Development of a new technique for recovery of cytokines from inflammatory sites in situ

• Published in: Journal of immunological methods Vol. 155; no. 1; p. 25
• Main Authors: Alam, R, Sim, T C, Hilsmeier, K, Grant, J A
• Format: Journal Article
• Language: English
• Published: Netherlands 19.10.1992
• Subjects: Allergens - immunology; Dust; Enzyme-Linked Immunosorbent Assay - methods; Filtration; Granulocyte-Macrophage Colony-Stimulating Factor - isolation & purification; Humans; Interleukin-1 - isolation & purification; Nasal Mucosa - immunology; Nasal Provocation Tests; Rhinitis - immunology
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(92)90267-W

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.