Interrelationships between interleukin (IL)-1, IL-6 and IL-8 in synovial fluid of various arthropathies

High levels of many cytokines, including interleukin (IL)-1, IL-6 and IL-8, were found in various arthropathies suggesting that they play a role in the pathogenesis of disease, although their relationship with the type and activity of disease is still not clear. The synovial fluid (SF) of 24 patients with rheumatoid arthritis (RA), 19 with psoriatic arthritis (PA) and 33 with osteoarthritis (OA) was analyzed for IL-1, IL-6 and IL-8. The highest concentration of the three cytokines was found in the SF of RA. IL- detectable levels (>-20 pg/ml) were observed in 8/24 (33.3%) patients with RA, in one patient with PA but in no patient with OA.IL-6 (mean±SD) (1610.37±1781.65 pg/ml) was higher in RA than in PA (672.47±867.40 pg/ml,p=0.043) and OA (89.45±120.52 pg/ml,p=0.0001). IL-8 (1042.72±698.64 pg/ml) was higher in RA than in PA (660.36±625.11 pg/ml,p=0.03) and OA (89.9±45.88 pg/ml,p=0.0001). A correlation between IL-1, IL-6 and IL-8 was found in RA. In all patients a correlation between IL-6 and IL-8 levels was found; moreover, these two cytokines were associated with SF indices of inflammation, such as white blood cells (WBC) count and total protein (TP) concentration.Out findings suggest that these interrelationships play a role in the evolution of more severe erosive arthropathy such as RA.     

Peptide inhibitor of interleukin-8 (IL-8) reduces staphylococcal enterotoxin-A (SEA) induced neutrophil trafficking to the lung

Staphylococcal enterotoxin A (SEA) is a superantigen, produced by some strains ofStaphylococcus aureus (S. aureus), which can cause a variety of clinical manifestations ranging from food poisoning to shock. SEA can also stimulate human alveolar macrophages to produce interleukin-8 (IL-8), a member of the -chemokine subfamily that activates and is chemotactic for neutrophils. In these studies we showed that in rabbits, intravenous SEA significantly decreased the circulating white blood cell population from a baseline value of 6409±2027×10³ cells/ml to 1943±862×10³ cells/ml in 7 h. There was a concommitent increase in IL-8 in the circulating plasma (baseline: 60±34 pg/ml, 7 h post SEA: 109±64 pg/ml). The increase in circulating IL-8 was accompanied by a much greater increase in the IL-8 concentration of the epithelial lining fluid (ELF) where the IL-8 increased from 0.05±0.08 ng/ml (control) to 13.8±9.3 ng/ml (SEA treated). The increase in IL-8 concentration in the alveolar spaces was paralleled by an increase in both the percentage of neutrophils (1.4±0.9% (control) to 26.0±10.8% (SEA treated)) and total number of neutrophils (0.04±0.02×10⁶/ml (control) to 4.8±3.3 10⁶/ml (SEA treated)) in the airspaces, and the numbers of neutrophils in the ELF correlated with the IL-8 concentration r=0.62, p=0.006). When antileukinate, a hexapeptide which inhibits the binding of IL-8 to neutrophils, was administered to animals receiving SEA, the IL-8 concentration in the ELF (14.8±10.7 ng/ml) was not significantly different from the concentration of IL-8 in those animals receiving SEA alone). However, both the percentage of neutrophils (9.5±3.2%), and the total number of neutrophils (1.3±1.0×10⁶/ml) in the ELF following SEA and antileukinate administration was significantly lower than in animals which only received SEA (p<0.05). The findings suggest that SEA released into the circulation during a Staphylococcal infection can cause an inflammatory reaction in the lung. Since this reaction is at least partially mediated by IL-8, antileukinate may have pharmacologic potential in reducing the inflammatory reaction.accepted by I. Ahnfelt-Rønne     

Endocrinology: Ovarian hyperstimulation syndrome: pre-ovulatory serum concentrations of interleukin-6, interleukin-1 receptor antagonist and tumour necrosis factor-{alpha} cannot predict its occurrence

