Macrophage inflammatory protein-2 (MIP-2) is a C-X-C chemokine that is important in recruiting neutrophils to inflammatory sites. Our previous reports demonstrated that lipopolysaccharide (LPS) or CpG-oligode-oxynucleotide (CpG-ODN) rapidly induce MIP-2 gene expression in the macrophage cell line, RAW 264.7. Here, we show that the DNA sequence of the MIP-2 promoter between -114 and +14 is sufficient for strong promoter activity in LPS- or CpG-ODN-stimulated RAW 264.7 cells. Importantly, comprehensive mutant analysis reveals that an Sp1 element in the promoter region between -114 and -94 is essential for synergistic MIP-2 promoter activation by NF-kappaB and c-Jun regardless of the presence of an AP-1 site. By combining deletion or site-specific mutant analysis with immunocomplex assays, we also confirmed that Sp1 mediates the recruitment of transcription factors NF- kappaB and c-Jun in LPS- or CpG-ODN-treated RAW 264.7 cells. Several lines of experimental evidence imply that the Sp1-binding element is an important determinant of MIP-2 promoter activity, and that NF-kappaB, c-Jun and Sp1 can functionally cooperate to elicit maximal activation of the promoter.

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A new medium selective for Bacteroides gracilis was developed. The medium is tryptic soy agar (Difco Laboratories, Detroit, Mich.) containing nalidixic acid, teicoplanin, sodium formate, sodium fumarate, and potassium nitrate. Ail 18 strains of B. gracilis tested grew with only minimal inhibition. Most of the other 214 organisms tested, including most Bacteroides species, other anaerobes, and a substantial number of facultative anaerobes, were significantly inhibited by the medium. In a diagnostic study of 49 clinical specimens (28 patients with intra-abdominal infection, mostly gangrenous or perforated appendicitis), four strains of B. gracilis were isolated (from 4 different patients) on B. gracilis selective agar but were not detected on standard media.

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