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Papers by Helene Henmar

Research paper thumbnail of High consistency in allergen composition of SQ house dust mite (HDM) tablet for sublingual immunotherapy (SLIT)

Journal of Allergy and Clinical Immunology, Feb 1, 2018

RATIONALE: Labyrithine nasal passages, the narrow nasal valve and complex static and dynamic aero... more RATIONALE: Labyrithine nasal passages, the narrow nasal valve and complex static and dynamic aerodynamics limit traditional intranasal steroids' (INS) ability to effectively deliver medication to superior/ posterior nasal regions. Exhalation delivery systems (EDS) exploit unique characteristics of nasal anatomy and aerodynamics to overcome these limitations and achieve superior/posterior drug delivery. We review published human in vivo gamma-scintigraphy deposition data for conventional INS (C-INS; e.g., Flonase/Nasonex), HFA-based pMDI's (e.g., QNASL/Zetonna), and EDS systems for liquids and powders. METHODS: Four recent gamma-deposition studies comparing different technologies for nasal delivery of topical steroids were included: (1) Conventional INS sprays (Flonase and Nasonex) versus pMDI (QNASL), (2) C-INS (Nasonex) versus pMDI (Zetonna), (3) C-INS versus EDSliquid, (4) C-INS versus EDS-powder and EDS-liquid. Data on regional deposition and clearance was compared. RESULTS: Qualitative deposition differences were large, though variability in segmentation methods prevents quantitative comparisons. In all studies, C-INS consistently deposit primarily anteriorly (in the valve region) with clearance along the nasal floor and little deposition in superior/posterior regions. Both pMDIs (QNASL/Zetonna) show a stationary ''hotspot'' in the non-ciliated vestibule, little delivery to superior/ posterior regions, and minimal clearance. EDS (liquid or powder) produce less deposition in the valve area and broad deposition to superior/posterior segments with a different clearance pattern. CONCLUSIONS: Human imaging data demonstrate poor drug deposition in superior/posterior sites with conventional nasal sprays, and greatly increased deposition throughout upper/posterior nasal passages with an EDS. In CRS, upper/posterior sites, including the middle and upper meatuses (ostiomeatal complex) where sinuses ventilate/drain and polyps originate, are the primary target for treatment.

Research paper thumbnail of Diverse and highly cross‐reactive T‐cell responses in ragweed allergic patients independent of geographical region

Allergy, 2019

Article type : Original Article: Basic and Translational Allergy Immunology Diverse and highly cr... more Article type : Original Article: Basic and Translational Allergy Immunology Diverse and highly cross-reactive T cell responses in ragweed allergic patients independent of geographical region

Research paper thumbnail of Variation of trypsin inhibitor and crude protein in chikpea cultivars

Food Chemistry, 1996

Data on the energy content of food products presented in Polish food composition tables is based ... more Data on the energy content of food products presented in Polish food composition tables is based on calculations including energy coefficients of proteins, fats and carbohydrates. According to present practice the total sum of carbohydrates is taken into account in these calculations without allowance for dietary fibre. As a consequence results of these calculations are overestimated. The same applies to the energy content of diets assessed with the use of Polish food composition tables. Following Eurofoods recommendations the energy content of foods was revised by taking into consideration available carbohydrates instead of total carbohydrates. This revision resulted in the reductions of the energy content of diets as well. While the energy supply per capita in Poland calculated with the use of the former methodology was 3370 kcal/day, an allowance for dietary fibre reduced this value to 3240 kcal, i.e. by approx. 4%. The extent of the reductions in the energy content of the diets of various population groups varied, depending on the importance of foods rich in dietary fibre present in these diets.

