Cerebrospinal fluid and peripheral blood leukocyte subsets in acute inflammation of the CNS

Leukocyte subsets in CSF and peripheral blood (PB) were determined in 21 patients with acute inflammation of the CNS using the monoclonal antibodies OKT3, OKT4, OKT8, Leu12, and OKM1 in an immunoperoxidase slide assay. There was a predominance of OKT3-positive cells in nearly all samples. Significant differences between acute aseptic and bacterial meningitis only were found in CSF and represented by a higher Leu12 and a lower OKT3 percentage in aseptic inflammation and a higher absolute amount of OKT4-, OKM1- and Leu12-positive cells in bacterial meningitis. Comparison between CSF and PB showed significant differences only in aseptic meningitis with a higher percentage of Leu12-positive cells and a lower percentage of OKT8-positive cells in CSF. The OKT4/OKT8 ratio seems to be generally lower in aseptic meningitis but significant differences only were found in comparison with healthy blood donors. In a case of herpes simplex encephalitis the ratio was strongly increased in CSF during the early phase of specific antibody production because of an absolute rise of OKT4-positive cells.     

Quantitation of lymphocyte subsets in cerebrospinal fluid and blood during the clinical course of aseptic and bacterial meningitis.

Mononuclear cell subsets in cerebrospinal fluid (CSF) and peripheral blood (PB) were monitored during the clinical course in 23 patients with acute meningitis using 6 monoclonal antibodies. Significant differences between aseptic and bacterial meningitis mainly consisted of a higher percentage of OKT4-positive cells in PB in the acute phase of bacterial meningitis. Significant differences between CSF and PB are found in the amount of most cell subtypes at all times except the acute phase of bacterial meningitis. The OKT4/OKT8 ratio was always significantly higher in CSF and correlated with the acuity of inflammation in bacterial meningitis.     

Lymphocyte subpopulations of the blood and cerebrospinal fluid in acute polyradiculoneuritis. Study of 14 cases using monoclonal antibodies

The T lymphocytes in blood and CSF of 14 patients with acute polyradiculoneuritis were studied using the OKT series of monoclonal antibodies. Results were compared with findings in two control groups. Three patients with cytomegalovirus infection showed elevation of circulating OKT 8. A significant increase in the mean ratio OKT 4/OKT 8 was noted in all other cases, corresponding in 4 of the 11 patients to a reduction in number of OKT 8 lymphocytes. The CSF was normal in 3 of the 14 cases.     

Lymphocyte subsets at different stages of subacute sclerosing panencephalitis: a study with monoclonal antibodies.

Lymphocyte subsets in cerebrospinal fluid (CSF) and peripheral blood were studied using monoclonal antibodies, in patients with subacute sclerosing panencephalitis, eight of whom were at stage 2 and seven at stage 4. Eighteen subjects affected with non immunological diseases constituted the controls. Regardless of the stage, patients with subacute sclerosing panencephalitis had lower percentages of OKT3+ (pan-T) cells in both CSF and peripheral blood, with an increase of OKIa+ cells (B cells, macrophages and active T cells) in peripheral blood. A difference was found in the proportion of OKT4+ (helper-inducer) and OKT8+ (suppressor/cytotoxic) cells in relation to the stage, the most striking finding being a significant decrease of OKT8+ with an increase of T4/T8 ratio in peripheral blood at an early stage.     

Mononuclear cell types in cerebrospinal fluid and blood of patients with multiple sclerosis. Quantitation by immunoenzyme microassay with panel of monoclonal antibodies

Phenotypic distribution of mononuclear cells in cerebrospinal fluid (CSF) and peripheral blood from patients with multiple sclerosis (MS) and, for reference, patients with acute aseptic meningoencephalitis (AM), and in blood only from healthy controls, was studied with an immunoenzymatic microassay enabling analysis even in the presence of a normal CSF cell count. In MS, increased CD5+ (pan-T) cell proportion in CSF compared with blood was not reflected by changes of CD4+ or CD8+ cells, while in AM, an increase of CD4+ cells was registered. Therefore, a population of CD5+, CD4-, and CD8- cells may be anticipated to exist in CSF of patients with MS. Numbers of OKB7+, OKM1+, or HLA-DR+ cells did not distinguish between MS and AM. Proliferating cells expressing transferrin receptors (OKT9+ cells) were generally few or absent in CSF and not useful as a marker of disease activity in either MS or AM.     

