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(Timisoara) The impact ragweed pollen and dust-mite allergens have on the respiratory mucosa
Romania (FASMR) - University of Medicine and Pharmacy "Victor Babes", Timisoara
Oncogen Center Timisoara
Prof. Carmen Panaitescu
Prof. Carmen Panaitescu, Assist. Prof. Laura Haidar (Marusciac)
Type of Research Project
- Basic science
What is the background of the project?
Worldwide, IgE-mediated allergy is the most important hypersensitivity disease, affecting up to 30% of the population in some countries. In Europe and especially in the Eastern parts of Europe including Romania, ragweed is one of the most important allergen sources, with a strong impact on human health. Ragweed (genus Ambrosia, family Asteraceae) is the most clinically relevant species known for its high potential to cause type I hypersensitivity reactions in late summer and fall. Another significant allergen source is represented by house dust mites (Dermatophagoides spp), which are arachnids with indoor habitat that induce respiratory allergies in a great number of people. There is an important part of the population sensitized to both pollen and house dust mite allergens. Currently there is no information on the added effect co-sensitization to perennial and seasonal allergens. Several studies have shown that protease allergens, such as the major allergens from Dermatophagoides spp (Der p 1, Der f 1) have a deleterious effect on the skin epithelium, destroying intercellular junctions and increasing skin permeability, thereby potentially facilitating the presentation of other allergens (such as ragweed pollen allergens) to the dendritic cells.
What is the aim of the project?
The project aims to evaluate the impact of ragweed pollen and house dust mite allergens on the respiratory mucosa that is damage to the epithelial cells or to the intercellular junctions.
What techniques and methods are used?
To this end a bronchial epithelial cell line will be used. The cells will be cultured and passaged in proper conditions, in order to maintain their characteristics. Then they will be incubated with different concentrations of allergens (house dust mite extract, major allergens from house dust mites, ragweed pollen extract, major allergens from ragweed pollen). The effect will be measured and therefore, the damage on the epithelial cells or the junctions will be quantified. - Cell culture methods: thawing and freezing, subculturing, cell counting, growth media preparation, etc - Preparation of HDM(house dust-mites) extract and measurement of protein concentration - Preparation of ragweed pollen extract and measurement of protein concentration - Measurement of transepithelial resistance with an Ohmvoltmeter - Interpretation of the transepithelial resistance obtained by incubation with different concentrations of allergens and combinations of allergens (Der p 1, house dust mite extract, Amb a 1, ragweed pollen extract), for different time durations - Real-time measurement of cell proliferation and adhesion with xCELLigence® system - Immunofluorescence staining
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student is expected to do the following tasks: - Observe the cells at the microscope - Count cells using the microscope and the cell counting chamber - Prepare solutions - weigh the materials needed, dissolve them and adjust the parameters (pH, viscosity etc.) - Prepare media for the cells. Mix the components of the media and adjust the parameters (pH, viscosity etc.) - Pipet reagents in order to prepare the solutions needed - Process cells – thaw, freeze, subculture, change media, etc - Centrifuge the cells using the ultracentrifuge - Stain cells – immunofluorescence for ZO-1, E-cadherin - Data interpretation ex: introduce in a database the results and compare them
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
There are introductory presentations regarding basic cell culture methods, basic immunology, as well as lab protocols for specific activities.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Subjects passed: biology, immunology
Are there any legal limitations in the student’s involvement
The student will have to sign: - confidentiality agreement - work protection training
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- Henriquez OA; Beste KD; Hoddeson EK; Parkos CA; Nusrat A; Wise SK. House dust mite allergen Der p 1 effects on sinonasal epithelial tight junctions. InInternational forum of allergy & rhinology 2013 Aug (Vol. 3; No. 8; pp. 630-635).
- Chen KW; Marusciac L; Tamas PT; Valenta R; Panaitescu C. Ragweed pollen allergy: burden; characteristics; and management of an imported allergen source in Europe. International archives of allergy and immunology. 2018;176(3-4):163-80.
- Waltl EE; Selb R; Eckl-Dorna J; Mueller CA; Cabauatan CR; Eiwegger T; Resch-Marat Y; Niespodziana K; Vrtala S; Valenta R; Niederberger V. Betamethasone prevents human rhinovirus-and cigarette smoke-induced loss of respiratory epithelial barrier function. Scientific reports. 2018 Jun 26;8(1):9688.
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