Harry G. Bluestein

Cerebrospinal fluid antibodies to neuronal cells: association with neuropsychiatric manifestations of systemic lupus erythematosus.

The validity of the hypothesis that some of the neuropsychiatric manifestations of systemic lupus erythematosus (SLE) are mediated by the direct effects of antibody binding to neuronal cell membranes is dependent on the demonstration of antineuronal activity within the central nervous system of patients with active central nervous system disease. Using a radiolabelled staphylococcal protein A assay, we tested cerebrospinal fluid from 27 patients with SLE and central nervous system manifestations, and cerebrospinal fluid from 18 additional patients with SLE but free of central nervous system disease for antibody reactive with the cultured human neuronal cell line SK-N-SH. Cerebrospinal fluid from 20 of 27 patients with active lupus central nervous system disease had increased immunoglobulin G (IgG) antineuronal activity compared with cerebrospinal fluid from two of 18 patients with SLE without central nervous system disease. Ninety percent of the patients with psychosis, organic brain syndrome or generalized seizures had increased IgG antineuronal activity as compared with only 25 percent of the patients who presented with hemiparesis or with chorea/hemiballismus. Antineuronal activity per microgram of IgG was concentrated eightfold in the cerebrospinal fluid of patients with active central nervous system disease as compared with the serum activity. Patients with or without active central nervous system disease did not differ significantly in the amount of serum antineuronal binding activity. The results are consistent with the hypothesis that the more diffuse central nervous system manifestations of SLE are a direct result of the interaction of antibody with neuronal cell membranes.

Neuropsychiatric disease in Sjögren's syndrome: Anti-ribosomal P and anti-neuronal antibodies

PURPOSE Patients with Sjögrens syndrome (SS) may develop nonfocal (i.e., psychiatric and/or cognitive dysfunction) as well as focal, neuropsychiatric disease (CNS-SS). Anti-ribosomal P and anti-neuronal antibodies have been associated with nonfocal neuropsychiatric disease in systemic lupus erythematosus (SLE), particularly psychosis and depression. This study examines the spectrum of psychiatric and cognitive dysfunction observed in SS patients with focal, as well as nonfocal, central nervous system (CNS) disease and relates these observations to the presence of serum and cerebrospinal fluid (CSF) anti-ribosomal and anti-neuronal antibodies. PATIENTS AND METHODS One hundred thirty-one patients--patients with primary SS (n = 91), patients with secondary SS (n = 34), and mothers of infants with neonatal lupus erythematosus (NLE) (n = 6)--were studied. Patients were referred to a large tertiary referral center and the population was highly selected for CNS disease. Patients were evaluated clinically for focal and nonfocal CNS disease. Sera from 131 patients and 34 paired sera/CSF samples were examined by enzyme-linked immunosorbent assay and radioimmunoassay for the presence of anti-ribosomal P and anti-neuronal autoantibodies, respectively. Clinical features were categorized and autoantibody profiles obtained and correlated independently for statistical significance. Data were analyzed using the two-tailed Fisher exact test. RESULTS Psychiatric or cognitive impairment, usually mild or moderate, occurred in over 80% (63 of 77) of this highly selected population of SS patients, and more than 60% of patients (48 of 77) had both. Anti-ribosomal P antibodies occurred in six (4.6%) patients with SS and related disorders. None of the patients with primary SS had anti-ribosomal P antibodies, whereas they were present in a small number of patients with secondary SS (i.e., 4 of 34 [12%]) and in 2 of 6 mothers of infants with NLE. There was no correlation between nonfocal CNS disease, including psychosis or severe depression, and the presence of anti-ribosomal P antibodies. Paired serum CSF samples from 34 SS patients with active CNS disease, including 6 with psychosis and 5 with severe depression, did not contain either anti-ribosomal P or anti-neuronal antibodies. Anti-ribosomal P and anti-neuronal antibodies were present in a control subset of SLE patients defined serologically by the presence of anti-nDNA antibodies. CONCLUSION Patients with primary SS associated with CNS disease, including psychosis and depression, do not have serum or CSF autoantibodies to ribosomal P peptide or neuronal antigens, detected by binding to neuroblastoma cells. Thus, autoantibodies associated with nonfocal or diffuse CNS disease in classical SLE (particularly psychosis and depression) are not present in CNS-SS. The observations suggest that nonfocal CNS disease in CNS-SS and CNS-SLE may be mediated by different immunopathologic mechanisms. Potentially, these observations may have diagnostic and therapeutic implications in the management of patients with CNS-SS and patients with CNS-SLE.

