αEβ7 Integrin Research Articles

A-213 αEβ7 Integrin (CD103) expression in both early and advanced stage of Cutaneous T cell Lymphoma's (CTCL)

A-213 αEβ7 Integrin (CD103) expression in both early and advanced stage of Cutaneous T cell Lymphoma's (CTCL)

Expression of integrin α4β1 and α4β7 on B cells correlates with autoimmune responses in Graves’ disease

BackgroundIntegrins are upregulated on endothelial cells and T-lymphocytes in autoimmune thyroid disease (AITD), potentially contributing to immune response localization. The role of integrins on B-cells in AITD remains unclear. MethodsPeripheral blood samples were collected from healthy controls (n = 56), patients with Graves’ disease (GD) (n = 37) and Hashimoto’s thyroiditis (HT) (n = 52). Ultrasound-guided fine-needle aspiration (FNA) of the thyroid was performed in patients with non-autoimmune thyroid disease (nAITD) (n = 19), GD (n = 11), and HT (n = 40). Integrins α4β7, α4β1, and αEβ7 in B cells were measured by flow cytometry. Serum zonulin levels were quantified via ELISA. Associations of integrins on B cells with thyroid hormones, thyroid autoantibodies, AITD duration, and zonulin were analyzed. ResultsHT patients exhibited lower α4β7 and higher α4β1 expression on B cells compared to healthy controls and GD patients. While α4β7 was predominant on circulating B cells, the dominant integrin expressed on intrathyroidal B cells varied with specific thyroid diseases. In GD patients, α4β7 and α4β1 expression on circulating B cells correlated positively and negatively with thyroid function and thyroid stimulating immunoglobulins (TSI) levels, respectively. Intrathyroidal α4β1+ B cells positively correlated with TSH levels in HT patients. Additionally, serum zonulin was elevated in HT patients, and intrathyroidal α4β7+ B cells and α4β1+ B cells correlated negatively and positively with zonulin levels, respectively. Integrin αEβ7 on B cells showed no significant association with AITD. ConclusionIntegrins expressed on B cells potentially play a role in the pathogenesis of AITD and might serve as immune biomarkers for the disease.

Old and New Biologics and Small Molecules in Inflammatory Bowel Disease: Anti Integrins

Recently, novel biologics or small molecular drugs have been introduced for overcoming the unmet needs associated with anti-tumor necrosis factor α agents for inflammtory bowel disease (IBD) treatment. Among these novel drugs, anti integrin agents block leukocyte trafficking to the intestine by blocking the interaction between integrin and cell adhesion molecules. Vedolizumab (anti-α4β7) is most widely used anti-integrin approved in both ulcerative colitis and Crohn's disease .It has been shown to be effective in both induction and maintenance therapy with a favorable safety profile due to gut selectivity. Several models incorporating clinical, genetic, immune and gut microbial markers to predict response to vedolizumab in IBD have been developed. Etrolizumab (anti-β7) blocks leukocyte trafficking via α4β7 and cell adhesion via αEβ7 integrins. In addition, the introduction of subcutaneous vedolizumab showed similar efficacy and safety with improved patients' convenience. Other investigational anti-integrin therapies include abrilumab (anti-α4β7 IgG2), PN-943 (orally administered and gut-restricted α4β7 antagonist peptide), AJM300 (orally active small molecule inhibitor of α4), and ontamalimab (anti-MAdCAM-1 IgG).

Cryo-EM structure of I domain-containing integrin αEβ7

The integrin family is a transmembrane receptor that plays critical roles in the cell-cell and cell-extracellular matrix adhesion, signal transduction such as cell cycle regulation, organization of the intracellular cytoskeleton, and immune responses. Consequently, dysfunction of integrins is associated with a wide range of human diseases, including cancer and immune diseases, which makes integrins therapeutic targets for drug discovery. Here we report the cryo-EM structure of the human α-I domain-containing full-length integrin αEβ7, which is expressed in the leukocytes of the immune system and a drug target for inflammatory bowel disease (IBD). The structure reveals the half-bent conformation, an intermediate between the close and the open conformation, while the α-I domain responsible for the ligand binding covers the headpiece domain by a unique spatial arrangement. Our results provide the structural information for the drug design targeting IBD.

Role of chitosan in intestinal integrity: TLR4 and IFNAR signaling in the induction of E-cadherin and CD103 in mice

Chitin and its derivative chitosan (Q) are abundant structural elements in nature. Q has modulatory and anti-inflammatory effects and also regulates the expression of adhesion molecules. The interaction between cells expressing the αEβ7 integrin and E-cadherin facilitates tolerogenic signal transmission and localization of lymphocytes at the frontline for interaction with luminal antigens. In this study we evaluated the ability of orally administered Q to stimulate E-cadherin and CD103 expression in vitro and in vivo. Our findings show that Q promoted epithelial cell migration, accelerated wound healing and increased E-cadherin expression in IEC-18 cells and isolated intestinal epithelial cells (IECs) after Q feeding. The upregulation of E-cadherin was dependent on TLR4 and IFNAR signaling, triggering CD103 expression in lymphocytes. Q reinforced the E-cadherin-αEβ7 axis, crucial for intestinal barrier integrity and contributed to the localization of lymphocytes on the epithelium.

Anti-β7 integrin treatment impedes the recruitment on non-classical monocytes to the gut and delays macrophage-mediated intestinal wound healing.

