Elevated levels of a soluble form of the T cell activation antigen CD27 in cerebrospinal fluid of multiple sclerosis patients.

The expression of the T cell membrane molecule CD27--a molecule that has recently been shown to belong to the nerve growth factor receptor superfamily--is strongly increased after activation of T lymphocytes via the T cell receptor/CD3 complex. In addition, activated cells release a 28-32 kDa soluble form of CD27 in their supernatant which can also be detected in serum and urine of healthy individuals. In this study we show that levels of soluble (s) CD27 are significantly elevated in cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients and of patients and of suffering from other inflammatory neurological diseases (OIND), whereas increased levels of sCD25 (soluble interleukin-2 receptor) were only found in CSF of patients with OIND. In MS patients, a significant correlation was found between CSF sCD27 titer and IgG index.     

Soluble CD4 and CD8 in the peripheral blood of patients with multiple sclerosis and HTLV-1-associated myelopathy

We evaluated the presence of soluble (s) CD4 and sCD8, released from activated T cells, in the sera of patients with multiple sclerosis (MS) and human T lymphotropic virus type 1 (HTLV-1)-associated myelopathy (HAM) using an enzyme-linked immunosorbent assay (ELISA). In addition, peripheral blood T cell subsets in patients with MS and HAM were analyzed by single and two color flow cytometry. The serum level of sCD8 was significantly elevated in MS patients as compared with controls (p less than 0.001). Sera from patients with an exacerbation of acute relapsing MS showed a higher sCD8 level than the patients in remission or controls (p less than 0.01 and p less than 0.001, respectively). The serum levels of both sCD4 and sCD8 were also significantly elevated in patients with HAM (p less than 0.001 and p less than 0.001, respectively). In addition, a significantly increased serum level of soluble interleukin-2 receptor (sIL-2R) was found in patients with HAM as compared with that of controls (p less than 0.001). These observations suggest that CD8 cells may be activated in the peripheral blood of patients with MS and sCD8 may be related to clinical activity, but that both CD4 and CD8 cells may be activated in the peripheral blood of patients with HAM.     

Impaired Fas-independent apoptosis of T lymphocytes in patients with multiple sclerosis

The homeostasis of the immune system is maintained by apoptotic (programmed cell death) elimination of potentially pathogenic, autoreactive mononuclear cells. There is emerging evidence that apoptosis mediated by the cell death receptor Fas is impaired in activated lymphocytes from patients with multiple sclerosis (MS), but other forms of apoptosis have not yet been fully evaluated. To further explore the dynamics of programmed cell death in MS, spontaneous and induced apoptosis of both peripheral and intrathecal mononuclear cells was investigated in clinically active MS patients and appropriate controls. In the MS group, spontaneous apoptosis of unfractionated mononuclear cells was significantly reduced, and activated intrathecal and peripheral T cells were found to be predominantly resistant to Fas-independent apoptosis. These results indicate that in clinically active MS, the reduced susceptibility of mononuclear cells to apoptosis is partly due to impairment of Fas-independent apoptotic pathways.     

Cerebrospinal fluid concentrations of soluble CD27 in HTLV-I associated myelopathy and multiple sclerosis

OBJECTIVES: Stimulation of T lymphocytes via the T cell receptor strongly enhances CD27 membrane expression and induces the release of a soluble 32 kDa form of CD27 (sCD27). CD27 is a member of the TNF receptor family, a group of molecules that have important roles in lymphocyte differentiation and survival. Raised concentrations of sCD27 have been reported in various immunopathological conditions and there is evidence that this molecule can serve as a marker of T cell activation in vivo. Concentrations of sCD27 in CSF were compared between patients with T cell mediated neurological disease and non-inflammatory controls. Also, the relation of CSF-sCD27 concentrations with clinical disease activity was investigated in patients with multiple sclerosis. METHODS: Four groups were studied: (1) eight patients with HTLV-1 associated myelopathy/ tropical spastic paraparisis (HAM)/TSP), (2) eight HTLV-I carriers, (3) 41 patients with multiple sclerosis, and (4) 43 patients with other neurological disease (OND). Concentrations of CSF-sCD27 were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Quantification of CSF-sCD27 differentiates patients with HAM/TSP from HTLV-I carriers (p<0.01) and from patients with OND (p<0.001). Moreover, the multiple sclerosis patient group was different from the OND group (p<0.0001). In patients with multiple sclerosis, CSF-sCD27 concentrations were higher in 24 patients with clinically active disease than in 17 with clinically stable disease. In addition, most of the patients with multiple sclerosis with high sCD27 concentrations showed an increase in EDSS, whereas none of the patients with low sCD27 had an EDSS increase. CONCLUSIONS: As a reliable marker of immunological disease activity in inflammatory white matter disease is still not available, it is proposed that quantification of CSF-sCD27 concentrations is a good candidate. Also, it may serve as a tool to stratify neurological diseases in inflammatory and non-inflammatory states.     

