title: Development of a new technique for recovery of cytokines from inflammatory sites in situ.
abstract: We attempted to recover cytokines from nasal mucosal surface following allergen challenge. Repeated lavage of nasal mucosa of seven allergic patients was done, but we failed to detect IL-1 beta in the lavage samples even in ten-fold concentrated materials. Therefore, we developed a new technique to recover cytokines using filter strips. Small filter strips were placed on nasal turbinates for 10 min at different time points after allergen challenge. The strips were air-dried, and stored. For recovery of cytokines individual strips were washed with small volumes of Hepes buffer containing 0.3% human serum albumin. Eluates were assayed for the presence of IL-1 beta and GM-CSF using commercially available ELISA. We were able to detect IL-1 beta and GM-CSF in eluates. Both cytokines were consistently detectable in the late phase allergic reaction peaking at 5 h. Nasal challenge with saline failed to detect any cytokine during the 7 h observation period. In standardization experiments known quantities of IL-1 beta and GM-CSF were applied to filter strips and the recovery ranged from 67 to 89%. Thus, we developed a simple technique of recovery of cytokines from inflammatory mucosa in situ.
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch, Galveston 77550.
authors.forename: R
authors.initials: R
authors.lastname: Alam
authors.forename: T C
authors.initials: TC
authors.lastname: Sim
authors.forename: K
authors.initials: K
authors.lastname: Hilsmeier
authors.forename: J A
authors.initials: JA
authors.lastname: Grant
mesh_terms: D000485:Allergens / Q000276:immunology; D004391:Dust; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005374:Filtration; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000302:isolation & purification*; D006801:Humans; D007375:Interleukin-1 / Q000302:isolation & purification*; D009297:Nasal Mucosa / Q000276:immunology*; D009299:Nasal Provocation Tests; D012220:Rhinitis / Q000276:immunology*
chemical_list: D000485:Allergens; D004391:Dust; D007375:Interleukin-1; D016178:Granulocyte-Macrophage Colony-Stimulating Factor