Monoclonal antibody analysis of blood T-cell subsets in multiple sclerosis

The present study deals with the characterization of peripheral blood T-cell subpopulations in multiple sclerosis (MS) patients during different stages of the disease. An indirect immunofluorescence assay was performed using monoclonal antibodies directed at lymphocyte surface antigens. Patients in exacerbation were found to have significantly (p < 0.001) reduced OKT ₈ + (T-suppressor) cells and a high helper/suppressor ratio (p < 0.001). Patients in remission showed a significant increase of suppressor T-cells compared to controls (p < 0.02) and patients during relapse (p < 0.001); H/S ratio was consequently low compared to acute MS (p < 0.001) and controls (p < 0.1). Patients with a progressive course showed an intermediate T-subset pattern. The results are discussed in the light of the most recent neuroimmunological approaches to MS.This study was supported by the C.N.R. grant n. 83/00635.52     

Search for retroviral sequences in peripheral blood mononuclear cells and brain tissue of multiple sclerosis patients

Summary DNAs from peripheral blood mononuclear cells (PBMCs) of 21 patients with multiple sclerosis (MS), 1 patient with tropical spastic paraparesis (TSP) as well as DNAs from brain and spinal cord of 5 MS cases and 3 controls were examined for human T-cell lymphotropic virus (HTLV)-related sequences by polymerase chain reaction. The primers used were derived from the HTLV-I gag, env and tax genes. Amplified products were separated on agarose gels, blotted onto nylon membranes and hybridized to specific radiolabelled oligonucleotides. The sensitivity of amplification and hybridization was one copy of target DNA in 10⁵ cellular genomes. None of the specimens was positive for HTLV-I sequences except the TSP probe. These negative data are all the more significant because brain material from MS patients was used in these studies. Our studies thus fail to support speculations that HTLV-I is involved in the aetiology of multiple sclerosis.     

T-cell subsets in spinal fluid of multiple sclerosis patients

CSF T-cells and T-cell subsets were characterized by monoclonal antibodies in 15 untreated multiple sclerosis (MS) patients, 17 immunosuppressed chronic progressive MS patients and 9 patients with other neurological diseases. A negative correlation was found between total cell numbers and T suppressor cell percentages in untreated and treated MS patients. A negative correlation (r = -0.71) was found between intrathecal IgG levels and T suppressor cell percentages in untreated MS patients. In peripheral blood, no correlation between T-cells and T-cell subsets with IgG levels was found. It is discussed that T-cell subsets and intrathecal IgG levels may be indicators of the activity of the inflammatory process in the brains of chronic progressive MS patients.     

Human spumaretrovirus antibody reactivity in multiple sclerosis

The role of human spumaretrovirus (HSRV) infections in the pathogenesis of multiple sclerosis (MS) was investigated with recombinant HSRV env-specific enzyme-linked immunosorbent assay. The presence of HSRV antibodies was determined in pairs of serum and cerebrospinal fluid (CSF) samples from 60 MS patients. In 7 of these patients serial serum and CSF samples were obtained in relation to the clinical activity of the disease during a period of 2 years. No increased antibody reactivity was demonstrable in the MS population compared with 14 aseptic meningitis patients, 50 blood donors and 16 healthy controls. Slightly elevated levels of antibodies were demonstrable in serum and/or CSF in 4 MS patients but also in 1 patient with aseptic meningitis, 1 blood donor and 1 child. No marked serum or CSF HSRV antibody fluctuation was observed in the MS patients followed longitudinally. Thus, this study does not support the involvement of HSRV in the pathogenesis of MS.     

Lymphocytoplasmapheresis in multiple sclerosis: one-year results in 6 patients

6 patients with definite MS underwent lymphocytoplasmapheresis for one year. Clinical data, evoked potential recordings and peripheral blood lymphocyte helper/suppressor ratio were assessed before and after the treatment and were compared with those of a control group of 10 multiple sclerosis patients. Lymphocytoplasmapheresis did not significantly modify clinical and laboratory findings compared with the control group.This work was supported by C.N.R. grant n^o 84-02427.56 Progetto finalizzato medicina preventiva e riabilitativa, Sottoprogetto SP3-Malattie del sistema nervoso.     

