Interleukin-2-induced tumor necrosis factor-alpha (TNF-alpha) gene expression in human alveolar macrophages and blood monocytes.
Recent investigations have demonstrated interleukin-2 receptor (IL-2R) expression on both human alveolar macrophages (AM phi) and blood monocytes (PBM), but the function of these receptors has not been fully elucidated. In this study, we demonstrate that human AM phi, as well as PBM, can be induced to express biologically active TNF-alpha after challenge with interleukin-2 (IL-2). Furthermore, we examined the expression of TNF-alpha at the mRNA level via Northern blot and in situ hybridization analysis. Normal AM phi, obtained by bronchoalveolar lavage, and PBM were stimulated with either IL-2 (2,000 U/ml) or lipopolysaccharide (LPS) (10 micrograms/ml) for 18 h. Specificity was demonstrated by neutralizing TNF-alpha activity with a polyclonal rabbit anti-human TNF-alpha antibody. PBM TNF-alpha biologic activity from 11 subjects challenged with either IL-2 or LPS was 19 +/- 6 and 85 +/- 15 U/ml/10(6) cells, respectively, which represented 5-fold and 21-fold increases over control values. AM phi TNF-alpha biologic activity from nine subjects was 110 +/- 28 (IL-2-mediated) and 304 +/- 69 (LPS-mediated) U/ml/10(6) cells, which represented 2- and 6-fold increases over controls. AM phi exhibited statistically greater (p less than 0.05) TNF production in response to both IL-2 and LPS as compared to PBM. IL-2 challenge resulted in an induction of TNF-alpha mRNA accumulation, as demonstrated by Northern blot and in situ hybridization analyses. TNF-alpha mRNA was quantitated by laser densitometry for Northern blots or by counting the number of silver grains/mononuclear phagocytic cell in the in situ hybridization analysis.(ABSTRACT TRUNCATED AT 250 WORDS)
Strieter
R M
RM
Department of Internal Medicine and Pathology, University of Michigan Medical School, Ann Arbor 48109-0360.
Remick
D G
DG
Lynch
J P
JP
Spengler
R N
RN
Kunkel
S L
SL
D000328:Adult; D001681:Biological Assay / Q000379:methods; D015152:Blotting, Northern / Q000379:methods; D002469:Cell Separation; D004247:DNA / Q000187:drug effects / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects; D008297:Male; D009000:Monocytes / Q000187:drug effects*; D009693:Nucleic Acid Hybridization / Q000187:drug effects; D011650:Pulmonary Alveoli / Q000166:cytology*; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000187:drug effects* / Q000235:genetics
D007376:Interleukin-2; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D004247:DNA