Interleukin-1 and interleukin-2 production by peripheral blood mononuclear cells in multiple sclerosis patients

The production of interleukin-1 (IL-1) and interleukin-2 (IL-2) by peripheral blood mononuclear cells (PBM) was assessed in multiple sclerosis (MS) patients in relapse, chronic progressive MS patients, patients with other neurological diseases (OND) and healthy subjects. Production was defined as the level of IL-1 and IL-2 in PBM supernatants. Neither spontaneous nor LPS-induced IL-1 production differed significantly in MS, OND patients or healthy individuals. On the other hand PHA-induced PBM IL-2 production was significantly less in MS patients in relapse (130 +/- 10.0 U/ml) than in chronic progressive MS patients (172 +/- 9.8 U/ml), OND patients (192 +/- 11.5 U/ml) and healthy subjects (215 +/- 13.8 U/ml) (P less than 0.02). Spontaneous IL-2 production was also diminished in MS patients in relapse (31 +/- 7.2 U/ml) as compared to chronic progressive MS patients (46 +/- 8.8 U/ml) and healthy subjects (49 +/- 11.1 U/ml) (P less than 0.01). Anti-Tac monoclonal antibody was used to study IL-2 receptor expression on the same sample of PBM that was used for IL-2 study. MS patients in relapse had significantly higher levels of IL-2 receptor-positive unstimulated PBM (6.0 +/- 2.2%) as compared to chronic progressive MS (2.0 +/- 0.9%), OND (2.5 +/- 1.1%) and healthy subjects (1.5 +/- 0.7%) (P less than 0.002). We postulate that reduced apparent IL-2 production by PBM of MS patients in relapse may result from immediate IL-2 binding to receptor expressed on activated T lymphocytes and internalization of IL-2-receptor complex.     

Elevated serum levels of interleukin-12 in chronic progressive multiple sclerosis

The serum levels of the heterodimeric cytokine IL-12 were measured by solid-phase ELISA in a group of healthy subjects, multiple sclerosis (MS) patients with secondary chronic progressive course of the disease and patients suffering from other neurological diseases (OND). Serum levels of IL-12 higher than 5 pg/ml (limit of sensitivity of the assay) were only found in 2/30 (6.7%) of the healthy subjects and none of the 8 subjects with OND. In contrast, IL-12 was found in the majority of CPMS patients' sera (10/15, 66.7%) with values ranging between 5.5 and 18.6 pg/ml. These results are suggestive for an up-regulated production of IL-12 in CPMS.     

Interleukin-12 is detectable in sera of patients with multiple sclerosis — association with chronic progressive disease course?

Multiple sclerosis (MS) is widely accepted as a systemic T- cell-mediated autoimmune disease with a T-helper type-1 (TH-1) profile of cytokine production. We addressed the question whether interleukin-12 (IL-12), as a central mediator of TH-1-cell activities, is detectable in sera of MS patients, and if there is any association with disease activity. We analysed 171 sera of patients with MS and meningitis, and healthy controls. IL-12 p40 protein was detectable at low levels in MS patients (median 43 pg/ml) and controls (median 49 pg/ml). Analysing different disease courses and activities, a significant elevation in stable primary progressive MS cases compared with controls (median 66 pg/ml) was found. IL-12 p40 protein was not detectable in cerebrospinal fluid probes of 10 patients. We conclude that the function of IL-12 in MS depends on expression and degradation of the different proteins. These could act proinflammatory as well as limiting the disease process. Copyright Lippincott Williams & Wilkins     

Immune deviation following pulse cyclophosphamide/methylprednisolone treatment of multiple sclerosis: Increased interleukin-4 production and associated eosinophilia

Multiple sclerosis (MS) is postulated to be a Th1-type cell-mediateds autiommune disease. Thus therapies that decrease T cell interferon (IFN)-γ production or increase interleukin (IL)-4 production would be expected to have an ameliorating effect on MS. Some progressive MS patients receiving pulse cyclophosphamide therapy developed periheral blood eosinophilia. We investigated whether cyclophosphamide-treated patients had immune deviation toward Th2 responses. We measured cytokine production in patients receiving either monthly intravenous methylprednisolone (MP), intravenous cyclophosphamide plus methylprednisolone (CY/MP), methotrexate, IFN-β1b, in untreated MS patients, and in healthy controls. Minimal IL-4 was secreted in untreated patients (129 · 62 pg/ml), methotrexate-treated patients (99 · 79 pg/ml), and healthy controls (50 · 13 pg/ml). A marked increase in IL-4 was observed in CY/MP patients (1,503 · 291 pg/ml). Patients treated with MP (418 · 160 pg/ml) or IFN-β1b (425 · 167 pg/ml) showed small increases. Eosinophilia in CY/MP-treated patients (6.0 · 0.7%) correlated with increased IL-4. IL-10 production was also increased in CY/MP-treated patients. Both CY/MP- and MP-treated groups had decreased production of IFN-γ compared with untreated MS. These findings demonstrate pronounced immune deviation favoring Th2-type responses after pulse cyclophosphamide therapy.     

