Sylwia Wrotek

Glutathione deficiency attenuates endotoxic fever in rats

Abstract Purpose: Glutathione constitutes the first line of the cellular defence mechanism against oxidative stress, and according to published data it is required by a number of factors that are involved in fever mechanism. The aim of the present study was to investigate whether or not glutathione deficiency can modulate a course of the fever induced by endotoxin (LPS). Material and methods: Intraperitoneal injection of LPS from Escherichia coli was used to provoke fever in Wistar rats. The level of liver glutathione was decreased by administration of phorone (Pho). Deep body temperature (Tb) in free running rats was recorded using a biotelemetry system. The concentration of TNF-α was estimated. Next, the supplementation of TNF-α was done using recombinant rat TNF-α. Results: Animals with decreased glutathione level responded with diminished fever after LPS injection (average Tb in Pho/LPS-treated and oil/LPS-treated animals were 36.90 ° ± 0.10 °C and 37.80 ° ± 0.15 °C, respectively). This response was accompanied by a significant attenuation of LPS-induced increase in TNF-α concentration (in the Pho/LPS-treated group it was 10.68 pg/mL ± 2.24, vs. 113.35 pg/mL ± 13.93 in oil/LPS-treated rats). Supplementation with TNF-α partially restored fever. Conclusion: Based on these data, we conclude that glutathione deficiency modifies the LPS-induced fever, in a TNF-α related manner.

Silver nanoparticles augment releasing of pyrogenic factors by blood cells stimulated with LPS

Silver nanoparticles (AgNPs) have cytotoxic properties via generation of reactive oxygen species which are involved in the generalized sickness behavior of the host, including fever and lethargy among others. The aim of the present study was to investigate the impact of AgNPs on the ability of rat peripheral blood mononuclear cells (PBMCs) to release fever mediating factors after stimulation with lipopolysaccharide (LPS). Body temperature and motor activity of the Wistar rats were measured by biotelemetry system. Rat PBMCs were stimulated with LPS and after that the cells were washed and incubated alone or with AgNPs. The final supernatants were injected intraperitoneally. The levels of endogenous pyrogens such as interleukin-1β (IL−1β), IL-6 and tumor necrosis factor-α (TNF-α) released from the PBMCs into the final supernatants were also estimated. The results indicated that injection of the supernatants from the cells stimulated with LPS induced fever and inhibited motor activity. These effects were potentiated by the presence of AgNPs during the final incubation. The presence of the AgNPs also resulted in significant increases in levels of endogenous pyrogens. The augmentation of fever in the rats by the AgNPs treatment of the cultures seemed to be primarily associated with the changes in interleukin-1β levels.

Polysaccharide peptide from Coriolus versicolor induces interleukin 6-related extension of endotoxin fever in rats

Abstract Purpose: Polysaccharide peptide (PSP) extracted from the Coriolus versicolor mushroom is frequently suggested as an adjunct to the chemo- or radiotherapy in cancer patients. In a previous study we showed that PSP induced a tumour necrosis factor-α (TNF-α)-dependent anapyrexia-like response in rats. Thus, PSP appears to be a factor which modifies a number of pathophysiological responses. Because of this, PSP is suggested as a potential adjuvant for cancer therapy during which cancer patients frequently contract microbial infections accompanied by fever. The aim of the present study was to investigate whether or not PSP can modulate the course of the fever in response to an antigen such as lipopolysaccharide (LPS). Materials and methods: Body temperature (Tb) of male Wistar rats was measured by biotelemetry. PSP was injected intraperitoneally (i.p.) at a dose of 100 mg kg−1, 2 h before LPS administration (50 µg kg−1, i.p.). The levels of interleukin (IL)-6 and TNF-α in the plasma of rats were estimated 3 h and 14 h post-injection of PSP using a standard sandwich ELISA kit. Results: We report that i.p. pre-injection of PSP 2 h before LPS administration expanded the duration of endotoxin fever in rats. This phenomenon was accompanied by a significant elevation of the blood IL-6 level of rats both 3 h and 14 h post-injection of PSP. Pre-treatment i.p. of the rats with anti-IL-6 antibody (30 µg/rat) prevented the PSP-induced prolongation of endotoxin fever. Conclusions: Based on these data, we conclude that PSP modifies the LPS-induced fever in IL-6-related fashion.

