Primary ciliary dyskinesia is characterized by chronic upper and lower respiratory infections which are caused by the grossly impaired ciliary transport. Since the cilia and neutrophils both utilize microtubular system for their movement, it has been speculated that neutrophil motility such as chemotaxis might be impaired in patients with primary ciliary dyskinesia. Neutrophils were purified from whole blood from 16 patients with primary ciliary dyskinesia and from 15 healthy controls. Chemotactic responses of neutrophils to leukotriene B4 (LTB4), complement 5a (C5a), and formylmethion-ylleucylphenylalanine (fMLP) were examined using the under agarose method. The chemotactic differentials in response to LTB4, C5a, and fMLP in neutrophils from the patient group were significantly lower than the corresponding values in neutrophils from the control group (p<0.05 for all comparisons). The difference in chemotactic index between the two groups was statistically significant for LTB4 and fMLP (p<0.05 for both comparisons), but not for C5a (p=0.20). Neutrophils from patients with primary ciliary dyskinesia showed a decreased chemotactic response as compared with those from normal subjects. It is concluded that the increased frequency of respiratory tract infection in patients with primary ciliary dyskinesia is possibly due to the defective directional migration of neutrophils, as well as to the defective mucociliary clearance of the airways.
Adolescent ; Chemotactic Factors/pharmacology ; Chemotaxis* ; Child ; Cilia/ultrastructure ; Comparative Study ; Complement 5a/pharmacology ; Dose-Response Relationship, Drug ; Dynein ATPase/chemistry ; Human ; Kartagener Syndrome/blood* ; Kartagener Syndrome/classification ; Leukotriene B4/pharmacology ; Male ; N-Formylmethionine Leucyl-Phenylalanine/pharmacology ; Neutrophils/physiology* ; Neutrophils/ultrastructure

Chemokine KC has been considered to be a murine homologue of human GRO/MGSA and was identified as chemoattractant for monocytes and neutrophils. This study examined the expression of KC mRNA in thioglycollate-elicited mouse peritoneal macrophages that were stimulated in vitro with Candida albicans (CA). Also examined were the inhibitory effects of IL-10 on the CA-induced expression of KC gene by Northern blot analysis. CA was found to induce chemokine gene expression in a gene-specific manner, CXC chemokine IP-10 mRNA expression was not detected in CA-stimulated macrophages. Maximum KC mRNA expression was observed approximately 2 hr after adding CA. The inhibitory action of IL-10 to CA-induced KC mRNA expression on mouse peritoneal macrophages was independent on concentration and stimulation time of IL-10 and was observed approximately one hour after adding IL-10 and CA simultaneously. IL-10 produced a decrease in the stability of KC mRNA, and CA-stimulated macrophages with cycloheximide blocked the suppressive effect of IL-10. These results suggest that CA also induces chemokine KC from macrophages, and IL-10 acts to destabilize CA-induced KC mRNA and de novo synthesis of an intermediate protein is a part of the IL-10 suppressive mechanism.
Animal ; Blotting, Northern ; Candida albicans/metabolism* ; Cells, Cultured ; Chemotactic Factors/genetics* ; Dactinomycin/pharmacology ; Dose-Response Relationship, Drug ; Gene Expression Regulation/drug effects* ; Growth Substances/genetics* ; Interleukin-10/pharmacology* ; Interleukin-10/metabolism ; Macrophages/physiology* ; Mice ; Mice, Inbred BALB C ; Nucleic Acid Synthesis Inhibitors/pharmacology ; RNA, Messenger/metabolism ; RNA, Messenger/drug effects

OBJECTIVETo investigate the regulation mechanism of apoptosis induced by the antisense bcl-2 treatment.

METHODSDNA content analysis and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) were adopted to detect apoptosis. Semi-quantitative reverse transcription-PCR was performed to detect the mRNA expression of bcl-2 c-myc survivin bax s100A(2) TNFalpha TGFbeta(1) and IL-6 in the small-cell lung cancer cell line NCI-H446 treated with antisense bcl-2 oligodeoxynucliotide.

RESULTSbcl-2 AS-PS-ODN treatment could induce apoptosis, accompanied with 72.71% up-regulation of IL-6 and 65.90% down-regulation of TNFalpha, whereas little or no effect was seen on c-myc survivin bax s100A(2) and TGFbeta(1).

CONCLUSIONIL-6 and TNFalpha may be involved in the regulation of apoptosis induced by antisense bcl-2 treatment.

Apoptosis ; drug effects ; genetics ; Chemotactic Factors ; genetics ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; In Situ Nick-End Labeling ; Inhibitor of Apoptosis Proteins ; Interleukin-6 ; genetics ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; S100 Proteins ; genetics ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1 ; Tumor Cells, Cultured ; drug effects ; Tumor Necrosis Factor-alpha ; genetics ; bcl-2-Associated X Protein