The pathogenesis of the ovarian hyperstimulation syndrome (OHSS)is poorly understood. Since significant elevations in cytokinesare found in 01155, our objective was to conduct a prospectivecase-controlled study to assess if preovulatory cytokine serumconcentrations can predict its occurrence. The study group wasselected from in-vitro fertilization patients who subsequentlydeveloped severe OHSS, along with a matched group who did notdevelop this complication (n = 20), and a healthy normal controlgroup (n = 10). Interleukin-6 (IL-6), interleukin-1 receptorantagonist (IL-1RA) and tumour necrosis factor- (TNF) measurementswere performed with sensitive immune-assays and confirmed withbioassays. Serum IL-6 (mean concentration ± SEM: 4.38± 0.36 pg/ml), IL-1RA (829 ± 292 pg/ml) and TNF(15.5 ± 132 pg/ml) concentrations did not show differencesthroughout the normal menstrual cycle group. Cytokine variabilityand pre-ovulatory values were similar in OHSS compared to controlledovarian hyperstiinulation (COH) patients. However, average follicularphase serum 1L-6 concentrations were higher in OHSS (8.71 ±0.41 pg/ml) and COH (7.66 ± 0.38 pg/ml) patients thanin normally menstruating women (4.34 ± 0.99 pg/ml) (P< 0.0001). Pre-ovulatory serum 1L-6 concentrations were alsohigher in OHSS (9 ± 0.94 pg/ml) and COH (73 ±0.97 pg/ml) patients than in controls (4.57 ± 1.1 pg/ml)(P < 0.01 and P < 0.04 respectively). IL-1RA and TNF concentrationswere comparable in all the groups. This study suggests thatcytokine measurements cannot be used to predict the occurrenceof OHSS prior to the administration of human chorionic gonadotrophin.     

Clinical significance of serum IL-18 determination in rheumatoid arthritis

Interleukin (IL)-18 is a novel cytokine expressing at inflammatory lesion. In this study, to evaluate the clinical significance of IL-18 determination, we have examined serum IL-18, inflammation markers, sFas, and sFas-ligand concentrations in patients with rheumatoid arthritis(RA). Serum was obtained from 56 RA patients aged 35-74 years, 16 osteoarthritis (OA) patients aged 36-78 years and 178 healthy subjects aged 20-72 years and IL-18 was measured by ELISA. Serum IL-18 concentrations in RA (240.1 +/- 15.6 pg/ml, mean +/- SE) were significantly higher than OA (151.8 +/- 12.7 pg/ml, p < 0.005) and healthy controls(141.5 +/- 26.1 pg/ml, p < 0.001). Serum IL-18 levels were significantly increased in II to IV stages of RA (stage II; 218.6 +/- 31.2 pg/ml, stage III; 258.7 +/- 38.4 pg/ml, stage IV; 231.6 +/- 13.1 pg/ml) than those in OA. Furthermore, a positive correlation was observed between serum IL-18 concentration and serum Fas level in patients with RA (r = 0.472), whereas there was no significant correlation between serum IL-18 and sFas-ligand or other inflammatory markers (CRP, RF, CA-RF, IL-6, and IL-8). The present study showed that serum IL-18 level increased in RA, but it is unknown how IL-18 is involved in the pathogenesis of RA. Further study will be necessary to clarify the role of IL-18 in RA.     

白细胞介素-37在类风湿性关节炎患者血清表达水平的研究

目的 通过检测类风湿性关节炎(RA)患者血清中细胞因子白细胞介素(IL)-37、肿瘤坏死因子-α(TNF-α)、IL-18、炎性指标红细胞沉降率以及C-反应蛋白(C-reactionprotein,CRP)水平,观察RA患者临床数据包括压痛关节数,肿胀关节数以及DAS28评分等,探讨RA患者血清IL-37水平升高的意义以其在RA发病机制中可能的作用.方法 80例RA患者、80例健康对照患者,通过酶联免疫吸附试验(ELISA)法检测其血清中细胞因子水平.结果 RA患者血清中IL-37[(40.33±11.25)pg/mL]、TNF-α[(110.41 ±35.37) pg/mL]、IL-18[(121.73±29.22) pg/mL]水平以及红细胞沉降率(ESR)[(42.31±15.02) mm./h]、CRP[(38.31±17.22) mg/L]水平明显高于对照组IL-37[(18.21±5.72) pg/mL]、TNF-α[(30.19±6.82) pg/mL]、IL-18[(55.47±7.29) pg/mL]水平,组间差异有统计学意义(P＜0.05).IL-37的表达与TNF-α、IL-18水平呈正相关(相关系数r=0.981,P=0.001).结论 IL-37在RA患者体内高表达并与其它几种炎性因子的表达具有相关性,IL-37可能作为炎性抑制因子参与了RA的发生、发展.     

