Isolation and characterization of lymphocytes from bovine intestinal epithelium and lamina propria

Clough, Ellen R.; Dean, Hansi

doi:10.1016/0165-2427(88)90045-1

Cited by 2 publications

(6 citation statements)

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“…The inguinal lymph nodes (IGLN) were cut into 2–3 mm 3 fragments and subjected to mechanical disruption and digestion in 5 mL of RP10 medium supplemented with 400 U/mL of V Collagenase, 0.1 mg/mL of DNase, and 2.5 U/mL of hyaluronidase at a temperature of 37 °C for a duration of 2 h [ 15 , 21 ]. The single-cell suspension obtained was subjected to cell counting and antibody staining.…”

Section: Methodsmentioning

confidence: 99%

“…The T-IEL isolation protocol for the abomasum was adapted from [ 21 ], and for the jejunum and ileum, it was based on [ 15 ]’s method for mucosal small intestine T-IEL isolation. Both procedures included minor modifications.…”

Section: Methodsmentioning

confidence: 99%

“…Intraepithelial T lymphocytes (T-IELs), which make up about 90% of all intestinal intraepithelial lymphocytes and 50–60% of total T lymphocytes in humans, play a critical role in pathogen clearance and gut homeostasis, and could be a central target for developing intervention strategies against GI diseases [ 1 , 2 , 3 , 4 , 5 , 6 , 7 ]. However, our understanding of the composition and function of bovine T-IELs is limited by technological constraints and the delayed availability of specific research tools [ 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

“…Historically, studies on T-IELs in cattle have been limited by resources and technology. Recent advancements and improved research tools provide a fresh opportunity to deepen our understanding of bovine T-IELs [ 10 , 11 , 15 , 16 , 17 , 18 , 19 , 20 , 21 ]. We investigated the T-IELs in the bovine small intestine, particularly in the jejunum, using flow cytometry.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of Bovine Intraepithelial T Lymphocytes in the Gut

Hada,

Li,

Kandel

et al. 2023

Pathogens

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Intraepithelial T lymphocytes (T-IELs), which constitute over 50% of the total T lymphocytes in the animal, patrol the mucosal epithelial lining to defend against pathogen invasion while maintaining gut homeostasis. In addition to expressing T cell markers such as CD4 and CD8, T-IELs display T cell receptors (TCR), including either TCRαβ or TCRγδ. Both humans and mice share similar T-IEL subsets: TCRγδ+, TCRαβ+CD8αα+, TCRαβ+CD4+, and TCRαβ+CD8αβ+. Among these subsets, human T-IELs are predominantly TCRαβ+ (over 80%), whereas those in mice are mostly TCRγδ+ (~60%). Of note, the majority of the TCRγδ+ subset expresses CD8αα in both species. Although T-IELs have been extensively studied in humans and mice, their profiles in cattle have not been well examined. Our study is the first to characterize bovine T-IELs using flow cytometry, where we identified several distinct features. The percentage of TCRγδ+ was comparable to that of TCRαβ+ T-IELs (both ~50% of CD3+), and the majority of bovine TCRγδ+ T-IELs did not express CD8 (CD8−) (above 60%). Furthermore, about 20% of TCRαβ+ T-IELs were CD4+CD8αβ+, and the remaining TCRαβ+ T-IELs were evenly distributed between CD4+ and CD8αβ+ (~40% of TCRαβ+ T-IELs each) with no TCRαβ+CD8αα+ identified. Despite these unique properties, bovine T-IELs, similar to those in humans and mice, expressed a high level of CD69, an activation and tissue-retention marker, and a low level of CD62L, a lymphoid adhesion marker. Moreover, bovine T-IELs produced low levels of inflammatory cytokines such as IFNγ and IL17A, and secreted small amounts of the immune regulatory cytokine TGFβ1. Hence, bovine T-IELs’ composition largely differs from that of human and mouse, with the dominance of the CD8− population among TCRγδ+ T-IELs, the substantial presence of TCRαβ+CD4+CD8αβ+ cells, and the absence of TCRαβ+CD8αα+ T-IELs. These results provide the groundwork for conducting future studies to examine how bovine T-IELs respond to intestinal pathogens and maintain the integrity of the gut epithelial barrier in animals.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of Bovine Intraepithelial T Lymphocytes in the Gut

Hada,

Li,

Kandel

et al. 2023

Pathogens

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“…Techniques for the isolation of the IELs of cattle have been developed (Nagi & Babiuk, 1987;Clough & Dean, 1988), and preliminary characterization of this population initiated (Nagi & Babiuk, 1989). However, the function of these cells in the host remains to be resolved.…”

Section: Introductionmentioning

confidence: 99%