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Multiple sclerosis (MS) is a central nervous disease thought to be elicited by an autoimmune process. Many studies in recent years have concentrated on finding the alterations in the peripheral blood immune profile in MS patients that would reflect disease activity. In the present study, we investigated surface antigen expression on lymphocytes and granulocytes from MS patients and control subjects. We have studied 29 patients suffering from relapsing-remitting or relapsing-progressive forms of MS. The disease was diagnosed in all patients at least 12 months before inclusion into the study. All patients had no attack at the study entry date or within a previous month. The control group included 29 age-matched subjects. Phenotyping of peripheral blood leukocytes was carried out with different fluorescence-conjugated murine monoclonal antibodies. The analysis was performed with three-color flow cytometry. The following antigens were determined [cluster of definition (CD)]: leukocyte common antigen (LCA) (B220, T 200, Ly-5), CD45; LPS-R (lipopolysaccharide receptor), CD14; found on all T cells, CD3; LFA-2 (lymphocyte function associated antigen, T 11), CD2; coreceptor for MHC class II molecules, found on helper T cells, CD4; coreceptor for MHC class I molecules, found on suppressor/cytotoxic T cells, CD8; B4, found on all human B cells, CD19; NCAM (neural cell adhesion molecule), CD56; integrin beta2 subunit, associated with CD11a (CD11a/CD18, LFA-1, alphaLbeta2) and CD11b (CD11b/CD18, Mac-1,CR3, alphaMbeta2), CD18; alphaL, alpha subunit of integrin LFA-1 (alphaLbeta2, CD11a/CD18), CD11a; alphaM, alpha subunit of integrin Mac-1 (CR3, alphaMbeta2, CD11b/CD18), CD11b; ICAM-1 (intercellular adhesion molecule), CD54; H-CAM, Hermes antigen, Pgp-1, CD44; AIM (activation inducer molecule), early activation antigen, CD69; T-cell receptor gammadelta, TCR gammadelta. In the MS group, we have found a significant increased expression of CD54 and CD44 antigens on lymphocytes, and higher percentage CD54(+) and CD11a+CD54(+) lymphocytes out of all lymphocytes compared with the control group. We have also found a significant increased expression of CD11a, CD18 and CD54 antigens on granulocytes, and higher percentage CD11b+CD18(+) granulocytes out of all granulocytes in MS patients compared with control. Higher levels of expression of the adhesion molecules may reflect the activation state of leukocytes in MS patients., (Copyright 1999 Lippincott Williams & Wilkins)

There is increasing evidence that inflammatory responses play an important role in the pathogenesis of atherosclerosis and cerebral infarct. The aim of this study was to determine the amount of immunocomplexes (i.c.) in patients with stroke in the early period of the disease. Studies were performed on 35 subjects. The concentration of immunocomplexes was determined with the precipitation method (3.5% polyethylenoglikol was used). Increased concentration of i.c. was found in patients with cerebral infarct (after 12 hours and 7 days). It suggests, that i.c. could be one of the markers for systemic inflammation and be important in the patogenesis of atherosclerosis and stroke.

The relationship between the central dopaminergic and the immune system is poorly understood. Experimental work suggest that damage of the nigrostriatal system may influence immunity. Immunological abnormalities have been described in Parkinson's disease and in a mouse model of this disorder induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In this report, we present evidence that reduced numbers of L3T4 T cells in blood, and diminished primary antibody response to sheep erythrocytes in MPTP treated mice can be restored by pargyline pretreatment. Since pargyline prevents dopamine depletion in the striatum in MPTP treated animals, our data extend previous experimental observations and support a possible role for dopamine in immune regulation.

Vascular diseases of CNS are usually a result of certain risk factors. In a number of cases vascular diseases are caused by inflammatory process within arteries. Establishing the intrathecal synthesis of immunoglobulins is an important indicator of the inflammatory process in the CNS. 16 patients with multifocal vascular lesions in neuroimaging examinations (MRI or CT), were analysed for the frequency of occurrence of inflammatory process within the CNS. The presence of intrathecal synthesis of IgG was established by mathematical formulas and presence oligoclonal IgG bands. In some patients presence of oligoclonal IgG bands was found, and regarded responsible for stroke. In this group the patients were younger, and mainly without other risk factors for vascular diseases. In the majority they were patients with systemic inflammatory process (e.g. SLE). The establishing of inflammatory process within the CNS sometimes has therapeutic implications.

