Cytotoxicity of Shiga toxin for primary cultures of human colonic and ileal epithelial cells

Moyer, Mary Pat; Dixon, Patricia; Rothman, Sara W.; Brown, J E

doi:10.1128/iai.55.6.1533-1535.1987

Cited by 32 publications

(15 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Histopathological studies of tissue samples from loops infected with SC502, the Inv-Tox+ mutant, revealed an important inflammatory infiltrate in the lamina propria and major alterations predominantly at the tips of shortened villi. These results confirmed the cytotoxicity of Shiga toxin for enterocytes in vivo (27). However, the most striking feature was infiltration of the epithelial lining of erythrocytes which were shed into the lumen along with large amounts of mucus.…”

Section: Discussionsupporting

confidence: 77%

“…However, none of the experiments which have been carried out have compared isogenic mutants in a relevant cell assay system. Recent evidence indicates that Shiga toxin is cytotoxic for primary cultures of human colonic cells (27). Tissue invasion requires additional chro-mosomally encoded products, among which smooth lipopolysaccharide (43,57), a product of the Kcp locus (8,43), and aerobactin (24, 28) have been implicated.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1

1988

View full text Add to dashboard Cite

A Toxmutant of Shigella dysenteriae 1, SC501, was genetically engineered by cloning the Shiga toxin operon, inserting a cassette into the A subunit gene, and exchanging this in vitro-mutagenized sequence with the wild-type gene. SC501 produced a low amount of residual cytotoxicity which was not neutralized by a rabbit immune serum directed against Shiga toxin. Invasion of cultured cells demonstrated that Shiga toxin had no effect on the rate of intracellular growth of bacteria or on the rapid killing of invaded host cells. On the other hand, several significant differences were observed in macaque monkeys infected intragastrically with either the wild-type strain or its mutant. The production of Shiga toxin by the invading strain was correlated with the presence of blood within stools, a sharp drop in blood polymorphonuclear cells, and histopathological alterations, such as the destruction of capillary vessels within the connective tissue of the colonic mucosa, severe inflammatory vasculitis of the peritoneal mesothelium, and major efflux of inflammatory cells to the intestinal lumen. It is proposed that Shiga toxin influences the severity of bacillary dysentery by inducing colonic vascular damage, which accounts for bloody stools, intestinal ischemia, and inflation of a polymorphonuclear intestinal compartment during the infectious process.

show abstract

Section: Discussionsupporting

confidence: 77%

mentioning

confidence: 99%

Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1

1988

View full text Add to dashboard Cite

show abstract

“…Analysis of the relative titre of LPS-specific faecal s-IgA revealed an increase and a prolonged response in the intermediate and severe disease groups even after the non-specific s-IgA level had dropped. The prolonged specific s-IgA produc- (36).…”

Section: Discussionmentioning

confidence: 99%

Semiquantitative estimation of Shigella antigen‐specific antibodies: correlation with disease severity during shigellosis

Islam

Wretlind

Hammarström

et al. 1996

APMIS

View full text Add to dashboard Cite

Systemic and mucosal antibody responses during shigellosis were analysed and correlated with clinical severity. Patients infected with Shigella dysenteriae type 1 (SDIP), and S. flexneri (SFIP) were grouped according to disease severity. The disease was more severe in SDIP than in SFIP. Higher levels of Shigella antigen-specific serum antibodies were associated with severe disease. In contrast, decreased serum levels of IgG, IgM and total protein were associated with severe disease. Faecal levels of total IgA, secretory IgA, albumin, and LPS specific s-IgA were increased in severe disease. The results of the study indicate that the level of humoral immune response correlates with the disease severity in shigellosis and that the local immune system in the mucosa is directly engaged in the response. This is also reflected at the systemic level by increased serum levels of specific antibodies and non-specific inflammatory parameters.

show abstract

“…Inhibition of protein synthesis. Protein synthesis was determined by measuring incorporation of [3H]L-leucine (27,29). Porcine aortic endothelial (PEC-15 [39]) and African green monkey kidney cells (Vero cells; ATCC CCL81; American Type Culture Collection, Rockville, Md.)…”

Section: Methodsmentioning

confidence: 99%

“…Monolayers from replicate plates were harvested with 0.05% trypsin-0.02% EDTA solution containing 0.04% (wt/ vol) trypan blue to determine cell death (26,27). Cell death was expressed as a percentage of that of the PBS control, and values were then adjusted so that the control value was 0%.…”

Section: Methodsmentioning

confidence: 99%

Physicochemical and biological properties of purified Escherichia coli Shiga-like toxin II variant

et al. 1991

View full text Add to dashboard Cite

Purified Escherichia coli Shiga-like toxin II variant (SLT-IIv) was characterized with regard to selected physical, chemical, and biological properties. N-terminal amino acid sequencing confirmed the identities of 33,000-, 27,500-, and 7,500-molecular-weight (MW) bands seen on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of purified SLT-IIv as the A subunit, Al fragment, and B subunit, respectively. The arginine-serine bond between amino acids 247 and 248 in the A subunit was determined to be the site for proteolytic cleavage into Al and A2 fragments. As with other SLTs, gel filtration chromatography of SLT-IIv gave estimates of the MW of holotoxin that were variable and less than predicted for a 1-A-subunit-5-B-subunit configuration. The MWs were estimated to be 40,000 and 43,000 by Sephacryl S-100 and Sephadex G-100 and <2,000 by Bio-Sil Sec-250 gel filtration chromatography. The isoelectric point of SLT-IIv holotoxin was 9.0. Cytotoxicity of SLT-IIv was destroyed by heating at 65°C for 30 min and by incubation with 2-mercaptoethanol and dithiothreitol, but it increased 30-fold by incubation with trypsin, chymotrypsin, or pepsin and 2-fold by incubation with thermolysin. SLT-IIv cytotoxic activity was stable at neutral and alkaline pH values but was lost at pHs 3, 4, and 5. SLT-IIv was stable in fluid from the anterior and posterior small intestines of pigs but was not enterotoxic in pig intestinal loops. The smallest doses of SLT-IIv that inhibited protein synthesis in porcine endothelial cells and Vero cells were 0.1 ng and 0.1 fg, respectively. 1300 on August 1, 2020 by guest

show abstract

Cytotoxicity of Shiga toxin for primary cultures of human colonic and ileal epithelial cells

Cited by 32 publications

References 20 publications

Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1

Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1

Semiquantitative estimation of Shigella antigen‐specific antibodies: correlation with disease severity during shigellosis

Physicochemical and biological properties of purified Escherichia coli Shiga-like toxin II variant

Contact Info

Product

Resources

About