Multiple sclerosis: activated cells in cerebrospinal fluid in acute exacerbations

Cerebrospinal fluid (CSF) lymphocytes from three young patients undergoing acute exacerbations of multiple sclerosis were studied by flow cytometry. Using a new method that simultaneously measures cell-surface antigens and the cell-cycle phase, we determined that the CSF lymphocytes in these patients were activated. The majority of the activated cells expressed the OKT4 phenotype, while very few expressed the OKT8 phenotype.     

Surface markers on lymphocytes from human cerebrospinal fluid : Identification by monoclonal antibodies

Lympocyte subpopulations in cerebrospinal fluid (CSF) and peripheral blood (PB) were studied using monoclonal antibodies and the common membrane markers. The results in three groups of patients were compared: 36 subjects with ‘non-immunological disorders’ (NID), 14 subjects with multiple sclerosis (MS) and 6 with subacute sclerosing panencephalitis (SSPE). It was found that, in patients with NID, (1) 90% of cells were T lymphocytes, reactive with OKT3; (2) the helper/suppressor (T4/T8) ratios were the same in the CSF and the PB; (3) the OKIa1 percentage was lower in the CSF than in the PB; and (4) only a few cells were ‘immature’, reacting with OKT10. Using the membrane markers (E rosettes, Fc IgG receptors and surface immunoglobulins), on the other hand, it was noted that the majority of cells in the CSF were identified as suppressor T lymphocytes and surface immunoglobulin-positive B cells were less common than the Ia1 marker suggested.There were no significant differences between the CSF results in patients with NID and MS but the OKT3 lymphocytes were reduced in CSF samples from patients with SSPE.     

A serial study of peripheral blood T lymphocyte subsets in relapsing-remitting multiple sclerosis

Twelve patients with relapsing-remitting multiple sclerosis and 3 control subjects were evaluated weekly for six months. Peripheral blood lymphocyte subsets were enumerated using monoclonal antibodies and analysis with a dual-laser fluorescence-activated cell sorter. Results were correlated with clinically assessed disease activity. Fluctuations occurred in the number of T lymphocytes (Leu 1+) and cells with the helper/inducer phenotype (Leu 3a/OKT4+) in both patients and controls. Fluctuations in the number of cells with the suppressor/cytotoxic (Ts/c) cell phenotype (OKT5, OKT8, Leu 2a+) also occurred. Variations in the relative numbers of cells labeled with all three Ts/c monoclonals did occur, but with OKT5 generally labeling fewer cells than OKT8 or anti-Leu 2a. When findings were correlated with disease activity, OKT5 was a more sensitive gauge of disease activity than either OKT8 or anti-Leu 2a. When peripheral blood lymphocyte subsets were correlated with disease activity, the following patterns were observed: a reduction in the number of Ts/c cells without evidence of clinical disease (four episodes), the development of new symptoms suggestive of an acute relapse but without a reduction in the number of Ts/c cells (two episodes), and a reduction in the number of Ts/c cells associated with acute relapse (two episodes). In the two patients with acute relapse and a reduction in the number of Ts/c cells, clinical disease preceded a reduction in the number of Ts/c cells in 1 patient, whereas the two events occurred simultaneously in the other patient. Taken together, these limited results indicate that in some cases a reduction in the number of Ts/c cells may be the result rather than the cause of disease activity and may represent an epiphenomenon.     

