[Interferon-alpha as treatment option in severe persistent uncontrolled bronchial asthma: an open label study]. / Interferon-alpha als Therapieoption bei der Behandlung des schwergradig persistierenden, unkontrollierten Asthma bronchiale: Eine offene Studie.

BACKGROUND: The purpose of this study was to evaluate the long-term safety and therapeutic effects of IFN-alpha in patients with severe persistent uncontrolled asthma on long-term oral glucocorticoid (GC) treatment. PATIENTS AND METHODS: The study included 16 patients (2 male, 14 female; age 39 years [range: 24 - 63]) with severe persistent asthma. Diagnosis and severity classification of asthma were established according to the guidelines of the "Deutsche Atemwegsliga". Eight patients stopped the therapy within 7 months due to side effects (n = 3), costs not covered by health insurance (n = 2), non-compliance (n = 2), and change of residence (n = 1). 8 patients (8 female, age 49 years [range: 35 - 68], duration of disease 16 years [range: 5 - 24]) were treated for at least 12 months with IFN-alpha (9 microg) 3 times/week. All patients were on oral glucocorticoids (GCs) for more than 5 years (average dose 17.5 [range: 5.0 - 64.0] mg/d). Clinical signs, lung function, need for reliever medication, number of emergency visits and hospitalisations and diary were assessed prior to and after 12 months of treatment. Data are given as percent of normal or median [range]. RESULTS: IFN-alpha improved lung function after 12 months: FEV1 64 vs. 75 %; FEV1/IVC 76 vs. 89 %; RV 153 % vs. 129 %; Rtot 193 vs. 111 % and morning PEF by 50 - 190 L/min. IFN-alpha also significantly reduced the use of reliever medication (10 [2 - 20] vs. 1 [0 - 3] puffs/d), nocturnal awakening (11 [4 - 30] vs. 1 [0 - 5]/month), emergency visits (7 [2 - 15] vs. 0 [0 - 5]/month) and hospitalisations (4 [1 - 8] vs. 0 [0 - 5]/year). In 5 patients the asthma attacks and nightly disturbances disappeared completely. The improvements were achieved despite a tapering of the oral GCs in all patients from 17.5 (5.0 - 64.0) to 2 (0 - 16) mg/d. In 5 patients GC treatment could be discontinued. The number of blood eosinophils decreased from 0.46 to 0.28 Gpt/L. Adverse events were transient and usually decreased within 3 to 4 weeks. Two patients developed an autoimmune thyreoiditis. CONCLUSION: In severe persistent, uncontrolled, and GC-dependent asthma, treatment with IFN-alpha leads to sustained clinical improvement and allows the reduction or discontinuation of oral GCs. Severe side effects may occur in isolated cases.

Additional benefit of a third year of specific grass pollen allergoid immunotherapy in patients with seasonal allergic rhinitis.

BACKGROUND: Aluminium-adsorbed six grass pollen allergoid therapy for 2 years was found to be efficacious and safe in patients with hay fever and seasonal asthma. Using high-dose, hypoallergenic allergen products (allergoids) enables a short-term pre-seasonal treatment regimen. However, it is not known whether further treatment beyond 2 yrs had any additional benefit. METHODS: Following an initial 2-year randomized, double-blind, multi-centre, placebo-controlled clinical trial in 154 grass pollen-allergic patients, an additional short course of specific immunotherapy with the high-dose, hypoallergenic grass pollen preparation Allergovit was performed in 61 patients of the active treatment group during the 3rd open-label treatment year. RESULTS: Further treatment of patients with the Allergovit 6-grass pollen preparation resulted in a further reduction of symptom medication score and improved quality of life in comparison to the first and second treatment year. Changes in allergen-specific IgG4 levels supported these results. CONCLUSIONS: Pre-seasonal short-term immunotherapy with the high-dose, hypoallergenic allergen preparation Allergovit has been shown to be efficacious and safe. A course of three years of 6-grass pollen SIT further improves allergic symptoms, quality of life and reduces the need for anti-allergic medication.

Further characterization of IgE-binding antigens from guinea pig hair as new members of the lipocalin family.