Expression of CD5 on B lymphocytes correlates with disease activity in patients with multiple sclerosis

There is growing evidence that implicates B lymphocytes and their products in the pathogenesis of multiple sclerosis (MS). A subpopulation of B lymphocytes expressing the CD5 antigen are involved in several autoimmune disorders through the release of autoantibodies. In this study, we used three-color flow cytometry to examine the expression of CD5 antigen on B lymphocytes from patients with relapsing-remitting MS, and correlated this expression with features of disease activity and circulating levels of autoantibodies against myelin basic protein. CD5 expression on B lymphocytes was significantly higher in patients with active MS when compared to patients with clinically stable MS or those with inflammatory or noninflammatory neurologic disorders. CD5(+) B lymphocytes from patients with active MS correlated significantly with the number of gadolinium-enhancing MRI lesions, and inversely with disease duration. The expression of CD5 on B lymphocytes in MS patients also correlated with circulating levels antibodies against myelin basic protein. Results presented here indicate that clinically active MS is associated with an expanded population of peripheral CD5(+) B lymphocytes.     

Soluble CD30 levels in plasma and cerebrospinal fluid in multiple sclerosis, HIV infection and other nervous system diseases

The CD30 molecule, a member of tumor necrosis factor superfamily, has been suggested to be preferentially expressed and released in soluble form by activated T cells that produce T helper 2 type (Th2) cytokines. To evaluate whether determination of soluble CD30 (sCD30) levels could have a diagnostic value in diseases associated with Th1 and Th2 cytokine involvement, we investigated sCD30 in plasma and cerebrospinal fluid (CSF) from patients with multiple sclerosis (MS), HIV infection and other nervous system diseases. There was no statistically significant difference for plasma sCD30 levels in these clinical groups. However, patients with HIV infection had higher levels of sCD30 in CSF than MS patients. The mean sCD30 values were 3 to 6 folds higher in plasma than in CSF in all patient groups. No relationships were found between sCD30 levels and different clinical variables of MS and HIV infection, except that higher plasma sCD30 levels in HIV-infected patients were found in those with higher CD4⁺ T cell counts (> 200 ± 10⁶) compared to the group with lower cell counts. The findings indicate that determinations of plasma and CSF sCD30 levels in MS and HIV infection have limited or no value as diagnostic or prognostic indicator.     

Correlation of interleukin-2 and soluble interleukin-2 receptor with clinical activity of multiple sclerosis.

Concentrations of interleukin-2 (IL-2) and soluble IL-2 receptor (sIL-2R) in serum and CSF samples were measured in 63 patients with multiple sclerosis (MS) to evaluate their usefulness as markers of disease activity. CSF concentrations of IL-2 and sIL-2R were significantly higher in MS relapse compared with MS patients in remission or with control subjects. These concentrations correlated with the clinical score by which disease severity was assessed, with the number of relapses per year, and with the total disease duration. Furthermore, there was evidence of intrathecal release of IL-2 and sIL-2R in clinically active MS. The results extend the notion that an activated cellular immune state parallels the evolution of the pathological process in MS and suggest that measurement of IL-2 and sIL-2R concentrations may provide an objective marker of disease activity in patients with MS.     

