sepharose and Hypersensitivity

Marine polysaccharides have been found as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. Due to numerous pharmaceutical properties of marine polysaccharides such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, and antihepatopathy activities, they have been applied in many fields of biomaterials, food, cosmetic, and pharmacology. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been evidenced as downregulators of allergic responses due to enhancement of innate immune system, alteration of Th1/Th2 balance forward to Th1 cells, inhibition of IgE production, and suppression of mast cell degranulation. This contribution, therefore, focuses on antiallergic properties of marine polysaccharides and emphasizes their potential application as bioactive food ingredients as well as nutraceuticals for prevention of allergic disorders.

Topics: Alginates; Anti-Inflammatory Agents; Antioxidants; Antiviral Agents; Chitin; Chitosan; Dietary Supplements; Glucuronic Acid; Hexuronic Acids; Hypersensitivity; Immunoglobulin E; Mast Cells; Oligosaccharides; Polysaccharides; Seaweed; Sepharose; Th1-Th2 Balance

Polysaccharides are macromolecules made up of many monosaccharides joined together by glycosidic bonds. Polysaccharides from marine sources are widely distributed as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. So far, marine polysaccharides have been used in many fields of biomaterials, food, cosmetic, and pharmacology. Especially, numerous pharmaceutical properties of marine polysaccharides have been revealed such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, antihepatopathy, antiuropathy, and antirenalpathy activities. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been found as modulators of allergic responses due to enhancing innate immune system, altering Th1/Th2 balance, inhibiting IgE production, and suppressing mast cell degranulation. This contribution, therefore, focuses specially on the immunomodulatory effect of marine polysaccharides and emphasizes their potential application as candidates of pharmaceuticals as well as nutraceuticals to prevent allergic disorders.

Topics: Alginates; Animals; Aquatic Organisms; Chitin; Drug Industry; Glucuronic Acid; Hexuronic Acids; Humans; Hypersensitivity; Immunity; Immunologic Factors; Polysaccharides; Sepharose

This work investigated the potential use of an alternative adsorbent to anti-immunoglobulin E (IgE)-agarose for IgE selective adsorption therapy. A screening of several commercially available adsorbents (Concanavalin A, Lens culinaris[Lc], d-tryptophan, poly-l-lysine, and aminohexyl immobilized on agarose) was done through batch system assays, considering some criteria, such as adsorption capacity, selectivity, and biocompatibility. In the Lc-agarose adsorbent, total IgE, and specific IgE--for the airborne allergens Dermatophagoides pteronyssinus and Blomia tropicalis--were significantly better removed (63, 58, and 59%, respectively) than immunoglobulin G (19%), immunoglobulin A (33%), immunoglobulin M (9%), and albumin (18%). This adsorbent was packed into a column and the effect of superficial velocity, ratio of plasma volume to bed volume, number of perfusions, and temperature on IgE adsorption were evaluated. In vitro simulation of therapeutic adsorption (single perfusion) indicated that about 50% of total IgE could be eliminated.

Topics: Adsorption; Blood Component Removal; Chromatography, Agarose; Complement C3-C5 Convertases; Extracorporeal Circulation; Humans; Hypersensitivity; Immunoglobulin E; Immunosorbent Techniques; Immunosorbents; Ligands; Plant Lectins; Polylysine; Sepharose; Tryptophan

The role of anti-IgE autoantibodies in IgE-related allergic diseases has not been elucidated sufficiently. For example, anti-IgE antibodies have been reported to cause both proallergic and antiallergic blocking reactions. Contrary to other authors, some authors revealed a positive correlation between total IgE and the amount of IgE/IgG complexes detected. By comparing the IgE levels of allergic patients with those of control persons and rheumatoid arthritis patients the present study contributes to our understanding of the role of anti-IgE autoantibodies. The sera were tested by means of ELISA concerning their content of free and complexed IgG and IgM anti-IgE. In addition, the amounts of IgE/IgG and IgE/IgM complexes were determined in the sera of allergic and control persons after Superose 6 column separation. We found that the allergic patients revealed significantly higher values of free IgM anti-IgE than patients with rheumatoid arthritis and than control persons (mean 0.470, p = 0.0164 and p = 0.0061, respectively). The corresponding values for IgE/IgM complexes also tend to be higher (mean 0.431, p = 0.0784 and p = 0.0601, respectively). However, the corresponding contents of IgG anti-IgE autoantibodies and IgE/IgG complexes did not differ significantly. After Superose 6 column separation, we detected IgE/IgG in fractions corresponding to MW of 150 to 180 kDa for the sera of both allergic and control persons. In contrast, the IgE/IgM complexes were found in fractions corresponding to MW 330 kDa. We conclude that the increased IgM anti-IgE autoantibody titer in the sera of allergic patients is not correlated with the high total IgE level. Moreover, we suggest that the IgE/IgG complexes form de nova during ELISA. Unlike the IgE/IgG complexes, the IgE/IgM complexes are assumed to occur already in circulating blood.

