Demonstration of auto-anti-idiotypic antibody cross-reacting with public idiotypic determinants in the serum of rye-sensitive allergic patients

The present study documents the presence, in the serum of one allergic individual, of auto-anti-idiotypic antibodies cross-reacting with public idiotypic determinants expressed on human IgE and IgG anti-Rye I antibodies. Sera from rye-sensitive patients were tested for specific IgG and IgE antibodies to Rye I by double antibody. The IgG fraction, isolated from the serum of a patient with a history of previous hyposensitization therapy, was repeatedly absorbed on Rye-I-Sepharose as well as on IgM- and IgG-Sepharose to remove anti-Rye I antibodies as well as any possible anti-heavy or light chain activity. This IgG fraction, named anti-idiotypic fraction (a-IdF), blocked in a dose-dependent fashion the reaction of IgG and IgE anti-Rye I antibodies with Rye I antigen. The a-IdF also blocked the binding of anti-rye antibodies to Rye I antigen in the serum of 20 unrelated allergic patients, indicating that these anti-Rye I antibodies bore public idiotypic determinants.     

Immunoenzyme test system with monoclonal antibodies to human IgG4 in the determination of allergen-specific antibodies in pollinosis

ELISA for determination of allergen-specific IgG4 antibodies was developed with the help of monoclonal anti-IgG4 antibodies obtained by classic hybridoma technique. Subclass specificity of antibodies were studied in sera of 108 patients suffering from pollinosis. Antibodies of this isotype were found in the majority of patients with tree pollen allergy but not in patients with grass pollen allergy. The level of IgG4 antibodies correlated with the severity of the disease but not with the intensity of skin tests. Specific hyposensitization resulted in significant increase of IgG4 antibody level in patients with tree pollen allergy. Determination of IgG4 antibodies is proved to be useful to reveal tree pollen allergy and to monitor hyposensitization therapy.     

Level of specific IgE and IgG in blood serum of patients with hay fever after several cycles of desensitization with pollinex vaccine]

Blood serum specific IgE and IgG levels have been assayed in a group of 54 patients allergic to grass pollen during the long-lasting desensitization with Pollinex Depot vaccine. Specific IgE and IgG levels were determined with RAST and IgG-RAST technique whereas a titre of the blocking antibodies with RAST inhibition test. It was found that cyclic administration of Pollinex vaccine, repeated every 3-4 years, significantly influences patients' immunologic system manifested by a decrease in specific IgE, and an increase in both IgG level and the titre of the blocking antibodies.     

Imbalances of T cell subpopulations in patients with atopic diseases and effect of specific immunotherapy.

Reduced numbers of T cells with Fc receptors for IgG (TG cells) are present in blood samples of patients with respiratory allergic disease, mainly those with severe symptoms. TG cells have been previously shown to be suppressor in the pokeweed mitogen- (PWM) dependent B cell differentiation. T cells with Fc receptor for IgM (TM cells), which help immunoglobulin production, are in a normal range. After specific hyposensitization, resulting in a sharp improvement of clinical symptoms, TG cell subset reached normal values.     

Control of allergic reactivity in human filariasis. Predominant localization of blocking antibody to the IgG4 subclass.

Patients with chronic helminth infections, despite having abundant basophils and mast cells specifically sensitized with antiparasite IgE and often exposed repeatedly to parasite Ag, rarely manifest allergic symptoms. This control of clinical allergic reactivity likely results from Ag-specific IgG "blocking antibodies" shown previously to be abundant in the sera of such patients. In the present study we used two approaches to determine in which of the four IgG subclasses this blocking activity was localized. First, specific antifilarial antibodies of each of the four IgG subclasses were quantified in the sera of 28 patients with Bancroftian filariasis and correlated with the levels of blocking activity in these sera (determined by histamine release assays). A significant correlation with blocking activity was seen only for antibodies of the IgG4 subclass, and, indeed, the correlation was especially strong in the group of totally asymptomatic patients (but with microfilariae circulating in the blood) in whom blocking antibody levels were highest. Interestingly, however, if the analysis excluded these asymptomatic microfilaremic patients and focused instead on those with lymphatic inflammatory pathology (who had relatively low levels of both serum blocking activity and specific IgG4 antibodies), then the small amount of blocking activity found in these sera correlated only with the levels of IgG1 subclass antibodies. The second approach utilized selective depletion of IgG4 (by anti-IgG4 affinity columns) from the sera of three microfilaremic patients with high levels of blocking activity and demonstrated clearly that removal of IgG4 abolished the majority of the blocking activity in these sera (53, 78, and 81%). These two sets of findings demonstrate a predominant role for specific IgG4 antibodies in blocking IgE-mediated allergic responses to the parasite Ag in vitro, but they also indicate that in some situations IgG1 antibodies can block such reactions. Furthermore, the correlation demonstrated between patients' clinical presentations and the levels of both their specific IgG4 antibodies and serum blocking activity suggests that these antibodies play a similar role in vivo as well.     

