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Abstract: PO1985

Human Bladder Tissues Express Gb3 and Are Targeted by Shiga Toxin

Session Information

Category: Pediatric Nephrology

  • 1700 Pediatric Nephrology

Authors

  • Liu, Yang, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Thaker, Hatim, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Tian, Songhai, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Zhang, Jie, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Manion, John, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Wang, Siyu, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States
  • Wu, Shan, Jilin University, College of Basic Medical Sciences, Department of Pathology, Changchun, Jilin, China
  • Xu, Zhonggao, The First Hospital of Jilin University, Department of Nephrology, Changchun, Jilin, China
  • Dong, Min, Boston Children's Hospital Department of Urology, Boston, Massachusetts, United States

Group or Team Name

  • Dong Lab
Background

Shiga-toxin (Stx) producing E. coli associated hemolytic uremic syndrome (STEC-HUS) is the main cause of acute kidney injury (AKI) in children. Glycosphingolipid globotriaosylceramide (Gb3) is the receptor for Stx and determines the tissue specificity of Stx. Previous studies have detected Gb3 in glomeruli, proximal tubules and collecting duct in human kidney, but whether Gb3 is also expressed in other urinary tissues such as bladders remains unknown.

Methods

We first established and validated two complementary detection methods for Gb3 on cultured cells, one with a monoclonal Gb3 antibody and the other detecting Stx bound to cells using an antibody against Stx. Wide-type (WT) and A4GALT (α-1, 4-galactose transferase, Gb3 synthase) knockout (KO) human bladder cancer 5637 cells were utilized as cell models. Using these two approaches, we examined Gb3 expression in the urinary system of human and different animal models. In addition, Stx was administered i.p. in WT and A4GALT KO C57 mice to evaluate the key role of Gb3 in Stx pathogenesis in kidney and bladder tissues. Finally, we examined the impact of Stx on bladder tissues with injection of Stx into the lumen of bladders.

Results

Gb3 is detected on the cell surface of WT 5637 cells, but not on A4GALT KO cells. Consistently, Stx binds to WT 5637 cells, but not A4GALT KO cells. We found that normal human bladder connective tissue and vascular endothelial cells express Gb3, which mediates binding of Stx. Gb3 expression were detected in Yorkshire pig, New Zealand white rabbit, CD1 and C57 mouse, but not in Dolly sheep. Exposure to Stx induced a large amount of inflammatory cells infiltration in the bladder submucosa in C57 mice, and bladder transitional cells necrosis were detected by pathological evaluation; while the transitional cells of A4GALT KO mice showed no corresponding changes.

Conclusion

Here we report the novel finding that Gb3 is expressed within bladder tissues in humans, suggesting that bladder tissues could be a key target of Stx in humans. Furthermore, we found that Gb3 expression varies among different animal models, which will guide the selection of proper animal models for investigating the impact of Stx on urinary tissues. Finally, our study revealed that Gb3 mediate bladder inflammatory cell infiltration and transitional cell necrosis in Stx treated C57 WT mice.

Funding

  • Other NIH Support