BACKGROUND: Guinea pigs are important sources of inhalant allergens in home and working environments. However, little is known about the molecular characteristics and the relevant epitopes of guinea pig allergens. Recently, several allergens have been identified in hair extract and urine, and the major allergen Cav p 1 (20 kDa) has been characterized. OBJECTIVE: The aim of the present study was to isolate and to characterize a further major allergen from guinea pig hair with 17 kDa. METHODS: Guinea pig hair extract was fractionated using anion exchange chromatography and reverse-phase high performance liquid chromatography. Analyses were carried out by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, 2D-PAGE, immunoblotting, immunoblot inhibition, glycoprotein detection, and N-terminal amino acid sequencing. RESULTS: The nonglycosylated 17 kDa allergen, which was named Cav p 2, was purified to homogeneity. On the basis its 15 N-terminal residues, there was 69% identity with a sequence of Bos d 2, an allergenic protein from cow dander belonging to the lipocalin family. The 2D-immunoblotting analyses of guinea pig hair extract demonstrated that Cav p 2 and Cav p 1, contained several isoforms with pI values ranging from 3.6 to 5.3. The 2D-immunoblot inhibition disclosed cross-reactive IgE epitopes on the allergens Cav p 2 and Cav p 1. Furthermore, Cav p 1 can form both monomers (20 kDa) and dimers (40-42 kDa). CONCLUSION: These studies provide important information on the isoallergen character of two relevant guinea pig allergens Cav p 1 and Cav p 2 as well as on their cross-reactive properties.

Purification and partial characterization of the major allergen, Cav p 1, from guinea pig Cavia porcellus.

BACKGROUND: Allergic reaction to guinea pig has been recognized as a problem in domestic settings and work environments for many years. Until recently, limited information was available on the properties of guinea pig allergen(s). In this study the major allergen Cav p 1 was characterized and the N-terminal amino-acid sequence was determined. METHODS AND RESULTS: Sera from 40 patients with IgE-mediated allergy to guinea pigs were investigated by means of immunoblotting using extracts prepared from guinea pig hair and urine. Three major allergens were identified within both sources with molecular weights (MW) of 8 kDa, 17 kDa and 20 kDa, respectively. From aqueous hair extracts the 20 kDa allergen (Cav p 1) was purified to homogeneity by anion exchange chromatography and reverse-phase HPLC and the N-terminal amino-acid sequence was determined. On the basis of the 15 residues, 57% identity was obtained from computer search with a sub-sequence of MUP (major urinary protein), a member of the lipocalin superfamily. Allergenic relationships among guinea pig allergens derived from various sources (hair and urine) or different animal species (mouse, rat, cat) were studied by ELISA inhibition assays. Neither urine of mouse, rat and cat, nor hair extracts of rat and cat produced appreciable inhibitions in guinea pig ELISA. CONCLUSION: Although the physicochemical characteristics of isolated Cav p 1 are very similar to those for other rodent allergens and furthermore partial sequence identity with Mus m 1 was found, it is clearly shown here to be an immunologically independent major allergen.

[Guidelines for conducting provocation tests with allergens. German Society of Allergology and Clinical Immunology and German Society for Immunology]. / Leitlinie für die Durchführung bronchialer Provokationstests mit Allergenen - Deutsche Gesellschaft für Allergologie und klinische Immunologie und Deutsche Gesellschaft für Pneumologie -

Guidelines for bronchial allergen provocation tests, first published in 1984, are updated by this paper. This version has been compiled in repeated meetings of a panel of experts after evaluation of the publications available. Precise statements regarding indications, contraindications, after safety measures are given. Three evidence-based protocols are submitted in detail.

[Asthma therapy for adults]. / Therapie des Asthma bronchiale im Erwachsenenalter.

The goal of asthma management is to achieve control of the condition. This essentially requires environmental control measures (allergen avoidance) and patient training and education. Drug treatment comprises anti-inflammatory (corticosteroids), and bronchodilatory controller therapy (long-acting beta 2-sympathomimetics, leukotriene receptor antagonists, retarded theophylline) as well as bronchodilatory medication as required (short-acting beta 2-sympathomimetics). The number and frequency of pharmacologic therapy relates to the severity of the clinical presentation. The combination of certain controller drugs (corticosteroids with long-acting beta 2-agonists, corticosteroids with leukotriene receptor antagonists, and beta 2-agonists with leukotriene receptor antagonists) yields a synergistic therapeutic effect as well as a compliance advantage.

Further characterization of IgE-binding antigens in kiwi, with particular emphasis on glycoprotein allergens.