During the last 12 years 4258 amniocenteses were performed between the 12th and 20th week of gestation (including 323 early amniocenteses carried out before 15th week). In every case, cytogenetical examination was performed and concentration of AFP was determined. In cases with elevated AFP level electrophoresis of AchE izoenzymes was performed. The results of the tests enabled us to calculate laboratory standard values of AFP in the amniotic fluid for 12th to 20th weeks of gestation. In 22 of 44 pregnancies with Down's syndrome the value of AFP concentration was below the 25th percentile of the laboratory normal value. In 5 of 10 pregnancies with Edward's syndrome AFP level exceeded significantly the 75th percentile of the laboratory norm. In two cases it was due to coexisting spina bifida and in one case due to omphalocele. In 28 amniotic fluid samples AFP concentration exceeded normal level and electrophoresis of AchE revealed additional band. In 26 cases increased values of AFP were due to open neural tube defect in the fetus: 13 cases of anencephaly and 13 cases of spina bifida; in the remaining two other cases omphalocele was found.

Unlabelled: The demonstration of the presence of intrathecal synthesis of immunoglobulins as oligoclonal band (OB) in cerebrospinal fluid (CSF) is considered to be important as an aid to multiple sclerosis diagnosis. We used the Phast System equipment for OB detection in the CSF. Separation was performed on polyacrylamide gels pI 3-9.7. After separation proteins were visualised with silver staining according to manufacturer instruction or transferred electrophoretically on to nitro-cellulose membrane and developed by immunostaining technique. We also calculated indexes of intrathecal synthesis of IgG. We tested CSF of patients with clinically defined multiple sclerosis (n = 29) according to Poser criteria. In all cases IgG indexes were normal. OB were present in 83% with silver staining and in 93% when we used immunoblotting technique., Conclusions: 1. Detection of OB with Past System is easily performed in routine diagnostic process. 2. Immunoblotting technique increases sensitivity of detection of OB and needs small amounts of CSF. 3. The demonstration of OB in CSF is a method of choice in diagnostic process of inflammatory diseases of nervous system, especially in cases of MS.

The role of the central dopaminergic system in modulating immune response is not completely established. We examined the influence of central dopamine depletion on selected parameters of immune functions in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated and untreated mice. IgM antibody production of splenocytes to sheep red blood cells was reduced in MPTP-treated mice (P < 0.001). Proliferation of splenocytes in response to a wide range of mitogen concentrations (Concanavalin A, phytohaemagglutinin, lipopolysaccharide) was also significantly diminished in MPTP-treated mice. Production of migration inhibition factor (MIF) was diminished only in low mitogen concentration. Our results obtained in the experimental model of Parkinson's disease provide evidence that the damage of the central dopaminergic pathways induces alterations of some immune functions in mice.

TNF alpha production by peripheral blood monocytes was studied in seventeen patients with a recent exacerbation of MS, thirteen with remission of MS and fourteen patients with SSPE. Monocytes from both MS groups spontaneously secreted high amounts of TNF alpha in vitro. Addition of lipopolisaccharide could not stimulate further synthesis of TNF alpha. In SSPE spontaneous and stimulated TNF alpha release did not differ from that in the control group. The observed changes in TNF alpha release in MS patients could play a role in the pathogenesis of the disease.

The immunological status of patients with Huntington's disease was studied and compared with that of an age-matched control group. No remarkable abnormalities in lymphocyte subpopulations were observed. The percentage of B cells, CD4+, CD8+, DR+ cells and stimulated cells bearing Tac receptors remained unchanged. The proliferative response to mitogens and the production of interleukin-1 (Il-1) were decreased, whereas the IgG level was increased. It is possible that changes in the levels of neurotransmitters affect the immunological function in basal ganglia disease.