多发性硬化患者病情演变与血浆和脑脊液趋化因子CXCL10水平的相关性

目的 观察急性期多发性硬化(MS)患者血浆和脑脊液中趋化因子CXCL10水平的动态变化规律及其与临床神经功能障碍的相关性,探讨其对疾病活动性的判定价值.方法 收集急性期MS患者、缓解期MS患者及健康对照各53例,神经系统非炎性疾病(NIND)32例,采用酶联免疫吸附试验法检测血浆和脑脊液中CXCL10水平,并进行扩展残疾状况评分量表(EDSS)评分.结果 (1)与急性期初期相比,急性期MS组患者第2、4周血浆CXCL10水平[(601±365)、(575±297)pg/ml]明显升高(t=-2.898、-2.651,P=0.001、0.003);第4周脑脊液中CXCL10水平[(1807±803)pg/ml]与急性期初期比较差异无统计学意义.(2)急性期初期MS组血浆CXCL10水平明显高于缓解期MS组[(287±118)pg/ml,t=3.555,P=0.001]和健康对照组[(248±130)pg/ml,t=4.895,P=0.000].(3)急性期MS组脑脊液CXCL10水平[(1774±604)pg/ml]明显高于NIND组[(122±114)pg/ml,t=15.192,P=0.000].(4)急性期MS组患者血浆与脑脊液中CXCL10水平间存在相关性(r=0.792,P=0.001);脑脊液CXCL10水平与同期EDSS评分之间存在相关性(r=0.526,P=0.002).结论 (1)MS患者血浆中CXCL10水平对判断疾病活动性有一定的参考价值.(2)急性期MS患者血浆CXCL10水平能在一定程度上反映其在脑脊液中的水平.(3)检测急性期MS患者脑脊液CXCL10水平对判断临床功能障碍程度有一定的参考价值.

Abstract：

Objective To investigate the evolution of CXCL10 in blood plasma and cerebrospinal fluid (CSF) during relapses of multiple sclerosis (MS),and the correlation between these and the clinical neurological dysfunction.Methods Fifty-three patients with definite MS during relapsing state (relapsing MS group) diagnosed by the McDonald criteria;fifty-three patients with definite MS during remitting state ( remitting MS group);thirty-two patients with non-inflammatory neurologic disease ( NIND group) and fiftythree healthy controls (NC group) were enrolled in the study.Each patient clinical status was evaluated with the Expanded Disability Status Scale ( EDSS).Plasma and CSF levels were analyzed by enzyme-linked immunoassay.Results ( 1 ) The CXCL10 level in plasma in relapsing MS group elevated significantly between the 2nd ( (601 ± 365 ) pg/ml,t = - 2.898,P = 0.001) and the 4th ( (575 ± 297 ) pg/ml,t = -2.651,P=0.003) week after relapsing;GXL10 in CSF (n =32) did not changed significantly in the 4th week after relapsing( (1807 ±803) pg/ml).(2) The CXCL10 level in plasma in relapsing MS group were significantly higher than that in the healthy control group ((248±130) pg/ml,(=4.895,P=0.000) and remitting MS group ((287 ±118) pg/ml,t = 3.555,P = 0.001 ).( 3 ) The CXCL10 level in CSF in relapsing MS group (( 1774 ± 604) pg/ml) was significantly higher than that in NIND group ( ( 122 ± 114) pg/ml,t= 15.192,P =0.000).(4) The CXCL10 level in plasma in relapsing MS group had correlation with that in CSF (r=0.792,P=0.001).The CXCL10 level in CSF in relapsing MS group had correlation with EDSS scores (r = 0.526,P = 0.002 ).Conclusions The CXCL10 level in plasma might be implemented as a paraclinical marker of disease activity in MS.The CXCL10 level in plasma of MS may be relevant to that in CSF.The CXCL10 level in CSF of MS may indicate the clinical neurological dysfunction.     