Effects of histamine andα-fluoromethylhistidine on brain tumor necrosis factor levels in rats

Besides the role of histamine (HA) as a neurotransmitter, a new concept has emerged presenting HA as an immunomodulator. Several studies have demonstrated interactions among HA, interleukin-1 (IL-1) and tumor necrosis factor (TNF), suggesting a possible bidirectional communication among them. In this study we have investigated the effects of i.p. administrations of HA diphosphate (6 µmol/kg) and-fluoromethylhistidine (FMH; 100 mg/kg) on TNF- levels in the hippocampus, hypothalamus, and posterior hypothalamic region of the rat brain. The concentrations of TNF- at 0 (Control, C) and 30 min after i.p. administration of HA were 0.26 ± 0.02 pg/mg and 0.32 ± 0.02 pg/mg in the hippocampus, 0.46 ± 0.04 pg/mg and 0.09 ± 0.006 pg/mg (p < 0.01) in the hypothalamus, and 0.47 ± 0.05 pg/mg and 0.26 ± 0.05 pg/mg in the posterior hypothalamic region. Three hours after FMH administration, an increase in the hippocampal levels of TNF- was observed (0.43 ± 0.04 pg/mg; p < 0.01), while in the hypothalamus (0.11 ± 0.02 pg/mg; p < 0.01) and in the posterior hypothalamic region (0.21 ± 0.04 pg/mg; p < 0.05) a decrease in TNF- levels was detected. These results suggest that changes in the histaminergic system influence TNF- production in the brain in an area-specific fashion.     

Elevated TNF receptor plasma concentrations in patients with rheumatoid arthritis

Summary Two types of tumor necrosis factor receptors have been characterized, both capable of transmitting the signal and exerting the biological functions of TNF and lymphotoxin. We measured the plasma concentrations of two types of TNF binding proteins (sTNFR-A and sTNFR-B) in patients with rheumatoid arthritis (RA) and spondylarthropathies (SpA) using an enzyme-linked binding assay. In normal controls (n = 43), mean plasma concentrations were 1030 ± 55 and 1461 ±59 pg/ml for sTNFR types A and B, respectively. In 67 patients with moderate RA, mean levels were 1422 ± 82 pg/ml (type A) and 2088 ± 109 pg/ml (type B); in 34 patients with severe RA, 2588 ±279 pg/ml and 4494 ± 550 pg/ml, respectively, were measured (P < 0.0001 compared to normal controls). Concentrations of both type A and type B sTNFR were highly correlated in severe RA (R² = 0.7) but not in SpA or normal controls. T lymphocytes in synovial fluid of patients with RA expressed predominantly type A TNF receptors on their surface; in some patients a weaker expression of type B receptors was also detectable. Soluble TNF binding proteins in patients with RA were able to neutralize TNF in a cytotoxiity assay, demonstrating their ability to act as TNF-inhibiting factors. We conclude that both types of TNF receptors are parameters of disease activity in RA and may also act as TNF antagonists.Abbreviations TNF tumor necrosis factor - TNFR tumor necrosis factor receptor - sTNFR soluble tumor necrosis factor receptor - RA rheumatoid arthritis - SpA Spondylarthropathy - ELIBA Enzyme linked binding assay - mAb monoclonal antibody     

Schistosome larvae stimulate macrophage cytokine production through TLR4-dependent and -independent pathways