Soluble CD8 and ICAM-1 in serum and CSF of MS patients treated with 6-methylprednisolone

Objective - We studied the effects of large doses of 6-methylprednisolone (6-MP) on serum and cerebrospinal fluid (CSF) soluble CD8 (sCD8) and intercellular adhesion molecule-1 (sICAM-1) levels in clinically active multiple sclerosis (MS) patients.
Material and methods - Paired serum and CSF samples were from 16 patients with definite MS, treated with 6-MP (1 g daily for 6 d) during an active phase of the disease. sCD8 and sICAM-1 levels were determined with ELISA before and after the therapy.
Results - Before 6-MP treatment, sCD8 levels in CSF were higher in MS patients than in patients with noninflammatory neurological disease and in healthy controls; sICAM-1 levels in serum and in CSF were higher in MS patients than in the two control groups. Ten of the 16 patients showed clinical improvement at the end of the treatment. After the therapy, serum and CSF sCD8 levels increased, whereas serum and CSF sICAM-1 levels decreased. There was no correlation between clinical improvement and laboratory parameters. We evaluated sCD8 and sICAM-1 in serum samples from 10 patients 6 months after the 6-MP treatment, when the disease was clinically silent. Neither sCD8 nor sICAM-1 levels differed from those of the control groups.
Conclusions - Our results suggest that high doses of 6-MP can influence serum and CSF sCD8 and sICAM-1 levels in active MS. At least part of the efficacy of corticosteroid treatment in MS might be ascribed to its effect both on the suppressive circuits of immune response, and on the expression of an adhesion molecule that favours lymphocyte trafficking across the blood-brain barrier.     

The role of the Epstein-Barr virus receptor CD21 in multiple sclerosis

Multiple Sclerosis (MS) is characterised by a chronic inflammation and demyelination of brain and spinal cord with a yet unknown aetiology. Based on multiple epidemiological and immunological studies, which suggest a role of Epstein–Barr virus (EBV) infection in the pathogenesis of MS, we investigated CD21 (CR2, complement receptor type 2), which serves as the EBV receptor. Serum concentrations of soluble CD21 receptor (sCD21) were determined in MS patients and controls. In accordance with findings in other autoimmune disorders decreased sCD21 levels were found in MS patients. On ß-IFN treatment serum sCD21 concentrations further decreased. To explore the role of the CD21 gene for MS susceptibility and the altered CD21 levels in MS patients we performed exon sequencing of the CD21 gene. While we identified new single nucleotide polymorphism (SNP) and confirmed previously reported SNPs, none of the SNPs was associated with MS. Our findings demonstrate that sCD21 expression is altered in MS patients similar to other autoimmune diseases although no evidence was found for a specific role of the CD21 gene in MS.     

Serum levels of soluble CD95 are not associated with amelioration of multiple sclerosis during pregnancy

Multiple sclerosis (MS) is considered an autoimmune disease of the central nervous system. Like various other autoimmune disorders, MS normally improves during pregnancy. Pregnant MS patients experience a significant reduction in relapse rates and magnetic resonance (MR) disease activity. How sex steroid hormones affect disease course remains unclear. We hypothesized that hormonal changes during pregnancy might modulate the autoimmune response by enhancing apoptosis of autoreactive T lymphocytes. One of the most important effectors of apoptosis in T cells is the CD95/CD95L system. We have previously reported that the soluble form of CD95 (sCD95) can block CD95-mediated apoptosis and that MS patients show elevated levels of sCD95. Therefore, we considered whether gravidity might influence serum levels of sCD95 in patients, and analyzed the concentration of sCD95 in the sera of 61 patients with relapsing-remitting (RR) MS before, during and after pregnancy. We found no association between serum levels of sCD95 and pregnancy-related immune suppression in MS patients. Thus, sex steroid hormones do not seem to affect the production of anti-apoptotic sCD95.     

