dinitrobenzenes and Hypersensitivity

Guidelines have been presented for the optimal selection and interpretation of various laboratory tests related to rheumatologic, immunologic, and allergic disorders. The last decade has witnessed many technologic changes in the performance of screening tests for autoantibodies, and an expeditious approach to the detection of the clinically useful autoantibody system has been outlined. When both an immunofluorescent assay using an actively dividing substrate such as HEp-2 and an immunodiffusion test for precipitin autoantibodies are performed on patient sera, it is likely that most of the significant diagnostic and prognostic systems will be detected. Although exact pathogenetic roles have not been defined for all autoantibodies, it is apparent that highly specific markers for certain rheumatic diseases exist and the pertinent clinical aspects of each individual autoantibody-autoantigen system has been summarized. Why so many autoantibodies are cross-reactive with seemingly unrelated antigens is a perplexing aspect of autoimmunity, a disease process for which the necessary and sufficient conditions of development have not been defined even after decades of research. One recent hypothesis regarding the generation of diverse autoantibodies is the tenet that production is interrelated in an auto-anti-idiotypic network. Support for this hypothesis comes from the recent successful use of anti-idiotypes to DNA autoantibodies in an attempt to abrogate pathogenetic effects of target organ damage in murine models of SLE. A similar approach might be undertaken with some of the other autoantibody systems in an attempt at a novel therapy for disorders of immunoregulation.

Topics: Animals; Antibodies, Antinuclear; Autoantibodies; Blood Sedimentation; C-Reactive Protein; Clinical Laboratory Techniques; Counterimmunoelectrophoresis; Creatine Kinase; Dinitrobenzenes; DNA; DNA, Single-Stranded; Hemagglutination Tests; Histones; Humans; Hypersensitivity; Immune System Diseases; Immunodiffusion; Immunoglobulins; Mice; Rheumatic Diseases; Rheumatoid Factor; RNA

Topics: Animals; Antibody Specificity; Cell Communication; Dinitrobenzenes; Dose-Response Relationship, Immunologic; Epitopes; Hemocyanins; Humans; Hypersensitivity; Immunization, Passive; Immunoglobulin E; Lymphocytes; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; T-Lymphocytes; Time Factors; X-Rays

The immunological mechanisms underlying the role of mast cells in the pathogenesis of inflammatory bowel disease (IBD) are poorly defined. In this study, non-IgE mediated colonic hypersensitivity responses in BALB/c mice induced by skin sensitization with dinitrofluorobenzene (DNFB) followed by an intrarectal challenge with dinitrobenzene sulfonic acid featured as a model to study the role of mast cells in the development of IBD. Vehicle- or DNFB-sensitized mice were monitored for clinical symptoms and inflammation 72 h after dinitrobenzene sulfonic acid challenge. DNFB-sensitized mice developed diarrheic stool, increased colonic vascular permeability, hypertrophy of colonic lymphoid follicles (colonic patches), and showed cellular infiltration at the microscopic level. Increased numbers of mast cells were found in the colon of DNFB-sensitized mice located in and around colonic patches associated with elevated levels of mouse mast cell protease-1 in plasma indicating mast cell activation. Colonic patches of DNFB mice, stimulated in vitro with stem cell factor indicated that an increase in TNF-alpha levels in the colon is mainly mast cell originated. Finally, neutrophil infiltration was observed in the colon of DNFB-sensitized mice. Induction of this model in mast cell-deficient WBB6F(1) W/W(v) mice shows a profound reduction of characteristics of the colonic hypersensitivity reaction. Reconstitution with bone marrow-derived mast cells in WBB6F(1) W/W(v) mice fully restored the inflammatory response. This study demonstrates the importance of mast cells in the development of clinical symptoms and inflammation in the presented murine model for IBD.