Thrombomodulin in the sera of patients with multiple sclerosis and human lymphotropic virus type-1-associated myelopathy

Damage to the blood-brain barrier, which mainly consists of cerebral endothelial cells, has been demonstrated in multiple sclerosis (MS) clinically and histochemically. To investigate the endothelial cell damage, we evaluated the presence of soluble thrombomodulin in the sera of patients with MS and human T lymphotropic virus type-1-associated myelopathy (HAM) using an enzyme-linked immunosorbent assay. Serum thrombomodulin levels were significantly increased in patients with acute relapsing MS during an exacerbation and chronic progressive MS as compared with those of controls (P < 0.001, respectively). Patients with HAM also had higher serum levels of thrombomodulin than did controls (P < 0.001). There was significant difference between patients with HAM and seropositive non-HAM carriers (P < 0.01). These results suggest that the detection of serum thrombomodulin could be used as a marker of endothelial cell damage in inflammatory diseases such as MS and HAM.     

Altered blood T-cell subsets in patients with multiple sclerosis

We have found an alteration in T-cell subsets in patients with active multiple sclerosis, specifically an increase in the T4:T8 ratio. These findings have been reproducibly obtained over the past four years, occurring in the majority of acute patients tested early in the course of an attack and in between 25 and 40% of chronic progressive patients, depending on their stage of illness. These changes correlate with pleocytosis in spinal fluid and with other abnormalities of immune function, such as spontaneous immunoglobulin production. They have been helpful in assessing disease activity in patients being treated on a variety of protocols and as part of research studies of immunoregulatory abnormality in multiple sclerosis, but have not been helpful as a diagnostic test for multiple sclerosis. The decrease of these cells in the peripheral blood of patients with active disease may be secondary to migration of these cells to the central nervous system, where they are sequestered.     

多发性硬化合并癫癎临床特点分析

目的探讨多发性硬化(multiplesclerosis,MS)患者中合并癫的情况。方法回顾1981—2004年北京协和医院收治的临床确诊和实验室支持确诊的MS患者119例,分析其中合并癫患者的临床特点。结果MS患者发生癫的患病率为5.0%±0.1%,明显高于普通人群患病率的0.5%～1.0%。但是起病年龄、额叶皮质下以及侧脑室旁病灶、年复发率及扩展功能障碍状态量表(EDSS)评分与多发性硬化患者是否出现癫无统计学明显相关。颞叶皮质下病灶与多发性硬化患者出现癫存在统计学相关(P=0.035)。结论MS患者较普通人群易患癫,其中存在颞叶皮质下病灶的MS患者更易出现癫。     

Multiple sclerosis and migration in Italy: A case/control study

The present study was performed on Milan residents, in order to evaluate the risk of acquiring multiple sclerosis between patients born in Milan and patients born in Southern Italy. For this purpose, 216 consecutive subjects with definite or probable multiple sclerosis, inmates of the Neurological Institute C. Besta (Milan) from 1968–1976, were matched with 216 controls, uniform as regards age, sex, profession and residence, who were hospitalized at the same time for other neurological diseases. Comparing the case/control pairs for the migration, we observed that the North-Centre born had a relative risk of acquiring the disease significantly superior to one (RR = 1.9; P = 0.01) as compared to migrants from the Southern regions of Italy.     

Multiple sclerosis in G: genes and geography

Multiple sclerosis (MS) shows uneven geographic distribution globally as well as within countries. In epidemiological studies we have previously demonstrated that there is a high-risk focus for MS in the southern Ostrobothnian region of western Finland. In genetic studies we recently identified haplotypes that associate with MS specifically in patients originating from southern Ostrobothnia suggesting a founder effect. Such haplotypes can be used as molecular tools for tracing common ancestry between patients in different geographic locations. In addition to providing clues to the historical origin, such a genetic archeological approach should help narrow the size of the shared haplotype, thus facilitating the identification of etiological variants and possibly define a superfamily of MS patients with common pathogenetic mechanisms.     

The influence of human allergic encephalitogenic peptide on cell-mediated cytotoxicity reactions in patients with multiple sclerosis

Multiple sclerosis patients and normal control persons were assayed for cell-mediated cytotoxicity against target cells coated with human allergic encephalitogenic peptide. Coating of different types of target cells resulted in increased cytotoxicity, most clearly seen against homologous lymphocytes and virus-infected fibroblasts. Both patients and controls showed reactivity against coated targets. A possible role for this type of reaction in the pathogenesis of multiple sclerosis is proposed.     

