Fluctuations in T helper subpopulations in relapsing-remitting multiple sclerosis

Patients with active multiple sclerosis (MS) have been reported to have a depletion of CD4+ CD45R+ cells, the immature resting CD4+ subpopulation. Using Leu 3a(anti-CD4) and Leu 18(anti-CD45R), the frequencies and absolute numbers of CD4+ CD45R+ and CD4+ CD45R- subsets were measured in 30 patients with MS and 17 healthy controls. These subsets were monitored every 6 weeks over a 6 month period. CD4+ CD45R- cells were found to be increased in relapse compared to remission (p less than 0.005) while CD4+ CD45R+ levels were not significantly altered in relapse. However, the CD4+ subset ratio (CD4+ CD45R-/CD4+ CD45R+) was significantly higher in relapse compared with remission (p less than 0.002). Furthermore, these findings were upheld when data from the same 6 patients in relapse and remission was compared. Increased disease activity was not associated with changes in any of the other parameters measured (total T cells, total CD4+ cells, suppressor cells or activated T cells). These results suggest that relapse in MS is accompanied by the conversion of CD4+ CD45R+ resting cells to CD4+ CD45R- primed cells.     

Immune system-associated antigens on the surface of peripheral blood lymphocytes in patients with Alzheimer's disease.

We investigated the immune-associated antigens of peripheral lymphocytes from 13 patients with Alzheimer's disease (AD) and 13 age-matched healthy control subjects using two-color analysis with flow cytometry. Four ratios of immune-related antigens, T/B lymphocytes, CD4/CD8, CD4/CD45R and CD4/HLA-DR, were compared for the AD and control groups. The T/B and CD4/CD8 ratios did not differ between the groups, the ratio of CD4+CD45R+ subset in the AD group was lower than the ratio in the control group, and the ratios of CD4+ CD45R- and CD4+HLA-DR+ subsets in the AD group were significantly higher. Further, in the AD group, the CD4+ CD45R+/CD4+ ratio was lower and the CD4+ CD45R- CD4+ ratio was higher than in the control group.     

CD4+ lymphocyte subsets in the cerebrospinal fluid of multiple sclerosis and non-inflammatory neurological diseases

Summary We studied paired cerebrospinal fluid (CSF) and peripheral blood (PB) samples from 18 inactive multiple sclerosis (MS) patients and 10 with non-inflammatory neurological diseases. By means of a dual-colour cytofluorimetric micromethod we were able to count 1500 cells on average in each CSF sample. We found a significant reduction of CD45RA+ and CD4+CD45RA+ cells in the CSF of MS patients. Similarly, CD45RA+ and CD4+CD45RA+ CSF/PB ratios were lower compared with controls. The reduction of suppressor-inducer T-cells did not correlate with CD8+ cell levels in the CSF. The CD4+ subset ratio (CD4+CD45RA–/CD4+CD45RA+) was significantly increased in the CSF of MS patients. Our data suggest that the reduction of CD4+CD45RA+ cells in the PB is not due to a segregation of such cells in the CSF. Conversely, CSF changes reflect changes in the PB similar to these found for other T-cell subsets.     

Decrease in memory (CDw29-high) cerebrospinal fluid cells from acute MS patients

The CD45R (2H4) and CDw29 (4B4) molecules are probably involved in the regulation of immunological memory: CD45R functionally defines naive or virgin cells (prior to activation by exposure to antigen), while CDw29 characterizes previously activated (memory) cells. By means of two-colour fluorescence analysis, we studied CD4 and CD8, CD45R and CD4 and CD8 CDw29 cells from the cerebrospinal fluid (CSF) of 27 acute (MSa), and 10 stable (MSs) multiple sclerosis (MS) patients. We also compared 17 patients affected with various non-inflammatory (OND) diseases of the central nervous system (CNS). The most striking finding was the increased percentage of CD4+CD29-, CD4+CD45R- and CD8+CD45R- cells and the decreased percentage of CD4-CDw29+ and CD8- CDw29+ subsets in MSa patients compared to OND and MSs populations. These data suggest a decrease in memory cells (CDw29+) during the acute phase of MS. The modulation of CDw29 receptor could play a role in the genesis of acute MS attacks.     