Topics: Animals; Colitis; Diarrhea; Dinitrobenzenes; Hypersensitivity; Hypertrophy; Intestinal Absorption; Lymph Nodes; Male; Mast Cells; Mice; Mice, Inbred BALB C; Tumor Necrosis Factor-alpha

We investigated the effect of spirulina on mast cell-mediated immediate-type allergic reactions. Spirulina dose-dependently inhibited the systemic allergic reaction induced by compound 48/80 in rats. Spirulina inhibited compound 48/80-induced allergic reaction 100% with doses of 100-1000 microg/g body weight, i.p. Spirulina (10-1000 microg/g body weight, i.p.) also significantly inhibited local allergic reaction activated by anti-dinitrophenyl (DNP) IgE. When rats were pretreated with spirulina at a concentration ranging from 0.01 to 1000 microg/g body weight, i.p., the serum histamine levels were reduced in a dose-dependent manner. Spirulina (0.001 to 10 microg/mL) dose-dependently inhibited histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP in RPMC, when spirulina (10 microg/mL) was added, transiently and significantly increased about 70-fold at 10 sec compared with that of control cells. Moreover, spirulina (10 microg/mL) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production. These results indicate that spirulina inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro.

Topics: Animals; Bacterial Proteins; Cyanobacteria; Dinitrobenzenes; Histamine; Histamine Release; Hypersensitivity; Immunoglobulin E; Mast Cells; p-Methoxy-N-methylphenethylamine; Peritoneal Cavity; Rats; Spirulina; Tumor Necrosis Factor-alpha

Human foods are usually prepared by cooking. Boiling of chicken egg-white (EW) led to decreased allergenicity, and abrogated intestinal uptake of immunoreactive ovalbumin (OVA) when fed to mice. Therefore, the effects of oral administration of boiled EW were examined further in BALB/c mice. Specific IgE, IgG1 and IgG antibody responses were suppressed by raw EW, but not by EW boiled for 5 or 60 min, fed prior to sensitization with 10 microg OVA or 1 microg DNP-OVA in alum. Similar results were obtained when mice were sensitized with 10 microg conalbumin, ovomucoid or lysozyme in alum. BALB/c spleen cell proliferation and secretion of Th2 cytokines IL-4 and IL-5 during in vitro stimulation with OVA were also suppressed by feeding raw EW, but not by boiled EW. Although heat denaturation of proteins can minimize allergenicity, the present results suggest that over-cooking of proteins may affect their intestinal antigen processing and thus prevent the induction of oral tolerance.

Topics: Animals; Cell Division; Conalbumin; Dinitrobenzenes; Egg Proteins; Female; Heating; Hypersensitivity; Immune Tolerance; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Muramidase; Ovalbumin; Ovomucin; Protein Denaturation; Rats; Rats, Sprague-Dawley; Spleen; Th2 Cells

Contact sensitivity (CS), anti-DNP (dinitrophenyl) IgG and IgE type antibodies were induced by skin applications of the contact sensitizer dinitrofluorobenzene. CS response was suppressed by treatment with the monovalent hapten dinitrobenzene sulfonate (DNBS) but was not affected by treatment with the DNP-conjugates (DNP22-Ficoll, DNP17-polyethylene glycol and DNP2-polyvinylpyrrolidone). On the other hand, IgE type response was depressed by treatment with the various DNP conjugates but was not affected by DNBS. The IgG type response was not appreciably affected by any of the treatments.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Dose-Response Relationship, Immunologic; Hypersensitivity; Immunity, Cellular; Immunoglobulin E; Immunoglobulin G; Immunosuppression Therapy; Mice; Mice, Inbred BALB C; Nitrobenzenes; Skin

Intravenous administration of syngeneic spleen cells (SPC), briefly pulsed with antigen in vitro, resulted in a profound state of IgE antibody unresponsiveness. One of the mechanisms of this unresponsiveness is responsible for an immediate tolerance which is induced without any suppressor cells. Characteristics of this immediate tolerance were investigated. Administration of antigen-pulsed spleen cells 4 h before the immunization, suppressed the production of IgE antibody triggered by the subsequent immunization. Pretreatment with cyclophosphamide had no effect on this rapid suppression, and this suppressive state could not be transferred to normal syngeneic recipients by the injection of spleen cells from the tolerant mice used in our experiment. These observations suggest that suppressor cells do not play an important role in immediate tolerance. The extent of this immediate tolerance induced by the injection of antigen-pulsed SPC depends on the number of antigen-pulsed SPC and the dose of antigen to which SPC had been exposed. Injection route of antigen-pulsed SPC has a great influence on the induction of immediate tolerance. The order of suppressive extent is intravenous, greater than intraperitoneal greater than subcutaneous. This suppression is specific to the antigen pulsed to SPC. Carrier-specific T cells are the major target of suppression in immediate tolerance. Antigen-pulsed T cells induce immediate tolerance most effectively in the subpopulations of SPC.

