Cytokines in asthma: effects on human pulmonary fibroblasts (835.8)

Abstract

In the asthmatic patient, CD4+ T lymphocytes elevate chemokines in airway fluids and blood. This study assayed effects of serum from asthmatic adults on the metabolism of healthy human lung fibroblasts in culture, comparing effects to those seen with sera from lung‐healthy volunteers. WI38 human lung fibroblasts (ATCC) were cultured in MEM + 15% fetal calf serum. At assay, 1500 cells in 100 µL were added to 96‐well plates, and after attachment, media was replaced with serum‐free MEM with 15 µL asthma or control sera. After 24 hr incubation, MTT formazan cell proliferation was assayed at 570 nm. Tests were repeated 3x.MTT of 24 asthmatics averaged 1.37 ± 0.12 vs 1.28 ± 0.11 for 16 controls, p=0.03 with 3 replications. Sera with high IgE (ELISA) were compared to low by MTT values and were not different. Serum and breath condensate have recently been assayed for multiple chemokines but not for effects of these effluents on healthy lung cells. Our tests, while not identifying components demonstrate the expression of substances in asthma that raise the inflammatory response of healthy cells (measured by MTT upregulation). We showed the upregulation was significantly greater by asthmatic sera than by control sera. The use of asthma serum‐sensitized cells in culture offers high possibility of clarifying the disease of asthma and its humoral processes beyond cell‐related triggers.Grant Funding Source: Sarah Morrison Student Grant to SL