Cell surface markers for diagnosis of central nervous system involvement in lymphoproliferative diseases

To diagnose lymphoproliferative central nervous system (CNS) involvement we have used monoclonal antibodies in an immunocytochemical method for differentiation of cells in cerebrospinal fluid (CSF) and peripheral blood. The cell distribution in 9 patients with B-cell lymphoma and 7 patients with chronic lymphatic leukemia was compared to that in a group of patients with aseptic meningitis. Most patients with neoplastic CNS involvement showed a high proportion of CSF B cells (OKB2+ and/or OKB7+) and a concurrently low proportion of CSF T cells (anti-Leu 1+). Proliferating cells expressing transferrin receptor (OKT9 labeled) were increased in the CSF of 2 patients with neoplastic CNS involvement. In 2 patients with infectious CNS complications, the cell distribution in CSF did not differ from that in patients with aseptic meningitis. Patients with leukemia who had no CNS symptoms, and also 1 patient with meningitis and blood-brain barrier damage, showed a normal cell distribution in CSF despite high B-cell numbers in the peripheral blood. This indicates a selective passage of leukocytes into the CNS and/or local proliferation.     

CSF cells in multiple sclerosis: monoclonal antibody analysis and relationship to peripheral blood T-cell subsets

Mononuclear cells were analyzed in CSF and blood of 102 patients with MS. In CSF, the majority (78%) of cells were T lymphocytes (T3+), and the ratio of inducer (T4+) to suppressor/cytotoxic (T8+) cells was 2:1. No characteristic alterations in CSF phenotypes could be related to changes in circulating T8 cells or to disease activity. In a group of 75 patients, CSF cell count was higher in patients with low numbers of circulating T8 cells than in those with normal T8 cells. Thus, decreases in suppressor cells in the blood of MS patients are associated with CSF pleocytosis but not with fluctuations in the ratio of different subsets in CSF. Furthermore, large numbers of T8 cells are not sequestered in CSF when these cells are decreased in peripheral blood.     

Peripheral blood and CSF T-cell subsets in Japanese MS patients

We analyzed peripheral blood and CSF T-cell subsets in Japanese patients with MS, using an OK series of monoclonal antibodies in an immunofluorescent technique. In the peripheral blood, suppressor T cells were increased, with a concomitant decrease in helper T cells, after recovery from acute exacerbation, but there was no relationship between T cells in peripheral blood and CSF. In the CSF, acute exacerbation was associated with an increase in Ia-positive cells and a decrease in T cells. CSF pleocytosis (with more than 30% of Ia-positive cells) probably indicated active disease.     

Leukocyte types in cerebrospinal fluid and peripheral blood enumerated immunoenzymatically in aseptic meningitis and the Guillain-Barré syndrome

To study celltype distribution simultaneously in peripheral blood (PB) and cerebrospinal fluid (CSF) from patients with aseptic meningitis (AM) (n = 14) and Guillain-Barré syndrome (GBS) (n = 9) we used an immunoenzymatic method that enabled the use of several monoclonal antibodies, also in CSF samples with normal cellcounts. In both patient groups a different cell-distribution in CSF compared to PB was found with regard to pan T cells (CD5+/anti-Leu1+), T cell subpopulations (CD4+/anti-Leu3+, CD8+/anti-Leu2+), B cells (OKB2+, OKB7+), monocytes/macrophages (CD11+/OKM1+) and HLA/DR expressing cells, whereas the distribution of HLA/DC+ cells was similar in CSF and PB. Thus, the CSF cell distribution does not reflect the distribution in PB. The proportion of T cells was higher and the proportion of B-cells was lower in CSF than in PB in both patient groups, which is a finding similar to that in patients with multiple sclerosis. The OKT9 marker, labelling proliferating cells expressing the transferrin receptor, was not useful as marker of local proliferation.     