Topics: Animals; Antibody Formation; Dinitrobenzenes; Dose-Response Relationship, Immunologic; Female; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Mice; T-Lymphocytes

The effect of a water-soluble fraction (CEF) that was prepared from an extract of Corynebacterium equi on primary reaginic antibody formation was studied in Balb/c mice. Mice were immunized with a hapten carrier (DNP-OVA) and received intraperitoneal injections of CEF 7 and 2 days prior to, or 2 and 7 days after the immunization. PCA titers of both antihapten (DNP) and anticarrier (OVA) antibodies of IgE class were reduced significantly by the CEF treatment. Evidence was presented in adoptive transfer experiments that the number of IgE-producing cells in the CEF-treated mice was lower than that of controls. Suppression of IgG1 anti-DNP antibody formation was also achieved by the CEF treatment. Formation of IgG1 anti-OVA antibodies, however, was not suppressed significantly by the treatment. The suppressive activities of CEF were shown to be dose-dependent, but timing of CEF administration did not appear critical.

Topics: Aerobiosis; Animals; Antibody Formation; Antibody-Producing Cells; Carrier Proteins; Chemical Fractionation; Corynebacterium; Dinitrobenzenes; Epitopes; Haptens; Hypersensitivity; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Reagins; Time Factors

Experiments presented in this paper were designed to test a new concept concerning the possible pathogenesis of the allergic phenotype. This concept, termed "allergic breakthrough" considers that one of the avenues toward the allergic phenotype involves coincidental sensitization combined with an imbalance in the normal damping mechanism that serves to limit IgE antibody production. The three predictions of this concept that can be tested experimentally are: 1) manipulations that are effective in heightening or re-establishing the damping mechanism should manifest persistence insofar as IgE antibody synthesis to the relevant allergen is concerned; 2) once allergic breakthrough has occurred, the height of production of IgE antibodies specific for the sensitizing agent should remain elevated at levels characteristic of the allergic phenotype, even after the threshold of damping activity has returned to a normal level; and 3) allergic breakthrough should display specificity in that breakthrough would occur in response to subsequent exposure to the specific antigen to which coincidental sensitization initially occurred, but not for other unrelated antigens. The studies presented herein confirm each one of these predictions, thereby providing substantial support for the validity of this concept as one possible distinguishing feature between individuals manifesting the nonallergic and allergic phenotypes, respectively.

Topics: Animals; Ascaris; Blood; Dinitrobenzenes; Dose-Response Relationship, Radiation; Epitopes; Freund's Adjuvant; Hypersensitivity; Immunity, Maternally-Acquired; Immunoglobulin E; Mice; Phenotype; Time Factors

Two biologically active serum molecules manifesting precisely opposite biologic effects, both of which are selective for IgE antibody synthesis, can be detected in the serum and ascites fluids of CFA-immune mice. One activity, described previously, is suppressive and hence termed suppressive factor of allergy (SFA); the other, reported for the first time herein, is enhancing and has been termed enhancing factor of allergy (EFA). The ability to detect one vs the other activity requires certain special manipulations such as different doses of low dose x-irradiation. Conclusive evidence for the existence of two distinct factors mediating these two opposing biologic effects was obtained in studies demonstrating that affinity chromatography on concanavalin A-Sepharose segregated the two molecular entities. Thus, SFA binds poorly or not at all to Con A-Sepharose, whereas EFA binds to Con A and can be recovered in the eluate eluted with the competitive sugar alpha-methyl-D-glucopyranoside.

Topics: Animals; Antibody Formation; Ascaris; Blood; Chromatography, Affinity; Dinitrobenzenes; Dose-Response Relationship, Immunologic; Dose-Response Relationship, Radiation; Freund's Adjuvant; Hemocyanins; Hypersensitivity; Immunity, Maternally-Acquired; Immunoglobulin E; Immunosuppression Therapy; Mice; Mice, Inbred BALB C; Time Factors

Dinitrophenyl derivatives of differing molecular weights and degrees of substitution have been contrasted with respect to their ability to elicit immediate type allergic responses and their capacity to induce antibody formation in the guinea pig. In contradistinction to dinitrophenyl-proteins, bis-DNP-lysine and DNP-polylysines (including a 100,000 molecular weight derivative) failed to induce antibody detectable by guinea pig passive cutaneous anaphylaxis. Dinitrophenyl-polylysines evoked urticarial responses non-specifically, but after succinylation were about as effective as dinitrophenyl-proteins in eliciting specific cutaneous reactions. An important factor influencing the effectiveness of bis-DNP-lysine in evoking specific wheal-and-erythema responses is antibody affinity for the dinitrophenyl-lysyl determinant.

