Antibacterial peptides are active defense components of innate immunity. Several studies confirm their importance at epithelial surfaces as immediate barrier effectors in preventing infection. Here we report that early in Shigella spp. infections, expression of the antibacterial peptides LL-37 and human beta-defensin-1 is reduced or turned off. The downregulation is detected in biopsies from patients with bacillary dysenteries and in Shigella- infected cell cultures of epithelial and monocyte origin. This downregulation of immediate defense effectors might promote bacterial adherence and invasion into host epithelium and could be an important virulence parameter. Analyses of bacterial molecules causing the downregulation indicate Shigella plasmid DNA as one mediator.

We have compared the influence of sample preparation upon the level of surface expression of T, B, and NK cell-related antigens as assessed by flow cytometry. Lysed whole blood (WBL), Ficoll-Paque separated peripheral blood lymphocyte (F-PBL), and frozen peripheral blood lymphocyte (Fr-PBL) were analyzed via single-and multicolor flow cytometry. The percentage of positive cells expressing the individual cell surface markers was not affected by the procedure for preparation of WBL, F-PBL, and Fr-PBL. In contrast, the fluorescence intensity level of individual cell surface markers varied depending on cell preparation. By using Quantum Simply Cellular (QSC) microbeads, the antibody binding capacity (ABC) of single-color stained cells was quantified and compared. The amount of monoclonal antibody (MAb) anti-CD3-FITC bound to Fr-PBL (mean ABC = 137,040) was significantly higher (P < 0.001) that the amounts bound to WBL (mean ABC = 112,410) and F-PBL (mean ABC = 107,738). In multicolor analysis, the fluorescence intensity of CD3-FITC and CD4-FITC was significantly higher on Fr-PBL than on WBL and F-PBL; CD8-PE and CD20-PerCP was significantly higher on WBL. Furthermore, the intensity of CD3 and CD4 was different on T-cell subsets. The intensity of CD3 staining in three-color analysis was lower than with single-color staining using the same fluorochrome.We conclude that particularly the method of cell preparation but also the selection of MAb combinations may influence the level of staining of certain lymphocyte antigens. This may be of relevance in the analysis of cellular activation and regulation of differentiation.

One hundred and thirty-three colonic biopsies of proven cases of Shigella colitis were examined and post-mortem examinations were carried out on 29 fatal cases at the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B) hospital between 1988 and 1992. The distribution of pathological lesions and the spectrum of histopathological changes in the intestinal tract of these patients, and the features of intestinal and extra-intestinal complications of shigellosis are presented. Septicaemia, hyponatraemia, hypokalaemia and hypoglycaemia were present in a high percentage of these cases. All but two patients were malnorished at the time of autopsy. Shigellosis patients rapidly became hypoproteinaemic and were susceptible to other infections including opportunistic infections. Mortality amongst shigellosis patients admitted to our hospital continues to be high in spite of adequate antibiotic and supportive therapy.

Shigella species cause bacillary dysentery in humans by invading epithelial cells of the colonic mucosa leading to colonic epithelial cell destruction and inflammation. For further analysis of local gut inflammation, morphological changes and the potential involvement of mediators in regulatory mechanisms of cell activation and cell proliferation were studied immunohistochemically in rectal mucosal biopsies taken from patients during the acute phase of shigellosis and at convalescence. Rectal biopsies from 25 Shigella dysenteriae-1 and 10 Shigella flexneri-infected patients and from 40 controls were studied. The frequencies of proliferative cells (Ki67-positive cells), p53-immunostaining cells, and cells coexpressing Ki67 with CD3 or with p53 were analyzed. Immunostaining for the inducible nitric oxide synthase (iNOS) and the endothelial NOS was assessed. In addition, the frequencies of apoptotic cells and CD68 ؉ cells that engulf apoptotic cells were assessed. By morphological grading, 20% of the patients had advanced inflammation (grade 3) in the acute phase; mild inflammation (grade 1) was seen in 37% of the patients at convalescence as well as in 10% of the controls. The findings in the present study suggest that in the acute phase of shigellosis inflammation is characterized by increased cell turnover in the lamina propria (LP) and the epithelium, increased iNOS expression in the surface epithelium, and apoptosis, which seems to be associated with LP macrophages. The findings also suggest that neither p53 nor iNOS are important factors for the induction of apoptosis in shigellosis. Expression of p53 may be related to early cell activation in crypt epithelium. Moreover, there is an indication of an active, low-level inflammatory process at convalescence. The results thus indicate that Shigella-induced inflammation is associated with a complex series of cellular reactions in the rectal gut mucosa which persist long after clinical symptoms have resolved.