Tumor necrosis factor and interleukin-1 in the CSF and sera of patients with multiple sclerosis

Serum and cerebrospinal fluid (CSF) from 31 patients with multiple sclerosis (MS) were examined to determine the levels of tumor necrosis factor (TNF) and interleukin (IL)-1 alpha (or IL-1 beta) by an enzyme-linked immunosorbent assay. TNF was detected in 29 (93.5%) of CSF from 31 cases of MS. TNF was also detectable in 100% of CSF from patients with acute relapsing MS in exacerbation. Patients with acute relapsing MS in exacerbation showed significantly higher CSF levels of TNF as compared with either those in remission or the controls (P less than 0.001 and P less than 0.0001, respectively). Increased levels of TNF were also detected in 35.5% of the MS sera, and especially in those with acute relapsing MS in exacerbation. Increased TNF levels were also frequent in the CSF and sera of patients with Guillain-Barré syndrome (GBS), which is also a demyelinating disease. No IL-1 alpha (or IL-1 beta) was detected in either CSF or sera of 31 MS patients. It is considered likely that TNF CSF levels may reflect disease activity in MS.     

Interferon-beta regulates cytokines and BDNF: greater effect in relapsing than in progressive multiple sclerosis

The mechanism of action of interferon (IFN)-beta therapy in multiple sclerosis (MS) is only partially known, and its efficacy changes with disease stage. In different forms of MS, we determined how IFN-beta regulates mononuclear cell production of the important anti-inflammatory Th2 cytokine - IL-10, the Th1 cytokine - IFN-gamma, and the brain-derived neurotrophic protein - BDNF. Activated T cells and monocytes from therapy-na?ve patients secreted more IL-10 than healthy controls. During IFN-beta therapy, however, T cells produced less IL-10. In vitro, IFN-beta stimulated IL-10 production by activated T cells, but inhibited IL-10 secretion by activated monocytes, a richer source of IL-10 than T cells. The form of MS also affected cytokine production. IL-10 and BDNF levels in MNC were high during relapsing/remitting (RR) MS, but low in progressive MS. Surprisingly, IFN-beta therapy increased BDNF levels in antidepressant-na?ve patients, but BDNF was lower during concurrent antidepressant drug therapy, suggesting an interaction between MS, depression, and neurodegeneration. IFN-beta in vitro strongly induced IL-10 and IFN-gamma in activated T cells in RRMS, but not in progressive MS, suggesting IFN resistance. IFN-beta effects are specific for disease state and immune subsets, possibly explaining why IFN-beta therapy is most effective in early T cell-regulated RRMS, but less beneficial in progressive MS, where chronic plaques contain few T cells and high numbers of monocytes.     

Interleukin-17 in transverse myelitis and multiple sclerosis

CSF IL-6 is elevated in transverse myelitis (TM) and predicts disability. Since IL-17 regulates cytokines (TNFalpha, IL-1beta and IL-6) known to stimulate IL-6 production by astrocytes, we sought to determine whether IL-17 was increased in TM and MS compared to healthy controls (HC) and other neurologic diseases (OND). IL-17 and IL-6 levels were measured in stimulated peripheral blood mononuclear cell (PBMC) supernatants from HC, MS, TM and OND. IL-17 was increased in TM compared to HC, MS, and OND (mean pg/ml+/-standard error; HC: 36.1+/-11.7, MS: 89.4+/-23.3, TM: 302.6+/-152.5, OND: 41.2+/-13.0, p=0.01). IL-6 was increased in TM relative to MS and HC (HC: 2624 pg/ml+/-641, MS: 6129+/-982, TM: 12,536+/-2657, OND: 6920+/-1801, p<0.002). MS patients with early disease (<2 years) also had increased levels of IL-17 (p<0.04) and IL-6 (p<0.05). Cytokine neutralization experiments demonstrated that IL-6 was the main inducer of astrocyte IL-6 production. We conclude that IL-17 and IL-6 production from PBMC in TM and early MS are increased and induce astrocyte IL-6 production through IL-6.     

Increased lymphocyte beta-adrenergic receptor density in progressive multiple sclerosis is specific for the CD8+, CD28- suppressor cell.