In CSF of patients with acute aseptic meningitis (AAM) an increase in the percentage of total and active T lymphocytes was observed, whereas in SSPE a decrease in T cells was noted. In SSPE there was the increase in the relative number of lymphocytes bearing "avid" FcIgG receptor and decrease in alpha-naphthyl nonspecific esterase-positive cells, but in AAM the cell proportions were not disturbed. The changes in the peripheral blood lymphocytes subsets were less evident. The results seem to suggest that in AAM there is a strong T response in CSF with well preserved proportions of T-cell subsets, while in SSPE a low T-cell level may be accompanied by disturbances in T-cell function.

Cell-mediated and humoral immunity were studied in 30 patients with subacute sclerosing panencephalitis (SSPE). Marked changes in cell-mediated immunity were observed, manifested as a decrease in total lymphocyte count, decrease of proportion of T cells forming late E rosettes, depression in lymphocyte blastogenesis, production of migration inhibition factor and delayed-type skin reactivity. The humoral immune responses was not so markedly changed and only increased levels of serum IgG, IgM and the C3c fragment of complement were noticed. The changes in immunity were more pronounced in patients with a more advanced stage of the disease. 19 patients were investigated 3 times at 7-week intervals during treatment with isoprinosine. In the group of patients (10 cases) whose clinical status was rapidly deteriorating, a marked decline of cell-mediated immunity was observed. In the group of patients showing a stationary course of the disease, cellular immunity was improving. The mechanism of the disturbances of non-specific immunity in SSPE and the influence of isoprinosine on the immune answer are discussed.

Cerebrospinal fluid (CSF) and peripheral blood (PB) lymphocyte sensitization to rabbit myelin basic protein (MBP) in 44 multiple sclerosis (MS) patients, 21 patients with other neurological diseases (OND) and 14 persons with neurosis was studied with the antigen-active rosette forming cells (Ag-ARFC) assay. The frequency of sensitization of CSF lymphocytes to MBP in groups of MS patients in the relapse stage and the chronic progressive stage was higher than in the group of MS patients in the stable stage and the OND patients. None of the healthy subjects showed a positive reaction with MBP. In BP there were no differences in the incidence of sensitization to MBP between patients in various stages of the disease, but it was higher than in the group of patients with OND and neuroses. In the patients who had suffered from MS for less than 4 years, sensitization to MBP was more common in CSF lymphocytes than in BP lymphocytes. The results suggest that primary sensitization to MBP occurs in CSF, and is probably secondary to myelin damage. However at present it is difficult to determine the extent to which sensitization of CSF and PB lymphocytes to MBP play a role in further demyelination processes.

In 20 patients with head trauma associated with loss of consciousness without massive damage to the tissues total count of lymphocytes T in the peripheral blood was determined, and test of lymphocyte blastic transformation and leucocyte migration inhibition were done. The investigations were done one time only, usually 5 days after trauma. In the patients the total count of lymphocytes and T-cells was decreased in the peripheral blood and production of the factor inhibiting leucocyte migration was reduced. The immune disturbances observed in these patients were due probably to the action of stress triggered by trauma.

Immunological investigations were carried out assessing the humoral and cell-mediated immunity in 22 patients with epilepsy before starting treatment and at least one year after the onset of treatment with phenytoin or carbamazepine. Sixteen patients were treated with phenytoin and 6 with carbamazepine. In the group treated with phenytoin a statistically significant fall was observed in the serum level of IgG and a decrease in the value of blastic transformation of lymphocytes after stimulation with PHA and PPD with a rise in the number of lymphocytes forming total rosettes with sheep erythrocytes. In the group treated with carbamazepine only a statistically significant fall was observed in blastic transformation of lymphocytes stimulated with PHA, and a trend was noticed for increasing serum IgG and IgM levels. The observed changes in immunological reactivity caused by administration of phenytoin and carbamazepine show that these drugs exert an immunomodulating effect, but changes produced by them are slight and caused no immune deficit in the observed group.

Using the test of active E rosettes stimulated with antigen the sensitization was studied of lymphocytes from the cerebrospinal fluid and blood to basic protein of myelin, galactocerebroside and cerebral gangliosides. The investigations were conducted in groups of patients with SSPE and with other non-inflammatory neurological diseases and in a control group without evidence of organic brain damage. It was shown that in SSPE sensitization of these lymphocytes from the CSF and from the blood to the studied antigens occurred with a frequency approaching that in the group of non-inflammatory neurological diseases and less frequently than this was formerly observed in multiple sclerosis.