Presence of T-cell subset abnormalities in newly diagnosed cases of multiple sclerosis and relationship with short-term clinical activity

Abnormalities of T-cell subsets in patients with multiple sclerosis are well known; in order to assess whether immunological abnormalities are relevant in the pathogenesis of the disease after its clinical onset, peripheral blood lymphocyte subsets (CD3⁺, CD4⁺, CD4⁺ CD45RA⁺, CD4⁺CD45RA^–, CD8⁺, CD8⁺CD57⁺, CD57⁺, CD25⁺) were analysed serially in 25 patients at the first clinical episode suggestive of inflammatory demyelinating disease and in an equal number of age- and sex-matched controls. During the follow-up period (12–18 months, mean 14) 6 of 25 patients presented new relapses: in this subgroup of patients, significant changes in CD4⁺ ratio (% CD4⁺CD45RA^–/%CD4⁺CD45RA^–) were detected in comparison both with healthy controls and with clinically stable patients. Patients clinically stable at follow-up did not display immunological abnormalities, regardless of the presence or absence of cerebrospinal fluid and/or magnetic resonance imaging alterations consistent with multiple sclerosis. These findings suggest a possible prognostic role of early T-cell subset imbalance in multiple sclerosis.     

T cell subsets in multiple sclerosis: a serial study

The relevance of abnormalities in the distribution of peripheral blood T lymphocyte subsets to the clinical manifestations of multiple sclerosis is not firmly established. A clinical and immunological follow-up of relapsing-remitting multiple sclerosis patients was performed in order to study the relationship of immune changes with the clinical course of the disease. Twenty patients were monitored monthly during a mean time of nine months for peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD19), including the immunoregulatory subsets CD4CD29 (helper-inducer), and CD4CD45RA (suppressor-inducer) and activated T helper cells (CD4CD25) by flow cytometry. A total of 14 untreated relapses was included. The most significant observations were a decrease in T suppressor-inducer CD4⁺ CD45RA⁺ subset during clinical relapses ( P = 0.028) that was also detectable one month before ( P = 0.020) and the lack of changes in CD4⁺ CD29⁺ and CD8⁺ T cells. In addition, variations in the percentage of CD4⁺CD25⁺ activated T helper cells were not associated with clinical exacerbations. These results indicate the existence of a temporal association of immune changes in peripheral blood, but not activation, with the clinical manifestations of multiple sclerosis.     

Circulating dendritic cells subsets and regulatory T-cells at multiple sclerosis relapse: differential short-term changes on corticosteroids therapy

Glucocorticoids remain the treatment of choice for MS relapses. However, little is known on the effect of intravenous methylprednisolone (IVMP) on dendritic cells (DCs) and regulatory T-cells (TReg). Our main goal was to quantify circulating myeloid and plasmacytoid DCs (mDCs and pDCs), and TReg at MS relapse versus healthy controls; and to analyse the short-term changes after IVMP for MS relapse. MS patients at relapse compared to controls showed higher %CD4+CD25high+ TReg (p<0.01). After 5-days of IVMP, activated T-lymphocytes (p=0.001), pDCs (p<0.0001), and CD11c+ mDCs (p<0.0001) decreased. By contrast, CD4+CD25+ and CD4+CD25high+ TReg further increased (p<0.0001 both). Changes on these subsets may play a relevant role in the immunosuppressive activity of this drug.     