Beta-adrenergic receptor density on T cells from healthy humans is greatest on suppressor cells (CD8+, CD28-) and the effect of catecholamines, secreted by the sympathetic nervous system, predominates on this subset. The sympathetic skin response, a measure of sympathetic nervous system function, is absent in most patients with chronic progressive multiple sclerosis (MS). We measured beta-adrenergic receptor density on suppressor cells, cytotoxic cells, and monocytes from patients with chronic progressive MS and healthy control subjects. Control receptor density on suppressor cells was 2.8 +/- 0.3 fmol/10(6) cells versus a density of 5.1 +/- 0.7 fmol/10(6) cells for patients. Cytotoxic cell (CD8+, CD28+) receptor density was 1.4 +/- 0.4 fmol/10(6) cells in control subjects and 0.9 +/- 0.3 fmol/10(6) cells in the patients. Monocytes displayed beta-adrenergic receptor densities of 2.6 +/- 0.4 fmol/10(6) cells in normal individuals and 2.7 +/- 0.4 fmol/10(6) cells in the patient group. CD8 lymphocyte beta-adrenergic receptor densities in patients with relapsing-remitting and those with stable MS were not different from control values, yet were significantly less than the values for patients with chronic progressive MS. We find that mononuclear cells from healthy control subjects and patients with chronic progressive MS proliferate in response to 200 units/ml of recombinant human interleukin-2 (IL-2) similarly. However, IL-2 treatment increased beta-adrenergic receptor density on normal mononuclear cells, but failed to increase it on mononuclear cells from patients with chronic progressive MS.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum and CSF levels of MCP-1 and IP-10 in multiple sclerosis patients with acute and stable disease and undergoing immunomodulatory therapies

The two chemokines, monocyte chemoattractant protein (MCP)-1 and gamma-interferon inducible protein (IP)-10, are thought to be involved in the pathogenesis of multiple sclerosis (MS). We measured MCP-1 and IP-10 levels in serum and CSF samples from 38 acute and 25 stable MS patients and from 40 controls. The latter consisted in patients with other inflammatory neurological diseases (OIND) or with non-inflammatory neurological diseases, and healthy controls. CSF MCP-1 levels exceeded those found in serum in all the patients studied as well as in healthy controls. CSF MCP-1 levels were significantly lower in acute MS [468+/-(S.E.M.) 18 pg/ml] than in stable MS (857+/-104 pg/ml). When detectable, serum and CSF IP-10 levels were significantly higher in acute MS (serum 331+/-66 pg/ml; CSF 118+/-16 pg/ml) than in stable MS (serum 69+/-7 pg/ml; CSF 25+/-2 pg/ml). Among OIND patients, those with HIV-1-associated dementia showed high serum and CSF levels of both MCP-1 and IP-10. Those with encephalitis showed high serum and CSF levels of IP-10 and CSF mononuclear pleiocytosis. We also evaluated the effects of 6-methylprednisolone or IFN-beta1a therapy on circulating MCP-1 and IP-10 levels. Neither MCP-1 nor IP-10 post-therapy levels varied significantly from baseline values. Our findings suggest that (a) MCP-1 could be constitutively produced within the brain; (b) MCP-1 and IP-10 CSF levels in acute MS vary significantly from those in stable MS, and these variations are inverse; and (c) current MS therapies do not modify circulating levels of MCP-1 and IP-10.     

Serum and cerebrospinal fluid concentration of inflammatory proteins MIP-1-alpha and MIP-1-beta and of interleukin 8 in the course of borreliosis

