Heparan sulfate plays a central role in a dynamic in vitro model of protein-losing enteropathy.
Protein-losing enteropathy (PLE), the loss of plasma proteins through the intestine, is a symptom in ostensibly unrelated diseases. Emerging commonalities indicate that genetic insufficiencies predispose for PLE and environmental insults, e.g. viral infections and inflammation, trigger PLE onset. The specific loss of heparan sulfate (HS) from the basolateral surface of intestinal epithelial cells only during episodes of PLE suggests a possible mechanistic link. In the first tissue culture model of PLE using a monolayer of intestinal epithelial HT29 cells, we proved that HS loss directly causes protein leakage and amplifies the effects of the proinflammatory cytokine tumor necrosis factor alpha (TNFalpha). Here, we extend our in vitro model to assess the individual and combined effects of HS loss, interferon gamma (IFNgamma), TNFalpha, and increased pressure, and find that HS plays a central role in the patho-mechanisms underlying PLE. Increased pressure, mimicking venous hypertension seen in post-Fontan PLE patients, substantially increased protein leakage, but HS loss, IFNgamma, or TNFalpha alone had only minor effects. However, IFNgamma up-regulated TNFR1 expression and amplified TNFalpha-induced protein leakage. IFNgamma and TNFalpha compromised the integrity of the HT29 monolayer and made it more susceptible to increased pressure. HS loss itself compromises the integrity of the monolayer, amplifying the effects of pressure, but also amplifies the effects of both cytokines. In the absence of HS a combination of increased pressure, IFNgamma, and TNFalpha caused maximum protein leakage. Soluble heparin fully compensated for HS loss, providing a reasonable explanation for patient favorable response to heparin therapy.
Bode
Lars
L
Burnham Institute for Medical Research, Glycobiology and Carbohydrate Chemistry Program, La Jolla, California 92037, USA.
Murch
Simon
S
Freeze
Hudson H
HH
D000418:Albumins / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000378:metabolism; D005434:Flow Cytometry; D020022:Genetic Predisposition to Disease; D006493:Heparin / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D007249:Inflammation; D007371:Interferon-gamma / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D018613:Microscopy, Confocal; D015233:Models, Statistical; D011312:Pressure; D011485:Protein Binding; D011487:Protein Conformation; D011504:Protein-Losing Enteropathies / Q000378:metabolism*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation
D000418:Albumins; D016207:Cytokines; D006493:Heparin; D006497:Heparan Sulfate; D006861:Hydrogen Peroxide; D007371:Interferon-gamma; D014409:Tumor Necrosis Factor-alpha