Brain-reactive lymphocytotoxic antibody in the cerebrospinal fluid of patients with systemic lupus erythematosus: Correlation with central nervous system involvement

Abstract Lymphocytotoxic activity was measured in the cerebrospinal fluids (CSF) of 21 patients with systemic lupus erythematosus (SLE) and the relationship between that activity and the presence of central nervous system (CNS) involvement was assessed. Lymphocytotoxicity was determined in a dye exclusion microcytotoxicity assay without knowledge of the patients clinical status. Lymphocytotoxic activity was detected in 10 21 SLE patients. The lymphocytotoxicity was complement dependent, neutralized by anti-immunoglobulin antiserum, and absorbed by normal human brain tissue. CNS involvement was diagnosed in 11 patients, 8 of whom had CSF lymphocytotoxicity. The presence of brain-reactive lymphocytotoxic antibody in CSF correlated with the clinical presence of CNS manifestations (χ2 = 3.82, p

BEN, a novel surface molecule of the immunoglobulin superfamily on avian hemopoietic progenitor cells shared with neural cells

BEN is a novel molecule of the immunoglobulin superfamily that we previously identified by means of a monoclonal antibody on neural cell populations during avian development and epithelial cells of the bursa of Fabricius. In this paper, we describe the expression of BEN by hemopoietic cells during ontogeny. In the thymus, BEN is expressed as early as E9, and from E12 until just after hatching 30-60% of thymocytes are BEN positive. Thus the cells expressing BEN are immature thymocytes and not yet differentiated T cells. In the spleen, BEN expression parallels the myelopoietic activity. It is present on 75% of splenocytes during embryonic development and falls rapidly to 20% of cells during the first week after hatching when the spleen is becoming a secondary lymphoid organ. BEN is also found on a large proportion (about 80% positive cells) of bone marrow cells during ontogeny. Post hatching, BEN is present on 40-50% of bone marrow cells. The population of BEN-positive cells in the bone marrow includes myeloid and erythroid progenitor cells, identified by their ability to form colonies in vitro. BEN expression is lost as progenitor cells proliferate and differentiate to develop mature colonies in the clonal assay. Mature myeloid cells, such as macrophages, granulocytes, thrombocytes, and erythrocytes do not express the BEN antigen. Taken together, these data demonstrated that BEN is a stage-specific rather than a lineage-specific differentiation antigen expressed by immature hemopoietic cells.

An antigen expressed by avian neuronal cells is also expressed by activated T lymphocytes

A monoclonal antibody, anti-BEN, initially characterized by its reactivity with an epitope present on the surface of avian bursa epithelial cells and neurons, also reacts with membrane molecules on some hemopoietic cells. In this study we examine BEN expression on lymphoid cells in thymus, spleen, and blood. We demonstrate that BEN is an activation antigen on mature T lymphocytes. It is not expressed on peripheral blood or splenic lymphocytes, but following mitogenic or allogeneic stimulation of blood lymphocytes it appears rapidly on a T cell subpopulation in parallel with the appearance of IL-2 receptors. BEN is also expressed on III-C5 cells, an avian IL-2-dependent permanent T cell line, and on immature CD4+CD8+ thymocytes. BEN is not expressed by resting or actively proliferating B cells. Biochemical analyses of the BEN protein on T lymphoblasts shows that the molecule is similar in size to the BEN molecules on bursa epithelial cells and on neurons. The physicochemical properties of the BEN protein and its tissue distribution differs from other known avian and mammalian T cell activation markers, differentiation antigens, and integrins. Thus BEN is a novel marker of activated T cells in birds.

Antibodies reactive with central nervous system antigens

The pathogenesis of the neuropsychiatric manifestations of systemic lupus erythematosus (SLE) remains an enigma. The observation that many of the lymphocytotoxic antibodies in SLE are also brain-reactive has led to the hypothesis that central nervous system (CNS) lupus, like the autoimmune hematologic manifestations of SLE, is due to the direct effects of autoantibodies to cell membrane antigens. Studies of neuron-reactive antibodies in SLE sera and cerebrospinal fluid support that hypothesis and suggest that the diffuse neuropsychiatric manifestations require the co-existence of serum antibodies to nerve cells and an alteration in the blood-brain barrier that allows those antibodies to enter the CNS.

Utility of the Ultrasound Examination of the Hand and Wrist Joints in the Management of Established Rheumatoid Arthritis

To investigate the usefulness of point‐of‐care hand and wrist joint ultrasound (US) examination in patients with established rheumatoid arthritis (RA).