Chemokines constitute a group of cytokines with a strong chemotactic action, playing an important role in the pathogenesis of inflammatory responses, including infectious meningitis. The results of in vitro experiments suggest synthesis of chemokines during Borrelia burgdorferi infection. The aim of this study was to investigate serum and cerebrospinal fluid (CSF) concentrations of the following chemokines: interleukin-8 (Il-8) and macrophage inflammatory protein 1 alpha and 1 beta (MIP-1 alpha and MIP-1 beta) in patients with neuroborreliosis. The study group consisted of 20 patients admitted to Neuroinfections and Infectious Diseases Department of the Medical University in Bia?ystok. The control group consisted of 12 healthy persons from whom blood samples were obtained, and 10 patients without meningitis, from whom CSF samples were taken for diagnostic purposes. Chemokine concentrations were measured with ELISA kits before treatment (baseline) and after 2 weeks of antibiotic therapy (post-treatment). Mean serum concentrations of chemokine were elevated in neuroborreliosis patients at baseline (Il-8--mean +/- SD = 668.25 +/- 661.51 pg/ml, MIP-1 alpha--124.90 +/- 89.37 pg/ml, MIP-1 beta--233.40 +/- 298.40 pg/ml) as compared to these in the control group (Il-8-23.72 +/- 7.68 pg/ml, MIP-1 alpha--36.81 +/- 4.74 pg/ml, MIP-1 beta--70.41 +/- 16.41 pg/ml). Post-treatment mean concentrations of Il-8 (197.70 +/- 285.56 pg/ml) and MIP-1 beta (102.70 +/- 42.56 pg/ml) remained significantly elevated, while the mean concentration of MIP-1 alpha (53.65 +/- 38.50 pg/ml) was insignificantly higher than that in the control group. The Il-8 mean concentration was the most elevated comparing to the controls and has decreased most significantly during the treatment. CSF concentrations of chemokines were significantly elevated both at baseline (Il-8--754.95 +/- 535.83 pg/ml, MIP-1 alpha--24.35 +/- 4.88 pg/ml, MIP-1 beta--27.6 +/- 8.38 pg/ml) and post-treatment (Il-8--98.20 +/- 74.74 pg/ml, MIP-1 alpha--18.60 +/- 2.87 pg/ml, MIP-1 beta--16.90 +/- 4.38 pg/ml) in comparison with the controls (Il-8--10.43 +/- 2.70 pg/ml, MIP-1 alpha--8.17 +/- 1.54 pg/ml, MIP-1 beta--7.27 +/- 1.58 pg/ml). MIP-1 alpha and MIP-1 beta CSF concentrations were significantly lower than their concentrations in serum. The Il-8 CSF concentration did not differ significantly from its serum concentration. However, in some patients Il-8 CSF concentration was much higher than that in the serum, which suggests its significant synthesis within the cns and its role in the pathogenesis of B. burgdorferi meningitis. Chemokine CSF concentrations were not correlated with cytosis and CSF protein concentration. The results indicate the induction of Il-8, MIP-1 alpha and MIP-1 beta synthesis in the course of neuroborreliosis and a decrease of their concentrations during 2 weeks of treatment, however, without reaching the normal values.     

Immunoreactive leukotriene C4 levels in CSF of MS patients

Using a sensitive and specific radioimmunoassay levels of leukotriene (LT)C4-like material were estimated in lumbar cerebrospinal fluid (CSF) samples from patients with multiple sclerosis (MS) in comparison to control patients with or without inflammatory processes in the central nervous system (CNS). Levels of LTC4-like material were significantly elevated (p less than 0.01) in CSF from patients with inflammatory diseases such as meningitis, polyradiculitis or meningoencephalitis (57 +/- 53 pg/ml, n = 16) as compared to those from control patients without inflammatory or immunological CNS diseases (21 +/- 16 pg/ml, n = 42). By contrast, LTC4-like material was 16 +/- 7 pg/ml in first manifestations of MS (n = 7). 21 +/- 16 pg/ml in remitting-relapsing MS (n = 15) and 10 +/- 6 pg/ml in chronic progressive MS (n = 8). These results argue against a significant pathophysiological role of cysteinyl-LT in MS.     

Increased plasma levels of 8-iso-PGF2alpha and IL-6 in an elderly population with depression

Oxidative damage and immune-inflammatory activation have been suggested to play a role in depression. The purpose of the study was to investigate possible associations and interactions of these pathophysiological mechanisms in geriatric depression by determining the levels of plasma 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) and interleukin-6 (IL-6) in elderly depressed individuals. Subjects over 60 years of age with depression and controls were randomly selected from a population in the community after screening with the Geriatric Depression Scale. Plasma concentrations of 8-iso-PGF2alpha and IL-6 were measured in both groups. Depressed patients had significantly higher mean (+/-S.D.) 8-iso-PGF2alpha levels compared to healthy controls (245.01+/-179.92 pg/ml vs 97.64+/-42.72 pg/ml, respectively). Similarly, the same groups demonstrated significantly elevated IL-6 levels compared with controls (58.73+/-39.90 pg/ml vs 15.41+/-9.27 pg/ml). This study indicates an association between increased levels of plasma 8-iso-PGF2alpha and IL-6 with depressive symptomatology in elderly individuals and indicates the necessity for further investigation, possibly within the framework of an integrated involvement of oxidative damage and inflammation in the pathophysiology of depression in the elderly.     

Soluble E-selectin in multiple sclerosis: raised concentrations in patients with primary progressive disease.