Specific hyposensitization of microbial allergy by using a synthetic artificial polyelectrolyte

In this work the effectiveness of complex immunotherapy, including specific inhalation hyposensitization with the introduction of artificial synthetic polyelectrolite, was studied. Specific allergen produced a good effect (75%) in the inhalation hyposensitization of hemophilic allergy. The intramuscular injection of artificial synthetic polyelectrolite, made in addition to the inhalation of allergen, produced a better desensitizing effect than the separate administration of allergen or polyelectrolite. The multiple administration of polyelectrolite produced a desensitizing effect on allergic reactions of type I with the tendency towards the decrease of reactions of type IV. Good prospects for the development of methods for special treatment with homologous allergen in combination with NA-5 in cases of microbial sensitization under the control of immunocompetent cells were shown.     

The Treatment of Allergic Rhinitis

Allergic inflammation of the nasal mucous membranes, like other atopic disorders, occurs primarily as the result of an antigenantibody reaction between external allergens and circulating skin-sensitizing antibodies. In addition, the disease process is frequently complicated by bacterial or viral infection. Effective treatment of allergic rhinitis, therefore, consists of: (1) changing the patient''s environment in order to remove the offending allergens, (2) removing the patient from his environment, (3) altering the patient''s response to environmental allergens by means of hyposensitization injections, (4) suppressing the allergic reaction with drugs, and (5) eliminating bacterial infection. Usually more than one of these therapeutic measures is required for the individual patient.     

The effect of hyposensitization on alternaria-induced lymphocyte blastogenesis

Twenty-one atopic children were treated with alternaria hyposensitization therapy. Leukocytes from these children were cultured with and without alternaria and phytohemagglutinin prior to treatment, and after 6 and 12 months of treatment. Patients demonstrating positive immediate cutaneous hypersensitivity showed significant lymphocyte transformation prior to hyposensitization whereas the skin test-negative patients did not. The skin test-negative patients developed significant lymphocyte transformation after 6 months of hyposensitization. Twelve months after the initiation of therapy both skin test-negative and skin test-positive patients showed a decreased lymphocyte reactivity, and there was no longer any significant change in the concomitant phytohemagglutinin responses indicating that hyposensitization had a specific effect on alternaria-induced in vitro lymphocyte reactivity. Plasma factors were found to have modulating effects on the lymphocyte reactivity of hyposensitized patients.     

Antibodies to Yersinia enterocolitica and Proteus mirabilis in blood sera of rheumatoid arthritis patients

Data on specific antibodies to the bacterial antigens of Yersinia enterocolitica (serovars 03, 09) and Proteus mirabilis in rheumatoid arthritis (RA) patients are presented. According to the data of some researchers, these organisms play some role in the pathogenesis of RA. The highest level of IgG antibodies to Y. enterocolitica antigens of both serovars was noted. The activity of specific IgG antibodies to serovar 03 was higher than that to serovar 09. In the sera of RA patients with P. mirabilis antgens the activity of IgM antibodies was higher.     

Production of anti-Candida antibodies in mice with gut colonization of Candida albicans

BACKGROUND: Production of antibodies that are specific for allergens is an important pathological process in inflammatory allergic diseases. These contain the antibodies against antigens of Candida albicans, one of the normal microbial flora in an intestinal tract. We studied the effects of the prednisolone administration on the production of anti-Candida antibodies in the gastrointestinally C. albicans-colonized mice. METHODS AND MATERIALS: BALB/c mice, treated with antibacterial antibiotics to decontaminate indigenous intestinal bacterial flora, were inoculated intragastrically with C. albicans. The mice, in which C. albicans grows intestinally, were administered prednisolone to induce temporary immunosuppression. The Candida growth in their intestinal tract and their antibody response to Candida were examined. RESULTS: Antibiotic treatment allowed establishment of C. albicans gastrointestinal colonization, but did not cause subsequent systemic dissemination of C. albicans in all the animals. When these animals received an additional treatment with prednisolone, they showed a significantly higher population of C. albicans in their feces than those of animals treated with antibiotics alone, and the organisms were recovered even from their kidney. This systemic dissemination by C. albicans appeared to be temporal, because all the mice survived without any symptoms for more than 2 months. Examination of the serum titers of total immunoglobulin (Ig)E antibodies and specific IgE and IgG antibodies against Candida antigens demonstrated that titers of total IgE increased, partially by day 14 and clearly at day 27, in prednisolone-treated Candida-colonized mice. Without prednisolone treatment, an increment of the serum titer was scarcely observed. By day 27, corresponding to the increase of total IgE, the anti-Candida IgE and IgG titer increased in mice of the prednisolone-treated group. CONCLUSION: Administration of prednisolone to Candida-colonized mice can induce production of the IgG, IgE antibodies against Candida antigens, perhaps through temporal systemic dissemination of Candida from the intestinal tract.     