Research paper thumbnail of Contents Vol. 144, 2007

International Archives of Allergy and Immunology, 2007

Access to full text and tables of contents, including tentative ones for forthcoming issues: www.... more Access to full text and tables of contents, including tentative ones for forthcoming issues: www.karger.com/iaa_issues 79 Endogenous Interleukin-10 Produced by Antigen-Irrelevant Cells Promotes the Development of Experimental Murine Allergic Conjunctivitis

Research paper thumbnail of Subject Index Vol. 144, 2007

International Archives of Allergy and Immunology, 2007

Research paper thumbnail of Consistent composition and potency of freeze dried tablets for sublingual immunotherapy

Journal of Allergy and Clinical Immunology

Research paper thumbnail of Fractionation of Source Materials Leads to a High Reproducibility of the SQ House Dust Mite SLIT-Tablets

International archives of allergy and immunology, 2016

The production of house dust mite (HDM) allergen products for allergy immunotherapy has tradition... more The production of house dust mite (HDM) allergen products for allergy immunotherapy has traditionally been based on purified mite bodies or whole-mite culture, which are quite different source materials with a limited possibility for adjusting the chemical composition. The SQ HDM SLIT-tablet is a fast-dissolving pharmaceutical formulation that has been developed for sublingual immunotherapy (SLIT) of HDM respiratory allergic disease. The objective of the present study was to establish a process for the production of drug substances for the SQ HDM SLIT-tablet offering a high reproducibility and independent control of the major allergens. Process controls were documented in a comprehensive process parameter qualification. The analyses comprised composition by crossed immunoelectrophoresis, protein content by BCA, total IgE binding potency by Centaur assay, quantitative major allergen determination by radial immunodiffusion and ELISA, and the ranking of emPAI scores generated by mass s...

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of an Intact Allergen Extract (Alutard SQ®) and 4 Commercially Available Allergoid Products for Birch Pollen Immunotherapy

Data Revues 00916749 V117i2ss S009167490503592x, Aug 16, 2011

Background: When breastfeeding is impossible or insufficient, the use of cow's milk-based hypoall... more Background: When breastfeeding is impossible or insufficient, the use of cow's milk-based hypoallergenic infant formulas is an option for infants suffering from or at risk of developing cow's milk allergy. As the Camelidae family has a large evolutionary distance to the Bovidae family and as camel milk differs from cow's milk protein composition, there is a growing interest in investigating the suitability of camel milk as an alternative to cow's milk-based hypoallergenic infant formulas. Methods: The aim of the study was to compare the allergenicity and immunogenicity of camel and cow's milk as well as investigating their cross-reactivity using a Brown Norway rat model. Rats were immunised intraperitoneally with one of four products: camel milk, cow's milk, cow's milk casein or cow's milk whey fraction. Immunogenicity, sensitising capacity, antibody avidity and cross-reactivity were evaluated by means of different ELISAs. The eliciting capacity was evaluated by an ear swelling test. Results: Camel and cow's milk showed similarity in their inherent immunogenicity, sensitising and eliciting capacity. Results show that there was a lower cross-reactivity between caseins than between whey proteins from camel and cow's milk. Conclusions: The study showed that camel and cow's milk have a low cross-reactivity, indicating a low protein similarity. Results demonstrate that camel milk could be a promising alternative to cow's milk-based hypoallergenic infant formulas.

Research paper thumbnail of Meaurement of trypsin inhibitor in chickpea variants using monoclonal antibodies; comparison with enzymic assay

Research paper thumbnail of Die klinische Wirksamkeit und immunologische Antwort einer SQ-standardisierten Gras-Allergie-Immuntherapie- Tablette (AIT) ist nach einer Behandlungssaison vergleichbar zwischen Kindern und Erwachsenen – Post-hoc- Analyse zwei randomisierter Studien

Research paper thumbnail of Sublingual Immunotherapy with Fast-Dissolving Grass Tablets Induces Comparable IgG4 Antibody and IgE-blocking Responses in Children and Adults