Topics: Adult; Antibodies, Anti-Idiotypic; Antibody Specificity; Arthritis, Rheumatoid; Biomarkers; Chromatography, Gel; Enzyme-Linked Immunosorbent Assay; Female; Germany; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunoglobulin M; Male; Middle Aged; Sepharose; Time Factors

Specificities of IgE and IgG4 antibodies in 12 cat-allergic patients were compared with respect to their reactivity towards 3 IgE-binding synthetic peptides of Felis domesticus allergen 1 (Fel dI): peptides 25-38 and 46-59 of chain 1 and peptide 15-28 of chain 2. Peptides were coupled to Sepharose and anti-Fel dI antibodies were isolated by affinity chromatography. Fel dI-specific IgE- and IgG4 antibody activity in the peptide eluates was measured using Fel dI binding assays. Fel dI-specific IgE/IgG4 ratios in the eluates from peptide-Sepharose were determined and compared with the IgE/IgG4 ratios in the eluates from Fel dI-Sepharose. The mean ratio Fel dI-specific IgE/IgG4 in the peptide eluates (0.84; range 0.06-4.6) was significantly higher than the mean ratio in the eluates from Fel dI-Sepharose (0.31; range 0.13-1.1), demonstrating a higher reactivity of IgE antibodies with the peptides, compared to IgG4. These results indicate differences between the B cells producing IgE antibodies and the B cells producing IgG4 antibodies.

Topics: Allergens; Amino Acid Sequence; Animals; Antibodies; Cats; Chromatography, Affinity; Glycoproteins; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Molecular Sequence Data; Peptides; Protein Binding; Radioallergosorbent Test; Sepharose

In this report a procedure is described to calculate the affinity constant of an antibody for solid-phase Ag. KSP (KASS solid phase) was defined as the reciprocal of the concentration of Ag required for half saturation of the Ab-binding sites, extrapolated to an infinitely small concentration of Ab (semi-saturation plot). Using this procedure, the affinity of IgE antibodies can be measured without interference from 'invisible' IgG antibodies. As a model system, mAbs against the cat allergen Fel d I were used. Serial dilutions of Fel d I-Sepharose were incubated with serial dilutions of mAb Fd1a and Fd1b, with or without rabbit antibodies as 'invisible' antibodies. The Ab-binding capacity of Sepharose-coupled Fel d I was low: 4.15% and 2.13% of the expected value for mAb Fd1a and Fd1b, respectively, and this must be taken into account when calculating KSP. The K values of mAb Fd1a and Fd1b, calculated from the gamma axis intercept of the semi-saturation plot, were 85 (pmol/test)-1 and 65 (pmol/test)-1 respectively. Using the semi-saturation plot, KSP of mAb Fd1a was not affected by the presence of rabbit antibodies against Fel d I, confirming the applicability of the procedure for measuring the KSP of IgE in patient sera. For one cat-allergic patient the KSP of IgE and IgG4 for Fel d I were calculated and found to be 62 (pmol/test)-1 and 147 (pmol/test)-1 for IgE and IgG4 respectively.

Topics: Allergens; Animals; Antibodies, Monoclonal; Antibody Affinity; Antigen-Antibody Reactions; Cats; Glycoproteins; Humans; Hypersensitivity; Immunoglobulins; Kinetics; Mice; Mice, Inbred BALB C; Rabbits; Radioallergosorbent Test; Radioimmunoassay; Sepharose

Topics: Chromatography, Affinity; Humans; Hypersensitivity; Immunoglobulin E; Immunosorbents; Polymers; Sepharose

Horseradish peroxidase (HRP) or bovine serum albumin (BSA) were covalently linked to polyacrylamide or agarose beads and were injected into control Syrian hamsters and hamsters previously immunized with either HRP or BSA. Animals sensitized to soluble antigen and subsequently challenged intravenously with the same antigen immobilized on beads developed an acute focal inflammatory response within 2 to 6 hours after injection. The acute response involved local deposition of IgG and complement (beta1A/beta1C globulin), polymorphonuclear leukocyte exudation, and variable amounts of hemorrhage. A focal vasculitis was occasionally present. Within 72 hours the reaction had become largely mononuclear or granulomatous in nature, and giant cell formation was seen within 4 days after immobilized antigen injection. Severe reactions developed only upon recognition of specific antigenic determinants; thus hamsters immunized against soluble HRP developed characteristic lesions only upon intravenous challenge with HRP-coated beads but not with beads coated with unrelated antigen (BSA). The beads elicited only a mild foreign body reaction in the control hamsters at 5 to 7 days after injection which was temporally and histopathologically distinct from the lesions in immunized hamsters. Thus, the state of existing immunity can influence the character and severity of the local pulmonary inflammatory response.

Topics: Acrylamides; Animals; Complement System Proteins; Cricetinae; Fluorescent Antibody Technique; Foreign-Body Reaction; Hypersensitivity; Immunoglobulin G; Inflammation; Injections, Intramuscular; Injections, Intravenous; Leukocytes; Lung; Lung Diseases; Microspheres; Peroxidases; Pulmonary Artery; Sepharose; Serum Albumin, Bovine