Fruit allergy is frequently associated with birch pollinosis. The aim of this study was to investigate which kiwi allergens were involved in subjects allergic to fruit alone and in patients allergic to both fruit and birch pollen. Sera of nine patients (five with both kiwi and birch pollen allergy and four with isolated kiwi allergy) were studied by immunoblot of kiwi extract. Eight of the nine sera reacted with the 30 kDa protein. Furthermore, IgE-binding proteins were seen at approximately 23 kDa (detected by five sera), 43 kDa and 80 kDa (four sera), and > 80 kDa (two sera). One serum showed no IgE binding to any kiwi allergen. The 30 kDa is the major allergen in kiwi and was purified by anion-exchange chromatography and characterized by isoelectrofocusing and amino acid sequencing. The comparison of its partial amino acid sequence with data from the Swiss Protein Bank revealed that this protein is actinidine. The carbohydrate structures in kiwi and birch pollen extracts were investigated with seven lectins. On kiwi blot, Aleuria aurantia agglutinin showed strong reactivity (indicating fucose residues) to the components of 35 to 92 kDa, while concanavalin A (indicating mannose, glucose or N-acetylglucosamine residues) showed weak binding at 67 kDa. In contrast, strong binding of Galanthus nivalis agglutinin (indicating mannose residues) and concanavalin A was found on birch pollen blots. The presence of IgE against carbohydrate structures was determined by means of enzyme-linked immunosorbent assay (ELISA) after periodate treatment of kiwi extract. The IgE binding was reduced by periodate treatment of kiwi coated microtiter plates, but not by sera reacting exclusively with the 30 kDa protein. Furthermore, selected sera were treated with proteinase K-digested kiwi and birch pollen extracts as the sources of crossreactive carbohydrate determinants. In accordance with the results of sodium periodate treatment, significant levels of anti-cross-reactive carbohydrate determinant IgE were found in sera from patients allergic to both kiwi and birch pollen. Our results show that the major allergen for kiwi allergy is the 30 kDa protein and additionally that the cross-reaction between kiwi and birch pollen allergy is mainly due to carbohydrate moieties.

Effect of hyposensitization for tree pollinosis on associated apple allergy.

Twenty patients suffering from birch pollen allergy received two or three courses of immunotherapy in successive years. In 9 patients, the fruit allergy improved; 4 patients reported no improvement. In 3 patients, the fruit allergy developed after beginning the immunotherapy. At the end of the 3 years, 16 of these patients were allergic to fruit, 13 of them to apple. After each preseasonal course of immunotherapy with tree pollen extract, a temporal and parallel increase in the titers of IgE antibodies to birch pollen allergens and apple allergens were observed. In contrast, only the titers of birch pollen allergen specific IgG and IgG4 increased, whereas apple allergen specific IgG and IgG4 did not, or only very slightly. In Western blot studies, IgG4 antibodies bound to more components of apple extract than birch pollen extract. On the average, IgG4 antibodies recognize more components of apple and birch pollen extracts than do IgE antibodies. In histamine release studies, the sensitivity of washed leukocytes to birch pollen extract decreased significantly during the observation time. However, the difference between apple extract-induced histamine release before and after immunotherapy was not significant. None of the immunological parameters investigated here correlate well with severity or prognosis of the fruit (apple) allergy. The clinical improvement of pollinosis was associated with a rise in birch pollen specific IgG4 antibody titers and a decrease of allergen-induced histamine liberation. Beside improvement of the fruit allergy in 56% of the cases, the courses of apple specific IgE and IgG4 antibody titers seem to indicate a slight sensitization against apple allergens.

Investigation of the stability of apple allergen extracts.

To determine optimal conditions for allergen preservation, we investigated the influence of different stabilizing additives and of storage temperature on the allergen activity of apple protein preparations, obtained by extraction in phosphate buffer or by precipitation in diacetone alcohol and resolubilization in phosphate buffer in the presence or absence of enzyme inhibitors. For this purpose, the extracts were stored for 6 months either in frozen state at -20 degrees C or in lyophilized state at -20 degrees C, 4 degrees C, or room temperature and were characterized by SDS-PAGE, immunoblot, ELISA inhibition, and prick test. The highest stability revealed the extracts that were prepared by precipitation in the organic solvent in the presence of enzyme inhibitors, lyophilized, and stored at -20 degrees C. For storage of extract solutions at 4 degrees C, PBS/glycerol and cysteine/sodium citrate/glycerol were found to be the most effective stabilizing additives.

Optimization of apple allergen preparation for in vivo and in vitro diagnostics.

The aim of our investigation was to obtain a well-characterized active apple extract suitable for both in vivo and in vitro diagnostics by a technically simple method. For this purpose, apple extracts were prepared by homogenization in potassium phosphate buffer or by precipitation in organic solvents and resolubilization in potassium phosphate buffer in the presence or in the absence of enzyme inhibitors. These extracts were comparatively investigated by means of SDS-PAGE, two-dimensional electrophoresis, immunoblotting, RAST inhibition, and prick test. The in vitro investigations indicated that extracts prepared by precipitation in organic solvents (diacetone alcohol) at -20 degrees C have a higher allergen activity than those prepared by extraction in aqueous solutions. From the in vivo tests (prick test), it was concluded that application of inhibitors of cytoplasmic enzymes (phenol oxidases, peroxidases, proteases) already during extraction is an essential precondition for active prick test solutions. Correspondingly, the extract obtained by solvent precipitation in the presence of enzyme inhibitors appeared to be most suitable for clinical application.

IgE rheumatoid factor. Occurrence and diagnostic importance in comparison with IgM rheumatoid factor and circulating immune complexes.

