Human small-intestinal epithelium contains functional natural killer lymphocytes

León, Francisco; Roldán, Ernesto; Sánchez, Lidia; Camarero, C.; Bootello, A; Roy, Garbiñe

doi:10.1016/s0016-5085(03)00886-2

Cited by 101 publications

(74 citation statements)

References 73 publications

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“…Intraepithelial lymphocytes (IEL), mainly consisting of CD8 + T cells, but also NK cells, reside within this epithelial layer, and a variety of leukocytes, including B cells, T cells, APC, and NK cells can be found in the underlying lamina propria (Fig. 1) [38][39][40]. Furthermore, the intestinal mucosa contains organized clusters of lymphoid tissue such as cryptopatches, isolated lymphoid follicles (ILF), and Peyer's patches [29].…”

Section: The Intestinal Mucosamentioning

confidence: 99%

Natural killer (NK) and NK-like cells at mucosal epithelia: Mediators of anti-microbial defense and maintenance of tissue integrity

Fuchs

Colonna

2011

European Journal of Microbiology and Immunology

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Section: The Intestinal Mucosamentioning

confidence: 99%

Natural killer (NK) and NK-like cells at mucosal epithelia: Mediators of anti-microbial defense and maintenance of tissue integrity

Fuchs

Colonna

2011

European Journal of Microbiology and Immunology

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“…In addition to the human surgical specimens reported here, we have also utilized this procedure with small bowel biopsies, obtaining a sufficient amount of IEL for molecular biology and functional assays with a minimum of three biopsies. 13 Figure 1 Schematic representation of the isolation procedure. The first two dot plots in the upper left corner illustrate the epithelial mixture of cells ('Whole Epithelial Cell Suspension') obtained after DTT/EDTA treatment of an ileal mucosa specimen.…”

Section: Isolation Of Iel By Annexin-beadsmentioning

confidence: 99%

“…13 The majority of CD3 þ IEL are CD8 þ Tc. 14 The presence of CD19 þ Bc (42%) or a high proportion of CD4 þ Tc (415%) in the IEL preparation after DTT/EDTA treatment is suggestive of contamination with lamina propria lymphocytes, and such cases were not included in our study.…”

Section: Phenotype and Functionality Of Iel After Separationmentioning

confidence: 99%

“…As it can be observed in Figure 4, the incorporation of propidium iodide (PI) by K562 cells, a sign of cell death, was enhanced in a dose-dependent fashion by coculture with viable purified IEL. This type of cytolysis is mediated by perforins produced by IEL after contact with K562 6,13 and constitutes an indication of the functional capacity of IEL.…”

Section: Phenotype and Functionality Of Iel After Separationmentioning

confidence: 99%

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Isolation of human small bowel intraepithelial lymphocytes by Annexin V-coated magnetic beads

León

Roy

2004

Laboratory Investigation

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Human intestinal intraepithelial lymphocytes (IEL) are important effector cells of the mucosal immune system and their study is hampered by the difficulty of their isolation. The molecular study of enriched samples of IEL is mandatory in the diagnosis of enteropathy-associated T-cell lymphoma and refractory celiac sprue. In order to isolate human small bowel IEL, we took advantage of the stress that intestinal epithelial cells (IEC) suffer during the conventional initial steps of IEL isolation, which induces their apoptosis but not that of IEL. After cell individualization by dithiothreitol and ethylenediamine tetraacetic acid, two-thirds of human IEC can be stained with Annexin-V due to their surface exposure of phosphatidyl serine, a sign of apoptosis. This percentage increases to 95% after performing a density gradient to enrich for IEL. This allows for the use of Annexin-Vcoated magnetic beads, originally designed for the removal of dead cells from cell cultures, to obtain 495% pure, 99% viable and untouched IEL after two rounds of depletion. This simple procedure has proven useful for the isolation of human IEL for functional and molecular studies and can conceivably facilitate the diagnosis of intestinal lymphoid malignancies that rely upon the study of pure IEL preparations.

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“…A family of NK cells is definitely innate lymphocyte effector cells, but is distinct from a classic family of T cells in that functional cytolysistriggering receptors such as NKp46, NKp44 and NKp30 are apparently expressed [5][6][7]. Despite such puzzling phenotypes, a general consensus is that NK cells are composed of a large family of lymphocytes, which includes NK-T, NK-CTL, CD8αα+ NK, CD3+ NK, decidual and intestinal NK cells [1,3,4,[8][9][10]. Because of their ability to purge transformed cells in vitro, the viable NK cells and their genetically engineered variants [for example, ref [11][12][13] may have potential values in clinical applications such as biological therapies.…”

Section: Introductionmentioning

confidence: 99%

Genomic analysis of CD8+ NK/T cell line, ‘SRIK-NKL’, with array-based CGH (aCGH), SKY/FISH and molecular mapping

et al. 2008

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We performed aCGH, SKY /FISH, molecular mapping and expression analyses on a permanent CD8+ NK/T cell line, 'SRIK-NKL' established from a lymphoma (ALL) patient, in attempt to define the fundamental genetic profile of its unique NK phenotypes. aCGH revealed hemizygous deletion of 6p containing genes responsible for hematopoietic functions. The SKY demonstrated that a constitutive reciprocal translocation, rcpt(5;14)(p13.2;q11) is a stable marker. Using somatic hybrids containing der(5) derived from SRIK-NKL, we found that the breakpoint in one homologue of no. 5 is located upstream of IL7R and also that the breakpoint in no. 14 is located within TRA@. The FISH analysis using BAC which contains TRA@ and its flanking region further revealed a ~231 kb deletion within 14q11 in the der(5) but not in the normal homologue of no. 14. The RT-PCR analysis detected mRNA for TRA@ transcripts which were extending across, but not including, the deleted region. IL7R was detected at least at mRNA levels. These findings were consistent with the immunological findings that TRA@ and IL7R are both expressed at mRNA levels and TRA@ at cytoplasmic protein levels in SRIK-NKL cells. In addition to rept(5;14), aCGH identified novel copy number abnormalities suggesting that the unique phenotype of the SRIK-NKL cell line is not solely due to the TRA@ rearrangement. These findings provide supportive evidence for the notion that SRIK-NKL cells may be useful for studying not only the function of NK cells but also genetic deregulations associated with leukemiogenesis.

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Human small-intestinal epithelium contains functional natural killer lymphocytes

Cited by 101 publications

References 73 publications

Natural killer (NK) and NK-like cells at mucosal epithelia: Mediators of anti-microbial defense and maintenance of tissue integrity

Natural killer (NK) and NK-like cells at mucosal epithelia: Mediators of anti-microbial defense and maintenance of tissue integrity

Isolation of human small bowel intraepithelial lymphocytes by Annexin V-coated magnetic beads

Genomic analysis of CD8+ NK/T cell line, ‘SRIK-NKL’, with array-based CGH (aCGH), SKY/FISH and molecular mapping

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