Cellular immunoregulatory mechanisms in the central nervous system: characterization of noninflammatory and inflammatory cerebrospinal fluid lymphocytes

Dual-label flow cytometric analysis of cerebrospinal fluid (CSF) and blood lymphocytes with combinations of monoclonal antibodies such as CD4 plus CD45R or Leu8, and CD8 plus CD11b was performed in 37 patients with noninflammatory neurological diseases (NINDs) to clarify the differences in cellular immunoregulatory mechanisms present in the central nervous system (CNS) and in the systemic circulation. In the CSF of patients with NINDs, the paucity of CD4+CD45R+ and CD8+CD11b+ cells was striking, whereas the same subsets accounted for substantial proportions in the blood. CD4+CD45R- and CD4+Leu8- cells as well as CD8+CD11b- cells increased in the CSF when compared with those in the blood. Seven patients with active multiple sclerosis (MS) and 10 patients with other inflammatory diseases in the CNS (CNS-infl) were also studied. Patients with active MS were characterized by a consistent increase in percentage of CD4+CD45R- cells in the CSF, whereas an increase of CD4- CD45R+ cells in the CSF was a feature of the patients with CNS-infl, when compared with patients with NINDs. These findings indicate that the CNS is routinely surveyed by particular subsets of lymphocytes different from those in the blood, and cellular immune reaction in the CNS varies according to the types of CNS inflammatory conditions.     

Immunoregulatory T-cells and lymphocytotoxic antibodies in active multiple sclerosis: weekly analysis over a six-month period

Immunoregulatory T-cell subsets were measured at weekly intervals over a 4 to 6 month period using monoclonal antibodies (anti-T4 = inducer cell; anti-T8 = suppressor/cytotoxic cell) in a group of 6 patients with multiple sclerosis (MS) and in 4 age- and sex-matched controls. Decreases in the T8 subset and increases in the T4:T8 ratio were present in 4 of the patients with MS but not in controls. Two patients who were neurologically stable during the study period had no changes in the T4:T8 ratio; 2 patients with intermediate disease activity of the relapsing-remitting type had elevated ratios on 3 and 4 occasions respectively; the patient with the most clinically active MS had an abnormal ratio 12 of 27 times. One patient with chronic-progressive MS had an elevated ratio on each occasion tested. No abnormalities in T-cell subsets were present in any of the controls. On three occasions an elevated T4:T8 ratio appeared to precede an acute relapse by 1.5 to 7 days. Lymphocytotoxic antibodies (LCA) against whole lymphocytes or against isolated T-cell subsets were measured in these patients and in a larger group of MS patients, and were not found to correlate with changes in T-cell subsets. This report extends previous findings linking changes in T-cell subsets to disease activity in patients with MS.     

T-cell subsets in the cerebrospinal fluid and peripheral blood of multiple sclerosis patients treated with high-dose intravenous methylprednisolne

To determine the effects of high-dose intravenous methylprednisolone (MP) on lymphocytes and lymphocyte subpopulations in the cerebrospinal fluid (CSF) and peripheral blood (PB) in multiple sclerosis (MS) patients, we studied 67 patients with definite MS treated with MP. They were classified according to the disease course: 32 chronic progressive (CP) patients, 25 relapsing-remitting (RR) patients, and 10 patients with a chronic progressive disease course accompanied by relapses and remissions (CP + RR). MS patients were treated with 1000 mgr intravenous MP daily for 10 consecutive days. Before and after MP treatment we simultaneously studied CSF and PB CD3 +, CD4 +, CD8 +, CD20 +, and Ial + cells subsets. Kurtzke's Expanded Disability Status Scale (EDSS) was used for clinical evaluation. Progression rate was defined as the ratio of EDSS to disease duration. Thirteen patients with lumbar disk herniation were investigated as controls. Before MP, we found in MS patients, especially in the CP group, significantly lower CD4 + T-cell percentages in the PB with respect to controls (P<0.05). The percentage of CD4 + T-cells in the CSF of MS patients was significantly higher compared with PB (p = 0.0001), and tended to be higher than in controls (p = 0.072). The CSF mononuclear cell counts were significantly correlated with higher percentages of CSF CD3 + (r = 0.40) and CD4 + (r = 0.47) T-cells and lower CSF CD8 + (r = -0.33) T-cell percentages. B-cell percentages in the CSF were significantly elevated compared with controls for all MS groups. No relation could be obtained between T- or B-cell subsets and EDSS or progression rate. After MP, a significant decrease in PB CD8 + T-cell percentage and simultaneously an increase of the percentage CD8 + T-cells in CSF was noted in the entire MS group and in the CP and RR MS patients. Except for the CP + RR MS patients, CD4 + T-cell percentages in the PB or CSF showed insignificant changes. Our findings support the view that in MS MP might affect the inflammatory process of demyelination by a selective and dissociative effect on T-suppressor/cytotoxic cells in the PB and CSF.     