Exposure of the mammalian host to infective larvae of Schistosomamansoni causes an acute inflammatory response in the skin andthe activation of several cell types of the innate immune responseincluding macrophages. Using an in vitro model of macrophageactivation, we show that schistosome larvae possess moleculesthat directly stimulate both thioglycollate-elicited macrophages(tM) and IFN-activated tM in vitro to produce several cytokinesincluding IL-6, IL-12p40 and IL-10. The parasite-derived moleculesare enriched within the material released by the parasite followingtransformation [0- to 3-h released larval preparation (0-3hRP)]but not within soluble preparations of whole larvae. Cytokineproduction was maintained in the presence of polymyxin B, confirmingthat contaminating endotoxin was not responsible. IL-12p40 andIL-10 production was much lower by cells from C3H/HeJ mice,which have defective Toll-like receptor 4 (TLR4), but IL-6 productionwas unaffected. Experiments using TLR4^–/– mice confirmedthat IL-12p40 production by tM in response to 0-3hRP was partlydependent upon functional TLR4, whereas IL-6 production wasentirely independent. In contrast, tM from MyD88^–/–mice failed to secrete either IL-12p40 or IL-6, underlininga pivotal role of TLR signalling in cytokine production by macrophagesin response to stimulation with 0-3hRP. Finally, we show thatglycan components of 0-3hRP are required for optimal cytokineproduction since protease treatment of 0-3hRP had no effecton IL-12p40 production and only a slight effect on IL-6, whilesodium meta-periodate treatment almost completely abolishedproduction of both cytokines.     

Evaluation of {beta}-endorphin and interleukin-6 in seminal plasma of patients with certain andrological diseases

Human semen contains large amounts of opioid peptides and cytokines.We have measured the concentrations of interleukin (IL)-6 in140 semen samples and of -endorphinin 77 semen samples. Themedian concentration of endorphinin seminal plasma from normozoospermicmen(n = 23) was 154.7 pg/ml (10th—90th percentiles, 42.0—774.6),and there was no significant difference in the -endorphin concentrationamong normozoospermic, oligozoospermic (n= 28), asthenozoospermic(n= 15), azoospermic(n= 4) and post-vasectomy (n= 7) samples.There was no correlation between -endorphin concentration andsperm characteristics, nor with blood hormones. Endorphinconcentration was lower in cases with immunelogical infertility,as revealed by a positive direct mixedantiglobulin reactiontest (n = 12) ( > 0.01), than inmatched controls. The medianconcentration of IL-6 insamples with normal sperm concentration,motility andmorphology with or without white blood cells (n=39) was 26.1 pg/ml (10th–90th percentiles, 7.3–172.3),and there was no significant difference in the IL-6 concentrationamong normozoospermic, oligozoospermic (n= 46),asthenozoospermic(n= 32), azoospermic (n= 13) and post-vasectomy (n= 10) samples.The IL-6 concentration was significantly higher in cases ofvaricocele (n= 22)without white blood cells in semen (P <0.001) than in matched controls without varicocele (n= 23).In addition, the IL-6 concentration was elevated (P < 0.0001)in cases with accessory sex gland inflammation (n= 40). IL-6concentration was positively correlated with white blood cellsin semen (n= 60, r = 0.59, P < 0.0001), but there was nocorrelation with -endorphin concentration. The IL-6 concentrationchosen to differentiate between cases with and without accessorygland inflammation was 45.3 pg/ml, with a specificity of 80.6%and a sensitivity of 92.5%. It is concluded that -endorphinin seminal plasma playsan immune suppressive role, and thatincreased IL-6 concentration may be related to testicular dysfunctionincases with varicocele. Furthermore, IL-6 is an accurate markerof accessory sex gland inflammation.     

Elevated serum interleukin-6 levels in patients with acute hepatitis

To study the mechanisms of hepatocyte injury, we examined serum interleukin-6 (IL-6) level in acute hepatitis patients. Based on their clinical features, these patients were divided into three groups, acute hepatitis (AH), severe acute hepatitis, and fulminant hepatic failure (FHF). The present study demonstrated that, in association with their clinical status, their serum IL-6 levels were gradually increased (16.5±14.5 pg/ml in AH, 26.3±19.0 pg/ml in severe AH, and 470.2±261.4 pg/ml in FHF; control level, 5.2±0.6 pg/ml). Furthermore, we found that a significant correlation between serum IL-6 level and prothrombin time existed in these patients and that the elevated serum IL-6 returned to a normal range after recovery from their hepatocyte injury. Thus, our study demonstrates that the serum IL-6 level is a possible marker for identifying the clinical status in acute hepatitis and that this cytokine may have some roles in hepatocyte injury.     

Which is more important after total knee arthroplasty: Local inflammatory response or systemic inflammatory response?