Journal of Allergy and Clinical Immunology, 2009

The benefits of treating multi-sensitized allergic rhinitis (AR) patients with a single allergen ... more The benefits of treating multi-sensitized allergic rhinitis (AR) patients with a single allergen immunotherapy tablet (AIT) have not been fully elucidated. We studied the effect of treating mono-and multi-sensitized AR patients, in whom symptoms were largely confined to the grass pollen season, with grass AIT. METHODS: Post-hoc analysis of data from 568 patients who participated in yr 1 of a 3-yr placebo-controlled trial evaluating the efficacy of grass AIT (75,000 SQ-T/2,800 BAU, Phleum-pratense, ALK-Abello´). Daily treatment started 16 weeks prior to grass-pollen season and continued throughout the season. Responses were analysed in patients sensitized to: a) grass only (mono-sensitized); b) grass 1 tree 1 any other allergen (tree multisensitized); and c) grass 1 any allergen except tree allergens (non-tree multi-sensitized). Sensitivity was determined by skin-prick testing. Daily symptom (runny, blocked, itchy nose, sneezing, gritty, red, itchy, watery eyes; 0 5 none, 1 5 slight, 2 5 moderate, 3 5 severe symptoms) and medication scores were compared. RESULTS: 161 patients were mono-sensitized, 191 tree multi-sensitized, and 216 non-tree multi-sensitized. Treatment with grass AIT was found to reduce symptom and medication scores in mono-and multi-sensitized patients regardless of presence or absence of tree-related sensitivities. Compared to placebo, grass AIT reduced median daily symptoms by 31%, 44%, and 30% (p < 0.0001) as well as median daily medication scores by 49%, 27%, and 60% (p < 0.0001) in mono-, tree multi-sensitized, and non-tree multi-sensitized patients, respectively. CONCLUSIONS: Treatment with a single-grass-allergen tablet yielded significant symptom and medication reductions in AR patients mono-or multi-sensitized to inhalant allergens regardless of the existence/absence of sensitivities to tree allergens.

Research paper thumbnail of The allergen-specific IgE reactivity pattern of Chinese house dust mite allergic patients

World Allergy Organization Journal, 2007

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of Three Intact Allergen Vaccines with Four Allergoid Vaccines for Subcutaneous Grass Pollen Immunotherapy

Journal of Allergy and Clinical Immunology, 2008

Research paper thumbnail of Mapping of Der p 2 antibody binding epitopes by site directed mutagenesis

Journal of Allergy and Clinical Immunology, 2004

MethodsrDer p 2 was subjected to site directed mutagenesis at six selected surface positions (K6,... more MethodsrDer p 2 was subjected to site directed mutagenesis at six selected surface positions (K6, K15, H30, E62, H74, K82) distributed over the entire molecular surface. rDer p 2 mutants containing one (n=6), two (n=2), three (n=1), four (n=1) and six (n=1) mutated amino acids were expressed in Pichia pastoris and analyzed by IgE inhibition and monoclonal antibody (mAb) binding

Research paper thumbnail of Biochemical Characterization of Intact Allergen Extracts and Chemically Modified Allergoids Representing Grass, Birch and House Dust Mite Allergen Extracts

Journal of Allergy and Clinical Immunology, 2006

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of an Intact Allergen Extract (Alutard SQ®) and 4 Commercially Available Allergoid Products for Birch Pollen Immunotherapy

Journal of Allergy and Clinical Immunology, 2006

Research paper thumbnail of Expression of the house dust mite allergen Der p 2 and mutants in the yeast pichia pastoris