T-cell subsets in the cerebrospinal fluid and blood of patients with multiple sclerosis

The absolute numbers and ratios of helper/inducer (T4) and cytotoxic/suppressor (T8) T-cells were determined in cerebrospinal fluid (CSF) and blood of patients with multiple sclerosis (MS) and various other neurologic diseases (OND). In patients with MS, the T4:T8 ratio was higher in both blood and CSF, and the increase was significantly greater in CSF than in blood. These findings were due to an increased proportion of T4-lymphocytes in the CSF and to a decreased proportion of T8-cells in blood. These results indicate the need for additional studies of CSF lymphocytes in patients with MS.     

T-cell subsets in multiple sclerosis: relationships between peripheral blood and cerebrospinal fluid

We contemporarily studied cerebrospinal fluid (CSF) and peripheral blood (PB) T-cell subsets, defined by monoclonal antibodies, in 29 patients with multiple sclerosis (MS) and 10 patients with other neurological diseases (OND). All subjects showed a clear-cut prevalence of CSF T-cells. Similarly, T-helper and T-suppressor subsets tended to show higher percentages in CSF in almost all subjects except relapsing MS, who were characterized by low percentages of T-suppressors in PB and even much lower percentages in CSF. Helper/suppressor ratios were found to be almost similar in the two body compartments of OND patients, lower in CSF than in PB of chronic progressive MS, always higher in CSF than in PB of relapsing MS. MS patients in remission showed both patterns of progressive MS and OND patients. Our results demonstrate that the loss of PB T-suppressor in relapsing MS is not due to a migration of such cells into CSF. Furthermore, regarding T-lymphocyte subsets, a typical CSF/PB pattern characterizes relapsing MS from other patients.     

Production of viral antibodies in vitro by CSF cells from mumps meningitis and multiple sclerosis patients

Cerebrospinal fluid (CSF) cells from 4 mumps meningitis and 11 multiple sclerosis (MS) patients were cultured in vitro for 7 days with and without pokeweed mitogen (PWM) stimulation. The cells produced varying amounts of IgG without stimulation and no significant increase of IgG synthesis was observed after PWM stimulation. Antibodies against mumps, measles, rubella, herpes simplex, and adeno viruses were measured in the supernatants of the cultures by a sensitive enzyme immunoassay. In the mumps meningitis patients, the largest amount of antibody was against mumps virus but low amounts of antibodies with other specificities were also synthesized by CSF cells of one patient. The most commonly detected specificities in MS patients were against measles and rubella viruses, whereas antibodies against adeno and mumps viruses were detected in only one CSF cell supernatant. No antibodies produced against herpes simplex virus in vitro were detected in any of the supernatants. The amounts of viral antibodies produced in vitro and intrathecally were only partially correlated.     

Multiple sclerosis and the HLA-D region: linkage and association studies

Inheritance patterns of multiple sclerosis (MS) in multiplex families suggest a complex aetiology involving environmental and genetically determined components. The association between the HLA class II DR15, DQ6, Dw2 haplotype and MS has been well documented in patients with ancestral origins in Northern Europe. Conversely, linkage analysis of this region in multiplex families, derived from a population base, has generated negative results. Thus, given the Dw2 specificity association, evidence implicating this locus in disease susceptibility appears contradictory. We have collected and determined the HLA-DR and -DQ haplotypes of 115 sibling pairs with multiple sclerosis, and confirm a significant association with the Dw2-associated haplotype, both in index cases and their affected siblings compared with controls. However, using a sibling pair linkage analysis that restricts haplotype sharing probabilities to defined genetic models, we have not observed linkage of this region to susceptibility in MS. We discuss the basis for association and linkage and conclude that the DR15, DQ6, Dw2 haplotype does represent a susceptibility locus but its contribution to the pathogenesis is small; although it may interact epistatically with other susceptibility genes, this haplotype is not necessary for disease expression.     

