TNFSF13 insufficiency disrupts human colonic epithelial cell growth and associated B cell dynamics
Xianghui Ma, Shaneice K. Nettleford, Yuhua Tian, Noor Dawany, Ayano Kondo, Yalan Li, Kelly Maurer, Tatiana A. Karakasheva, Rawan Shraim, Patrick A. Williams, Louis R. Parham, Lauren A. Simon, Charles H. Danan, Maire A. Conrad, David A. Piccoli, Marcella Devoto, Neil Romberg, Kathleen E. Sullivan, Klaus H. Kaestner, Judith R. Kelsen, Kathryn E. Hamilton
Xianghui Ma, Shaneice K. Nettleford, Yuhua Tian, Noor Dawany, Ayano Kondo, Yalan Li, Kelly Maurer, Tatiana A. Karakasheva, Rawan Shraim, Patrick A. Williams, Louis R. Parham, Lauren A. Simon, Charles H. Danan, Maire A. Conrad, David A. Piccoli, Marcella Devoto, Neil Romberg, Kathleen E. Sullivan, Klaus H. Kaestner, Judith R. Kelsen, Kathryn E. Hamilton
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Research Article Gastroenterology Inflammation

TNFSF13 insufficiency disrupts human colonic epithelial cell growth and associated B cell dynamics

Abstract

Cytokines mediating epithelial and immune cell interactions modulate mucosal healing—a process that goes awry with chronic inflammation as in inflammatory bowel disease. TNFSF13 is a cytokine important for B cell maturation and function, but roles for epithelial TNFSF13 and putative contribution to inflammatory bowel disease are poorly understood. We evaluated functional consequences of a novel monoallelic TNFSF13 variant using biopsies, tissue-derived colonoids and induced pluripotent stem cell (iPSC)-derived colon organoids. TNFSF13 variant colonoids exhibited a >50% reduction in secreted TNFSF13, increased epithelial proliferation, and reduced apoptosis, which was confirmed in iPSC-derived colon organoids. Single cell RNA-sequencing and flow cytometry suggested FAS as the predominant colonic epithelial receptor for TNFSF13, which was confirmed by co-immunoprecipitation and binding assays. Imaging mass cytometry revealed an increase in epithelial-associated B cells in TNFSF13 variant colon tissue sections. Finally, TNFSF13 variant colonoids co-cultured with memory B cells demonstrated a reduction in immunoglobulin-producing plasma cells compared to control colonoid cocultures. Our findings support a role for epithelial TNFSF13 as a regulator of colonic epithelial growth and epithelial crosstalk with B cells.

Authors

Xianghui Ma, Shaneice K. Nettleford, Yuhua Tian, Noor Dawany, Ayano Kondo, Yalan Li, Kelly Maurer, Tatiana A. Karakasheva, Rawan Shraim, Patrick A. Williams, Louis R. Parham, Lauren A. Simon, Charles H. Danan, Maire A. Conrad, David A. Piccoli, Marcella Devoto, Neil Romberg, Kathleen E. Sullivan, Klaus H. Kaestner, Judith R. Kelsen, Kathryn E. Hamilton

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Figure 6

Epithelial-secreted TNFSF13 modulates differentiation of memory B cells to plasmablasts and plasma cells.

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Epithelial-secreted TNFSF13 modulates differentiation of memory B cells ...
(A) Schematic of human memory B cell and colonoid coculture experimental model. (B) Percentage of plasmablasts differentiated from sorted human memory B cells at day 8 via co-culture with equal numbers of control, VEO-IBD, or variant colonoids in IntestiCult:B cell media (1:1). (C) Percentage of plasmablasts at day 8 cultured in B cell media:conditioned media (1:1). (D and E) Percentage of (D) plasma cells and (E) IgA+ plasma cells differentiated at day 14 in B cell media:conditioned media (1:1). (F) IgA ELISA from differentiated B cell culture media at day 14. (G and H) Percentage of (G) plasma cells and (H) IgA+ plasma cells at day 14 when conditioned media mixture was added starting day 7 (cells cultured in B cell media alone days 0–6. (I) IgA ELISA from B cell culture media at day 14 with conditioned media added starting day 6 (n = 3 independent replicates). (J) ELISA for IgG in media from differentiated human memory B cells at day 14 and (K) at day 14 with delayed (day 6) conditioned media addition. (L and M) Percentage of (L) plasma cells and (M) IgA+ plasma cells at day 14 from IgG or FAS neutralizing antibody–treated (nFAS-treated) B cells cultured in control colonoid-conditioned media:B cell media (1:1). Three independent control colonoid lines used. Unless otherwise noted, n = 3 patient lines for control and VEO-IBD; n = 3 passages for variant colonoids and iPSC-organoids; n = 11 independent B cell donors. One-way ANOVA with multiple comparisons or 2-tailed Student’s t test used for statistical analysis. P values shown unless P > 0.05.