OBJECTIVE--To determine whether concentrations of soluble E-selectin (sE-selectin), an immunological marker of endothelial activation, were correlated with gadolinium-DPTA enhancement on MRI in patients with multiple sclerosis. METHODS--Serial sE-selectin concentrations were measured in 28 patients with multiple sclerosis undergoing monthly gadolinium (Gd) enhanced MRI of the brain and spinal cord, and in 10 control subjects. C reactive protein (CRP), von Willebrand factor (vWF), and tumour necrosis factor-alpha (TNF alpha) were also determined. RESULTS--Primary progressive patients had significantly increased sE-selectin concentrations compared with the relapsing remitting and secondary progressive patients who had normal sE-selectin concentrations (22.2 (SD1 6.1) ng/ml v 9.8 (SD2.1) ng/ml and 7.7 (SD2.7) ng/ml, respectively, P = 0.03). This difference was attributable to five of the 10 primary progressive patients who had persistently raised sE-selectin concentrations, with relatively inactive MRI studies. No correlation could be found between sE-selectin concentrations and Gd enhancement on MRI, but a close correlation existed between mean concentrations of sE-selectin and TNF alpha (r = 0.71, P < 0.001). Despite raised sE-selectin and TNF alpha concentrations, primary progressive patients had normal CRP concentrations (1.03 (SD1.14) mg/l), which were significantly lower than the relapsing remitting (3.16 (SD2.54) mg/l) and secondary progressive patients (2.28 (SD2.1) mg/l, P = 0.03). Raised CRP concentrations did correlate with infectious episodes, clinical relapse, and Gd enhancement, and were significantly raised when no MRI activity was found. Concentrations of vWF were normal in all patient groups. CONCLUSIONS--The results further high-light the differences between patients with primary progressive and those with relapsing remitting/secondary progressive multiple sclerosis.     

Platelet activating factor is elevated in cerebral spinal fluid and plasma of patients with relapsing-remitting multiple sclerosis.

Platelet-activating factor (PAF) is a phospholipid mediator of inflammation with a wide range of biological activities, including the alteration of barrier function of endothelium. A biological assay combined with high pressure liquid chromatography-tandem mass spectrometry showed that plasma and cerebral spinal fluid (CSF) PAF levels in 20 patients with relapsing/remitting or secondary progressive multiple sclerosis (MS) studied by magnetic resonance imaging (MRI) were significantly higher than in healthy controls (plasma: 3.29+/-4.52 vs. 0.48+/-0.36 ng/ml, p < 0.002; CSF: 4.95+/-6.22 ng/ml vs. 0.01+/-0.04 ng/ml, p < 0.0001). Values were also significantly higher in relapsing/remitting than in secondary progressive (plasma: 5.10+/-4.97 vs. 0.52+/-0.85 ng/ml, p < 0.005; CSF: 8.59+/-6.39 vs. 0.55+/-0.68 ng/ml, p < 0.002). It was also found that both plasma (R2: 0.65) and CSF (R2:0.72) levels were correlated with the MRI number of gadolinium enhancing lesions, which are markers of blood-brain barrier (BBB) injury, whereas their peaks were not correlated with the MRI number of white matter lesions, nor with the expanded disability status score (EDSS) according to Kurtze [Kurtze, J.F., 1983. Rating neurological impairment in multiple sclerosis: an expanded disability scale (EDSS). Neurology 33, 1444-1452]. Both plasma and CSF in patients with relapsing/remitting MS and marked gadolinium enhancement contained the two major molecular species of PAF: 1-0-hexadecyl- (C16:O) and 1-0-octadecyl-sn-glycero-3-phosphocholine (C18:O). The ratio of the two molecular species was different in the two biological fluids, being PAF C18:0 more abundant in CSF and PAF C16:0 in plasma, indicating a different cellular origin of PAF or different enzymatic processing. These findings suggest that PAF is a significant mediator of BBB injury in the early stages of MS, rather than a marker of its progression and severity.     

