ABSTRACT
Triple-color flow cytometry with a panel of antibodies comprising GD2, CD56, and CD45 was performed to analyze cerebrospinal fluids (CSF) from a patient with retinoblastoma who was suspicious of meningeal metastasis based on clinical presentation. Our results showed that the cells in CSF demonstrated the immunophenotype positive for GD2 and CD56 but negative for CD45 antigen, which suggested the presence of CSF metastasis of retinoblastoma. At the end of eight cycles of intrathecal chemotherapy, CSF specimen was analyzed with Flow cytometry immunophenotyping (FCI) again and the result showed no detectable malignant cells with the same immunophenotype. Our conclusion is that FCI can be a quick and reliable method for the diagnosis of CSF metastasis of retinoblastoma and the immunophenotype (GD2+, CD56+, and CD45-) can be used to recognize residual retinoblastoma cells in CSF.
Subject(s)
CD56 Antigen/cerebrospinal fluid , Gangliosides/cerebrospinal fluid , Immunophenotyping , Leukocyte Common Antigens/cerebrospinal fluid , Retinoblastoma/cerebrospinal fluid , CD56 Antigen/immunology , Child, Preschool , Flow Cytometry , Gangliosides/immunology , Humans , Leukocyte Common Antigens/immunology , Male , Retinoblastoma/immunologyABSTRACT
We investigated CD45RA and CCR7 expression in CD4+ and CD8+ subsets of cerebrospinal fluid (CSF) lymphocytes, both immediately ex vivo and after stimulation, from 134 patients with a variety of inflammatory and non-inflammatory neurological diseases. Most inflammatory diseases had a higher CD4+:CD8+ ratio and higher percentage of effector memory T cells (T(EM)) than non-inflammatory controls, excluding active infection. Moreover, we found that patients with highly elevated cell counts in the CSF tended to have a lower percentage of central memory T cells (T(CM)) than patients with low or absent pleocytosis, with a concomitant increase in T(EM). We also found that samples with elevated IgG index or presence of oligoclonal bands had a significantly higher CD4+:CD8+ ratio than normal samples, consistent with increased CD4+ help for intrathecal IgG synthesis by B cells.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Central Nervous System Diseases/cerebrospinal fluid , T-Lymphocyte Subsets/immunology , Adolescent , Adult , Aged , Aged, 80 and over , CD4-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/metabolism , Central Nervous System Diseases/immunology , Central Nervous System Diseases/metabolism , Child , Child, Preschool , Female , Humans , Immunoglobulin G/cerebrospinal fluid , Infant , Inflammation/cerebrospinal fluid , Inflammation/immunology , Inflammation/metabolism , Leukocyte Common Antigens/biosynthesis , Leukocyte Common Antigens/cerebrospinal fluid , Leukocyte Common Antigens/immunology , Male , Middle Aged , Receptors, CCR7/analysis , Receptors, CCR7/biosynthesis , Receptors, CCR7/immunology , T-Lymphocyte Subsets/metabolism , Young AdultABSTRACT
Recent studies suggest inflammatory mechanisms involved in the pathogenesis of major psychiatric disorders (MPD). T cells play a major role during inflammation, but little is known about T cell subpopulations in the cerebrospinal fluid (CSF). We investigated the frequency of cells positive for the surface markers CD4, CD8, CD25, CD45, CD69, and CD127 in 45 paired cerebrospinal fluid (CSF) and peripheral blood (PB) samples by multiparameter flow cytometry from patients with MPD of the schizophrenic and affective spectrum with normal CSF cell counts and compared them with those from patients with non-inflammatory (NIND), chronic inflammatory (CIND) neurological disorders, and meningitis (MEN). In MEN patients, CD4+ cell frequency in PB, but not in CSF, was significantly increased as compared to CIND and NIND. No difference between patient groups was observed for CD8+. CD4+CD45RO+ double positive cells in PB were significantly lower in CIND than in MEN or NIND. The frequency of CD4+CD25+ cells in PB was significantly higher in MEN than in MPD or CIND. For CSF, the percentage of CD4+CD127(dim) cells was significantly lower in MEN than in MPD. CD4+CD127(dim) in PB and CSF showed overlapping characteristic clusters between MPD and CIND and MEN patients. Overall, the hypothesis of low degree inflammation in a subgroup of MPD is supported. The analysis of lymphocyte subsets in PB and CSF constitutes a novel promising tool to understand underlying pathomechanisms in psychiatric and neurological disorders on an individual case level.
Subject(s)
Flow Cytometry/methods , Mental Disorders/immunology , Nervous System Diseases/immunology , T-Lymphocyte Subsets/immunology , Adolescent , Adult , Affective Disorders, Psychotic/blood , Affective Disorders, Psychotic/cerebrospinal fluid , Affective Disorders, Psychotic/immunology , Aged , Antigens, CD/blood , Antigens, CD/cerebrospinal fluid , Antigens, Differentiation, T-Lymphocyte/blood , Antigens, Differentiation, T-Lymphocyte/cerebrospinal fluid , CD4 Antigens/blood , CD4 Antigens/cerebrospinal fluid , CD8 Antigens/blood , CD8 Antigens/cerebrospinal fluid , Female , Humans , Immunophenotyping/methods , Interleukin-2 Receptor alpha Subunit/analysis , Interleukin-2 Receptor alpha Subunit/blood , Interleukin-7 Receptor alpha Subunit/analysis , Interleukin-7 Receptor alpha Subunit/blood , Lectins, C-Type , Leukocyte Common Antigens/blood , Leukocyte Common Antigens/cerebrospinal fluid , Male , Meningitis/blood , Meningitis/cerebrospinal fluid , Meningitis/immunology , Mental Disorders/blood , Mental Disorders/cerebrospinal fluid , Middle Aged , Nervous System Diseases/blood , Nervous System Diseases/cerebrospinal fluid , Schizophrenia/blood , Schizophrenia/cerebrospinal fluid , Schizophrenia/immunology , T-Lymphocyte Subsets/cytology , T-Lymphocytes/cytology , T-Lymphocytes/immunology , Young AdultABSTRACT
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.
Subject(s)
CD4-Positive T-Lymphocytes/physiology , CD8-Positive T-Lymphocytes/physiology , Lymphocyte Activation/physiology , Optic Neuritis/pathology , Acute Disease , Adolescent , Adult , Evoked Potentials, Visual/physiology , Female , Flow Cytometry/methods , Gene Expression Regulation/physiology , HLA-DR Antigens/blood , HLA-DR Antigens/cerebrospinal fluid , Humans , Leukocyte Common Antigens/blood , Leukocyte Common Antigens/cerebrospinal fluid , Male , Middle Aged , Optic Neuritis/blood , Optic Neuritis/cerebrospinal fluid , Optic Neuritis/physiopathology , Vision, Ocular/physiologyABSTRACT
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.