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B Are Involved in TNF-
Responses to Group B Streptococci1
* Department of Experimental Pathology and Microbiology, University of Messina, Messina, Italy; and
Department of Pharmacology and Neuroscience, Medical University of South Carolina, Charleston, SC 29425
TNF-
is a mediator of lethality in experimental infections by
group B streptococcus (GBS), an important human pathogen. Little is
known of signal transduction pathways involved in GBS-induced TNF-
production. Here we investigate the role of mitogen-activated protein
kinases (MAPKs) and NF-
B in TNF- production by human monocytes
stimulated with GBS or LPS, used as a positive control. Western blot
analysis of cell lysates indicates that extracellular signal-regulated
kinase 1/2 (ERK 1/2), p38, and c-Jun N-terminal kinase MAPKs, as well
as IB, became phosphorylated, and hence activated, in both LPS-
and GBS-stimulated monocytes. The kinetics of these phosphorylation
events, as well as those of TNF- production, were delayed by 30–60
min in GBS-stimulated, relative to LPS-stimulated, monocytes. Selective
inhibitors of ERK 1/2 (PD98059 or U0126), p38 (SB203580), or NF-B
(caffeic acid phenetyl ester (CAPE)) could all significantly reduce
TNF- production, although none of the inhibitors used alone was able
to completely prevent TNF- release. However, this was completely
blocked by combinations of the inhibitors, including PD98059-SB203580,
PD98059-CAPE, or SB203580-CAPE combinations, in both LPS- and
GBS-stimulated monocytes. In conclusion, our data indicate that the
simultaneous activation of multiple pathways, including NF-B, ERK
1/2, and p38 MAPKs, is required to induce maximal TNF- production.
Accordingly, in septic shock caused by either GBS or Gram-negative
bacteria, complete inhibition of TNF- release may require treatment
with drugs or drug combinations capable of inhibiting multiple
activation pathways.
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