sepharose and Autoimmune-Diseases

Our investigation explores the immuno-efficiency of sulphated polysaccharides enriched Porphyra vietnamenis. Isolated polysaccharide fraction (17.1-25.8%) was characterized by FTIR and NMR spectroscopy which showed the presence of typical linear backbone structure called as porphyran. Oral administration of porphyran (200-500 mg/kg) evoked a significant (P ≤ 0.05) increase in weight of the thymus, spleen and lymphoid organ cellularity. The total leucocyte and lymphocyte count was increased significantly (P<0.005). The increase in the percent neutrophil adhesion to nylon fibres as well as a dose-dependent increase in antibody titre values was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen and significant changes were observed in the formation of formazone crystals. It also prevented myelosuppression in cyclophosphamide drug treated rats. The results indicated that P. vietnamenis possesses potential immunomodulatory activity and has therapeutic potential for the prevention of autoimmune diseases.

Topics: Administration, Oral; Animals; Antibody Formation; Autoimmune Diseases; Cell Adhesion; Immunologic Factors; Lymphocyte Count; Mice; Neutrophils; Porphyra; Rats; Rats, Wistar; Sepharose

Topics: Animals; Autoimmune Diseases; Capsules; Diabetes Mellitus, Type 1; Graft Survival; Islets of Langerhans Transplantation; Mice; Mice, Inbred NOD; Sepharose; Time Factors; Transplantation, Isogeneic

A quantitative assay with microSepharose was used to determine GAD65Ab and IA-2Ab levels in 771 population-based patients diagnosed with diabetes mellitus at 15 to 34 years of age, and in 828 matched controls. Among the patients, 587 (76%) were classified with type I, 108 (14%) with type II, and 76 (10%) with unclassifiable diabetes. The levels above normal demonstrated a prevalence of GAD65Ab in 66% of type I diabetes, 50% of type II diabetes and 54% of unclassifiable patients and for IA-2Ab in 40%, 17% and 21%, respectively. Among the autoantibody-positive sera, the LADA patients had a lower GAD65Ab index (median 0.19, p < 0.0001) and IA-2Ab index (median 0.28, p < 0.0001) than the type I patients (median 0.37 and 0.66). Patients with unclassifiable diabetes had a GAD65Ab (median 0.43) or IA-2Ab (median 0.63) index which was not different from the type I diabetes patients. Our data demonstrate that young adult new-onset LADA patients have low level GAD65Ab and IA-2Ab. The low-level autoantibodies may signify a less aggressive beta-cell autoimmunity, which may explain why these patients are often classified with type II or non-insulin-dependent diabetes.

Topics: Adolescent; Adult; Autoantibodies; Autoimmune Diseases; Biomarkers; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Female; Glutamate Decarboxylase; Humans; Islets of Langerhans; Male; Sepharose

Macrophages (m phi) from prediseased autoimmune-prone MRL/+ and MRL/ lpr mice have a marked defect in endotoxin (LPS)-induced expression of several cytokines including interleukin 1 (IL-1). The progressive nature of this defect over time suggests that it may develop in response to specific extracellular stimuli. In this report, we show that adhesion is an essential factor for the development of aberrant IL-1 expression by m phi from autoimmune-prone MRL mice. Thus, when MRL/+ m phi were allowed to adhere before being stimulated with LPS, they demonstrated a striking defect in expression of both IL-1 message and protein in comparison with multiple normal strains. In marked contrast, when MRL/+ m phi were maintained in a non-adherent state by culture on agarose, the IL-1 defect was not evident and IL-1 expression was restored to nearly normal levels. Since an identical defect in IL-1 expression was found when MRL/+ m phi were cultured on a variety of extracellular matrix proteins (including laminin, fibronectin, type I collagen, and type IV collagen), it appears that IL-1 underexpression is dependent on the adhesive state per se rather than on engagement of any one specific adhesion receptor. Moreover, the cytoskeletal inhibitor cytochalasin D had no effect on the magnitude of the defect, indicating that the adhesion-dependent signaling events necessary to elicit IL-1 underexpression are independent of cytoskeletal rearrangement. Taken together, these results indicate that m phi from autoimmune prone MRL/+ mice have an adhesion-dependent signaling abnormality that leads to profound underexpression of the cytokine IL-1.

Topics: Animals; Autoimmune Diseases; Cell Adhesion; Cells, Cultured; Culture Media; Cytochalasin B; Cytoskeleton; Disease Susceptibility; Extracellular Matrix Proteins; Gene Expression Regulation; Heterozygote; Interleukin-1; Lipopolysaccharides; Lymphoproliferative Disorders; Macrophages, Peritoneal; Mice; Mice, Inbred MRL lpr; Mice, Inbred Strains; Sepharose; Signal Transduction

Autoantibodies from the MRL/lpr mice react with numerous proteins on neuronal cell surfaces. The purpose of this study was to isolate and characterize a population of autoantibodies reactive preferentially or exclusively with nervous system tissue. Using a purified plasma membrane preparation from brain cortex of balb/c mice coupled to diaminopropylamine agarose gel, we affinity-isolated antineuronal antibodies from pooled MRL/lpr immunoglobulins. The isolated immunoglobulins reacted with brain cortex plasma membranes and neuroblastoma cells (but not liver, kidney, or fibroblasts) by Western blot and indirect immunofluorescence with confocal microscopy. By Western blot, the epitopes in the brain cortex were proteins of apparent molecular weights 101, 63, 53, 43, 39, and 33, kd; the epitopes in the neuroblastoma cells were 63, 57, and 53 kd. Lectin column isolation revealed that the 101 and 63 kd epitopes were glycosylated. Indirect immunofluorescence revealed that the antibodies bound to the cell soma more intensely than to the cell processes of viable cultured neuroblastoma cells. The cell surface localization of this binding was confirmed by confocal microscopy. Within the central nervous system the antibodies bound more intensely to primary cultures of isolated neurons from fetal cortex than to hippocampal or neostriatal cells. With these antibodies we can begin studies of their potential pathogenic effects.