N-Acetyl-l-cysteine exacerbates generation of IL-10 in cells stimulated with endotoxin in vitro and produces antipyresis via IL-10 dependent pathway in vivo

N-Acetyl-l-cysteine (NAC) is a well-known medication, primarily used as a mucolytic agent in pulmonary disease. Recently, we have found that NAC possesses antipyretic properties. The aim of the present study was to investigate the mechanism by which NAC attenuates fever. The concentration of interleukin (IL)-10 and prostaglandin (PG) E2 were measured using ELISA kit in the supernatants aspirated after stimulation of peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS, 1μg/mL) and NAC (10mM). The body temperature of the Wistar rats was measured using biotelemetry system. To inhibit endotoxic fever, NAC (200mg/kg; i.p.) was injected into the rats one hour prior to the LPS administration (50μg/kg; i.p.). The pre-treatment of LPS-stimulated PBMCs with NAC resulted in a significant decrease in PGE2 concentration in comparison to the cells treated with LPS alone (PGE2 level was 386.1±61.9pg/mL vs. 2078.9±157.9pg/mL, respectively, p<0.001). Furthermore, in these cells we observed a significant increase in IL-10 level (142.1±2.62pg/mL in NAC+LPS stimulated cells vs. 54.4±0.6pg/mL in LPS stimulated cells, p<0.001). The injection of anti-IL-10 antibody into the rats abolished antipyretic properties of NAC. Body temperature in animals treated with anti-IL-10+NAC/LPS was 38.28±0.12°C vs. 37.73±0.06°C in IgG+NAC/LPS rats (p<0.001) and 38.31±0.20°C in NaCl/LPS-treated animals (n.s.). Based on these data, we conclude that NAC acts as an antipyretic via IL-10 stimulation. This finding provides a new insight into the immunopharmacology of NAC, and we believe that in a future it will contribute to the new and/or more accurate application of NAC in medicine.

Buthionine sulfoximine, a glutathione depletor, attenuates endotoxic fever and reduces IL-1β and IL-6 level in rats

Purpose The aim of our study was to investigate the effect of buthionine sulfoximine (BSO) ‐ a glutathione depletor ‐ on a course of endotoxic fever and IL‐1&bgr; and IL‐6 production. Material and methods Male Wistar rats were subjected to intraperitoneal injection of lipopolysaccharide (LPS) from E. coli (50 &mgr;g/kg, ip) to provoke fever. The level of spleen glutathione, plasma interleukin (IL)‐1&bgr;, IL‐6, and deep body temperature (Tb) were measured. Results The LPS administration provoked fever (the average Tb was 38.14 ± 0.05 °C in NaCl/LPS‐treated rats vs 37.10 ± 0.03 °C in control, not‐treated rats; p < 0.001). We observed that LPS injection induced a decrease in spleen glutathione level (7.67 ± 0.92 nM/g vs 13.27 ± 0.47 nM/g in not‐treated rats; p < 0.001). Furthermore, the injection of LPS provoked an elevation of plasma IL‐1&bgr; and IL‐6 concentration (from values below the lowest detectable standard in not‐treated animals to 199.99 ± 34.89 pg/mL and 7500 ± 542.21 pg/mL, respectively; p < 0.001). Pretreatment with BSO enhanced glutathione decrease in LPS‐treated rats (5.05 ± 0.49 nM/g), and significantly affected fever (maximal Tb was 37.81 ± 0.07°C in BSO/LPS‐treated rats vs 38.76 ± 0.11 °C in NaCl/LPS‐treated rats). BSO 4 h after LPS injection decreased IL‐1&bgr; and IL‐6 gene expression (about 1.5 fold, and 2 fold, respectively). In a consequence we observed a decrease in plasma IL‐6 concentration (4 h after LPS injection plasma IL‐6 was 4167.17 ± 956.54 pg/mL in BSO/LPS‐treated rats vs 7500 ± 542.21 pg/mL in NaCl/LPS‐treated rats; p < 0.001), and later IL‐1&bgr; (7 h after LPS injection the IL‐1&bgr; concentration was not detected). Conclusion Based on these data, we conclude that BSO, in addition to well‐known application as an inhibitor of glutathione synthesis, is an antipyretic agent which reduces both IL‐1&bgr; and IL‐6 concentration. HighlightsButhionine sulfoximine (BSO) decreases spleen glutathione level.BSO inhibits endotoxic fever.BSO attenuates expression of LPS‐stimulated IL‐1&bgr; and IL‐6 genes.BSO decreases plasma level of IL‐1&bgr; and IL‐6.

Analysis of the involvement of cytokines in allergy and breast cancer association

Aim of the study The existence of a correlation between allergy disorders and cancer diseases has been confirmed by several epidemiological studies. Although the molecular mechanism involved in this phenomenon remains unknown, there are data indicating that certain cytokines, engaged in allergic processes, have antineoplastic activities. The aim of the present study was to explore the association between advanced breast cancer and allergic state on the molecular level. Material and methods We determined and compared the mRNA and protein expression of interleukin-1β (IL-1β), IL-4, IL-6, and interferon-γ (IFN-γ), cytokines known for antitumor properties, in the blood of advanced breast cancer patients and individuals with allergic diseases related to type 2 response. In addition, we performed an in vitro assay of reactivity of peripheral blood mononuclear cells after exogenous antigen stimulation. As a preliminary to molecular analysis we conducted a questionnaire study concerning the incidence of allergy among breast cancer patients and healthy subjects without malignancy. Results The results of the survey study revealed a negative relation between breast cancer and allergy prevalence. Subsequent molecular analysis, however, did not show statistically significant differences in cytokines mRNA and protein expression levels between allergic patients and those with malignancy. The in vitro reactivity test also did not reveal marked differences between IL-1β, IL-4 and IL-6 production after PBMC triggering with exogenous antigen. Conclusions We concluded that the studied cytokines (IL-1β, IL-4, IL-6, and IFN-γ) are not engaged in breast cancer-allergy negative relation.