We aimed to determine the relationship between functional recovery after knee arthroplasty and systemic and local inflammatory responses. A prospective, clinical study of thirty patients who had osteoarthritis was conducted. After the total knee arthroplasty (TKA), intraarticular IL-6 levels, serum IL-6 levels and serum CRP levels were measured. The primary outcome measures for functional recovery after TKA were the the Knee Society Score (KSS) and Western Ontario and McMaster Universities Index (WOMAC). All patients were examined preoperatively and at 4, 8 and 24 weeks postoperatively. The mean postoperative intraarticular IL-6 level was 218355.1 pg/ml, the mean postoperative serum CRP level was 109.9 mg/L and the mean postoperative serum IL-6 level was 219.0 pg/ml. Preoperative and 4-, 8- and 24-week postoperative KSS and WOMAC scores were evaluated. Significant correlations were found between intraarticular IL-6 concentrations and KSS and WOMAC scores at the first month according to the Pearson correlation test, but no correlations were found between serum IL-6 and CRP levels and KSS and WOMAC scores. The local inflammatory response is more important than the systemic response for early postoperative functional recovery. After TKA, control of local inflammation is much more important than control of systemic inflammation.     

Physical activity decreases the number of β-adrenergic receptors on human lymphocytes

Summary On intact human lymphocytes a specific binding site (BS) for¹²⁵I-Cyanopindolol (¹²⁵I-CYP), a derivative of the -blocking drug pindolol, was characterized. Inhibition of binding for catecholamines in the following order of potency: l-isoprenaline > l-adrenaline > l-noradrenaline proves the BS as a ₂-receptor subtype. In 77 healthy persons (36 females, 41 males) the number of BS amounted to 2,639±125 BS/cell without any significant correlation to age (17–86 years) or sex. The dissociation constant (K_D) indicating the affinity of iodocyanopindolol to the BS on intact lymphocytes was K _d =1.9±1.1×10^–10 M. A change of the number of -adrenergic receptors on intact human lymphocytes has been measured previously in asthmatics and in the myocardium of patients with congestive heart failure. We investigated a possible change in the -receptors on lymphocytes by physical and mental activity (stress) in physicians going about their daily routine work. Persons left alone in a hospital room reading or sleeping were defined as inactive controls. The number of BS on intact lymphocytes was significantly higher in inactive persons at 8 a.m. (2,230±482 BS) compared to active persons (1,743±285 BS;P<0.05) and at 1 p.m. (2,394±253 BS vs 1,733±556;P<0.05) but not different at 6 p.m. (1,634±578 BS vs 1,768±588 BS;P<0.1). The K _d remained unchanged under all conditions. The serum noradrenaline and adrenaline levels were also measured during the day. The serum adrenaline levels were higher in the active group than in the inactive group at 8 a.m. (46±11 pg/ml vs 22±8 pg/ml;P<0.01) and at 1 p.m. (36±13 pg/ml vs 13±5 pg/ml;P<0.01) but not at 6 p.m. (37±28 pg/ml vs 13±6 pg/ml;P<0.1). Our data clearly show higher serum adrenaline levels but fewer BS on intact human lymphocytes in active persons than in inactive persons at 8 a.m. and 1 p.m. The high catecholamine serum levels may cause a down-regulation of the number of -receptors in the active persons.Abbreviations ¹²⁵I-CYP ¹²⁵Iodocyanopindolol - BS binding site - K _d dissociation constant - cAMP cyclic adenosine monophosphate With support of Deutsche Forschungsgemeinschaft Kr 679/3-I     

Immune-regulating Th1- and Th2-cytokines in chronic infections caused by hepatitis B and C viruses

The serum levels of Th1 (gamma-IFN and sIL-2r) and of Th2 (IL-10) cytokines were measured in 33 patients (23 males and 10 females, mean age 23.1 +/- 1.9) with chronic viral hepatitis (CVH) according to a disease etiology (6 patients with hepatitis B--CVHB, 15 patients with hepatitis C--CVHC, and 12 patients with a mixed form of chronic hepatitis B and C--HBV + HCV). Besides, the contents of the studied cytokines were compared with the traditional infection markers and the presence of viremia. The similar indices taken from 10 healthy persons served as controls. The concentration of gamma-IFN was found to be reliably higher (p < 0.05) in patients of all three groups (0.32 +/- 0.07, 0.34 +/- 0.09 and 0.25 +/- 0.06 pg/ml, respectively) regardless of a disease etiology and as compared with the control value (0.09 +/- 0.04) pg/ml). At the same time, the levels of gamma-IFN, sIL-2r and IL-10 (0.25 +/- 0.06 pg/ml, 166.5 +/- 31.3 IU/ml and 48.1 +/- 8.4 pg/ml, respectively) was found to be reliably (p < 0.05 and p < 0.01) higher, as compared to the controls (0.09 +/- 0.04, 57.1 +/- 5.6 and 10.8 +/- 7.8, respectively), only in patients with the mixed infection of hepatitis. Like in our previous study, a trend was established towards the growing mean values of the IL-r level from its lowest parameters in the group of CVHB patients towards its highest parameters in the group with the mixed hepatitis form. According to our data, the IL-2r level correlated reliably with the activity of AlAt (r = 0.452; p < 0.05), while the gamma-IFN content correlated reliably with the IL-10 concentration (r = 0.805; p < 0.05), and the gamma-IFN content correlated with the IL-10 concentration (r = 0.805; p < 0.01) irrespective of disease pathology.     

