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  • 1
    Electronic Resource
    Electronic Resource
    Non-anaphylactic surface-exposed peptides of the major birch pollen allergen, Bet v 1, for preventive vaccination (2004)
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 34 (2004), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Almost 100 million allergic patients are sensitized to the major birch pollen allergen, Bet v 1, a 17 kDa protein containing most of the IgE epitopes present in pollens of trees belonging to the Fagales order and plant-derived food.Objective Our aim was to develop an approach for the rational design of B cell epitope-derived, non-allergenic peptide allergy vaccines.Methods According to the three-dimensional (3-D) structure of birch pollen allergen, Bet v 1, six peptides comprising 25–32 preferably solvent-exposed amino acids were synthesized.Results Because of lack of secondary structure, the peptides showed no allergenic activity in allergic patients. In a mouse model of birch pollen allergy, peptide vaccination induced Bet v 1-specific IgG and prevented IgE-mediated allergic sensitization to Bet v 1. The protective role of peptide-induced blocking antibodies is demonstrated by inhibition of allergic patients IgE binding to the allergen and by blocking of allergen-induced basophil degranulation.Conclusion Our results indicate the mechanistic importance of blocking antibodies for allergy vaccination and present a B cell epitope-based approach for the rational design of safe peptide allergy vaccines whenever the structure of the disease-eliciting allergen is known.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2222.2004.02081.x
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  • 2
    Electronic Resource
    Electronic Resource
    Vaccines for birch pollen allergy based on genetically engineered hypoallergenic derivatives of the major birch pollen allergen, Bet v 1 (2004)
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 34 (2004), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background We have recently engineered recombinant derivatives of the major birch pollen allergen Bet v 1 (rBet v 1 fragments and trimer) with strongly reduced allergenic activity.Objective The aim of this study was the in vivo characterization of potential allergy vaccines based on Al(OH)3-adsorbed genetically modified rBet v 1 derivatives in mice.Methods BALB/c mice were immunized either with courses of nine injections of increasing doses of Al(OH)3-adsorbed rBet v 1 wild-type, rBet v 1 fragments, rBet v 1 trimer or Al(OH)3 alone in weekly intervals or with three high-dose injections applied in intervals of 3 weeks. Humoral immune responses to rBet v 1 wild-type and homologous plant allergens were measured by ELISA and Western blotting, and the ability of mouse antibodies to inhibit the binding of allergic patients IgE to Bet v 1 was studied by ELISA competition experiments.Results In both schemes, hypoallergenic rBet v 1 derivatives induced low IgE but high IgG1 responses against rBet v 1 wild-type. The IgG1 antibodies induced by genetically modified rBet v 1 derivatives cross-reacted with natural Bet v 1 and its homologues from alder (Aln g 1) as well as hazel (Cor a 1) and strongly inhibited the binding of birch pollen allergic patients' IgE to Bet v 1 wild-type.Conclusion Genetically modified hypoallergenic rBet v 1 derivatives induce blocking antibodies in vivo. Their safety and efficacy for the treatment of birch pollen and associated plant allergies can now be evaluated in clinical immunotherapy studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2222.2004.01857.x
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization of the protective and therapeutic efficiency of a DNA vaccine encoding the major birch pollen allergen Bet v 1a (2004)
    Oxford, UK : Munksgaard International Publishers
    Allergy 59 (2004), S. 0 
    Details
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background:  An estimated 100 million individuals suffer from birch pollen allergy. More than 95% of birch pollen-allergic subjects react with the major birch pollen allergen Bet v 1a, and almost 60% of them are sensitized exclusively to this allergen.Objective:  DNA immunization using the Bet v 1a gene was evaluated with respect to its prophylactic and therapeutic efficacy.Methods:  A DNA vaccine containing the entire Bet v 1a cDNA under the control of a CMV-promoter was constructed. In order to estimate the protective efficiency, animals received three injections of this vaccine prior to sensitization with recombinant Bet v 1a. Vice versa, in a therapeutic approach, sensitization was followed by treatment with the DNA vaccine.Results:  The Bet v 1a DNA vaccine induced strong Bet v 1-specific antibody responses with a Th1-biased response type. Animals which received the DNA vaccine were protected against a following allergic sensitization with Bet v 1a. The protective effect was characterized by suppression of Bet v 1-specific immunoglobulin (Ig)E production, lack of basophil activation and enhanced interferon (IFN)-γ expression. In a therapeutic situation, treatment of sensitized animals with DNA vaccines decreased IgE production, IgE-mediated basophil release and drastically reduced anaphylactic activity as measured by passive cutaneous anaphylaxis assays. Concerning the cellular immune response, DNA immunization induced a sustaining and dominant shift from a Th2 type response towards a balanced Th1/Th2 type response as indicated by increased IFN-γ but unchanged IL-5 levels in lymphoproliferation assays.Conclusion:  The results demonstrate the allergen-specific protective and therapeutic efficacy of a DNA vaccine encoding the clinically highly relevant allergen Bet v 1a indicating the suitability of this concept for the treatment of allergic diseases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1398-9995.2003.00335.x
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  • 4
    Electronic Resource
    Electronic Resource
    Optimization of codon usage is required for effective genetic immunization against Art v 1, the major allergen of mugwort pollen (2003)
    Oxford, UK : Munksgaard International Publishers
    Allergy 58 (2003), S. 0 
    Details