T cell subsets in multiple sclerosis. A longitudinal study of exacerbating-remitting cases

Twenty-four untreated MS patients with exacerbating-remitting disease were longitudinally studied for T-cell subset distribution within peripheral blood, using monoclonal antibodies OKT3, OKT4 and OKT8. A decreased percentage of OKT8 reactive cells, with a correlative increase of the OKT4/OKT8 ratio, was detected in relapsing MS patients, in most cases within 2 weeks before and 1 week after the onset of relapse. Longitudinal analysis of individual fluctuations allowed to detect during relapse an increase of OKT4/OKT8 ratio over the value recorded during remission in 78% of MS patients. Only 50% of patients however exhibited an OKT4/OKT8 ratio exceeding the 5% confidence upper limit of healthy control values. Relapse was more often associated with T-cell subset abnormalities in patients who suffered several attacks during the period of study. MS patients in remission, when considered as a group did not show significant abnormalities of the T-cell subset balance, although some individuals did present with wider T-cell subset fluctuations than healthy controls.     

Lymphocyte subpopulations in patients with multiple sclerosis.

Using monoclonal antibodies, peripheral blood helper/inducer (OKT4) and cytotoxic/suppressor (OKT8) lymphocytes were measured in 14 normal controls, 36 patients with multiple sclerosis at different stages of the disease and 15 patients with isolated optic neuritis. Thirty-four of these individuals were studied on two or more occasions at intervals up to 340 days. Patients with multiple sclerosis in relapse had low levels of OKT8 cells (14.07% +/- 3.79) compared with controls (29.42% +/- 4.69) and this abnormality returned to normal within approximately one month of the onset of new symptoms. Further changes occurred with new relapses. Low OKT8 cells were also found in patients with isolated optic neuritis (18.76 +/- 3.71) or progressive multiple sclerosis (19.91% +/- 7.96); the same pattern of recovery was seen in these two groups as in patients with multiple sclerosis in relapse. In 25% patients studied on two or more occasions after an episode of demyelination abnormalities of lymphocyte subpopulations occurred which were not accompanied by new clinical symptoms or signs. Fluctuations of this kind did not occur in controls. The findings have implications for the pathogenesis and management of patients with multiple sclerosis.     

Subsets of lymphoid cells in blood and thymus in myasthenia gravis : Monoclonal Antibody Analysis

Lymphoid cell subpopulations in peripheral blood and thymus were analyzed in patients with myasthenia gravis (MG) using monoclonal antibodies. The proportion of lymphocytes of T lineage (OKT 3 +, OKT 4 +, OKT 8 + cells) in peripheral blood of 11 MG patients, was significantly decreased in comparison with controls, but the ratio of OKT 4+/OKT 8+ cells was not different. Thymus cells were studied in 9 patients. The percentage of OKIa 1 + cells was significantly higher in MG thymus than in control thymus (P < 0.0005). There were no significant differences in the proportions of T lymphocyte subsets between MG and control thymuses.     

The behaviour of OKT3-, OKT4- and OKT8-positive cells during phases of elevated spontaneous chemiluminescence activity (CL-A) in multiple sclerosis patients

Summary The chemiluminescence activity (CL-A; synonym=burst activity, BA) and the percentage of OKT3-, OKT4-and OKT8-positive peripheral blood cells were serially examined in four control persons and in eight patients with multiple sclerosis. When the OKT values obtained in phases of increased CL-A (clinical remission) were compared with those of the control group, the percentage of OKT3-positive cells was reduced (P=0.014), and that of OKT4-positive cells increased (P=0.014); there were no significant changes in the percentage of OKT8-positive cells (P=0.171). After the CL-A had returned to normal values, the OKT4-positive cells remained elevated (P=0.029), whereas the OKT3- (P=0.342) and OKT8-positive cells (P=0.443) showed no significant changes. When in the same patients, phases of increased CL-A were compared with phases in which values were not elevated, a reduced percentage of OKT3-positive cells was found in phases with increased CL-A (P=0.031); however, the OKT4-positive and OKT8-positive cells did not differ significantly (P=0.156 and 0.281).     

The inflammatory process in polymyositis: monoclonal antibody analysis of muscle and peripheral blood immunoregulatory lymphocytes.