Hyposensitization with Dermatophagoides pteronyssinus Antigen: Trial in Asthma Induced by House Dust

A double-blind clinical trial of hyposensitization with aqueous extracts of Dermatophagoides pteronyssinus (the house-dust mite) and human skin scales showed a substantial improvement in symptoms in 11 asthmatics allergic to house dust treated with the D. pteronyssinus extract and a reduction in their need for other therapy. Five patients were well for a year but six relapsed. These results contrasted with the generally unfavourable course of the patients treated with the extract of human skin scales. Asthma due to house-dust allergy may be substantially improved by hyposensitization with D. pteronyssinus extract.     

Pilot study of IFN-gamma-induced specific hyposensitization for house dust mites in atopic dermatitis: IFN-gamma-induced immune deviation as a new therapeutic concept for atopic dermatitis

IFN-gamma/IL-4 imbalance is a central immunologic defect which is responsible for increased IgE antibody response in atopic dermatitis. Effects of hyposensitization were controversial in atopic dermatitis. Reversed IFN-gamma/IL-4 balance was induced using IFN-gamma in atopic dermatitis and specific hyposensitization with house dust mites (HDM) was tried in the status of IFN-gamma-induced immune deviation. A total of 58 atopic dermatitis patients who had obvious allergy to HDM were selected in this study. IFN-gamma-induced hyposensitization for HDM was tried in 10 patients. Twenty-two patients received IFN-gamma therapy and six were treated by simple hyposensitization. Twenty were enrolled as control subjects. The clinical severity scores decreased effectively only by IFN-gamma-induced hyposensitization for HDM. Specific hyposensitization for HDM in the status of IFN-gamma-induced immune deviation successfully improved atopic dermatitis. HDM might play a pathogenic role in subpopulation of atopic dermatitis.     

Bacterial flora from bronchoalveolar lavage and occurrence of class IgG and IgA antibodies to Chlamydia pneumoniae in patients with chronic obstructive pulmonary disease (COPD)

Bronchoalveolar lavage taken from 46 patients (ranging in age from 21 to 71 years, mean 50.6 +/- 13.9) was examined for aerobic and anaerobic bacterial flora. Sera taken from 39 of patients as well as sera taken from 25 healthy blood donors of similar age (P = 0.99) were examined to determine IgG and IgA antibodies to C. pneumoniae. Bacterial flora was routinely cultured and determined using ATB computer system (bioMérieux,). IgG and IgA antibodies were tested by the enzyme immunoassays (Labsystems, Finland, Helsinki). Sera containing anti -C. pneumoniae IgG antibodies with titers of 45 EIU or higher and IgA with titers of 12 EIU or higher were considered positive. 143 of aerobic and 74 of anaerobic bacterial strains were cultured. Streptococci group viridans, pneumococci, enteric bacilli, Haemophilus spp., Prevotella spp., Actinomyces spp., Bifidobacterium spp. and Veilonella spp. were most often cultured. 66.6% of patients had IgG or IgA antibodies, in contrast, to the control group in which 60.0% and 44.0% of examined blood donors had IgG and IgA antibodies respectively. COPD patients were more frequently positive for specific anti-C. pneumoniae antibodies than the healthy donors (p = 0.003). The difference in a seropositivity rate of specific IgA and IgG antibodies was significant (p = 0.00002 and p = 0.003 respectively). Bronchoalveolar lavage of patients suffering from COPD can be contaminated with high number of aerobic and anaerobic bacterial species, and immunological status of the patients indicated persistent infection caused by C. pneumoniae more often than in controls.     

Measurement of IgG-blocking antibodies: development and application of a radioimmunoassay.