Journal of Allergy and Clinical Immunology, 2002

sequences from TLPs. A cDNA clone was produced by RT-PCR using as a primer a sequence from the kn... more sequences from TLPs. A cDNA clone was produced by RT-PCR using as a primer a sequence from the known TLP of tomato. The original N-terminal sequence of the bell pepper TLP was obtained by 5'-RACE. Sequence analysis of the cDNA clone revealed similarity to thaumatin (54,1%) and to TLPs, such as PR P23 from tomato (92,8%), Pru av 2 (44,3%), or Mal d 2 (42%). The 23 kDa protein from bell pepper was designated Cap a 1. To produce recombinant (r) Cap a 1, its His-tagged coding sequence was ligated into the plant expression vector potato virus X (PVX) pgR106. N. benthamiana plants were infected with A. tumefaciens GV 3101 harbouring the modified virus within an agrobacterial plasmid. RT-PCR was performed with RNA isolated from newly formed upper leaves one week post inoculation. Sequencing of the RT-PCR products confirmed the presence and the correct sequence of Cap a 1 mRNA transcribed from a viral subgenomic promoter. Two weeks after inoculation, plants were harvested and tested for the expression of rCap a 1. The protein was purified by Ni-NTA chromatography. In immunoblot analysis, rCap a 1 exhibited IgE-binding capacity identical to the natural protein, indicating that the recombinant protein was expressed in its correct conformation. IgE cross-reactivity with tbaumatin and other allergenic TLPs was demonstrated. House dust mites (HDM) are reportedly a major cause of allergic diseases, such as asthma and rhinitis. Group 2 allergens are recognized as major allergens in several mite species. The genes for several of these allergens have been cloned, and isoforms within each genus have been identified. We have cloned 7 isoforms of the Dermatophagoides pteronyssinus allergen, Der p 2. Our results are in agreement with previously reported data suggesting a limited degree of sequence polymorphism. Each clone differed from the others by 2-4 residues. The amino acid changes at residues 40, 47, 76 and 111 were all conservative. However, substitution of asparagine 114 to an aspartic acid resulted in a change of the polypeptides calculated pI. We have expressed Der p 2 in the yeast, Pichia pastoris. Der p 2 is expressed as a fusion to the alpha-factor signal sequence from Saccharomyces cerevisiae. The signal peptide is cleaved off by the constitutively expressed Kex 2 proteinase, leading to the secretion of mature Der p 2 into the culture medium. Purification of the recombinant allergen was performed by sequential ammonium sulfate precipitation followed by size exclusion chromatography. Recombinant Der p 2 showed IgE-reactivity towards serum from house dust mite sensitive patients in SRID analysis. To explore the potential use of recombinant allergens in specific allergy vaccination, we have sought to engineer mutant allergens that display reduced IgE-binding properties but maintain an authentic three dimensional folding patterm as well as a full complement of T cell epitopes. The circular dichroism spectra, 260 to 184 nm range, was used to monitor the structural integrity of natural and recombinant Der p 2. Amino acid residues for site-directed mutagenesis were selected based on identification of residues conserved among various mite species, HDM (Der p/f and Eur m) and storage mites (Tyr p, Lep d and Gly d), as well as an analysis of the molecular surface of Der p 2 revealed by its NMR structure (#l.9, 1A9V.PDB). Hev b 3, a major allergen for latex allergic spina bifida patients, has been characterized on its molecular and biochemical level and is available as a recombinant protein. The aim of the study was to generate a human recombinant single chain fragment directed against Hev b 3 using the ETH2 human antibody phage library and to characterize its binding ability to natural Hev b 3 in latex C serum and to recombinant Hev b 3 in ELISA and immunoblot experiments. We obtained several specific clones after three rounds of panning the ETH2 human antibody phage library against rHev b 3. Clone G4 was expressed as a soluble protein with an N-terminal hexahistidyl tag in E. coli HB2152 cells and purified by Ni-NTA affinity chromatography. G4 produces comparable reactivity to natural and recombinant Hev b 3 in ELISA and immunoblot experiments. No crossreactivity was observed with Hev b 1, another spina bifida associated latex allergen with high sequence similarity to Hev b 3. In immunoblot and ELISA inhibition experiments rHevb 3 was able to abolish binding of G4 to nHev b 3 in a dose-dependent manner. This Hev b 3 specific single-chain antibody represents a useful tool to control latex extracts used for in vivo diagnosis for their Hev b 3 content and to trace Hev b 3 in latex devices.

Research paper thumbnail of Purification and characterization of various pollen profilins

Journal of Allergy and Clinical Immunology, 2002

The Journal of Allergy and Clinical Immunology, Volume 109, Issue 1, Pages S132, January 2002, Au... more The Journal of Allergy and Clinical Immunology, Volume 109, Issue 1, Pages S132, January 2002, Authors:Helene Henmar; Henrik Ipsen; Thomas Lenhard; Peter Thorsted. ...