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background:  As the major allergen of mugwort pollen, Art v 1 is an important target for specific immunotherapy. However, both recombinant protein as well as a gene vaccine for Art v 1 failed to be immunogenic in mice. In order to improve immunogenicity we focused on genetic immunization because interspecific differences of codon usage have been shown as an obstacle for effective induction of immune responses with gene vaccines encoding infectious pathogens.Objective:  In order to find out, whether codon usage might also be used to improve genetic immunization with allergen genes, the response against a gene vaccine expressing the wild-type gene of Art v 1 (pCMV-wtArt) was compared with a synthetic codon-optimized vector with human codon usage (pCMV-humArt).Methods:  Balb/c mice were injected intradermally with pCMV-wtArt or pCMV-humArt. In vitro expression levels of both constructs were compared in transfection experiments. Total immunoglobulin G (IgG), IgG1, IgG2a and IgE antibodies were analyzed by enzyme-linked immunosorbent assay and the anaphylactic activity of the sera was determined by allergen-specific degranulation of rat basophil leukemia-2H3 cells.Results:  No immune response was detectable with the gene vaccine expressing the wildtype Art v 1, but immunization with pCMV-humArt revealed a strong and allergen-specific induction of antibody responses. The antibodies recognized both the recombinant as well as the purified natural (glycosylated) Art v 1 molecule. The response type was Th1-biased, as indicated by high levels of IgG2a antibodies. Expression analysis with B16 mouse melanoma cells transfected with pCMV-humArt or pCMV-wtArt revealed an impaired expression of the wild-type vector but normal translation after recoding.Conclusion: The results demonstrate that optimization of codon usage offers a simple way to improve immunogenicity and therefore should be routinely considered in the development of gene vaccines for the treatment of allergy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1398-9995.2003.00170.x
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  • 5
    Electronic Resource
    Electronic Resource
    The role of the spleen and splenic autotransplants in clearing experimental bacteremia caused by the gram-negative bacteriumEscherichia coli (1986)
    Springer
    Research in experimental medicine 186 (1986), S. 229-238 
    Details
    ISSN: 1433-8580
    Keywords: Spleen ; Experimental bacteremia ; E. coli ; Sepsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Several clinical and laboratory studies have demonstrated the risk of sepsis in connection with encapsulated bacteria. The importance of clearing these organisms by the spleen is now well accepted. In contrast, the present work deals with the clearance of non-encapsulated gram-negative bacteria(Escherichia coli). The animals used for the experiments (a heterogenous population of rabbits) were divided into a control group, a splenectomized group, and an autotransplanted group. The histological examination of the splenic transplants revealed typical splenic tissue including lymphatic follicles with germinal activity 42 days after transplantation. For estimation of the clearance capacity different amounts of bacteria were injected i.v. into the rabbits, and colony-forming bacterial cells in the blood were counted at certain intervals. In the control group no bacteria could be detected in the blood after 7 min. All animals of the splenectomized and autotransplanted groups showed a remarkable decrease in clearance efficiency (no bacteria in the blood after 19 min). No difference in the clearance kinetics could be shown between splenectomized and autotransplanted animals. Measuring the uptake of bacterial cells into different organs elicited low incorporation in the spleen as compared to non-immunocompetent organs, but no difference between normal spleen and splenic replants. However, saturation withE. coli cells reached higher limits in the normal spleen than in autotransplants. The immunologic capacity with respect to IgM-producing lymphocytes was measured by the hemolytic plaque assay. The results showed a severe malfunction of the autotransplants as compared to the normal spleen (only 2% of the activity of the control group). Vaccination againstE. coli increased the clearance efficiency in all three groups. The data presented in this paper point out that several functions of the spleen cannot be carried out by autotransplants. The reasons, therefore, may be limited transplant mass and/or decreased specific functions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01852049
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  • 6
    Electronic Resource
    Electronic Resource
    81. Immunologische Kapazität und Durchblutung von Milzautotransplantaten — Experimentelle Ergebnisse am Schwein (1986)
    Springer
    Langenbeck's archives of surgery 369 (1986), S. 399-403 
    Details
    ISSN: 1435-2451
    Keywords: Splenic autotransplantation ; Immune reaction ; Perfusion ; Pig ; Autologe Milztransplantation ; Immunreaktion ; Durchblutung ; Schwein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Diese Studie beschreibt quantitativ Immunvorgänge und die Durchblutungsrate von Milzregeneraten nach Autotransplantation von Milzgewebe in das Omentum majus. Das Gewicht und die Perfusion der Transplantate 12 Monate postop. waren deutlich reduziert. Die Anzahl der Antikörper-produzierenden Zellen in den Transplantaten war im Vergleich zur normalen Milz nur gering vermindert, während die Milzregenerate das Serum-IgM-Defizit nach Splenektomie nach 4 Monaten ausglichen. Weiters fanden wir verschiedene Proliferationsraten für B- und T-Lymphocyten in den Transplantaten und der normalen Milz nach Mitogenstimulation.
    Notes: Summary The study deals with the immunological capacity and perfusion rate of splenic grafts in pigs after autologous transplantation into the omentum majus. 12 months after operation the weight and relative blood flow of the regenerated splenic tissue were clearly reduced. The number of antibody forming cells in the grafts was only slightly decreased compared to the normal spleen, whereas the autotransplants could balance a serum-IgM-deficiency, observed due to splenectomy, 4 months after grafting. Furthermore the mitogenic stimulation of T- and B-lymphocytes showed different proliferation rates for the autotransplants compared to the spleen.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01274398
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