Increased CD8+ central memory T cells in patients with multiple sclerosis

A T-cell-mediated autoimmune process against central nervous system myelin is believed to underlie the pathogenesis of multiple sclerosis (MS). Formation of immunological memory is based on the differentiation of na?ve T cells to memory T cells after exposure to antigens and specific cytokines. The aim of this study was to analyse peripheral blood mononuclear cells in patients with MS for different T-cell subsets including na?ve and memory T cells. Flow cytometry and enzyme-linked immunosorbent assay were used to analyse memory T-cell subsets and plasma concentration of interleukin-15 (IL-15) in peripheral blood of MS patients, patients with other neurological disorders and healthy controls. MS patients had a skewed distribution of T cells with an increased level of CD8+/CCR7+/CD45RA - central memory T cells (TCM) compared to healthy controls. In addition, MS patients showed significantly higher levels of plasma IL-15 than healthy controls did. Upregulated CD8+ TCM in MS patients may reflect a persistent chronic inflammatory response that may have been induced during early stages of the disease. This derangement may be important for maintaining chronic inflammation in MS.     

CD4(+) memory T cells with high CD26 surface expression are enriched for Th1 markers and correlate with clinical severity of multiple sclerosis

An aberrant immune activation is believed to be important in the pathogenesis of multiple sclerosis (MS). Expression of CD4(+) T lymphocyte surface molecules indicative of immune activation and effector functions has been correlated with disease severity and activity. CD4(+) CD45R0(+) CD26(high) memory T lymphocytes contained the high levels of markers of Th1, activation, and effector functions and cell counts of this subset correlated with MS disease severity. This subset had lower expression of PD-1, CCR4, and L-selectin in MS than in controls. These changes were only partially normalised by treatment with interferon-beta. We point to this subset as a putative target for immunological monitoring of MS disease activity and of treatment efficacy.     

Lymphocyte subset proportions in Guillain-Barré syndrome patients treated with plasmapheresis

We examined the changes in the proportions of certain T cell subsets in 18 Guillain-Barré syndrome patients treated with plasmapheresis (PP). PP was performed within 8 days after the onset using a continuous-flow blood cell separator or immunoadsorption. The proportions of CD3+, CD4+ and CD4+CD45RA+ cells at the onset were decreased and that of CD19+ was increased compared with those of 25 normal controls. However, the proportions of CD4+CD45R-, CD4+HLADR+, CD4+CD25+ cells and the CD8+ subpopulations were similar in patients and controls. After PP, the altered subsets tended to normalize, particularly in the group that improved after PP.     

Preferential increase of IL-2R+ CD4+ T cells and CD45RB- CD4+ T cells in the central nervous system in experimental allergic encephalomyelitis.

