Characterization of T-cell Receptor Repertoire in Inflamed Tissues of Patients with Crohnʼs Disease Through Deep Sequencing

Chapman, Christopher G.; Yamaguchi, Rui; Takagi, Kenji; Weidner, Jerome G.; Imoto, Seiya; Kwon, John H.; Fang, Hua; Yew, Poh Yin; Marino, Susana R.; Miyano, Satoru; Nakamura, Yusuke

doi:10.1097/mib.0000000000000752

Cited by 39 publications

(27 citation statements)

References 27 publications

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“…Indeed, a break in tolerance to commensal antigens is thought to induce chronic intestinal inflammation in humans 65 . Increased expansion and skewing of T cells has been reported in patients with IBD 39,66 and in the CD4 + CD45RB hi adoptive transfer mouse model 67 . The clonal expansion that we observed here in adoptive transferred Rag -/mice fed with diet 2019 suggest that these expanded clones are pathogenic, but further studies are required to test this hypothesis.…”

Section: Discussionmentioning

confidence: 99%

Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Chen

Iuga

Filho

et al. 2018

Preprint

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A wealth of experimental data points to immunological and environmental factors in the pathogenesis of inflammatory bowel disease (IBD). Here we study the role of IL-23, the microbiome, and the diet in the development of colitis. To promote IL-23 expression in vivo,we generated a mouse model in which IL-23 was conditionally expressed by CX 3 CR1 + myeloid cells, upon cyclic administration of tamoxifen in a specific diet (diet 2019). IL-23 expression induced an intestinal inflammatory disease that resembled ulcerative colitis in humans with cycles of acute disease and remission. The relapses were caused by the diet switch from the conventional diet used in our facility (diet 5053) to the diet 2019, and were not dependent on tamoxifen after the first cycle. The switch in the diet modified the microbiota, but did not alter the levels of IL-23. Colitis induction depended on the microbiota and required CD4 T lymphocytes. Colitis-inducing CD4 + T cells were found in the mesenteric lymph node and large intestine during remission and were able to trigger disease when transferred to lymphopenic mice, but only upon diet modification. The CD4 TCR repertoire in the diseased recipient Rag -/-mice had reduced diversity associated with the expansion of dominant T cell clones. These findings reveal a critical role for IL-23 in generation of a CD4 + T cell population in mice that is sensitive to a modification of intestinal bacterial flora subsequent to a dietary manipulation. Dietary changes occurring in the context of altered IL-23 expression may contribute to the onset and progression of IBD.

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Section: Discussionmentioning

confidence: 99%

Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Chen

Iuga

Filho

et al. 2018

Preprint

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“…Earlier studies have documented increased clonality of lamina propriaresiding T cells [13] and differential TCR-β chain usage between IBD subjects and non-IBD controls [13][14][15][16][17]. In recent years, only few studies using HTS of the TCR repertoire were performed in IBD patients, showing restricted TCR repertoires in CD and UC patients, although these analyses focused on the clones present in lamina propria [18][19][20][21]. We hypothesized that pediatric patients with UC possess unique T cell clones, and their expansion may correlate with degree of intestinal inflammation.…”

Section: Clinical and Experimental Immunologymentioning

confidence: 99%

Altered T cell receptor beta repertoire patterns in pediatric ulcerative colitis

Werner

Nunberg

Rechavi

et al. 2019

Clinical &Amp; Experimental Immunology

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The antigenic specificity of T cells occurs via generation and rearrangement of different gene segments producing a functional T cell receptor (TCR). High-throughput sequencing (HTS) allows in-depth assessment of TCR repertoire patterns. There are limited data concerning whether TCR repertoires are altered in inflammatory bowel disease. We hypothesized that pediatric ulcerative colitis (UC) patients possess unique TCR repertoires, resulting from clonotypical expansions in the gut. Paired blood and rectal samples were collected from nine newly diagnosed treatmentnaive pediatric UC patients and four healthy controls. DNA was isolated to determine the TCR-β repertoire by HTS. Significant clonal expansion was demonstrated in UC patients, with inverse correlation between clinical disease severity and repertoire diversity in the gut. Using different repertoire variables in rectal biopsies, a clear segregation was observed between patients with severe UC, those with mild-moderate disease and healthy controls. Moreover, the overlap between autologous blood-rectal samples in UC patients was significantly higher compared with overlap among controls. Finally, we identified several clonotypes that were shared in either all or the majority of UC patients in the colon. Clonal expansion of TCRβ-expressing T cells among UC patients correlates with disease severity and highlights their involvement in mediating intestinal inflammation.

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“…FCB has been used in combination with antibody staining for phosphoflow analysis and cytokine detection in T cell subpopulations . In our work, we sought to optimize a two‐dye FCB staining for routine phenotyping and analyze Vβ usage in CD4 + and CD8 + T cells, as oligoclonal expansion of Vβ groups occurs in autoimmune and malignant diseases . Antibodies to surface markers were tested using Combo 1 concentrations of DyLight 800 plus DyLight 350 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Optimization and standardization of fluorescent cell barcoding for multiplexed flow cytometric phenotyping

et al. 2017

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Fluorescent cell barcoding (FCB) is a cell-based multiplexing technique for high-throughput flow cytometry. Barcoded samples can be stained and acquired collectively, minimizing staining variability and antibody consumption, and decreasing required sample volumes. Combined with functional measurements, FCB can be used for drug screening, signaling profiling, and cytokine detection, but technical issues are present. We optimized the FCB technique for routine utilization using DyLight 350, DyLight 800, Pacific Orange, and CBD500 for barcoding six, nine, or 36 human peripheral blood specimens. Working concentrations of FCB dyes ranging from 0 to 500 μg/ml were tested, and viability dye staining was optimized to increase robustness of data. A five-color staining with surface markers for Vβ usage analysis in CD4+ and CD8+ T cells was achieved in combination with nine sample barcoding. We provide improvements of the FCB technique that should be useful for multiplex drug screening and for lymphocyte characterization and perturbations in the diagnosis and during the course of disease.

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Characterization of T-cell Receptor Repertoire in Inflamed Tissues of Patients with Crohnʼs Disease Through Deep Sequencing

Cited by 39 publications

References 27 publications

Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Altered T cell receptor beta repertoire patterns in pediatric ulcerative colitis

Optimization and standardization of fluorescent cell barcoding for multiplexed flow cytometric phenotyping

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Characterization of T-cell Receptor Repertoire in Inflamed Tissues of Patients with Crohnʼs Disease Through Deep Sequencing

Cited by 39 publications

References 27 publications

Diet Modifies Colonic Microbiota and CD4+ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Diet Modifies Colonic Microbiota and CD4+ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Altered T cell receptor beta repertoire patterns in pediatric ulcerative colitis

Optimization and standardization of fluorescent cell barcoding for multiplexed flow cytometric phenotyping

Contact Info

Product

Resources

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Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23

Diet Modifies Colonic Microbiota and CD4⁺ T Cell Repertoire to Trigger Flares in a Novel Model of Colitis Induced by IL-23