• Title/Summary/Keyword: Intestinal cells

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Segmented Filamentous Bacteria Induce Divergent Populations of Antigen-Specific CD4 T Cells in the Small Intestine

  • Yi, Jaeu;Jung, Jisun;Han, Daehee;Surh, Charles D.;Lee, You Jeong
    • Molecules and Cells
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    • v.42 no.3
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    • pp.228-236
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    • 2019
  • CD4 T cells differentiate into $ROR{\gamma}t/IL$-17A-expressing cells in the small intestine following colonization by segmented filamentous bacteria (SFB). However, it remains unclear whether SFB-specific CD4 T cells can differentiate directly from naïve precursors, and whether their effector differentiation is solely directed towards the Th17 lineage. In this study, we used adoptive T cell transfer experiments and showed that naïve CD4 T cells can migrate to the small intestinal lamina propria (sLP) and differentiate into effector T cells that synthesize IL-17A in response to SFB colonization. Using single cell RT-PCR analysis, we showed that the progenies of SFB responding T cells are not uniform but composed of transcriptionally divergent populations including Th1, Th17 and follicular helper T cells. We further confirmed this finding using in vitro culture of SFB specific intestinal CD4 T cells in the presence of cognate antigens, which also generated heterogeneous population with similar features. Collectively, these findings indicate that a single species of intestinal bacteria can generate a divergent population of antigen-specific effector CD4 T cells, rather than it provides a cytokine milieu for the development of a particular effector T cell subset.

Inhibitory effects of calcium against intestinal cancer in human colon cancer cells and $Apc^{Min/+}$ mice

  • Ju, Jihyeung;Kwak, Youngeun;Hao, Xingpei;Yang, Chung S.
    • Nutrition Research and Practice
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    • v.6 no.5
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    • pp.396-404
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    • 2012
  • The aim of the study was to investigate the inhibitory effects of calcium against intestinal cancer in vitro and in vivo. We first investigated the effects of calcium treatment in HCT116 and HT29 human colon cancer cells. At the concentration range of 0.8-2.4 mM, calcium significantly inhibited cell growth (by 9-29%), attachment (by 12-26%), invasion (by 15-31%), and migration (by 19-61%). An immunofluorescence microscope analysis showed that the treatment with calcium (1.6 mM) for 24 h increased plasma membrane ${\beta}$-catenin but decreased nuclear ${\beta}$-catenin levels in HT29 cells. We then investigated the effect of dietary calcium on intestinal tumorigenesis in $Apc^{Min/+}$ mice. Mice received dietary treatment starting at 6 weeks of age for the consecutive 8 weeks. The basal control diet contained high-fat (20% mixed lipids by weight) and low-calcium (1.4 mg/g diet) to mimic the average Western diet, while the treatment diet contained an enriched level of calcium (5.2 mg calcium/g diet). The dietary calcium treatment decreased the total number of small intestinal tumors (by 31.4%; P < 0.05). The largest decrease was in tumors which were ${\geq}$ 2 mm in diameter, showing a 75.6% inhibition in the small intestinal tumor multiplicity (P < 0.001). Immunohistochemical analysis showed significantly reduced nuclear staining of ${\beta}$-catenin (expressed as nuclear positivity), but increased plasma membrane staining of ${\beta}$-catenin, in the adenomas from the calcium-treated groups in comparison to those from the control group (P < 0.001). These results demonstrate intestinal cancer inhibitory effects of calcium both in human colon cancer cells and $Apc^{Min/+}$ mice. The decreased ${\beta}$-catenin nuclear localization caused by the calcium treatment may contribute to the inhibitory action.

Characterization of valacyclovir transport mechanism across the intestinal epithelium

  • Han, H.;Covitz, M.;Surendran, N.;Stewart, B.;Amidon, G.L.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.119-119
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    • 1997
  • Valacyclovir is a L-valyl ester prodrug of acyclovir which is a highly effective and selective antiviral agent in the treatment of herpes virus diseases. Valacyclovir is rapidly and almost completely converted to acyclovir and increases the oral bioavailability of acyclovir three to five fold. However, the intestinal absorption mechanism of valacyclovir is not clear. If the improved absorption mechanism of valacyclovir is fully understood, it will provide a rationale of designing the amino acid ester prodrugs of polar drugs containing hydroxyl group. The main objective of our present study is to characterize the membrane transport mechanism of valacyclovir. Methods : Intestinal absorption of valacyclovir was investigated by using in-situ rat perfusion study and its wall permeability was estimated by modified boundary layer model. The membrane transport mechanism was also investigated through the uptake study in Caco-2 cells and in CHO-hPepTl cells. Results : In the rat perfusion study, the wall permeability of valacyclovir was ten times higher than acyclovir and showed concentration dependency, Valacyclovir also demonstrated a D,L stereo-selectivity with L-isomer having an approximately five-fold higher permeability than D-isomer. Mixed dipeptides and cephalexin, which are transported by dipeptide carriers, strongly competed with valacyclovir for the intestinal absorption, while L-valine did not show any competition with valacyclovir. This indicated that the intestinal absorption of valacyclovir could be dipeptide carrier-mediated. In addition, the competitive uptake study in Caco-2 cells presented that dipeptides reduced the valacyclovir uptake but valine did not. Also, in IC$\sub$50/ study, valacyclovir showed strong inhibition on the $^3$H-gly-sar uptake in CHO-hPepTl cells over-expressing a human intestinal peptide transporter. Taken together, the result from our present study indicated that valacyclovir utilized the peptide transporter for the intestinal absorption.