Elevated Serum Levels of IL-21 in Kawasaki Disease

Purpose

The serum level of immunoglobulin (Ig)E has been reported to be elevated in patients with Kawasaki disease (KD). We investigated whether interleukin (IL)-21, rather than IL-4, could be related to elevated serum levels of IgE in KD.

Methods

Sera from 48 patients with KD and 12 controls with high fever were collected to determine the level of IgE using an immunoassay system and the levels of IL-4 and IL-21 were determined using enzyme-linked immunosorbent assay kits.

Results

The median IL-21 level of KD patients was significantly elevated, at 499.5 pg/mL (range: <62.5-1,544 pg/mL), whereas that of controls was <62.5 pg/mL (<62.5-825 pg/mL; P<0.001). The median IL-4 level of KD patients was not elevated (4.0 pg/mL; 2.1-7.6 pg/mL). The median level of total IgE in KD patients was 58.0 IU/mL (5-1,109 IU/mL). No statistically significant correlation was found between IL-21 and total IgE levels (Spearman''s R=0.2; P=0.19).

Conclusions

Patients with KD have elevated levels of IL-21 in the serum. IL-21 may play a role in the pathogenesis of KD.     

Serum interleukin-17 levels are associated with nephritis in childhood-onset systemic lupus erythematosus

OBJECTIVES:

To determine the serum interleukin-17 (IL-17) levels in childhood-onset systemic lupus erythematosus patients and to evaluate the association between IL-17 and clinical manifestations, disease activity, laboratory findings and treatment.

METHODS:

We included 67 consecutive childhood-onset systemic lupus erythematosus patients [61 women; median age 18 years (range 11-31)], 55 first-degree relatives [50 women; median age 40 years (range 29-52)] and 47 age- and sex-matched healthy controls [42 women; median age 19 years (range 6-30)]. The childhood-onset systemic lupus erythematosus patients were assessed for clinical and laboratory systemic lupus erythematosus manifestations, disease activity [Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)], cumulative damage [Systemic Lupus International Collaborating Clinics/American College of Rheumatology (ACR) Damage Index] and current drug use. Serum IL-17 levels were measured by an enzyme-linked immunosorbent assay using commercial kits.

RESULTS:

The median serum IL-17 level was 36.3 (range 17.36-105.92) pg/mL in childhood-onset systemic lupus erythematosus patients and 29.47 (15.16-62.17) pg/mL in healthy controls (p=0.009). We observed an association between serum IL-17 levels and active nephritis (p=0.01) and migraines (p=0.03). Serum IL-17 levels were not associated with disease activity (p=0.32), cumulative damage (p=0.34), or medication use (p=0.63).

CONCLUSION:

IL-17 is increased in childhood-onset systemic lupus erythematosus and may play a role in the pathogenesis of neuropsychiatric and renal manifestations. Longitudinal studies are necessary to determine the role of IL-17 in childhood-onset systemic lupus erythematosus.     