Intrathecal synthesis of soluble HLA-G and HLA-I molecules are reciprocally associated to clinical and MRI activity in patients with multiple sclerosis

The aim of this study was to provide further insight into the effective contribution of classical soluble HLA-A, B and C class Ia (sHLA-I) and non-classical soluble HLA-G class Ib (sHLA-G) molecules in immune dysregulation occurring in multiple sclerosis (MS). We evaluated by enzyme-linked immunosorbent assay (ELISA) technique intrathecal synthesis and cerebrospinal fluid (CSF) and serum levels of sHLA-I and sHLA-G in 69 relapsing-remitting (RR), 21 secondary progressive (SP) and 13 primary progressive (PP) MS patients stratified according to clinical and magnetic resonance imaging (MRI) evidence of disease activity. We also tested, as neurological controls, 91 patients with other inflammatory neurological disorders (OIND) and 92 with non-inflammatory neurological disorders (NIND). Eighty-two healthy volunteers served as further controls for sHLA-I and sHLA-G determinations. An intrathecal production of sHLA-I and sHLA-G detected by specific indexes was significantly more frequent in MS patients than in controls (P<0.01). An intrathecal synthesis of sHLA-I was prevalent in clinically (P<0.02) and MRI active (P<0.001) MS, whereas a CSF-restricted release of sHLA-G predominated in clinically (P<0.01) and MRI stable (P<0.001) MS. sHLA-I levels were low in the serum of clinically active (P<0.001) and high in the CSF of MRI active (P<0.01) MS. Conversely, sHLA-G concentrations were decreased in the serum of clinically stable MS (P<0.01) and increased in the CSF of MRI inactive MS (P<0.001). The trends towards a negative correlation observed between CSF and serum concentrations and intrathecal synthesis of sHLA-I and sHLA-G in patients without evidence of clinical and MRI activity confirmed that intrathecal production and fluctuations in CSF and serum concentrations of sHLA-I and sHLA-G were reciprocal in MS. Our results suggest that, in MS, a balance between classical sHLA-I and non-classical sHLA-G products modulating both MRI and clinical disease activity in opposite directions may exist.     

Proteolytic shedding of the macrophage scavenger receptor CD163 in multiple sclerosis

The scavenger receptor CD163 is selectively expressed on tissue macrophages and human monocytes. CD163 has been implicated to play a role in the clearance of hemoglobin and in the regulation of cytokine production by macrophages. Membrane CD163 can be cleaved by matrix metalloproteinases (MMP) resulting in soluble CD163 (sCD163). In the present report the shedding of CD163 was investigated in multiple sclerosis (MS). An upregulation of plasma sCD163 and a down regulation of membrane CD163 in MS patients compared to healthy controls was observed. The levels of plasma sCD163 correlated with plasma MMP-9 levels in controls, but not in MS patients. Moreover, evidence was obtained for CD163-cleaving MMP activity in plasma of MS patients. Finally, the increased proteolytic shedding of CD163 correlated to reduced plasma levels of circulating inflammatory cytokines. Collectively, our results provide evidence for proteolytic shedding of CD163 in MS and suggest a possible link to cytokine production.     

CD95-mediated apoptosis and DNA fragmentation in MS

OBJECTIVE: To investigate possible associations of soluble CD95 (sCD95) serum levels and DNA defragmentation with different MS disease stages and activities. METHODS: Sera of 114 patients were analysed by an ELISA technique for sCD95. In a subgroup of 18 relapsing-remitting MS patients and controls we studied DNA fragmentation by the TUNEL-method in CSF cytospins. RESULTS: sCD95 was detectable in sera of MS patients, healthy controls and meningitis patients without significant differences. CSF specimens showed modest amounts of apoptotic cells in MS and controls. CONCLUSION: We could not demonstrate an association of MS disease course or activity with the expression of sCD95 in sera. DNA fragmentation in the CSF was not significantly enhanced compared to controls. Thus the analysed markers of programmed cell death appear not suitable to monitor MS disease courses.     

Heightened expression of survivin in activated T lymphocytes from patients with multiple sclerosis

The perpetuation of the inflammatory process in multiple sclerosis (MS) may arise from the failure to eliminate potentially pathogenic autoreactive lymphocytes by programmed cell death (apoptosis). Such impairment may be caused by multiple abnormalities of apoptosis regulatory proteins. In this study, we investigated the expression of survivin, a recently described cell cycle-regulated antiapoptosis protein, in lymphocytes from patients with active relapsing-remitting MS and appropriate controls. Survivin reactivity was detected in intrathecal lymphocytes from some MS patients, but not in resting peripheral lymphocytes. However, mitogen stimulation of resting lymphocytes induced survivin expression, which was significantly higher in stimulated intrathecal and peripheral T lymphocytes from MS patients when compared to controls. In contrast, cellular expression of the antiapoptosis protein Bcl-2 was relatively similar between MS patients and the control groups. Moreover, heightened survivin expression in MS patients correlated with T lymphocyte resistance to apoptosis, and was independent of cellular expression of the death receptor Fas. These findings suggest that upregulation of the antiapoptotic protein survivin in mitogen-stimulated T lymphocytes is a feature of multiple sclerosis.     