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Immunohistochemistry of Gastrointestinal Endocrine Cells in the Meckel′s Diverticulum of the Bean Goose, Anser fabalis Latham

  • Ku, Sae-Kwang;Lee, Hyeung-Sik;Park, Ki-Dae;Lee, Jae-Hyun
    • Animal cells and systems
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    • v.4 no.4
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    • pp.375-379
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    • 2000
  • The appearance of some gastrointestinal endocrine cells in the Meckel's diverticulum (MD) of the bean goose, Anser fabalis Latham was observed using specific antisera against serotonin, gastrin, cholecystokinin (CCK)-8, glucagon, secretin, somatostatin and human pancreatic polypeptide (HPP) with the peroxidase antiperoxidase (PAP) method. Among these specific antisera, serotonin-, gastrin-, CCK-8-, somatostatin- and HPP-immunoreactive cells were demonstrated in this study. Serotonin-, gastrin- and somatostatin-immunoreactive cells were detected at moderate frequency and CCK-8- and HPP-immunoreactive cells was rare and low frequencies, respectively. These immunoreactive cells were located in the superficial epithelium, intestinal crvpt and intestinal glands with spherical or spindle shaped cells having long cytoplasmic processes (open typed-cell). Mucosal layer of MD was composed of simple columnar epithelium and numerous intestinal glands. In addition, numerous lymphatic tissues were also demonstrated. In conclusion, histological profiles of MD were similar to any parts of the large intestine, especially the cecum, but the appearance, distribution and relative frequency of gastrointestinal endocrine cells were similar to those of upper parts of the small intestine. Although the exact digestive functions were unknown, the finding that the appearance, distribution and relative frequency of gastrointestinal endocrine cells in MD is similar to small intestine may be considered as distinct evidence that this organ may have some digestive functions.

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Establishing porcine jejunum-derived intestinal organoids to study the function of intestinal epithelium as an alternative for animal testing

  • Bo Ram Lee;Sun A Ock;Mi Ryung Park;Min Gook Lee;Sung June Byun
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.2-11
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    • 2024
  • Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Phosphorylation of tyrosine-14 on Caveolin-1 enhances lipopolysaccharide-induced inflammation in human intestinal Caco-2 cells

  • Gong Deuk Bae;Kyong Kim;Se-Eun Jang;Dong-Jae Baek;Eun-Young Park;Yoon Sin Oh
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.311-319
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    • 2023
  • Caveolin-1 (Cav-1) is the main structural component of the caveolae on the plasma membrane, which regulates various cellular processes, including cell growth, differentiation, and endocytosis. Although a recent study demonstrated that Cav-1 might be involved in diabetes-associated inflammation, its exact role in the intestine was unclear. In this study, we examined the intestinal expression of Cav-1 in diabetic conditions. We also investigated its effect on lipopolysaccharide (LPS)-induced inflammation by expressing this protein in human intestinal Caco-2 cells lacking Cav-1. We observed that increased Cav-1 levels and decreased expression of tight junction proteins affected intestinal permeability in high-fat diet-induced diabetic mice. When Caco-2 cells were treated with LPS, Cav-1 enhanced the NF-κB signaling. Moreover, LPS reduced the expression of tight junction proteins while it increased cell-cell permeability and reactive oxygen species generation in Caco-2 cells and this effect was amplified by cav-1 overexpression. LPS treatment promoted phosphorylation of tyrosine-14 (Y14) on Cav-1, and the LPS-induced NF-κB signaling was suppressed in cells expressing non-phosphorylatable Cav-1 (tyrosine-14 to phenylalanine mutant), which reduced intestinal barrier permeability. These results suggest that Cav-1 expression promotes LPS-induced inflammation in Caco-2 cells, and phosphorylation of Y14 on Cav-1 might contribute to the anti-inflammatory response in LPS-induced NF-κB signaling and cell permeability.