Research paper thumbnail of Determination Of Group 1 and 2 Isoforms From House Dust Mite Using Different Antibody Based Assays

Journal of Allergy and Clinical Immunology, 2011

Research paper thumbnail of Biochemical Characterization Of Intact Allergen Extracts Vaccine (Alutard SQ®) And Chemically Modified Allergoid Vaccines Representing Birch, Grass And House Dust Mite

Journal of Allergy and Clinical Immunology, 2007

Research paper thumbnail of High consistency in allergen composition of SQ house dust mite (HDM) tablet for sublingual immunotherapy (SLIT)

Journal of Allergy and Clinical Immunology, Feb 1, 2018

RATIONALE: Labyrithine nasal passages, the narrow nasal valve and complex static and dynamic aero... more RATIONALE: Labyrithine nasal passages, the narrow nasal valve and complex static and dynamic aerodynamics limit traditional intranasal steroids' (INS) ability to effectively deliver medication to superior/ posterior nasal regions. Exhalation delivery systems (EDS) exploit unique characteristics of nasal anatomy and aerodynamics to overcome these limitations and achieve superior/posterior drug delivery. We review published human in vivo gamma-scintigraphy deposition data for conventional INS (C-INS; e.g., Flonase/Nasonex), HFA-based pMDI's (e.g., QNASL/Zetonna), and EDS systems for liquids and powders. METHODS: Four recent gamma-deposition studies comparing different technologies for nasal delivery of topical steroids were included: (1) Conventional INS sprays (Flonase and Nasonex) versus pMDI (QNASL), (2) C-INS (Nasonex) versus pMDI (Zetonna), (3) C-INS versus EDSliquid, (4) C-INS versus EDS-powder and EDS-liquid. Data on regional deposition and clearance was compared. RESULTS: Qualitative deposition differences were large, though variability in segmentation methods prevents quantitative comparisons. In all studies, C-INS consistently deposit primarily anteriorly (in the valve region) with clearance along the nasal floor and little deposition in superior/posterior regions. Both pMDIs (QNASL/Zetonna) show a stationary ''hotspot'' in the non-ciliated vestibule, little delivery to superior/ posterior regions, and minimal clearance. EDS (liquid or powder) produce less deposition in the valve area and broad deposition to superior/posterior segments with a different clearance pattern. CONCLUSIONS: Human imaging data demonstrate poor drug deposition in superior/posterior sites with conventional nasal sprays, and greatly increased deposition throughout upper/posterior nasal passages with an EDS. In CRS, upper/posterior sites, including the middle and upper meatuses (ostiomeatal complex) where sinuses ventilate/drain and polyps originate, are the primary target for treatment.

Research paper thumbnail of Diverse and highly cross‐reactive T‐cell responses in ragweed allergic patients independent of geographical region

Allergy, 2019

Article type : Original Article: Basic and Translational Allergy Immunology Diverse and highly cr... more Article type : Original Article: Basic and Translational Allergy Immunology Diverse and highly cross-reactive T cell responses in ragweed allergic patients independent of geographical region

Research paper thumbnail of Variation of trypsin inhibitor and crude protein in chikpea cultivars

Food Chemistry, 1996

Data on the energy content of food products presented in Polish food composition tables is based ... more Data on the energy content of food products presented in Polish food composition tables is based on calculations including energy coefficients of proteins, fats and carbohydrates. According to present practice the total sum of carbohydrates is taken into account in these calculations without allowance for dietary fibre. As a consequence results of these calculations are overestimated. The same applies to the energy content of diets assessed with the use of Polish food composition tables. Following Eurofoods recommendations the energy content of foods was revised by taking into consideration available carbohydrates instead of total carbohydrates. This revision resulted in the reductions of the energy content of diets as well. While the energy supply per capita in Poland calculated with the use of the former methodology was 3370 kcal/day, an allowance for dietary fibre reduced this value to 3240 kcal, i.e. by approx. 4%. The extent of the reductions in the energy content of the diets of various population groups varied, depending on the importance of foods rich in dietary fibre present in these diets.