A radioimmunoassay for measuring blocking antibodies has been developed. We used the ragweed antigen E system to show that the same blocking antibodies (IgG) measured by inhibition of antigen-induced leukocyte histamine release were precipitated in the binding assay (r8 = 0.96, p less than 0.001), thus validating a widely applicable technique for measuring blocking antibodies. Binding of phospholipase-A (Phos-A), the major allergen in honey bee venom, was also shown to correlate significantly with inhibition of histamine release. Hymenoptera (insect) hypersensitivity was used as a model to demonstrate application of the binding assay. Sera obtained from patients undergoing whole body extract therapy contained negligible amounts of specific blocking antibodies. Significantly higher blocking antibody titers to both whole honey bee venom and Phos-A were measured in sera drawn from patients immunized with whole venom. The use of the binding radioimmunoassay should facilitate management of allergic disease processes in which blocking antibodies are thought to be protective.     

Study of the effect of antibodies in the intestinal tract of germ-free baby pigs

The effect of antibodies in the intestinal tract was studied in germ-free baby pigs whose intestinal barrier was closed to macromolecules by the peroral administration of modified cow's milk for the first 72 hours after birth. They were then all contaminated with the pathogenic strainEscherichia coli 055 in amounts of 10⁹ bacterial cells per animal. The controls, which were not given any antibodies, all died within 24 hours. All the experimental animals given 12.5–50ml immune colostrum or serum survived, while of those given 50ml normal serum or colostrum containing natural antibodies reacting with theEscherichia coli test strain, 50% survived. No circulating antibodies were found in the serum of the experimental animals after the administration of serum or colostrum. The antibodies present in colostrum thus appear to protect the newborn organism directly in the intestinal tract, which is the first site of bacterial invasion, as well as after infiltration into the blood stream.     

Increased Levels of IgG Antibodies against Human HSP60 in Patients with Spondyloarthritis

Spondyloarthritis (SpA) comprises a heterogeneous group of inflammatory diseases, with strong association to human leukocyte antigen (HLA)-B27. A triggering bacterial infection has been considered as the cause of SpA, and bacterial heat shock protein (HSP) seems to be a strong T cell antigen. Since bacterial and human HSP60, also named HSPD1, are highly homologous, cross-reactivity has been suggested in disease initiation. In this study, levels of antibodies against bacterial and human HSP60 were analysed in SpA patients and healthy controls, and the association between such antibodies and disease severity in relation to HLA-B27 was evaluated.Serum samples from 82 patients and 50 controls were analysed by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (Ig)G1, IgG2, IgG3 and IgG4 antibodies against human HSP60 and HSP60 from Chlamydia trachomatis, Salmonella enteritidis and Campylobacter jejuni. Disease severity was assessed by the clinical scorings Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI) and Bath Ankylosing Spondylitis Metrology Index (BASMI).Levels of IgG1 and IgG3 antibodies against human HSP60, but not antibodies against bacterial HSP60, were elevated in the SpA group compared with the control group. Association between IgG3 antibodies against human HSP60 and BASMI was shown in HLA-B27⁺ patients. Only weak correlation between antibodies against bacterial and human HSP60 was seen, and there was no indication of cross-reaction.These results suggest that antibodies against human HSP60 is associated with SpA, however, the theory that antibodies against human HSP60 is a specific part of the aetiology, through cross-reaction to bacterial HSP60, cannot be supported by results from this study. We suggest that the association between elevated levels of antibodies against human HSP60 and disease may reflect a general activation of the immune system and an increased expression of human HSP60 in the synovium of patients with SpA.     

Eye muscle antibodies in endocrine exophthalmos. Clinical and serological observations

Serum samples were obtained from 65 patients with endocrine exophthalmos class I-V. In 33/65 patients who were treated either with prednisone or with ciclosporin, blood was sampled before, during and after therapy. Antibodies against eye muscle were determined during the course of immunosuppressive therapy in order to have an objective parameter of the therapeutic effect. To ascertain the specificity of the reaction both eye and abdominal muscles were used as antigens in an ELISA system. Both IgG and IgM antibodies were detected. In 45/65 patients (71%) eye muscle antibodies were positive before starting therapy. Antibodies were mostly detected in patients with active disease. Patients with exophthalmos of recent onset always had IgM antibodies whereas patients with chronic exophthalmos were mostly IgG positive. Patients with relapse showed mostly IgG but also IgG and IgM positivity in 2 cases. In 58% of cases only IgG antibodies were found whereas in 34% both IgG and IgM were detected and in 8% only IgM antibodies. There was no association between antibodies directed against eye muscle and thyroid microsomal and thyroglobulin antibodies or with the state of thyroid function. Furthermore there was no correlation between exophthalmos classes and eye muscle antibody binding activity. The antibody level declined during therapy with prednisone or with ciclosporin but rose again 8-12 weeks after stopping the drug in patients with progressive disease.     

