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After Stimulation with Cell Walls of Streptococcus pneumoniae and Regulate Inducible Nitric Oxide Synthase and ICAM-1 Expression Via Autocrine Loops1
*
Department of Neurology,
Institute of Medical Immunology, and
Institut für Mikrobiologie und Hygiene, Universitaetsklinikum Charité, Humboldt University Berlin, Germany; and
§
Institut für Klinische Pharmakologie und Toxikologie, Free University Berlin, Berlin, Germany
TNF-
, inducible NO synthase (iNOS), and ICAM-1 are considered to
be key proteins in the inflammatory response of most tissues. We tested
the hypothesis that cell walls of Streptococcus
pneumoniae (PCW), the most common cause of adult bacterial
meningitis, induce TNF-, iNOS, and ICAM-1 expression in rat primary
brain microvascular endothelial cell cultures. We detected TNF- mRNA
by RT-PCR already 1 h after stimulation with PCW, while TNF-
protein peaked at 4 h (9.4 ± 3.6 vs 0.1 ± 0.1 pg/µg
protein). PCW induced iNOS mRNA 2 h after stimulation, followed by
an increase of the NO degradation product nitrite (18.1 ± 4 vs
5.8 ± 1.8 at 12 h; 18.1 ± 4 vs 5.8 ± 1.8
pmol/µg protein at 72 h). The addition of TNF- Ab
significantly reduced nitrite production to 62.2 ± 14.4%
compared with PCW-stimulated brain microvascular endothelial cells
(100%). PCW induced the expression of ICAM-1 (measured by FACS), which
was completely blocked by TNF- Ab (142 ± 18.6 vs 97.5 ±
12.4%; 100% unstimulated brain microvascular endothelial cells).
Cerebral endothelial cells express TNF- mRNA as well as iNOS mRNA
and release the bioactive proteins in response to PCW. PCW-induced NO
production is mediated in part by an autocrine pathway involving
TNF-, whereas ICAM-1 expression is completely mediated by this
autocrine loop. By these mechanisms, cerebral endothelial cells may
regulate critical steps in inflammatory blood-brain-barrier disruption
of bacterial meningitis.
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