Research paper thumbnail of Contents Vol. 144, 2007

International Archives of Allergy and Immunology, 2007

Access to full text and tables of contents, including tentative ones for forthcoming issues: www.... more Access to full text and tables of contents, including tentative ones for forthcoming issues: www.karger.com/iaa_issues 79 Endogenous Interleukin-10 Produced by Antigen-Irrelevant Cells Promotes the Development of Experimental Murine Allergic Conjunctivitis

Research paper thumbnail of Subject Index Vol. 144, 2007

International Archives of Allergy and Immunology, 2007

Research paper thumbnail of Consistent composition and potency of freeze dried tablets for sublingual immunotherapy

Journal of Allergy and Clinical Immunology

Research paper thumbnail of Fractionation of Source Materials Leads to a High Reproducibility of the SQ House Dust Mite SLIT-Tablets

International archives of allergy and immunology, 2016

The production of house dust mite (HDM) allergen products for allergy immunotherapy has tradition... more The production of house dust mite (HDM) allergen products for allergy immunotherapy has traditionally been based on purified mite bodies or whole-mite culture, which are quite different source materials with a limited possibility for adjusting the chemical composition. The SQ HDM SLIT-tablet is a fast-dissolving pharmaceutical formulation that has been developed for sublingual immunotherapy (SLIT) of HDM respiratory allergic disease. The objective of the present study was to establish a process for the production of drug substances for the SQ HDM SLIT-tablet offering a high reproducibility and independent control of the major allergens. Process controls were documented in a comprehensive process parameter qualification. The analyses comprised composition by crossed immunoelectrophoresis, protein content by BCA, total IgE binding potency by Centaur assay, quantitative major allergen determination by radial immunodiffusion and ELISA, and the ranking of emPAI scores generated by mass s...

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of an Intact Allergen Extract (Alutard SQ®) and 4 Commercially Available Allergoid Products for Birch Pollen Immunotherapy

Data Revues 00916749 V117i2ss S009167490503592x, Aug 16, 2011

Background: When breastfeeding is impossible or insufficient, the use of cow's milk-based hypoall... more Background: When breastfeeding is impossible or insufficient, the use of cow's milk-based hypoallergenic infant formulas is an option for infants suffering from or at risk of developing cow's milk allergy. As the Camelidae family has a large evolutionary distance to the Bovidae family and as camel milk differs from cow's milk protein composition, there is a growing interest in investigating the suitability of camel milk as an alternative to cow's milk-based hypoallergenic infant formulas. Methods: The aim of the study was to compare the allergenicity and immunogenicity of camel and cow's milk as well as investigating their cross-reactivity using a Brown Norway rat model. Rats were immunised intraperitoneally with one of four products: camel milk, cow's milk, cow's milk casein or cow's milk whey fraction. Immunogenicity, sensitising capacity, antibody avidity and cross-reactivity were evaluated by means of different ELISAs. The eliciting capacity was evaluated by an ear swelling test. Results: Camel and cow's milk showed similarity in their inherent immunogenicity, sensitising and eliciting capacity. Results show that there was a lower cross-reactivity between caseins than between whey proteins from camel and cow's milk. Conclusions: The study showed that camel and cow's milk have a low cross-reactivity, indicating a low protein similarity. Results demonstrate that camel milk could be a promising alternative to cow's milk-based hypoallergenic infant formulas.

Research paper thumbnail of Meaurement of trypsin inhibitor in chickpea variants using monoclonal antibodies; comparison with enzymic assay

Research paper thumbnail of Die klinische Wirksamkeit und immunologische Antwort einer SQ-standardisierten Gras-Allergie-Immuntherapie- Tablette (AIT) ist nach einer Behandlungssaison vergleichbar zwischen Kindern und Erwachsenen – Post-hoc- Analyse zwei randomisierter Studien

Research paper thumbnail of Sublingual Immunotherapy with Fast-Dissolving Grass Tablets Induces Comparable IgG4 Antibody and IgE-blocking Responses in Children and Adults