Topics: Animals; Antibody Specificity; Antigen-Antibody Reactions; Autoantibodies; Autoimmune Diseases; Blotting, Western; Brain Neoplasms; Cell Membrane; Cerebral Cortex; Chromatography, Affinity; Disease Models, Animal; Female; Fibroblasts; Fluorescent Antibody Technique; Gels; Immunosorbent Techniques; Kidney; Liver; Liver Neoplasms, Experimental; Lupus Erythematosus, Systemic; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Molecular Weight; Nerve Tissue Proteins; Neuroblastoma; Neurons; Organ Specificity; Sepharose

Naturally occurring autoantibodies against native DNA (nDNA) in SLE sera showed diverse antigen binding characteristics. The antibodies isolated by affinity chromatography using nDNA linked to Sepharose 4B exhibited specificity towards nDNA and showed strong reactivity with DNA-psoralen photoadduct by direct binding assay and competitive ELISA. The anti-DNA antibody belong to both IgG and IgM immunoglobulin classes and their ratio was 5:1. The possible significance of altered conformation of nDNA in the etiology of SLE has been discussed.

Topics: Antibodies, Antinuclear; Antibody Diversity; Antibody Specificity; Autoimmune Diseases; Binding, Competitive; Chromatography, Affinity; Cross Reactions; DNA; DNA, Single-Stranded; Enzyme-Linked Immunosorbent Assay; Ficusin; Humans; Immunoglobulin G; Immunoglobulin M; Lupus Erythematosus, Systemic; Nucleic Acid Conformation; Sepharose

A rapid method has been developed for enrichment of Jo-1 antigen (histidyl-tRNA synthetase) from HeLa cells. The enzyme has been prepared from post-ribosomal supernatant by successive chromatography with Blue Sepharose and Poly-U-Sepharose, followed by DEAE-high performance liquid chromatography (HPLC). By this method, enzyme could be obtained within 4 days of HeLa cell harvesting, with 40% recovery of the enzymatic activity. The apparent native molecular size of the enzyme as determined by HPLC-size exclusion column chromatography was approximately 120 kDa. Under denaturing conditions using SDS-polyacrylamide gel electrophoresis the enzyme subunit size was approximately 55 kDa. The antigen preparation, although not homogeneous, was found to react only with anti-Jo-1 positive antisera when tested by immunoblotting with many patient sera of defined autoantibody specificities, making the preparation useful for immunologic studies of anti-Jo-1 antibodies.

Topics: Amino Acyl-tRNA Synthetases; Antibodies, Antinuclear; Autoantigens; Autoimmune Diseases; Chromatography, Gel; Chromatography, High Pressure Liquid; Collodion; Electrophoresis, Polyacrylamide Gel; HeLa Cells; Histidine-tRNA Ligase; Humans; Molecular Weight; Rheumatic Diseases; Sepharose

Agarose acrobeads were produced by encapsulating polyacrolein microspheres (acrobeads) of 0.2 micron average diameter within an agarose matrix. Crosslinked agarose acrobeads of diameters ranging from 0.5 to 0.8 mm were found to be optimal spheres for specific hemoperfusion purposes. Agarose provides the biocompatibility and mechanical strength of the agarose acrobeads. Acrobeads contain a high aldehyde-group content through which various amino ligands, i.e., proteins, antigens, antibodies, enzymes, and so on, can be covalently bound in a single step under physiological pH (or other pH). Thus, antibodies, antigens, or toxic materials may be directly removed from whole blood by hemoperfusion. During in vitro and in vivo hemoperfusion trials, the content of erythrocytes, leukocytes, and thrombocytes was essentially unaltered. Likewise, a battery of the soluble blood components (Cl-, K+, Na+, Ca2+, PO3/4-), total proteins, albumin, and C'4 component of the complement cascade, as well as the enzymes SGOT, LDH, and alkaline phosphatase, remained constant within narrow limits during the hemoperfusion procedure. The chemical and physical structure of the beads is stable; neither acrolein nor bead fragments were detected in hemoperfusion trials. Similarly, leakage of antibody bound to the agarose acrobeads into the blood is insignificant. Thus far, we have demonstrated the efficacy of the crosslinked agarose acrobeads for extracorporeal removal of "unwanted" substances from whole blood in the following systems: (a) removal of specific antigens (digoxin or paraquat removal with antidigoxin or antiparaquat antibodies bound to the acrobeads, respectively), (b) removal of specific antibody (antiBSA) removal with BSA bound to the beads), (c) removal of immune complexes (BSA-antiBSA complex removal with C1q bound to acrobeads), and (d) removal of specific metals (removal of iron with deferoxamine bound to the agarose acrobeads).

Topics: Animals; Antigen-Antibody Complex; Autoantibodies; Autoimmune Diseases; Biocompatible Materials; Glutaral; Hemoperfusion; Humans; Indicators and Reagents; Paraquat; Protein Binding; Sepharose