B cell epitopes of the major timothy grass pollen allergen, phl p 1, revealed by gene fragmentation as candidates for immunotherapy.

Group 1 grass pollen allergens are recognized by IgE antibodies of almost 40% of allergic individuals and therefore belong to the most important elicitors of Type I allergy worldwide. We have previously isolated the cDNA coding for the group 1 allergen from timothy grass, Phl p 1, and demonstrated that recombinant Phl p 1 contains most of the B cell as well as T cell epitopes of group 1 allergens from a variety of grass and corn species. Here we determine continuous B cell epitopes of Phl p 1 by gene fragmentation. IgE antibodies of grass pollen allergic patients identified five continuous epitope-containing areas that on an average bound 40% of Phl p 1-specific IgE antibodies and were stably recognized in the course of disease. In contrast to untreated patients, patients undergoing grass pollen immunotherapy started to mount IgG(4) antibodies to the recombinant IgE-defined fragments in the course of immunotherapy. The protective role of these IgG(4) antibodies is demonstrated by observations that 1) increases in rPhl p 1 fragment-specific IgG(4) were in parallel with decreases in Phl p 1-specific IgE, and 2) preincubation of rPhl p 1 with patients sera containing rPhl p 1 fragment-specific IgG(4) blocked histamine release from basophils of an untreated grass pollen allergic patient. We propose to use recombinant Phl p 1 fragments for active immunotherapy in order to induce protective IgG responses against IgE epitopes in grass pollen allergic patients. This concept may be applied for the development of allergy vaccines whenever the primary sequence or structure of an allergen is available.     

Measurement of IgG blocking antibodies by interference in the radioallergosorbent test

We previously found that sera of patients immunized with ragweed pollen extract contained a factor that interfered with the binding of IgE antibodies to solid-phase allergens in the radioallergosorbent test (RAST). We now describe an assay, RAST interference, to measure this factor, and we present evidence that the factor is IgG blocking antibody. Sera from immunized allergic patients were heated at 56 degrees C for 4 hr to destroy heat-labile Fc determinants on IgE and were tested for their ability to prevent binding of additional IgE antibody to solid-phase allergens in the RAST. Eight of 10 sera from allergic immunized patients gave RAST interference dose-response curves that did not differ from the arbitrary standard. The factor causing interference showed specificity for the immunizing antigen, was heat-stable, eluted from Sephadex G-200 in the 7S peak, was present only in sera of immunized patients, and rose after initiation of immunization. These results indicated that RAST interference can be used to measure IgG blocking antibodies with the same reagents employed for the measurement of IgE antibodies, provided the antiserum to IgE is specific for the heat-labile FC determinants on IgE.     

A quantitative analysis of cedar pollen-specific immunoglobulins in nasal lavage supported the local production of specific IgE, not of specific IgG

Many studies have proved the relevance of local immune responses, rather than systemic immunity, to the pathogenesis of allergic rhinitis. Indeed, allergen-specific B lymphocyte undergoes class switching to IgE in situ. However, the relative contribution of in situ production to the amount in serum is still ambiguous. Here, a quantitative comparison of the local concentration of allergen-specific IgE with the systemic concentration was explored for the estimation. Among seasonal rhinitis patients, total and Japanese cedar pollen (JCP)-specific IgE, IgA and IgG antibodies were quantified in nasal lavage fluid (NLF) and serum with the time-resolved fluorescence immunosorbent assay. Although the total amounts of IgE and IgG classes in the NLF, which were apparently passive discharge from the mucosal tissue, were smaller and variable, the relative proportions of JCP-specific antibodies could be quantitatively compared between NLF and serum or between subjects. The proportions of specific IgE in the NLF were remarkably higher than in serum (average 13.2-fold) in most subjects, which strongly supported the predominant in situ production of the specific IgE and subsequent dilutions in the systemic circulations. Similar but smaller values were obtained for IgA (average 3.7-fold). In contrast, the specific proportions of IgG in the NLF were surprisingly consistent with serum (average 1.0-fold), suggesting that the specific IgG was mostly produced in the downstream lymphoid organs. The local productions of specific IgE would encourage the topical therapies and the usage of the NLF for the diagnosis of allergic rhinitis.