Production and secretion of calcitonin gene-related peptide from human lymphocytes

Programmed cell death (apoptosis) is critical for the normal development and homeostasis of the immune system. There is increasing evidence that dysregulations of apoptotic pathways are associated with autoimmune disease, including multiple sclerosis (MS). Cellular commitment to apoptosis is partly regulated by the Bcl-2 family proteins, which includes the death antagonists Bcl-2 and Bcl-X(L), and death agonists Bax and Bad. Since the role of these proteins in the pathogenesis of MS is currently unknown, we analyzed their expression profile in peripheral and intrathecal lymphocytes from MS patients and appropriate controls. We observed a significant reduction in the expression ratios of pro-apoptotic to anti-apoptotic Bcl-2 members in both peripheral and intrathecal lymphocytes from MS patients when compared to corresponding ratios in patients with inflammatory or noninflammatory neurologic controls, or healthy individuals. The relative coexpression ratios of these pro- and anti-apoptotic Bcl-2 family proteins in MS were more significant than the expression of individual members. The low cellular expression ratios of pro-apoptotic proteins in MS were confirmed in vitro activated T lymphocytes. Cellular expression of Bcl-2, Bcl-X(L), Bax or Bad in MS patients was independent of the expression of other apoptotic regulatory molecules, such as Fas receptor protein or FLIP. Our findings suggest that the abnormal expression patterns of Bcl-2 family proteins in MS may promote apoptotic resistance of potentially pathogenic, autoreactive lymphocytes, and may allow for continuing cellular proliferation and tissue destruction within the central nervous system.     

Increased serum levels of soluble CD14 indicate stable multiple sclerosis

The aim of this study was to evaluate a possible role of soluble CD14 (sCD14) in multiple sclerosis (MS). We found that sCD14 serum levels measured by ELISA were higher in MS patients compared to neurological and healthy controls. Within the MS group sCD14 levels were increased in relapsing-remitting and secondary progressive MS compared to primary progressive MS. Furthermore, sCD14 concentrations were increased during stable disease. An increased expression of sCD14 was also detected after treatment with interferon-beta. In summary, we report evidence that serum sCD14 levels are increased in MS and correlate inversely with disease activity in relapsing MS patients.     

急性动脉粥样硬化性脑梗死患者外周血CD137的表达特点及其临床意义

目的探讨急性动脉粥样硬化性脑梗死(atherosclerotic cerebral infarction,ACI)患者外周血肿瘤坏死因子受体超家族(tumor necrosis factor receptor super family,TNFRSF)成员CD137的表达特点及其临床意义。方法入选发病72h以内住院患者共46例,其中ACI组27例,无症状性颈动脉狭窄(asymptomatic carotid stenosis,ACS)组19例,另选择同期健康体检者20名为正常对照组(normal control,NC)。采用流式细胞术和流式细胞磁珠微阵列法(CBA)检测各组观察对象外周血CD4~+T细胞及其CD4~+CD28-T细胞亚群表面CD137的表达和血浆可溶性CD137水平。结果 ACI组外周血CD4~+CD28-T细胞比例[(14.13±4.42)%]、CD4~+CD28-T细胞CD137的表达[(2.60±2.14)%]、血浆sCD137[(3.43±2.48)pg/mL]水平显著高于ACS组[(9.03±4.60)%、(0.70±0.66)%、(2.07±0.814)pg/mL]和NC组[(8.08±3.30)%、(0.45±0.31)%、(1.26±0.70)pg/mL](P0.01),且ACI组CD4~+T细胞的CD137表达及血浆sCD137水平与NIHSS评分(P0.05)和梗死体积(P0.01)呈正相关。结论 ACI患者发病早期CD4~+CD28-T细胞、CD4~+T细胞及其CD4~+CD28-T细胞表面CD137、血浆sCD137水平升高,且CD4~+T细胞的CD137表达及血浆sCD137水平与NIHSS评分呈正相关,提示其可能作为评估脑梗死临床严重程度的外周生物学标志。     