Context-Dependent Regulation of Type17 Immunity by Microbiota at the Intestinal Barrier

  • Begum Akuzum;June-Yong Lee
    • IMMUNE NETWORK
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    • v.22 no.6
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    • pp.46.1-46.25
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    • 2022
  • T-helper-17 (Th17) cells and related IL-17-producing (type17) lymphocytes are abundant at the epithelial barrier. In response to bacterial and fungal infection, the signature cytokines IL-17A/F and IL-22 mediate the antimicrobial immune response and contribute to wound healing of injured tissues. Despite their protective function, type17 lymphocytes are also responsible for various chronic inflammatory disorders, including inflammatory bowel disease (IBD) and colitis associated cancer (CAC). A deeper understanding of type17 regulatory mechanisms could ultimately lead to the discovery of therapeutic strategies for the treatment of chronic inflammatory disorders and the prevention of cancer. In this review, we discuss the current understanding of the development and function of type17 immune cells at the intestinal barrier, focusing on the impact of microbiota-immune interactions on intestinal barrier homeostasis and disease etiology.

Cells in the Respiratory and Intestinal Tracts of Chickens Have Different Proportions of both Human and Avian Influenza Virus Receptors

  • Kim, Jin-A;Ryu, Si-Yun;Seo, Sang-Heui
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.366-369
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    • 2005
  • Avian influenza viruses playa crucial role i,n the creation of human pandemic viruses. In this study, we have demonstrated that both human and avian influenza receptors exist in cells in the respiratory and intestinal tracts of chickens. We have also determined that primarily cultured chicken lung cells can support the replication of both avian and human influenza viruses.

Involvement of D2 Receptor on Dopamine-induced Action in Interstitial Cells of Cajal from Mouse Colonic Intestine

  • Zuoa, Dong Chuan;Shahia, Pawan Kumar;Choia, Seok;Jun, Jae-Yeoul;Park, Jong-Seong
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.218-226
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    • 2012
  • Dopamine is an enteric neurotransmitter that regulates gastrointestinal motility. This study was done to investigate whether dopamine modulates spontaneous pacemaker activity in cultured interstitial cells of Cajal (ICCs) from mouse using whole cell patch clamp technique, RT-PCR and live $Ca^{2+}$ imaging analysis. ICCs generate pacemaker inward currents at a holding potential of -70 mV and generate pacemaker potentials in current-clamp mode. Dopamine did not change the frequency and amplitude of pacemaker activity in small intestinal ICCs. On the contrary dopamine reduced the frequency and amplitude of pacemaker activity in large intestinal ICCs. RT-PCR analysis revealed that Dopamine2 and 4-receptors are expressed in c-Kit positive ICCs. Dopamine2 and 4 receptor agonists inhibited pacemaker activity in large intestinal ICCs mimicked those of dopamine. Domperidone, dopamine2 receptor antagonist, increased the frequency of pacemaker activity of large intestinal ICCs. In $Ca^{2+}$-imaging, dopamine inhibited spontaneous intracellular $Ca^{2+}$ oscillations of ICCs. These results suggest that dopamine can regulate gastrointestinal motility through modulating pacemaker activity of large intestinal ICCs and dopamine effects on ICCs are mediated by dopamine2 receptor and intracellular $Ca^{2+}$ modulation.

Functional Expression of a Dipeptide Transporter Obtained from Intestinal HT-29 Cells Using Xenopus Oocytes (장관세포인 HT-29에 존재하는 디펩티드수송체의 Xenopus oocyte에서의 발현)

  • Oh, Doo-Man;Yang, Chae-Ha
    • Journal of Pharmaceutical Investigation
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    • v.25 no.4
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    • pp.299-305
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    • 1995
  • Cloning the gene encoding a dipeptide transporter is necessary for understanding the absorption mechanism of peptides and peptide-like drugs in the gastrointestinal tract. Functional expression of a dipeptide transporter after microinjection into Xenopus laevis oocytes was performed using the mRNA purified from human intestinal HT-29 cells. Fifty nanoliters of purified mRNA (1 mg/mL) were microinjected into healthy oocytes followed by incubation for 4 days in order to express a dipeptide transporter. Functional expression was determined by a uptake assay using 10 Ci/mL $[^3H]-glycylsarcosine$, a dipeptide substate of the transporter. Seasonal variability and batch-to-batch variability were greater in summer. The usage of beveled micropipettes improves viability of oocytes at 4 days after microinjection. Expression of a dipeptide transporter in oocytes after microinjection of mRNA obtained from HT-29 cells was significantly larger than those after microinjection of water or mRNA obtained from the rabbit intestine.

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