Multiple sclerosis: genetic versus environmental aetiology: epidemiology in Israel updated

The incidence and prevalence of multiple sclerosis (MS) were compared, controlling for age, in native-born Israelis of different origins and in immigrants to Israel. This comparison was carried out in two populations, countrywide and in Jerusalem. In the countrywide population, ascertainment was based mainly on hospitalizations; it included 252 patients who were native-born and 150 who had immigrated from Africa-Asia (AA immigrants). The 89 MS patients of Jerusalem also included patients diagnosed in outpatient clinics. In native-born Israelis whose father was born in Europe-America (I-EA), the incidence and prevalence of MS were found to be as high as or even higher than that found previously in immigrants from Europe-America. Among native-born Israelis whose father was born in Africa or Asia (I-AA), the yearly age-adjusted incidence and prevalence rates were found to be 1.4- to 1.8-fold higher than among AA immigrants, pointing to environmental factors. The incidence and prevalence rates in the I-EA were 1.2- to 1.6-fold higher than in the I-AA, pointing to genetic factors. These results seem to point to both environmental and genetic factors in the aetiology of MS. Further research is needed, however, to disentangle the genetic factors from possible environmental differences in the two ethnic groups.     

Multiple sclerosis: Demyelination and myelination inhibition of organotypic tissue cultures of the spinal cord by sera of patients with Multiple Sclerosis and other neurological diseases

Summary Sera from 44 patients with Multiple Sclerosis, of three patients with neurological syndromes compatible with Multiple Sclerosis, of 34 patients suffering from other neurological diseases and of 25 pregnant healthy young women were tested for their demyelinating activity in myelinated tissue cultures. In order to leave the investigators unprejudiced, all sera were coded and intermixed with controls of rabbit EAE serum which had a potent demyelinating capacity. Demyelination was graded (from 0–4), heat lability at 56°C (complement dependency?) was also tested with each serum. Only demyelination of a degree of 2 and more, which was abolished by heating to 56°C, was counted as positive.Six of the 44 sera from MS patients (13.6%), 19 of 37 sera from neurological patients and none of the healthy young women demyelinated. Thus, serum demyelination of tissue cultures seems to be a nonspecific indicator of chronic disease of the nervous system and is of considerable general neurological interest, but does not indicate a demyelinating disease.Myelination inhibition was not observed with any of the human sera tested for it.

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Zusammenfassung Seren von 44 Patienten mit Multipler Sklerose (MS), 3 Patienten mit neurologischen Syndromen, bei welchen eine Multiple Sklerose nicht mit Sicherheit ausgeschlossen werden kann, von 34 Patienten mit anderen neurologischen Krankheiten und 25 gesunden schwangeren Frauen wurden auf ihre Entmarkungsaktivität in myelinisierten organotypischen Gewebskulturen geprüft. Um eine Voreingenommenheit der Untersucher möglichst zu vermeiden, wurden die Seren verschlüsselt und mit stark entmarkendem Serum eines Kaninchens mit Experimenteller Allergischer Encephalomyelitis — ebenfalls verschlüsselt — mituntersucht. Es wurden 5 Entmarkungsgrade unterschieden (0–4) und die Thermolabilität (Complement-Abhängigkeit?) bei 56°C getestet. Nur Entmarkungsgrade von 2 und mehr, die bei 56°C reduziert wurden, wurden als positiv gewertet.6 der 44 Seren von MS-Patienten (13,6%), 19 der 37 Seren von anderen neurologischen Patienten und keines von den gesunden jungen Frauen entmarkte. Somit scheint Serum-induzierte Entmarkung in Gewebskulturen ein unspezifischer Indikator chronischer neurologischer Krankheiten zu sein. Der Befund ist von erheblichem allgemeinem neurologischem Interesse, eignet sich aber nicht als Test für Entmarkungskrankheiten.Myelinationshemmung in Kulturen wurde mit keinem der getesteten Seren beobachtet.

     

Social support as a buffer in the relationship between treatment for depression and T-cell production of interferon gamma in patients with multiple sclerosis

OBJECTIVE: This study examined the buffering effects of social support on the relationship between depression and autoaggressive immune function in multiple sclerosis (MS). METHODS: Fourteen participants with comorbid diagnoses of MS and major depressive disorder received 16 weeks of psychotherapy or antidepressant medications. Depression and T-cell production of interferon-gamma (IFN-gamma), a lynchpin in MS pathogenesis, were assessed at baseline and posttreatment. Social support was assessed at baseline. RESULTS: Both depression and T-cell production of IFN-gamma were significantly reduced over the 16 weeks of treatment. There was a significant interaction between change in depression, change in IFN-gamma, and social support (R(2)=.26, P=.03) such that social support served as a buffer. CONCLUSION: These results support the hypothesis that social support buffers the effects of change in depression on IFN-gamma production. However, these findings should be viewed as preliminary due to the small sample size and the absence of a control condition.