Th1 and Th2 serum cytokine profiles characterize patients with Hashimoto's thyroiditis (Th1) and Graves' disease (Th2)

OBJECTIVES: The aim of this study was to document the pattern of immune response, assessed by the measurement of both Th1 and Th2 serum cytokines, in patients suffering from autoimmune thyroid disease and toxic nodular goiter. METHODS: Both Th1 and Th2 serum cytokine levels were assayed in patients suffering from Graves' disease (GD, n = 25), Hashimoto's thyroiditis (HT, n = 21), and toxic nodular goiter (TNG, n = 7) and compared with corresponding levels of 25 healthy controls. Serum concentrations of IL-2, IL-1 beta, INF-gamma, TNF-alpha, IL-12, IL-15, IL-10, IL-18, IL-4 and IL-5 were assayed in fasting serum samples. RESULTS: It was found that patients with HT had higher IL-2 serum levels (12.16 +/- 0.66 pg/ml) compared to patients with TNG (9.25 +/- 0.84 pg/ml), GD (7.86 +/- 0.30 pg/ml) and controls (7.36 +/- 0.45 pg/ml; p = 0.0001), higher INF-gamma levels (7.60 +/- 0.33 pg/ml) compared to patients with TNG (5.77 +/- 0.55 pg/ml), GD (5.74 +/- 0.24 pg/ml) and controls (5.09 +/- 0.27 pg/ml; p = 0.0009), higher IL-12 levels (3.57 +/- 0.19 pg/ml) compared to patients with TNG (2.57 +/- 0.21 pg/ml), GD (2.48 +/- 0.13 pg/ml) and controls (2.59 +/- 0.23 pg/ml; p = 0.004), and higher IL-18 levels (27.52 +/- 1.75 pg/ml) compared to patients with TNG (18.71 +/- 2.24 pg/ml), GD (15.44 +/- 1.39 pg/ml) and controls (15.16 +/- 1.62 pg/ml; p = 0.0002). In contrast, patients with GD had higher serum levels of IL-4 (4.11 +/- 0.33 pg/ml) compared to patients with HT (3.0 +/- 0.16; p = 0.02) and higher IL-5 levels (4.22 +/- 0.30 pg/ml) compared to patients with TNG (3.21 +/- 0.58 pg/ml), HT (2.75 +/- 0.16 pg/ml) and controls (2.0 +/- 0.19 pg/ml; p = 0.0001). Patients had lower IL-1 beta serum levels (TNG 2.45 +/- 0.20, HT 2.52 +/- 0.14, GD 2.68 +/- 0.12 pg/ml) compared to controls (3.6 +/- 0.20 pg/ml; p = 0.008). CONCLUSIONS: The above findings suggest that a Th1 pattern of immune response characteristic of cellular immunity is dominant in HT, whereas the predominance of Th2 cytokines in GD indicates a humoral pattern of immune reaction.     

Increased nocturnal interleukin-6 excretion in patients with primary insomnia: a pilot study

The aim of the present study was to investigate whether there is a difference in evening/nocturnal interleukin-6 (IL-6) serum excretion in patients with primary insomnia compared to controls. We hypothesized that in insomniac patients, the excretion of evening/nocturnal IL-6 is enhanced, like observed in aged adults and after sleep deprivation in healthy subjects. We studied IL-6 serum concentrations in 11 patients (two males and nine females) with primary insomnia and 11 age and gender-matched healthy controls. Sleep was monitored polysomnographically for three consecutive nights. The measurement of IL-6 (from 19:00 h to 09:00 h) in 2-h intervals were performed prior to and during the last laboratory night. Polysomnographically determined sleep parameters and subjective ratings of sleep demonstrated clear-cut impairments of sleep in the insomniac group. Nocturnal IL-6 secretion was significantly increased (p<.05) in insomniac patients for the whole measurement period (mean area under the curve+/-SD: 27.94+/-14.15 pg/ml x 2h) compared to controls (16.70+/-7.64 pg/ml x 2h). Total IL-6 secretion correlated inversely with subjectively perceived sleep quality and amount of slow wave sleep in the insomniac patients. Amount of Wake Time correlated positively with IL-6 excretion in insomniacs. The results of the present study demonstrate significantly increased nocturnal IL-6 secretion in insomniacs. It might be speculated that chronic primary insomnia with polysomnographically documented sleep impairments activates the production of IL-6 analogous to sleep deprivation studies in healthy subjects. This might also implicate a higher risk for inflammatory and cardiovascular diseases in patients with chronic insomnia.     