An analysis was made of the lymphocyte subpopulations in the muscle lesions and the peripheral blood of 25 patients with inflammatory myopathy, in the acute or chronic phase of the disease. Percentages of activated T lymphocytes (65% +/- 3.4), both helper and suppressor/cytotoxic, macrophages (25% +/- 3.2) and B cells (11% +/- 0.9) in the tissues were similar at all stages of the illness; T cells were, however, more common in acute polymyositis than in acute dermatomyositis, where B cells were significantly increased. A loss of circulating OKT8-positive lymphocytes in the peripheral blood was demonstrated, supporting other evidence of disturbed immunoregulation. It was concluded that the attack on muscle fibres is mediated by T cells, macrophages, and B cells, with the first two playing the major roles.     

Circulating T-cell subsets in Guillain-Barré syndrome

We compared the distribution of circulating T-cell subsets within 2 weeks of onset of symptoms in 14 patients with acute Guillain-Barré syndrome (GBS) and 37 normal controls. The levels of OKT4+ (putative helper-inducer) cells was definitely abnormal (decreased) in 3/13 tested. The levels of OKT8+ (putative suppressor-cytotoxic) cells were elevated in 3 and decreased in 2 of the 14 tested. Abnormal OKT4/OKT8 ratios were detected in 5 (2 elevated and 3 decreased) patients. Four of the 5 GBS patients with abnormal OKT4+/OKT8+ ratios were studied sequentially at least 4 times over 1-10 months; there was a return towards a normal ratio in all. Serial studies in 3 other GBS patients showed consistently normal values. In comparison, sequential studies over 5-24 months in 7 normals showed no abnormal OKT4+/OKT8+ ratios. Thus, abnormalities in OKT4+/OKT8+ ratios appear to be a marker of systemic events during symptomatic phases of GBS. It is not as yet known whether this is related to the cause or is secondary to the clinical manifestations of GBS.     

Cytotoxic activity of peripheral blood and cerebrospinal fluid lymphocytes from patients with multiple sclerosis and other neurological diseases : Analysis at the Single Cell Level Using Morphological and Surface Marker Phenotype Criteria

The large granular lymphocyte (LGL) has been identified in normal individuals' MNC population as the NK-K cell (Ault and Weiner 1978; Timonen et al. 1981); it bears the OKM1 surface antigen (Breard et al. 1981; Fast et al. 1981) and while negative for T cell antigens OKT4 and OKT8, is a low avidity E-rosette forming cell. However, in an unfractionated nylon wool passed peripheral blood lymphocyte (NWP PBL) population, we show that not more than 50% of NK activity in normal or OND control NWP PBL and 30% NK activity in MS NWP PBL can be attributed to this cell. Nevertheless, 100% of control K cell activity and 50% of MS K cell activity can be mediated by an LGL. MS patients have normal proportions of LGLs in their NWP PBL. The proportion of LGLs in CSF of MS and OND patients is too low to account for the number of CSF K cells. While in control NWP PBLs, the LGLs are OKM1⁺ and mediate NK and ADCC, in MS the LGL NK effectors are probably different from LGL-K cell effectors. In MS both populations include effector cells with T cell surface antigens. Thus, the OKM1⁺ LGL characteristics may not be used in analysis of NK and K cells in multiple sclerosis.     

Borrelia-induced meningoradiculitis--two different forms of the disease

The pattern of CSF cytological findings in the acute stage and after 10 days of antibiotic and steroid treatment was examined in 14 patients with meningoradiculitis due to an arthropode-transmitted infection with Borrelia duttoni (burgdorferi). At the acute onset of neurological symptoms, 12 patients revealed lymphocytic pleocytosis (cell count 50-500 X 10(6) cells/l) with numerous plasma cells (5-12% of the total cell count) and neutrophilic granulocytes (0.2-3.4%). 10-14 days after the onset of penicillin infusion and steroid treatment, cell count was reduced by at least 50% in 8 patients. Plasma cells then varied between 2 and 8%, whereas the neutrophilic granulocytes had disappeared. Two patients with slowly progressive neurological symptoms had a normal cell count, and inflammatory changes in CSF cytology were not observed.