Hypothesis: Grasses exposed to high levels of urban pollution express new allergens that may be responsible for symptoms in patients whose clinical sensitization has not yet been identified.
Objectives:
1) To obtain natural proteins from the LTP and TLP families, as well as recombinant proteins expressed in a heterologous system from Lolium perenne (wild grass), and to perform their immunological characterization using in vitro techniques.
2) To assess in vivo and in vitro sensitization to these pollution‑induced overexpressed proteins through skin prick tests and Basophil Activation Tests (BAT).
3) To determine the clinical response using a Nasal Provocation Test.
4) To compare sensitization and clinical allergy to LTP and TLP from L. perenne according to the degree of exposure and urban pollution.
Methodology:
We will obtain and produce LTP and TLP protein families that are overexpressed under high pollution conditions. Their IgE‑binding capacity will be characterized using immunoblotting and direct and/or inhibition ELISA with sera from allergic patients. Skin prick tests, Basophil Activation Tests (BAT), and Nasal Provocation Tests will be performed using natural and recombinant proteins, as well as L. perenne pollen collected from polluted versus non‑polluted areas, to assess sensitization and clinical symptoms in grass‑allergic patients. These evaluations will be conducted across the three participating centers with different levels of exposure and urban pollution (Madrid, Ciudad Real, and Alicante).
