Analysis of cytokine transcripts in the bronchoalveolar lavage cells of patients with asthma.
A panel of steady-state cytokine mRNAs was analyzed in the bronchoalveolar lavage (BAL) cells from asthmatic subjects or patients challenged with ragweed allergen. This was achieved by combining both qualitative and quantitative assays using the reverse transcription-polymerase chain reaction (RT-PCR). Analysis of BAL cells from six mild allergic asthmatic and five nonasthmatic, nonallergic subjects showed no qualitative differences in the profile of cytokine mRNAs (including interleukin [IL]-1 beta, IL-2, IL-5, IL-6, IL-8, and granulocyte/macrophage colony-stimulating factor), except for tumor necrosis factor-alpha, which was detected in three out of six asthmatic BAL samples but in none of the controls. A key cytokine, IL-5, has been implicated in the pathogenesis of allergic inflammation through the recruitment of eosinophils. We found a significant enhancement of steady-state IL-5 transcripts in the BAL cells from allergen-challenged as compared with the saline-challenged control sites of four asthmatic patients; furthermore, the cellular source for IL-5 mRNA was identified in the mononuclear cell fraction, but not in the purified eosinophils, of the allergen-challenged BALs. These results suggest that the significant increase of IL-5 transcripts is primarily from the infiltrating mononuclear cells. Our study also demonstrates the power of qualitative and quantitative PCR analysis in determining the molecular basis of allergic inflammatory diseases.
Johns Hopkins Asthma and Allergy Center, Baltimore, Maryland 21224-6801.
G
G
Krishnaswamy
M C
MC
Liu
S N
SN
Su
M
M
Kumai
H Q
HQ
Xiao
D G
DG
Marsh
S K
SK
Huang
D000328:Adult; D000485:Allergens / Q000378:metabolism; D001249:Asthma / Q000378:metabolism*; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA; D005260:Female; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*
D000485:Allergens; D016207:Cytokines; D015848:Interleukin-5; D012333:RNA, Messenger; D004247:DNA