Lymphocyte subpopulations were evaluated in the cerebrospinal fluid and peripheral blood in 22 patients in the initial stage of aseptic meningitis and during convalescence. In the acute stage of the disease the proportion of lymphocytes T, including their active forms, was increased, during convalescence a further significant rise was observed in the subpopulation of lymphocytes T with a fall of the proportion of lymphocytes bearing the high "avid" Fc IgG receptor. In patients with acute meningitis and during convalescence a rise was observed in the proportion of lymphocytes T in the peripheral blood. In the acute stage of the disease the number of lymphocytes with a high binding ability of the Fc receptor was decreased. Changes in lymphocytes subpopulations in aseptic meningitis suggest mobilization of lymphocytes engaged in the cell-mediated immune response.

Cell-mediated and humoral immunity was studied in 74 patients with acute cerebral vascular disease. During the first two days after the onset of disease marked changes of cell-mediated immunity were observed, manifested as a decrease in total lymphocyte count in the peripheral blood, decrease in number of T lymphocytes, depression in lymphocyte blastogenesis and production of the migration inhibition factor, and a delayed-type skin reactivity. The changes were most evident in patients with severe lesions of brain tissue resulting from primary cerebral haemorrhage and cerebral infarction with fatal outcome. In the group of patients with cerebral infarction with improvement of neurological symptoms the immunological changes were not so pronounced as in the two above-mentioned groups, the smallest changes being found in patients with subarachnoid haemorrhage. We suppose that the depression in the immunne function was caused by severe stress during the course of disease. Impairment of the immune function may increase susceptibility to infection. The humoral immune response was not so evidently changed, and the observed increase of IgA in the sera was probably present before the stroke. In cases with good clinical course some improvement in the immunological parameters was observed, but full recovery did not occur until 3 weeks after the onset of disease.

We studied the distribution of T, T helper (TH), and T suppressor (TS) cells in blood and CSF using an immunoperoxidase slide assay and monoclonal antibodies. In subacute sclerosing panencephalitis, we found an elevated level of TS and a decreased level of TH cells in both CSF and blood. In MS, the TH/TS ratio was increased.

Humoral and cell-mediated immunity was determined in 28 epileptics, in 22 patients observed and investigated because of an only one seizure, and 26 healthy controls. Anticonvulsants had not been used in the treatment of these cases up to that time. The total lymphocyte count and the count of lymphocytes forming spontaneously rosettes with sheep erythrocytes were found to be significantly lower in the group of patients with epilepsy of unknown aetiology in relation to controls. The quantitative changes in the composition of the lymphocyte population were not significant and caused, probably, no immunological deficit, but were only a reaction to organic damage to the nervous system. Slight disturbances in the functions of the immune system cannot be, however, ruled out since this group showed also a significantly higher IgG level which suggests long-standing antigenic stimulation. No defect in IgA synthesis and disorders of the cell-mediated immunity (blastic transformation test, leucocyte migration inhibition test, skin test) were demonstrated in this study, although they had been reported by some authors in cases of epilepsy, mainly, however, in patients treated with anticonvulsants. In the patients having started anticonvulsive treatment these investigations will be repeated for establishing the effect of drugs on the immune reactions.

Subpopulations of lymphocytes in the CSF and peripheral blood were studied in 30 patients with MS, 16 with other neurological diseases (OND) and 15 control subjects without any neurological abnormalities. In patients with relapse of MS, the absolute numbers of total lymphocytes, alpha-naphthyl acid esterase (ANAE) positive, E-rosette forming and bearing the "avid" FcIgG receptor lymphocytes were significantly increased in the CSF as compared with stable or slowly progressive MS patients, patients with other OND and control subjects. The relative number of ANAE-positive cells was higher, and "avid"FcIgG receptor bearing cells lower in the CSF of all patients with MS than in the two other groups. The significance of the finding is unclear. The imbalance between lymphocyte subpopulations may reflect a primary defect in MS, or may be secondary, due to the presence of circulating immune complexes. In peripheral blood no substantial differences in lymphocyte behavior were observed between MS patients and other groups.