We examined lymphocytes isolated from the spinal cord (SC), peripheral blood (PB) and lymph nodes (LN) draining the immunization site of Lewis rats with acute experimental allergic encephalomyelitis (EAE). Cells were analysed for T cell subset markers CD4 (mAb W3/25) and CD8 (mAb OX8), for IL-2R (mAb OX39), and for high molecular mass leukocyte common antigen (LCA, CD45RB) expression (mAb OX22). T cells expressing high (CD45RB+) or low (CD45RB-) molecular mass LCA are of different maturational stages and/or separate lineages. CD4+ T cells were more predominant in SC than in PB and LN; CD8+ T cells were scarce in SC but common in PB and LN. Activated CD4+ T cells (IL-2R+) were common in the SC and LN but infrequent in blood. CD4+ T cells that were CD45RB+ were scarce in the SC. In contrast, the majority of CD4+ T cells in the PB and LN were CD45RB+. The preferential accumulation of IL-2R+ CD4+ T cells and of CD45RB- CD4+ T cells in the central nervous system (CNS) indicates that a selective mechanism directs cell egress into CNS lesions in EAE.     

Lymphocyte subsets in relapsing-remitting Multiple Sclerosis: A longitudinal study of B lymphocytes and T lymphocytes

Abstract

Changes in lymphocyte subset populations may provide dues to the dysimmune mechanisms involved in relapsing remitting multiple sclerosis (RRMS). The lymphocyte subgroup CD4+CD45RA+, thought to be responsible for the induction of suppression is decreased in some patients with MS compared to controls. A possible role for another lymphocyte subset, CD19+CD5+ lymphocytes; has been proposed in autoimmune diseases and multiple sclerosis (MS). To expand this we studied CD4+CD45RA + (T) lymphocytes and CD19+CD5+ (B) lymphocytes in nine patients with relapsing-remitting MS (RRMS) and nine controls. The patients were examined monthly for an average of ten months and nine relapses were observed in seven patients. One patient underwent monthly gadolinium enhanced magnetic resonance imaging (MRI). Normal percentages CD4+CD45RA+ lymphocytes were found in patients with RRMS. No significant abnormalities in the CD19+CD5+ lymphocyte subpopulation were noted, although a tendency for higher percentages of this subset (approaching statistical significance, P = 0.056) was detected. [Neurol Res 1994; 16: 385-388]     

Changes in lymphocyte subsets in myasthenia gravis: correlation with level of antibodies to acetylcholine receptor and age of patient.

We report a detailed analysis of the subsets of lymphocytes in patients with myasthenia gravis (MG). There was a slight, nonsignificant increase in the level of CD5+ B lymphocytes among MG patients as compared with normal controls. The proportion of CD5+ T cells in MG was similar to that in controls. However, whereas age had no effect on the level of these cells in normal individuals, a significant age-related decrease of these cells was present in MG patients. The proportion of double-positive CD4+CD8+ T cells was significantly increased in MG. The level of the CD29+CD4+ (helper-inducer) subset was significantly higher in MG patients than in controls. There was no correlation between the titer of autoantibodies to acetylcholine receptor and the level of either CD29+CD4+ T cells or CD5+ B cells among MG patients. The only T-cell subset that correlated with the autoantibody titer was the CD45RA+CD4+ (suppressor-inducer) subset of CD4+ T cells.     

Isolation of myelin basic protein–specific T cells predominantly from the memory T-cell compartment in multiple sclerosis

Myelin antigen–reactive T cells have been implicated in the pathogenesis of multiple sclerosis (MS). Myelin-reactive T cells can be isolated from control subjects as well as individuals who have MS. Experimental models of MS indicate that recently stimulated, myelin-reactive T cells have greater encephalitogenic potential than resting T cells. Activation induces changes in T-cell surface antigens that may distinguish previously stimulated, memory T cells from naive T cells. Therefore, we examined 108 myelin basic protein (MBP)-reactive T-cell lines from 7 MS and 8 control subjects to determine whether MBP-reactive T cells originated in the memory T-cell subset or in the naive subset. Isotypes of CD45 were used that designate memory or naive T cells. In subjects having MS, 84% of the MBP-reactive T cells resided in the memory T-cell subset. However, in control subjects, only 13% of MBP-specific T cells originated from the memory T-cell subset. This result suggests that a substantial proportion of MBP-reactive T cells from some individuals with MS have been previously activated in vivo. This difference provides additional support for the hypothesis that myelin antigen–specific T cells are involved in the pathogenesis of MS. Ann Neurol 1999;45:33–39     