The sera of 80 patients suffering from rheumatoid arthritis (RA)--30 of them with extraarticular manifestations (EAM) and 50 patients with articular disease only--of 25 patients with other joint diseases and of 30 normal healthy subjects, were analyzed for the presence of 1) IgE rheumatoid factors (IgE RF) by means of a solid phase radioimmunoassay and an ELISA, 2) IgM rheumatoid factors by using solid phase radioimmune techniques, and 3) circulating immune complexes (CIC) with the C1q binding test (C1q BT) and the solid phase conglutinin binding test (SPCBT). The best technique to discriminate RA patients with EAM from RA patients without EAM and patients with other articular diseases was the determination of IgE RF (73.3%, 38.0%, 0%, resp.) compared to IgM RF (86.7%, 78.0%, and 56.0%) and CIC (C1q BT: 80.0%, 66.0% and 45.0%; SPCBT: 46.7%, 22.0%, and 20.0%). The results suggest a certain role of IgE RF for diagnosis and a possible development of extraarticular manifestations in rheumatoid arthritis.

[Experiences with desensitization of early spring pollen allergy using 2 tree pollen extracts]. / Erfahrungen bei der Desensibilisierung der Frühblüher-Allergie mit 2 verschiedenen Baumpollen-Extrakten.

Forty patients with tree pollen-induced allergy (rhinitis, conjunctivitis or combination from both with asthma) were hyposensitized with an extract from isolated birch pollen or a pollen mixture (hazel-, alder-, oak- and hornbeam pollen). The distribution of the different extracts was random. After 3-year treatment, no statistical differences were found between the two groups, especially regarding clinical effectiveness, duration of symptoms or symptom score. In conclusion, we recommend a hyposensitization with birch pollen extract in tree pollen-induced allergy. If evidence is present of sensitization against other tree pollens, especially hazel or alder, or in cases without therapeutic effect after two years of hyposensitization with birch pollen, an attempt with a tree pollen mixture extract is indicated.

[The quantitative determination of the IgM rheumatoid factor using a solid-phase radioimmunoassay. Comparison with the agglutination test and the radioimmuno-polyethyleneglycol precipitation test]. / Die quantitative Bestimmung des IgM-Rheumafaktors mittels eines Festphasen-Radioimmuntests Vergleich mit Agglutinationstests und dem Radioimmun-Polyethylenglykol-Präzipitationstest.

A solid-phase radioimmunoassay capable of detecting nanogram quantities of human IgM rheumatoid factor using a monoclonal anti-mu-chain antibody is described. Human IgG did not interfere with the detection of IgM RF by this method. The small nonspecific binding of nonRF IgM to the human IgG coated tubes utilized in the assay must be corrected for by assaying samples in parallel bovine serum albumin coated control tubes only in cases of deviation of IgM from normal range. 69 coded and randomly arranged sera from patients with rheumatoid arthritis (RA), nonrheumatic joint diseases and healthy adult control subjects were investigated by this method, agglutination techniques as well as RIPEGA. A good correlation between solid-phase radioimmunoassay and agglutination techniques was found. Patients with seropositive RA had significantly higher concentrations of IgM RF than seronegative RA patients or control subjects (mean +/- 1 SD = 133,3 +/- 187,2 micrograms/ml versus 4,7 +/- 6,5 micrograms/ml and 2,2 +/- 4,0 micrograms/ml; resp.).

[Etiopathogenetic aspects of psoriatic arthritis with special reference to immunologic factors]. / Atiopathogenetische Aspekte der Arthritis psoriatica unter besonderer Berücksichtigung immunologischer Faktoren.

On 21 patients with psoriasis and 22 with psoriatric arthritis examinations for the recognition of the immune regulation were performed and the results were compared with those of healthy control persons and patients with rheumatoid arthritis. The psoriatric arthritis shows humoral as well as cellular disturbances of immune regulations which in the group comparison with the psoriasis and the rheumatoid arthritis partly have communities, but partly also have differences. With regard to size and distinction of the disturbances the patients with rheumatoid arthritis have the highest degree of severity. With the help of the techniques used by us systemic cellular disturbances of the immune regulations could not be proved in the psoriasis. The psoriatic arthritis to a certain extent occupies a medium position. But a clear coordination of these disturbances into the pathogenetic process is not yet possible.

[Relationship between clinical parameters and labor (author's transl)]. / Beziehungen zwischen klinischen Parmetern und Wehentätigkeit.

In 200 cases of childbirth supervised by intra-amniotic tocometry the relations between various clinical parameters such as age, parity, gestosis, rise of temperature, birth weight, small circumference of the head, duration of the cervical dilatation and the expulsion period, and the exactly defined motility of the uterus were investigated during different phases. The results confirmed the very limited influence of selected clinical phenomena upon the multifactiorially regulated total complex of labour.