Multiple sclerosis: deficient in vitro responses of blood mononuclear cells to IFN-β

IFN-beta may modify the clinical course of multiple sclerosis (MS) but is not curative, and there are also patients whose disease does not respond to IFN-beta as currently administered. Tests are warranted with a capacity to early discriminate responders from non-responders, thereby altering treatment option for the individual patient. In vitro effects of IFN-beta on expression of activation-associated cell surface markers and cytokine production need to be explored in this context. Here we report on the influence in vitro of IFN-beta on blood mononuclear cells (MNC) prepared from MS patients and healthy controls. MNC were subjected to short-term culture in the presence of IFN-beta at concentrations of 100 U/ml and 1000 U/ml. Expression of cell surface molecules CD40, CD69, CD80, CD86, CD95 and HLA-DR was measured by flow cytometry. IL-10 and IL-12 p40 production in culture supernatants was measured by ELISA. MNC exposed to IFN-beta in vitro enhanced expression of the co-stimulatory CD80, CD86, the early activation antigen CD69 and the cell death receptor CD95. Expression of CD40 and HLA-DR was not influenced. IFN-beta increased IL-10 but suppressed IL-12 p40 production. In vitro effects of IFN-beta on MNC were similar in MS patients and in healthy subjects, except that IFN-beta-induced augmentation of CD86 and CD69 expression was less pronounced in MS, in particular in untreated MS patients. Individual MS patients clearly responded differently to IFN-beta in vitro in comparison with the majority of patients in this cross-sectional study. In conclusion, anti-inflammatory effects of IFN-beta on blood MNC include augmentation of IL-10 production and suppression of IL-12 p40 production, which are accompanied by enhancement of CD69, CD80, CD86 and CD95 expression. The less pronounced IFN-beta-induced effects on CD86 and CD69 expression in MS vs controls might reflect a defect in immunoregulation in MS. Larger groups should be evaluated, and follow-up studies performed in MS patients before/during IFN-beta treatment in relation to clinical outcome measures to evaluate the usefulness of these markers for possible differentiation between responders and non-responders to IFN-beta treatment.     

Increased level of salivary prostaglandins in patients with major depression

We quantified the amounts of salivary prostaglandin (PG) D2, PGE2, and PGF2 alpha by radioimmunoassay in 32 patients with major depressive disorder, 16 patients with minor depressive disorder, 24 patients with neurotic disorders (panic, generalized anxiety, phobic, somatization, and obsessive compulsive), and 28 healthy controls. In the saliva of patients with major depressive disorder, the concentrations of immunoreactive PGs (PGD2, 385 +/- 71 pg/ml; PGE2, 498 +/- 105 pg/ml; PGF2 alpha, 444 +/- 100 pg/ml) were significantly higher than those of the healthy controls (PGD2, 129 +/- 18 pg/ml; PGE2, 207 +/- 25 pg/ml; PGF2 alpha, 164 +/- 17 pg/ml). On the other hand, the salivary concentrations of immunoreactive PGs from patients with minor depressive disorder or neurotic disorders were comparable to those of the controls. These results suggest that the level of salivary PGs may be an indicator of major depressive disorder.     

Increased T-lymphocyte interleukin-6 binding in patients with multiple sclerosis.

Multiple sclerosis (MS) represents a T-cell-mediated autoimmune demyelinating disease of the central nervous system associated with altered immunoregulation. Interleukin (IL)-6 is a cytokine that has several effects on the neuroimmune system. Specific IL-6 receptors have been found in human lymphocytes and neuroglial cells. The aim of the present study was to assay IL-6 binding on peripheral blood T lymphocytes in MS patients. We found that T cells from MS patients had significantly more IL-6 receptors [Bmax: 279 +/- 7 vs. 246 +/- 8 (mean +/- SEM) receptors/cell, in patients and controls, respectively], whereas Kd values were similar to those of healthy subjects [26.8 +/- 0.7 vs. 25.4 +/- 0.6 (mean +/- SEM) pM, in patients and controls, respectively]. Significant (P < 0.05) differences in IL-6 binding values were observed between stable patients and those relapsing (272 +/- 9 vs. 300 +/- 12 (mean +/- SEM) receptors/cell, respectively). We found significantly (P < 0. 001) higher amounts of IL-6 receptors on CD4+ T cells from MS patients than on CD4+ lymphocytes from controls (434 +/- 11 vs. 363 +/- 9 (mean +/- SEM) receptors/cell, respectively); CD8+ T cells showed very few IL-6 receptors in both patients and controls. These data are discussed in terms of MS immune pathogenesis and pathophysiology, because T-cell activation seems to be linked to increased IL-6 binding. The upregulated IL-6 system might be involved in antibody-mediated demyelinating pathways, because IL-6 is well known to enhance humoral immune response.