Systemic T-cell activation in acute clinically isolated optic neuritis

We examined untreated 60 patients with acute monosymptomatic optic neuritis (ON). Patients examined early after onset showed increased expression of HLA-DR and CD45R0 on CD4 and CD8 T cells. Expression of HLA-DR on CD4 T cells was higher in patients without IgG oligoclonal bands. Expression of HLA-DR on CD4 and CD8 T cells correlated negatively with measures of disease activity and positively with measures of good visual function, and expression of CD45R0 on CD4 T cells correlated negatively with measures of disease activity. We hypothesize that HLA-DR expression may characterize a protective T-cell subset in ON.     

CD45RA+ ICAM-3+ lymphocytes in cerebrospinal fluid and blood as markers of disease activity in patients with multiple sclerosis

OBJECTIVES: Autoreactive T cells targeted against antigens of the myelin sheath are suggested to play an important role in the pathogenesis of multiple sclerosis (MS). Naive (CD45RA+) T cells and intercellular adhesion molecule-3 (ICAM-3) are markers for un-activated lymphocytes. This study was performed to investigate, whether the expression levels of these antigens both on cerebrospinal fluid (CSF) and peripheral blood lymphocytes can be used as activity markers in MS. MATERIALS AND METHODS: Corresponding blood and CSF samples were obtained from 31 patients with relapsing-remitting MS. Of the 31 MS patients 23 were suffering from acute relapses at the time of examination and all of them were treated with high-dose methylprednisolone (MP). Blood was collected again on the 10th day of therapy and after 3 months. The control group consisted of 12 healthy persons. Two-color flow cytometry was performed to evaluate the percentage of both CD45RA+ and ICAM-3+ cells within the lymphocyte population. RESULTS: The percentage of CD45RA+ ICAM-3+ cells in the CSF of MS patients with relapses was significantly increased compared to patients in remission (P<0.05). In blood, a significantly lower percentage of CD45RA+ ICAM-3+ lymphocytes was found in both patient groups compared to healthy controls (Relapse: P<0.05, Remission: P<0.10). Additionally, we found a significant increase (P < 0.01) in the percentage of CD45RA+ ICAM-3+ lymphocytes in blood of MS patients suffering from acute relapse on the 10th day of high-dose MP treatment. CONCLUSION: Our data suggest that the percentage of CD45RA+ ICAM-3+ lymphocytes in CSF can be used as marker of disease activity in MS patients.     

Migration of T-cell subsets in multiple sclerosis and the effect of interferon-beta1a

OBJECTIVES: Migration of inflammatory cells across the blood-brain barrier is a central event in the formation of multiple sclerosis (MS) lesions and is known to be enhanced in MS patients. This study investigates the migration of CD4+ and CD8+ T-cell subsets and the effects of interferon-beta1a (IFN-beta1a) treatment on migration and matrix metalloproteinase-9 (MMP-9) production of these T-cell subsets. MATERIALS AND METHODS: An ex vivo transwell system was established to compare the migratory behaviour of lymphocytes isolated from normal controls and untreated MS patients. In addition, MS patients were investigated longitudinally after initiation of IFN-beta1a treatment. RESULTS: Migration of CD4+ T cells (P < 0.05), but not of CD8+ T cells, was enhanced in untreated MS patients compared with controls and was normalized by treatment with IFN-beta1a. In addition, IFN-beta1a treatment reduced MMP-9 production of CD4+ but not CD8+ T cells. CONCLUSION: Our results indicate that CD4+ T cells, but not CD8+ T cells, contribute to the enhanced ex vivo migration observed in MS.