Generation of human cytomegalovirus-specific cytotoxic T lymphocytes in a short-term culture

A method to generate human cytomegalovirus (HCMV)-specific CTL (cytotoxic T lymphocytes) from human peripheral blood mononuclear cells is described. This assay is unique in comparison with other methods reported to date, because it only requires a short-term (6 days) coculture of PBM and autologous infected fibroblasts without the addition of exogenous IL-2 (interleukin-2) and nevertheless is sensitive enough to determine HCMV-specific killing in a short (6 h) 51Cr-release assay using autologous HCMV-infected fibroblasts as targets. The virus-specific killing is mediated by CTL of the CD8 phenotype and it can be inhibited by a HLA class I monoclonal antibody. The sensitivity of the assay can be significantly enhanced by pretreating the targets with interferon-gamma (IFN-gamma) prior to infection with HCMV. HCMV-specific 51Cr-release is more than doubled when the IFN-gamma pretreated targets are used. This increase is mostly due to enhanced sensitivity of the fibroblasts to killing mediated by CD8-positive CTL, but some killing can be attributed to CTL of the CD4 phenotype.

Lymphocyte reactivity of antisera to cryoproteins in systemic lupus erythematosus

Abstract Cryoglobulins from the sera of patients with systemic lupus erythamatosus (SLE) contain lymphocytotoxic antibodies. Since cryoprecipitation may be a property of antigen—antibody complexes, the presence of antibody to lymphocytes in SLE cryoprotein led us to examine the cryoprecipitated material for evidence of lymphocyte membrane antigens. Rabbit antisera to each of eight SLE cryoproteins were incubated with 51 Cr-labeled human peripheral blood lymphocytes. The lymphocytes were tested for their ability to function as antibody-sensitized targets in an antibody-dependent, cell-mediated cytotoxicity (ADCC) assay. Normal rabbit serum (NRS) and rabbit antiserum to human IgM, IgG, brain tissue (HBT), and thoracic duct lymphocytes (TDL) were used as control sera. Antibody binding to target lymphocytes resulting in greater than 5% 51 Cr-release occurred with SLE lymphocyte targets sensitized with anti-HBT ( 16 16 ). TDL ( 13 13 ), IgM ( 8 16 ), and IgG ( 2 14 ), but not with NRS ( 0 16 ). Normal lymphocytes were sensitized by anti-HBT ( 17 17 ), TDL ( 17 17 ), and IgM ( 2 11 ), but not with IgG ( 0 7 ) or NRS ( 0 17 ). Of the eight antisera to SLE cryoproteins, three sensitized both SLE and normal lymphocytes, whereas, the remaining five recognized cell-surface antigens contained only on SLE lymphocytes. The lymphocyte antigens detected by the anticryoprotein antisera remained on the target cell surfaces following overnight incubations. The demonstration of lymphocyte reactivity by antisera to SLE cryoprotein indicates that the SLE cryoprecipitates contain lymphocyte membrane antigens or, at least, antigens cross-reactive with cell-surface determinants. The SLE specificity of select antisera suggests the existence of SLE cell-associated lymphocyte antigens.

Epstein-Barr virus and rheumatoid arthritis

ConclusionThe immune system of patients with RA does not react normally to EBV infection. The control mechanisms that limit the effects of EBV infection are blunted in RA and, perhaps because of that, antibody responses to gBV-induced cellular antigens are significantly higher than in healthy individuals. There is little reason at present to suggest that EBV is a causative agent in RA. In fact, the recent studies by Sih,erman and Schumacher [9] of 12 patients with very early (less than 6 weeks) RA showed that they did not differ significantly from healthy controls in their antibody titers to VCA or EBNA, and none of them had detectable antibody to EA. Using immunofluorescence to detect anti-RANA, those investigators found a 70% incidence in their normal controls and a similar incidence in the patients with early onset RA. Thus, although patients with chronic RA exhibit abnormal serologic responses to EBVrelated antigens, those abnormalities do not appear to be a characteristic feature early in the disease, and the absence of antibodies to EA suggests that primary EBV infection or reactivation of latent infection is not a direct cause of RA.The EBV-related immunoregulatory defects in RA, although probably not etiologic, may contribute to the chronicity of the disease. RA subjects do not appear to harbor more infectious EBV particles in the orophaynx (the major EBV repository) than do healthy individuals [27]. However, the higher incidence of spontaneous in vitro outgrowth of EBV-containing lymphoblast cell lines from RA peripheral blood [13, 14] suggests that patients with RA may have an increased body burden of EBV-transformed cells. EBV-induced lymphoblasts are more potent stimulators of T cell activation than are normal B lymphocytes.