Specific Norovirus Interaction with Lewis x and Lewis a on Human Intestinal Inflammatory Mucosa during Refractory Inflammatory Bowel Disease

Abstract

Inflammatory bowel disease (IBD), which includes Crohn’s disease (CD) and ulcerative colitis (UC), is related to immunological and microbial factors, with the possible implication of enteric viruses. We characterized the interaction between human noroviruses (HuNoVs) and blood group antigens in refractory CD and UC using HuNoV virus-like particles (VLPs) and histological tissues. Immunohistochemistry was conducted on inflammatory tissue samples from the small intestine, colon, and rectum in 15 CD and 9 UC patients. Analysis of the regenerative mucosa of the colon and rectum revealed strong expression of sialylated Lewis a (sLea) and Lewis x (sLex) antigens and HuNoV VLP binding in the absence of ABO antigen expression in both UC and CD. Competition experiments using sialidase, lectins, and monoclonal antibodies demonstrated that HuNoV attachment mostly involved Lea and, to a lesser extent, Lex moieties on regenerative mucosa in both UC and CD. Further studies will be required to understand the implications of specific HuNoV binding to regenerative mucosa in refractory IBD. IMPORTANCE Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), are progressive diseases affecting millions of people each year. Flare-ups during IBD result in severe mucosal alterations of the small intestine (in CD) and in the colon and rectum (in CD and UC). Immunohistochemical analysis of CD and UC samples showed strong expression of known tumoral markers sialyl Lewis a (CA19.9) and sialyl Lewis x (CD15s) antigens on colonic and rectal regenerative mucosa, concurrent with strong human norovirus (HuNov) VLP GII.4 affinity. Sialidase treatment and competition experiments using histo-blood group antigen (HBGA)-specific monoclonal antibodies and lectins clearly demonstrated the implication of the Lewis a moiety and, to a lesser extent, the Lewis x moiety in HuNov recognition in regenerative mucosa of CD and UC tissues. Further studies are required to explore the possible implications of enteric viruses in the impairment of epithelial repair and dysregulation of inflammatory pathways during severe IBD.

Finding the sweet spot: glycosylation mediated regulation of intestinal inflammation

“…Importantly, increased CD44v6 expression in IBD correlates with increased inflammatory infiltrates and with increased mucosal tissue damage 42 . More recent immunohistological and liquid chromatography mass spectrometric glycomics analyses have demonstrated upregulated expression of sialylated Lewis family O-glycans on epithelial CD44v6 and on Muc2 in inflamed human and murine colonic mucosa 33 , 34 , 82 – 85 . Furthermore, recent functional studies have shown that antibody targeting of specific sialyl-Lewis glycans on epithelial CD44v6 blocks human neutrophil transepithelial migration (TEpM), protects against DSS-induced colitis in vivo and promotes epithelial wound closure following colonic injury 33 , 34 , 86 .…”

Epidemiological Impact of GII.17 Human Noroviruses Associated With Attachment to Enterocytes

“… b The values are given in resonance unit (RU). c For duodenal tissues, the intensity of the signal is graded based upon a positive control from a previous study ( Tarris et al, 2021 ). +++, strong; ++, intermediate; +, low/focal; and −, negative.…”

“…The samples were taken during routine surgical procedures performed at the University Hospital of Dijon (France). The detection of A, H, and Le b antigens and the experimental conditions for VLP binding on histological sections were described previously using VLPs diluted at 5 μg/ml ( Marionneau et al, 2002 ; Tarris et al, 2019 ); ( Tarris et al, 2021 ). Attached VLPs were detected with GII.17-specific polyclonal serum described above.…”

Intestinal norovirus binding patterns in non-secretor individuals

“…Four µm-thick microtome sections were cut from FFPE samples. Sections were deparaffinized, quenched and pretreated for endogenous peroxidase inhibition, as previously described (15). Five µg/ml of in-house purified GII.4, GII.3 or GII.17 VLP diluted in PBS with BSA 1% were incubated on each slide overnight at 4°C.…”

“…During flare ups, marked structural alterations are seen in the inflamed mucosae (16). Blood group ABO(H) antigens are usually not displayed at the surface of inflammatory and regenerative colonic and rectal mucosa, whereas a strong expression of Sialyl-Le a (sLe a ) (also known as CA19.9) and to a lesser extent Sialyl-Le x (sLe x ) (also known as CD15s) is observed (15,(22)(23)(24). The absence of sLe a antigen expression on quiescent mucosa contrasts with its reversible expression in regenerative mucosa and its persistent expression in dysplasia (22).…”