. Thus splenectomy increases the magnitude of the febrile response of guinea pigs and the uptake of systemically administered LPS.

Thermal response to zymosan: the differential role of complement.

OBJECTIVES: This study was designed to determine whether the complement (C) system may be involved in the febrile response to zymosan (Zym), a glycan derived from yeast cell walls. METHODS: Cobra venom factor (CVF) at 100 U/animal or its vehicle, pyrogen-free saline (PFS), was injected intravenously (i.v.) into guinea pigs to deplete serum C. Eighteen hours later, a low or high dose of Zym or its vehicle, PFS, was administered i.v. or intraperitoneally (i.p.) to these animals. The core temperature (T(c)) was measured continuously by thermocouples. Serum C levels were determined by sheep erythrocyte hemolytic assay. RESULTS: Zym at 1 mg/kg caused a 1 degrees C T(c) rise that was not significantly affected by CVF pretreatment. However, CVF-induced hypocomplementation converted the T(c) fall ( approximately 1.2 degrees C) produced by 100 mg/kg of Zym i.p. into a 1 degrees C T(c) rise. Similarly, CVF pretreatment did not affect the T(c) rise caused by 0.5 mg/kg of Zym i.v., but converted the T(c) fall induced by 25 mg/kg i.v. into a 1 degrees C T(c) rise. A separate experiment showed that 25, but not 0.5 mg/kg of Zym i.v., decreased serum C by 34% in 15 min; C did not recover over the next 6 h. A second i.v. injection of 25 mg Zym/kg 210 min later, when the T(c) had recovered but the serum C had not, yielded a smaller and briefer T(c) fall. CONCLUSION: These results suggest that Zym is inherently pyrogenic, but this effect is manifested only when the dose of zymosan is too small to activate C or when C has been reduced by prior activation.