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H Ipsen

Publications and source records attributed to H Ipsen.

43 records · Page 3Linked to original sources

Immunochemical characterization of reference alder (Alnus glutinosa) and hazel (Corylus avellana) pollen extracts and the partial immunochemical identity between the major allergens of alder, birch and hazel pollens.

Dialysed extracts of alder (Alnus glutinosa) and hazel (Corylus avellana) pollens were characterized by immunochemical methods. The alder pollen extract contained at least 33 distinct antigens of which one, antigen 5, was classified as a major allergen, three, antigens 4, 10, and 17, were classified as intermediate allergens and two, antigens 19 and 23, were classified as minor allergens. The molecular weight and pI of the major allergen were determined to be 19,000 D and 5.2, respectively. The hazel pollen extract contained at least 40 distinct antigens, two of which were classified as major allergens, antigens 8 and 13, three as intermediate allergens, antigens 12, 17, and 26, and seven as minor allergens, antigens 6, 11, 16, 18, 29, 39, and 40. The molecular weights and pI's of the major allergens of hazel pollen were determined to be: antigen 8; Mw = 12,000 D, pI = 5.1 and less than 3.5, antigen 13; Mw = 26,000 D, pI = 5.1. Further, crossed line immunoelectrophoresis and tandem crossed immunoelectrophoresis of alder, birch and hazel strongly indicate that there exists an immunochemical partial identity between the major allergens (antigens 5 (alder), 23 (birch), and 13 (hazel)) from these tree pollens.

Allergens↗

Immunochemical characterization of antigens of Parietaria judaica pollen. Identification of allergens by means of crossed radio immunoelectrophoresis.

A crude extract of Parietaria judaica pollen was obtained by means of extraction, centrifugation and dialysis, and studied by means of quantitative immunoelectrophoresis. Crossed immunoelectrophoresis, using a high-titer purified rabbit antibody fraction, showed that the pollen extract contained at least 26 antigens of which 18 moved towards the anode, 6 moved towards the cathode and 1 moved both towards the anode and the cathode. The allergens in the extract were identified by means of crossed radio immunoelectrophoresis. Nine of the 26 antigens were able to bind specific human IgE to their corresponding immunoprecipitates, and 4 of these antigens can be classified as major allergens. The apparent molecular weights of 17 antigens were determined by a combination of size exclusion chromatography and immunochemical analysis. Ten antigens had Kav values corresponding to molecular weights in the range of 10-40 kilodaltons, 1 antigen possessed an apparent molecular weight of less than 10 kilodaltons, and six antigens had molecular weights above 40 kilodaltons. Most of the antigens, as analysed by column isoelectric focusing, had pI values between 4 and 7. Crossed line immunoelectrophoresis using an extract of Parietaria officinalis shows that the antigens of this extract exhibit a high degree of identity with those of P. judaica.

Adult↗

Isolation and immunochemical characterization of the major allergen of birch pollen (Betula verrucosa).

The classification of some of the extractable birch pollen antigens as allergens was established by crossed radioimmunoelectrophoresis (CRIE). In CRIE the major allergen (antigen 23) exhibited the strongest "radiostaining," and only a few other components of birch pollen extract were visibly radiostained. The major allergen and a preparation containing mainly the minor allergens, antigens 25 and 19, were isolated from a crude aqueous birch pollen extract by a combination of anion-exchange, size-exclusion, and chelate chromatography. Antigen 23 was purified to near homogeneity. The molecular weights and the pIs of antigens 23, 25, and 19 were determined to be 17,000 daltons, pI 5.25 (5.5, 5.0); 25,000 daltons, pI 5.0 (4.9, 5.4); and 29,000 daltons, pI 6.2 (5.4), respectively. The classification of antigen 23 as the major allergen in birch pollen was supported by results of RAST inhibition experiments, RAST screening, and skin prick testing.

Allergens↗

Specific IgE response toward allergenic molecules during perennial hyposensitization: a three-year prospective double-blind study.

Allergenic extracts are multicomponent systems containing a number of potentially allergenic molecules. In a 3 yr prospective double-blind study, sera from 40 patients hyposensitized with either a two-component timothy extract or a 20-component timothy extract were evaluated by means of crossed radioimmunoelectrophoresis. The patients exhibited specific IgE response toward individual allergenic molecules, and IgE with new specificities did not develop after hyposensitization. A high degree of clinical protection could generally be obtained on treatment with the 20-component timothy extract, but this was true only with the two-component timothy extract containing the allergenic molecules that matched the individual IgE response.

Adolescent↗

Differential recognition of recombinant Phl p 5 isoallergens by Phl p 5-specific T cells.

BACKGROUND: At present, recombinant allergens are considered for the diagnosis and treatment of atopic allergies. To evaluate the theoretical impact of isoallergen variation on the selection of isoallergens for specific allergy vaccination, we characterized the T-cell response of allergic patients to the Phleum pratense major allergen, Phl p 5, and five of its recombinant isoallergens. METHODS: Phl p 5-specific T cell lines (TCLs) were isolated from the peripheral blood of 3 allergic rhinitis patients, and their reactivity patterns were studied in detail. RESULTS: The TCLs were highly crossreactive with related grasses. The crossreactivity with Poa pratensis was more extensive than with Lolium perenne, directly reflecting the sequence identity between Phl p 5, Poa p 5, and Lol p 5. The TCLs produced IFN-gamma and IL-4 simultaneously, resembling a Th0-like cytokine production profile. Interestingly, when T cell clones were tested with natural Phl p 5 and five rPhl p 5 isoallergens, a differential recognition pattern was found. One of the TCLs exclusively reacted with Phl p 5b, another reacted with all isoforms tested, and the third reacted strongly with native purified Phl p 5, but only weakly with all five recombinant isoallergens. CONCLUSION: These results indicate that Phl p 5-specific T cells are highly heterogeneous, and that they differentially recognize isoallergens. This indicates that when recombinant Phl p 5 is considered for allergy vaccination, a mixture of isoallergens representing the different isoallergen groups may clinically prove to be more effective than single isoallergens.

Allergens↗

The significance of isoallergenic variations in present and future specific immunotherapy.

The isoallergenic variation of the tree pollen major allergens has been studied by 2D gel electrophoresis, and by analysis of several recombinant clones. The studies have included both antibody-based and T cell stimulation assays. Bet v 1, the major allergen of birch, forms at least 24 spots when conventional extracts are analyzed by 2D gel electrophoresis. Comparison of Bet v 1-encoding DNA sequences reveals a considerable number of amino acid substitutions. This sequence variation can theoretically account for the number of spots observed in 2D gels. Whereas pools of serum from allergic individuals and monospecific antibodies raised in rabbits bind to most if not all spots in 2D gels, analyses of individual serum and/or murine monoclonal antibodies show individual patterns of reactivity with various subsets of spots. These observations point to a model in which amino acid substitutions induce local perturbations of the allergen surface, causing differences in epitope structure. Furthermore, analysis of pollen from individual trees shows that each tree produces individual subsets of Bet v 1 spots. When analyzed in stimulation assays, T cell clones also display differences in reactivity to different isoallergens. In conclusion, we have shown that Bet v 1 is heterogeneous, and that individual trees produce various subsets of isoallergens which display differences in reactivity both towards antibodies and T cells. A careful selection of isoform may therefore be of major importance if recombinant allergens or synthetic peptides are to be used for conventional immunotherapy.

Allergens↗