Thermal and motor behavior in experimental autoimmune encephalitis in Lewis rats

Abstract Thermoregulation in patients, who suffer from multiple sclerosis (MS) is impaired and may result in either increases or decreases in body temperature. Disturbances in body temperature correlate with acute relapses, and for this reason, it is an important issue in everyday life of those who suffer from MS. Although rat experimental autoimmune encephalitis (EAE) appeared useful for the examination of current therapies against MS, it has not been thoroughly investigated in terms of body temperature. The purpose of this study was to examine the effect of EAE induction on thermal and motor behavior in the rats. Subcutaneous injection of encephalitogenic emulsion into both pads of hind feet of the Lewis rats provoked symptoms of EAE. Body temperature (Tb) and motor activity of rats were measured using biotelemetry system. We report a significant increase in body temperature within 24 h prior to the EAE manifestation (12 h average of Tb for EAE induced animals was higher by 1.07 ± 0.06 °C during day-time and by 0.5 ± 0.05 °C during night time in comparison to the control rats). On the other hand, the onset of EAE symptoms was associated with gradual decrease of body temperature, and during the first night-time Tb was lower by 1.03 ± 0.08 °C in comparison to the control rats. The inhibition of the motor activity started from the night time, 2 days before EAE onset. On the basis of our data, we concluded that the pattern of body temperature changes after EAE induction may be considered as useful symptom (prodrom) to predict precisely the time of EAE onset. Furthermore, we suggest that EAE in rats may be a suitable model to study mechanism of body temperature alternations observed in MS patients.

Toward Antitumor Immunity and Febrile Infections: Gamma/Delta (γδ) T Cells Hypothesis

Retrospective as well as prospective clinical studies indicate that episodes of high fever as a typical reaction to an acute infection during the entire human life span are inversely related to cancer incidence. Laboratory data aimed at discovering why fever incidents appear beneficial in lowering the risk of cancer remain rudimentary at best. Several hypotheses have been presented thus far, and recent debates have pointed to the effect of fever on innate and adaptive immune functions. In this paper we focus on a particular mode of adaptive immune functioning that involves a type of T cells carrying a receptor composed of gamma/delta chain heterodimer (γδ T-cell receptor; γδ TCR). We present an argument for a key role of infectious fever in the recruitment and enhancement of immune antitumor competence of γδ T cells during the life span of a human being. The unique physiology of γδ T lymphocytes indicating a possible cellular link between innate and adaptive immunity—including modes of antigen recognition as well their presence in tissues prone to infections, metabolic stress, and neoplastic development—makes them a target for exploration in the context of fever and cancer risk, and for future cancer immunotherapy.

Fever as a factor contributing to long-term survival in a patient with metastatic melanoma: A case report

BACKGROUND Malignant melanoma is a cancer that arises from pigment cells in the skin called melanocytes. The long-term survival of a patient with advanced melanoma is rare. CASE We present a unique case of a female patient who has suffered from malignant melanoma for more than 13 years. The disease progressed quickly, and 19 months after diagnosis, the patient was classified as having stage IV melanoma. After several years, the patient had several episodes of fever that were not deliberately treated with medication. After each episode of fever, the patient observed the disappearance of tumours, which was confirmed by medical examination. Interestingly, since her initial diagnosis, the patient has refused most of the proposed medical treatments. Consequently, only some of the surgical procedures were performed. Currently, despite the initially poor prognosis, the patient only suffers symptoms that are the result of surgical resection of brain metastases. Most of her malignant tumours either disappeared or have stabilized without further growth. CONCLUSIONS The onset of fever has altered the typical and unfavourable course of melanoma, causing remission or at least stabilization. This observation, in accordance with others in this field, suggests that fever in cancer patients should not be treated immediately, but should be allowed to develop under the care of a physician.

Axonal regeneration inhibitors: emerging therapeutic options

For the most part, the central nervous system is unable to regenerate. The majority of injuries of vascular, inflammatory, degenerative and traumatic aetiology lead to an irreversible loss of central nervous system function. The paper presents the role of Nogo-A, MAG and OMgp proteins in the inhibition of central nervous system regeneration, and their potential clinical significance.