Cytokine-induced inhibition of bone matrix proteins is not mediated by prostaglandins

Interleukin-1 (IL-1) and tumor necrosis factor (TNF), two pleiotropic cytokines produced in inflammatory processes, inhibit bone matrix biosynthesis and stimulate prostanoid formation in osteoblasts. In the present study, the importance of prostaglandin formation in IL-1 and TNF-induced inhibition of osteocalcin and type I collagen formation has been examined. In the human osteoblastic cell line MG-63, IL-1 (10–1000 pg/ml), IL-1 (3–300 pg/ml) and TNF- (1–30 ng/ml) stimulated prostaglandin E₂ (PGE₂) formation and inhibited, 1,25(OH)₂-vitamin D3-induced osteocalcin biosynthesis as well as basal production of type I collagen. Addition of PGE₂ or increasing the endogenous formation of PGE₂ by treating the cells with arachidonic acid, bradykinin, Lys-bradykinin or des-Arg⁹-bradykinin, did not affect osteocalcin and type I collagen formation in unstimulated or 1,25(OH)₂-vitamin D3-stimulated osteoblasts. Four non-steroidal anti-inflammatory drugs, indomethacin, flurbiprofen, naproxen and meclofenamic acid, inhibited basal, IL-1 - and TNF--stimulated PGE₂ formation in the MG-63 cells without affecting IL-1 - or TNF--induced inhibition of osteocalcin and type I collagen formation. In isolated, non-transformed, human osteoblast-like cells, IL-1 and TNF- stimulated PGE₂ formation and concomitantly inhibited 1,25(OH)₂-vitamin D3-stimulated osteocalcin biosynthesis, without affecting type I collagen formation. In these cells, indomethacin and flurbiprofen abolished the effects of IL-1 and TNF- on prostaglandin formation without affecting the inhibitory effects of the cytokines on osteocalcin biosynthesis. These data show that IL-1 and TNF inhibit osteocalcin and type I collagen formation in osteoblasts independently of prostaglandin biosynthesis and that non-steroidal antiinflammatory drugs do not affect the effects of IL-1 and TNF on bone matrix biosynthesis.accepted by W. B. van den Berg     

Interleukin-10 (IL-10) secretion in systemic lupus erythematosus and rheumatoid arthritis: IL-10-dependent CD4+CD45RO+ T cell-B cell antibody synthesis

Interleukin-10 (IL-10) is a major immunoregulatory cytokine and has a multitude of immunomodulatory effects in the immune system. In this study, we have examined the secretion andin vitro function of IL-10 in B cell hyperactivity in antibody production in two common autoimmune diseases, systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). IL-10 was detectable in serum of all active SLE and serum and synovial fluid samples of all RA patients but in none of the normal controls. B cells and CD4+CD45RO+ memory T cells secreted highly enhanced levels of IL-10 in SLE and RA versus normals. Increased IgM and IgG production by B cells-CD4+CD45RO+ T cells in SLE and RA was IL-10 dependent, since neutralization of IL-10 cytokine by anti-IL-10 antibody drastically reduced Ig synthesis in these coculture experiments. B cell hyperactivity in autoantibody production in SLE and RA may be a function of IL-10-dependent CD4+CD45RO+ Th2 cell activation. Therefore, IL-10 may play an important role in highly disturbed immune system and B cell-T cell function in these immune disorders.     

The pro- and anti-inflammatory markers in patients with acute myocardial infarction and chronic stable angina

The aim of this study was to assess the plasma levels of VEGF and interleukin-10 in patients with acute myocardial infarction (AMI) and stable chronic angina (SA) and correlate the values with traditional CHD risk factors, left ventricular ejection fraction (LVEF) and established inflammatory marker hsCRP. Fifty patients with AMI and 30 with SA were enrolled. IL-10 levels in AMI patients were lower than in SA patients (9.81 +/- 5.0 versus 22.63 +/- 8.38 pg/ml, p < 0.00001). IL-10 levels were lower in AMI and SA patients with multiple CHD risk factors than in patients < or = 2 risk factors (SA: 19.48 +/- 2.94 versus 23.77 +/- 2.94 pg/ml; p < 0.005; AMI: 8.64 +/- 4.43 versus 11.85 +/- 4.09 pg/ml; p < 0.05) and patients with AMI and single-vessel than with multi-vessel disease (8.45 +/- 3.86 versus 10.72 +/- 3.95 pg/ml; p < 0.05). VEGF levels in AMI patients were higher than in SA patients (312.0 +/- 67.0 versus 221.0 + /- 50 pg/ml; p < 0.005). VEGF levels were higher in AMI patients with multi-vessel disease than in patients with single-vessel disease (348.74 +/- 45.23 versus 252.05 +/- 21.12 pg/ml; p < 0.005), with LVEF <40% and Killip class III-IV than in patients with LVEF >40% and Killip class I-II (338.8 +/- 51.59 versus 271.8 +/- 50.51 pg/ml; p < 0.005 and 340.71 +/- 52.94 versus 275.45 +/- 49.48 pg/ml; p < 0.05, respectively) and with chest pain > 6 h versus < 6 h (330.03 +/- 58.58 versus 292 +/- 57.53 pg/ml; p < 0.05). HsCRP concentrations in AMI patients were higher than in SA (1.24 +/- 0.47 versus 0.42 +/- 0.14; p < 0.0001). HsCRP was correlated with IL-10 (r = -0.413; p < 0.05) and VEGF (r = 0.319; p < 0.05). Acute myocardial infarction is associated with elevated VEGF levels and decreased concentration of IL-10. There is a significant correlation between levels of inflamatory markers and CHD risk factors and the function of the left ventricle on admission.     

