“Functional analysis of tropomyosin of parasitic nematodes” Dissertation

Mein persönlicher Dank gilt meiner Familie für Ihre Unterstützung und meiner Badminton-Truppe dafür, dass sie mir eine zweite Familie geworden sind. 1 1 ZUSAMMENFASSUNG 5 1 SUMMARY 7 2 INTRODUCTION 9 2.1 General issues 9 2.1.1 Parasitic nematodes 9 2.1.2 Tropomyosin 10 2.2 Part I – RNA interference in Heligmosomoides polygyrus bakeri 13 2.2.1 Genetic manipulation of parasitic nematodes 13 2.3 Part II – Influence of rAv-TMY on a murine model of allergic airway inflammation 16 2.3.1 Immune modulation and evasion by helminths 16 2.3.2 Asthma 18 2.3.2.1 Immunological reactions leading to airway hyper-reactivity 18 2.3.3 Worms and allergy 20 2.4 Aims 23 3 RESULTS 24 3.1 Part I – RNA interference in Heligmosomoides polygyrus bakeri 24 3.1.1 Target gene and sequence fragments 24 3.1.2 Feeding of Hp-tmy dsRNA does not induce a phenotype in H. p. bakeri, but in C. elegans 24 3.1.3 H. p. bakeri larvae ingest fewer bacteria compared to C. elegans larvae 26 3.1.4 Electroporation with siRNA does not induce a phenotype in adult H. p. bakeri 27 3.1.5 Soaking with Hp-tmy dsRNA does not induce a phenotype in H. p. bakeri 28 3.1.6 BLAST analysis to identify RNAi-related orthologous in helminths 30 3.2 Part II – Influence of rAv-TMY on a murine model of allergic airway inflammation 33 3.2.1 rAv-TMY modulates airway hyperresponsiveness 33 6.4 Immunological methods 80 6.4.1 Enzyme-linked immunosorbent assay (ELISA) 80 6.4.1.1 Serum levels of IgE (total and OVA-IgE), OVA-IgG1, OVA-IgG2a 80 6.4.1.2 Serum levels of IgG subclasses specific for OVA or rAv-TMY 80 6.4.1.3 Serum / BALF levels of total IgA 81 6.4.1.4 Cytokine detection 81