Journal of Allergy and Clinical Immunology, 2009

The benefits of treating multi-sensitized allergic rhinitis (AR) patients with a single allergen ... more The benefits of treating multi-sensitized allergic rhinitis (AR) patients with a single allergen immunotherapy tablet (AIT) have not been fully elucidated. We studied the effect of treating mono-and multi-sensitized AR patients, in whom symptoms were largely confined to the grass pollen season, with grass AIT. METHODS: Post-hoc analysis of data from 568 patients who participated in yr 1 of a 3-yr placebo-controlled trial evaluating the efficacy of grass AIT (75,000 SQ-T/2,800 BAU, Phleum-pratense, ALK-Abello´). Daily treatment started 16 weeks prior to grass-pollen season and continued throughout the season. Responses were analysed in patients sensitized to: a) grass only (mono-sensitized); b) grass 1 tree 1 any other allergen (tree multisensitized); and c) grass 1 any allergen except tree allergens (non-tree multi-sensitized). Sensitivity was determined by skin-prick testing. Daily symptom (runny, blocked, itchy nose, sneezing, gritty, red, itchy, watery eyes; 0 5 none, 1 5 slight, 2 5 moderate, 3 5 severe symptoms) and medication scores were compared. RESULTS: 161 patients were mono-sensitized, 191 tree multi-sensitized, and 216 non-tree multi-sensitized. Treatment with grass AIT was found to reduce symptom and medication scores in mono-and multi-sensitized patients regardless of presence or absence of tree-related sensitivities. Compared to placebo, grass AIT reduced median daily symptoms by 31%, 44%, and 30% (p < 0.0001) as well as median daily medication scores by 49%, 27%, and 60% (p < 0.0001) in mono-, tree multi-sensitized, and non-tree multi-sensitized patients, respectively. CONCLUSIONS: Treatment with a single-grass-allergen tablet yielded significant symptom and medication reductions in AR patients mono-or multi-sensitized to inhalant allergens regardless of the existence/absence of sensitivities to tree allergens.

Research paper thumbnail of The allergen-specific IgE reactivity pattern of Chinese house dust mite allergic patients

World Allergy Organization Journal, 2007

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of Three Intact Allergen Vaccines with Four Allergoid Vaccines for Subcutaneous Grass Pollen Immunotherapy

Journal of Allergy and Clinical Immunology, 2008

Research paper thumbnail of Mapping of Der p 2 antibody binding epitopes by site directed mutagenesis

Journal of Allergy and Clinical Immunology, 2004

MethodsrDer p 2 was subjected to site directed mutagenesis at six selected surface positions (K6,... more MethodsrDer p 2 was subjected to site directed mutagenesis at six selected surface positions (K6, K15, H30, E62, H74, K82) distributed over the entire molecular surface. rDer p 2 mutants containing one (n=6), two (n=2), three (n=1), four (n=1) and six (n=1) mutated amino acids were expressed in Pichia pastoris and analyzed by IgE inhibition and monoclonal antibody (mAb) binding

Research paper thumbnail of Biochemical Characterization of Intact Allergen Extracts and Chemically Modified Allergoids Representing Grass, Birch and House Dust Mite Allergen Extracts

Journal of Allergy and Clinical Immunology, 2006

Research paper thumbnail of Comparison of Allergenicity and Immunogenicity of an Intact Allergen Extract (Alutard SQ®) and 4 Commercially Available Allergoid Products for Birch Pollen Immunotherapy

Journal of Allergy and Clinical Immunology, 2006

Research paper thumbnail of Expression of the house dust mite allergen Der p 2 and mutants in the yeast pichia pastoris

