OBJECTIVETo investigate what role IL-11 plays in periodontal disease and to determine the level of IL-11 in HGFs stimulated with IL-1alpha and TNF-alpha.

METHODSHGFs were stimulated with IL-1alpha and TNF-alpha alone or in combination. The production of IL-11 was measured using enzyme-linked immunosorbent assay (ELISA). IL-11 and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) messenger RNA (mRNA) levels in HGFs were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSIL-1alpha significantly increased the levels of IL-11 in HGFs. TNF-alpha also significantly augmented IL-11 production in HGFs, and synergistically stimulated HGFs to produce IL-11 when combined with IL-1alpha. Indomethacin, an inhibitor of prostaglandin synthesis, significantly reduced IL-11 production by HGFs stimulated with IL-1alpha and TNF-alpha individually or in combination. IL-1alpha alone or combined with TNF-alpha enhanced the ratio of IL-11/GAPDH mRNA expression in HGFs, and the augmentation was abolished by indomethacin after co-incubation for 24 hs.

CONCLUSIONSProduction of IL-11 in HGFs stimulated with IL-1alpha and TNF-alpha was transcriptionally upregulated by the endogenous prostaglandin synthesis. Inhibition of prostaglandin might suppress the osteoclastogenesis by IL-11 in inflammatory periodontal diseases.

Cells, Cultured ; Cyclooxygenase Inhibitors ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Gingiva ; cytology ; metabolism ; Humans ; Indomethacin ; pharmacology ; Interleukin-11 ; biosynthesis ; Interleukin-1alpha ; pharmacology ; RNA, Messenger ; biosynthesis ; Tumor Necrosis Factor-alpha ; pharmacology

Because of an increased number of Acanthamoeba keratitis (AK) along with associated disease burdens, medical professionals have become more aware of this pathogen in recent years. In this study, by analyzing both the nuclear 18S small subunit ribosomal RNA (18S rRNA) and mitochondrial 16S rRNA gene loci, 27 clinical Acanthamoeba strains that caused AK in Japan were classified into 3 genotypes, T3 (3 strains), T4 (23 strains), and T5 (one strain). Most haplotypes were identical to the reference haplotypes reported from all over the world, and thus no specificity of the haplotype distribution in Japan was found. The T4 sub-genotype analysis using the 16S rRNA gene locus also revealed a clear sub-conformation within the T4 cluster, and lead to the recognition of a new sub-genotype T4i, in addition to the previously reported sub-genotypes T4a-T4h. Furthermore, 9 out of 23 strains in the T4 genotype were identified to a specific haplotype (AF479533), which seems to be a causal haplotype of AK. While heterozygous nuclear haplotypes were observed from 2 strains, the mitochondrial haplotypes were homozygous as T4 genotype in the both strains, and suggested a possibility of nuclear hybridization (mating reproduction) between different strains in Acanthamoeba. The nuclear 18S rRNA gene and mitochondrial 16S rRNA gene loci of Acanthamoeba spp. possess different unique characteristics usable for the genotyping analyses, and those specific features could contribute to the establishment of molecular taxonomy for the species complex of Acanthamoeba.
Acanthamoeba/classification/genetics/growth & development/*isolation & purification ; Acanthamoeba Keratitis/*parasitology ; Cell Nucleus/*genetics ; DNA, Mitochondrial/*genetics ; DNA, Protozoan/genetics ; Humans ; Japan ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 16S/*genetics ; RNA, Ribosomal, 18S/*genetics