Topics: Antibodies; Antibody Formation; Antigens; Dermatitis, Atopic; Dinitrobenzenes; Guinea Pigs; Haptens; Hypersensitivity; Hypersensitivity, Immediate; Indicators and Reagents; Lysine; Passive Cutaneous Anaphylaxis; Polylysine; Skin

When one or two drops of a dilute, non-irritating solution of 2,4-dinitrochlorobenzene (DNCB) is applied to a small area of skin of the intact guinea pig, about 20 per cent of the applied material, or some derivative of it, is soon excreted in urine. In normal, as well as in specifically sensitized guinea pigs, DNCB at the site of local application becomes rapidly bound to skin protein through primary chemical bonds. Twenty-four hours after application roughly half of the material present at the local skin site is still extractable with organic solvents. Of the non-extractable dinitrophenyl groups, about 99 per cent are in epidermis, and about 85 per cent are substituted in epsilon-NH(2) groups of lysine residues. Only traces of bound dinitrophenyl groups were observed in the corium. It is uncertain whether these are formed in situ, or are experimental contaminants, or are migratory epidermally formed conjugates. Even when DNCB is injected intradermally it combines predominantly with overlying epidermis and with epidermal components of hair follicles, but only slightly with corium. The 2,4-dinitrophenyl conjugates which are localized in the deeper, viable half of the epidermis, close to the epidermal-dermal junction, are inferred to be the agents responsible for specifically evoking the allergic response in sensitized animals. Conjugates which are situated in the outer, cornified half of the epidermis are shown to be incapable of eliciting the allergic response. The results furnish a basis for interpreting a common pattern of lesions in allergic contact dermatitis as it occurs spontaneously in man.

Topics: Animals; Dermatitis; Dermatitis, Allergic Contact; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Epidermis; Guinea Pigs; Humans; Hypersensitivity; Male; Nitrobenzenes; Proteins; Skin; Skin Physiological Phenomena

Topics: Blood Proteins; Dermatitis, Atopic; Dinitrobenzenes; Humans; Hypersensitivity; Nitrobenzenes; Skin

2,4-dinitrophenylsulfenyl chloride (DSCl) and 2,4-dinitrophenylthiocyanate (DSCN) elicited allergic reactions of the delayed type when applied to the skin of guinea pigs and of human beings who had been sensitized by prior exposure to 2,4-dinitrofluorobenzene (DF). DSCl and DSCN, together with 2,4-dinitrobenzene sulfonate (DSO(3)), constitute a clearly defined group of allergenic dinitrophenyl compounds in that they all combined with skin protein in vivo through reaction with cysteine or cystine. In vitro, these compounds combine with free SH groups, and with -S-S- groups of hair and epidermis, but not with -S-S- groups of oxidized glutathione or of bovine gamma globulins. DSO(3), DSCl, and DSCN did not react with amino groups in vivo, but did react with protein amino groups in vitro at pH values of about 10. Another group of dinitrophenyl compounds (DF, DCl, and DBr) previously had been shown to combine with lysine epsilon-NH(2) groups of epidermal proteins. In the present work it was found that these compounds do not react with the disulfide groups of these proteins, either in vivo or in vitro. Moreover, they did not seem to react with SH groups of viable skin, although they are highly reactive with sulfhydryl in vitro. This apparent discrepancy between reactivity with SH groups in vitro and in vivo may be due to the fact that the chromatographic technique employed was relatively insensitive for the sulfhydryl derivative. When a compound of either group was applied to the skin surface, dinitrophenyl-amino acids were recovered from the epidermis but not from the dermis. The results are discussed from the viewpoint of the epidermal localization of dinitrophenyl-protein conjugates.

Topics: Allergens; Animals; Cattle; Cysteine; Cystine; Dinitrobenzenes; Dinitrofluorobenzene; Disulfides; Guinea Pigs; Humans; Hypersensitivity; Immune System Diseases; Lysine; Molecular Weight; Nitrobenzenes; Proteins; Skin; Sulfenic Acids; Sulfhydryl Compounds