Journal of Allergy and Clinical Immunology, 2002

sequences from TLPs. A cDNA clone was produced by RT-PCR using as a primer a sequence from the kn... more sequences from TLPs. A cDNA clone was produced by RT-PCR using as a primer a sequence from the known TLP of tomato. The original N-terminal sequence of the bell pepper TLP was obtained by 5'-RACE. Sequence analysis of the cDNA clone revealed similarity to thaumatin (54,1%) and to TLPs, such as PR P23 from tomato (92,8%), Pru av 2 (44,3%), or Mal d 2 (42%). The 23 kDa protein from bell pepper was designated Cap a 1. To produce recombinant (r) Cap a 1, its His-tagged coding sequence was ligated into the plant expression vector potato virus X (PVX) pgR106. N. benthamiana plants were infected with A. tumefaciens GV 3101 harbouring the modified virus within an agrobacterial plasmid. RT-PCR was performed with RNA isolated from newly formed upper leaves one week post inoculation. Sequencing of the RT-PCR products confirmed the presence and the correct sequence of Cap a 1 mRNA transcribed from a viral subgenomic promoter. Two weeks after inoculation, plants were harvested and tested for the expression of rCap a 1. The protein was purified by Ni-NTA chromatography. In immunoblot analysis, rCap a 1 exhibited IgE-binding capacity identical to the natural protein, indicating that the recombinant protein was expressed in its correct conformation. IgE cross-reactivity with tbaumatin and other allergenic TLPs was demonstrated. House dust mites (HDM) are reportedly a major cause of allergic diseases, such as asthma and rhinitis. Group 2 allergens are recognized as major allergens in several mite species. The genes for several of these allergens have been cloned, and isoforms within each genus have been identified. We have cloned 7 isoforms of the Dermatophagoides pteronyssinus allergen, Der p 2. Our results are in agreement with previously reported data suggesting a limited degree of sequence polymorphism. Each clone differed from the others by 2-4 residues. The amino acid changes at residues 40, 47, 76 and 111 were all conservative. However, substitution of asparagine 114 to an aspartic acid resulted in a change of the polypeptides calculated pI. We have expressed Der p 2 in the yeast, Pichia pastoris. Der p 2 is expressed as a fusion to the alpha-factor signal sequence from Saccharomyces cerevisiae. The signal peptide is cleaved off by the constitutively expressed Kex 2 proteinase, leading to the secretion of mature Der p 2 into the culture medium. Purification of the recombinant allergen was performed by sequential ammonium sulfate precipitation followed by size exclusion chromatography. Recombinant Der p 2 showed IgE-reactivity towards serum from house dust mite sensitive patients in SRID analysis. To explore the potential use of recombinant allergens in specific allergy vaccination, we have sought to engineer mutant allergens that display reduced IgE-binding properties but maintain an authentic three dimensional folding patterm as well as a full complement of T cell epitopes. The circular dichroism spectra, 260 to 184 nm range, was used to monitor the structural integrity of natural and recombinant Der p 2. Amino acid residues for site-directed mutagenesis were selected based on identification of residues conserved among various mite species, HDM (Der p/f and Eur m) and storage mites (Tyr p, Lep d and Gly d), as well as an analysis of the molecular surface of Der p 2 revealed by its NMR structure (#l.9, 1A9V.PDB). Hev b 3, a major allergen for latex allergic spina bifida patients, has been characterized on its molecular and biochemical level and is available as a recombinant protein. The aim of the study was to generate a human recombinant single chain fragment directed against Hev b 3 using the ETH2 human antibody phage library and to characterize its binding ability to natural Hev b 3 in latex C serum and to recombinant Hev b 3 in ELISA and immunoblot experiments. We obtained several specific clones after three rounds of panning the ETH2 human antibody phage library against rHev b 3. Clone G4 was expressed as a soluble protein with an N-terminal hexahistidyl tag in E. coli HB2152 cells and purified by Ni-NTA affinity chromatography. G4 produces comparable reactivity to natural and recombinant Hev b 3 in ELISA and immunoblot experiments. No crossreactivity was observed with Hev b 1, another spina bifida associated latex allergen with high sequence similarity to Hev b 3. In immunoblot and ELISA inhibition experiments rHevb 3 was able to abolish binding of G4 to nHev b 3 in a dose-dependent manner. This Hev b 3 specific single-chain antibody represents a useful tool to control latex extracts used for in vivo diagnosis for their Hev b 3 content and to trace Hev b 3 in latex devices.

Research paper thumbnail of Purification and characterization of various pollen profilins

Journal of Allergy and Clinical Immunology, 2002

The Journal of Allergy and Clinical Immunology, Volume 109, Issue 1, Pages S132, January 2002, Au... more The Journal of Allergy and Clinical Immunology, Volume 109, Issue 1, Pages S132, January 2002, Authors:Helene Henmar; Henrik Ipsen; Thomas Lenhard; Peter Thorsted. ...

Research paper thumbnail of Determination Of Group 1 and 2 Isoforms From House Dust Mite Using Different Antibody Based Assays

Journal of Allergy and Clinical Immunology, 2011

Research paper thumbnail of Biochemical Characterization Of Intact Allergen Extracts Vaccine (Alutard SQ®) And Chemically Modified Allergoid Vaccines Representing Birch, Grass And House Dust Mite

Journal of Allergy and Clinical Immunology, 2007