Plasma endotoxin and cytokine levels in neutropenic and non-neutropenic bacteremic patients

Plasma endotoxin, tumor necrosis factor- (TNF-), interleukin 1 (IL-1), interleukin 1 receptor antagonist (IL-1ra), and interleukin 6 (IL-6) concentrations in 69 bacteremic patients were compared with those in 54 nonbacteremic patients suffering from suspected bacterial infections. Only three (11%) of the 27 patients with gram-negative bacteremia showed detectable levels of endotoxin. TNF- was detected in 6% of the bacteremic patients and in none of the nonbacteremic patients. Median IL-6 levels were significantly higher in bacteremic than in nonbacteremic patients (55 vs. 0 pg/ml, p=0.0008). IL-6 concentrations were similar in neutropenic and non-neutropenic bacteremic patients (median 55 vs. 74 pg/ml). In contrast, neutropenic bacteremic patients had significantly lower concentrations of tIL-1ra than non-neutropenic bacteremic patients (250 vs. 1,950 pg/ml, p<0.0001). Patients with fatal bacteremia had significantly higher concentrations of IL-6 and IL-1ra than the survivors (median, 450 vs. 40, p=0.012 and 7,600 vs. 420 pg/ml, p=0.0075, respectively). Determinations of endotoxin or TNF- in patients with suspected bacteremia failed to offer clinically relevant data on the prognosis of these patients. IL-6 levels correlated with both the presence of bacteremia and the risk of death. Granulocytopenic patients with bacteremia had lower levels of circulating IL-1ra than patients with normal granulocyte counts, and these levels correlated with poor outcome.     

血清抗中性粒细胞胞浆抗体与儿童闭塞性细支气管炎诊治的相关性研究

目的探讨血清抗中性粒细胞胞浆抗体(ANCA)与儿童闭塞性细支气管炎(BO)诊治的相关性。方法将80例感染后BO患儿作为研究对象,给予布地奈德混悬液+异丙托溴铵溶液雾化吸入,孟鲁司特+阿奇霉素口服等规范化治疗。选取同期门诊体检的60例健康儿童作为对照组。比较治疗前后患儿血清白介素-6(IL-6)、白介素-8(IL-8)、血C-反应蛋白(CRP)和ANCA表达水平,分析ANCA表达水平与血清炎症指标以及患儿转归的相关性。结果观察组患儿治疗有效6例(7.50%),好转71例(88.75%),无效3例(3.75%),治疗有效率为96.25%(77/80);观察组患儿的血清IL-6、IL-8、CRP和ANCA表达水平明显高于对照组,观察组患儿治疗后血清IL-6、IL-8、CRP和ANCA表达水平明显低于治疗前,差异均有统计学意义(均P<0.01);经Pearson相关性分析显示,观察组患儿血清ANCA表达水平与IL-6表达水平呈正相关(r=0.53,P=0.006),与IL-8表达水平呈正相关(r=0.46,P=0.001),与CRP表达水平呈正相关(r=0.61,P=0.012);出院1年再入院组(n=15)、门诊组(n=21)、缓解组(n=41)三组患儿出院时的血清IL-6、IL-8和ANCA表达水平呈明显递减趋势,差异均有统计学意义(均P<0.01),三组患儿的CRP表达水平无明显差异(P>0.05)。结论血清ANCA是一种BO患儿病情严重程度和转归的较好观测指标,值得临床推广应用。