Lack of association of the CD14/C -159T polymorphism with susceptibility and progression parameters in Turkish multiple sclerosis patients

Soluble (s) CD14, being a receptor for lipopolysaccharides (LPSs) may inhibit LPS-triggered apoptosis and T lymphocyte proliferation. C to T exchange at position -159 in the promoter region of the CD14 gene might lead to higher sCD14 levels. Limited number of groups have studied whether these polymorphisms might influence the development of organ specific autoimmunity and whether higher CD14 levels are associated with increased levels of cytokines trigerring inflammatory processes. However their data contradict each other. In this study serum levels of sCD14 based on ELISA were measured in 77 treatment-naive patients and in 67 healthy controls. As the C-159T proximal promoter region regulates sCD14 levels, we investigated whether C-159T polymorphism is related to progression index in 250 MS patients vs. 183 healthy controls. CD14 polymorphism frequency between the healthy controls and the MS patients were not significantly different. While TT genotype of MS patients demonstrated significantly lower sCD14 levels compared to CC genotype; this difference was not reflected on the disease progression index. Our study that extends the prior data of previous studies reflects that sCD14 do not appear to be a solely prominent element of innate immunity in MS.     

Immunologic variables in acute mania of bipolar disorder

Macrophages, lymphocytes and their products, may be involved in the pathophysiology of psychiatric disorders. The cell-mediated immune activation response of manic patients during pre-medication and medication stages remains unclear. The purpose of this case-control study was to investigate the plasma levels of immunologic variables, including interleukin (IL)-1 receptor antagonist (IL-1RA), soluble CD 4 (sCD4) and sCD8, and TH1 (interferon [IFN]-gamma and IL-2) and TH2 (IL-4 and IL-10) cytokines in patients with pre-medicated, medicated bipolar mania. The study subjects, aged 16-44 years, were physically healthy patients with Young Mania Rating Scale (YMRS) scores > or =26, and normal controls, aged 19-40 years, were matched for sex. The immune variables were measured in acute mania and in consequent remission (YMRS scores < or =12) among bipolar patients. The plasma levels of IL-1RA, sCD4, and sCD8 were found significantly increased in pre-medicated acute manic patients as compared to normal controls. But only IL-1RA and sCD8 were found different in remitted bipolar patients as compared to normal controls. For TH1 cytokines, culture supernatant level of IFN-gamma was found significantly lower in manic patients of both acute and remission stages as compared to normal controls. No significant difference was found in IL-2 level in pre-medicated acute manic patients compared to controls. For TH2 cytokines, no significant differences in IL-4 and IL-10 levels were observed. We showed that cell-mediated immune response was activated in patients with bipolar disorder during the pre-medication, medication, and the remission stages. Our study findings suggest that the immune-modulation in patients with bipolar disorder may be abnormal.     

Disease activity in multiple sclerosis correlates with T lymphocyte expression of the inhibitor of apoptosis proteins.

The pathogenesis of multiple sclerosis (MS) is thought to involve failure of programmed cell death (apoptosis) to eliminate potentially pathogenic, autoreactive T lymphocytes. This failure may be caused by multiple abnormalities of the cell death machinery. The inhibitors of apoptosis (IAP) proteins are central regulators of cell death that inhibit apoptosis induced by a variety of stimuli. In this study, we investigated the dynamics of cellular IAP-1, IAP-2, and X-linked IAP, in resting and mitogen stimulated T lymphocytes from MS patients and relevant controls. The expression of IAP proteins was significantly higher in mitogen stimulated T lymphocytes from patients with clinically active MS when compared to corresponding expressions from patients with stable MS or from other controls. Heightened expression of IAP proteins in patients with active MS correlated with clinical features of disease activity, and with T lymphocyte resistance to apoptosis. In contrast, cellular expression of the anti-apoptosis protein Bcl-2 did not differ between active and stable MS, and was relatively similar between MS patients and controls. These findings suggest that overexpression of IAP proteins in stimulated T lymphocytes is a feature of clinically active multiple sclerosis.