Closing mucosal defects to reach mucosal healing is an important goal of therapy in inflammatory bowel disease (IBD). Among other cells, monocyte-derived macrophages are centrally involved in such intestinal wound healing. We had previously demonstrated that the anti-α4β7 integrin antibody vedolizumab blocks the recruitment of non-classical monocytes as biased progenitors of wound healing macrophages to the gut and delays wound healing. However, although important for the interpretation of disappointing results in recent phase III trials in IBD, the effects of the anti-β7 antibody etrolizumab on wound healing are unclear so far. We analyzed the expression of etrolizumab targets on human and mouse monocyte subsets by flow cytometry and assessed their function in adhesion and homing assays. We explored wound-associated monocyte recruitment dynamics with multi-photon microscopy and compared the effects of etrolizumab and vedolizumab surrogate (-s) antibodies on experimental wound healing and wound-associated macrophage abundance. Finally, we investigated wound healing macrophage signatures in the large intestinal transcriptome of patients with Crohn's disease treated with etrolizumab. Human and mouse non-classical monocytes expressed more αEβ7 integrin than classical monocytes and were a target of etrolizumab-s, which blocked non-classical monocyte adhesion to MAdCAM-1 and E-Cadherin as well as gut homing in vivo. Intestinal wound healing was delayed on treatment with etrolizumab-s along with a reduction of peri-lesional wound healing macrophages. Wound healing macrophage signatures in the colon of patients with Crohn's disease were substantially down-regulated on treatment with etrolizumab, but not with placebo. Combined blockade of αEβ7 and α4β7 with etrolizumab seems to exceed the effect of anti-α4β7 treatment on intestinal wound healing, which might help to inform further investigations to understand the recent observations in the etrolizumab phase III trial program.

The Underappreciated Role of Secretory IgA in IBD.

Eighty percent of antibody secreting cells (ASCs) are found in the intestine, where they produce grams of immunoglobulin (Ig) A daily. immunoglobulin A is actively transcytosed into the lumen, where it plays a critical role in modulating the gut microbiota. Although loss of immune tolerance to bacterial antigens is the likely trigger of the dysregulated immune response that characterizes inflammatory bowel disease (IBD), little effort has been placed on understanding the interface between B cells, IgA, and the microbiota during initiation or progression of disease. This may be in part due to the misleading fact that IgA-deficient humans are mostly asymptomatic, likely due to redundant role of secretory (S) IgM. Intestinal B cell recruitment is critically dependent on integrin α4β7-MAdCAM-1 interactions, yet antibodies that target α4β7 (ie, vedolizumab), MAdCAM-1 (ie, ontamalimab), or both β7 integrins (α4β7 and αE [CD103] β7; etrolizumab) are in clinical use or development as IBD therapeutics. The effect of such interventions on the biology of IgA is largely unknown, yet a single dose of vedolizumab lowers SIgA levels in stool and weakens the oral immunization response to cholera vaccine in healthy volunteers. Thus, it is critical to further understand the role of these integrins for the migration of ASC and other cellular subsets during homeostasis and IBD-associated inflammation and the mode of action of drugs that interfere with this traffic. We have recently identified a subset of mature ASC that employs integrin αEβ7 to dock with intestinal epithelial cells, predominantly in the pericryptal region of the terminal ileum. This role for the integrin had not been appreciated previously, nor the αEβ7-dependent mechanism of IgA transcytosis that it supports. Furthermore, we find that B cells more than T cells are critically dependent on α4β7-MAdCAM-1 interactions; thus MAdCAM-1 blockade and integrin-β7 deficiency counterintuitively hasten colitis in interleukin-10-deficient mice. In both cases, de novo recruitment of IgA ASC to the intestinal lamina propria is compromised, leading to bacterial overgrowth, dysbiosis, and lethal colitis. Thus, despite the safe and effective use of anti-integrin antibodies in patients with IBD, much remains to be learned about their various cell targets.

Etrolizumab as induction and maintenance therapy in patients with moderately to severely active Crohn's disease (BERGAMOT): a randomised, placebo-controlled, double-blind, phase 3 trial

Etrolizumab as induction and maintenance therapy in patients with moderately to severely active Crohn's disease (BERGAMOT): a randomised, placebo-controlled, double-blind, phase 3 trial

The multifunctional nature of CD103 (αEβ7 integrin) signaling in tissue-resident lymphocytes.

Intestinal tissue-resident lymphocytes are critical for maintenance of the mucosal barrier and to prevent enteric infections. The activation of these lymphocytes must be tightly regulated to prevent aberrant inflammation and epithelial damage observed in autoimmune diseases, yet also ensure that antimicrobial host defense remains uncompromised. Tissue-resident lymphocytes express CD103, or αE integrin, which dimerizes with the β7 subunit to bind to E-cadherin expressed on epithelial cells. Although the role of CD103 in homing and retention of lymphocytes to and within peripheral tissues has been well characterized, the molecular signals activated following CD103 engagement remain understudied. Here, we highlight recent studies that elucidate the functional contribution of CD103 in various lymphocyte subpopulations, either as an independent signaling molecule or in the context of TCR co-stimulation. Finally, we will discuss the gaps in our understanding of CD103 biology and the therapeutic potential of targeting CD103 on tissue-resident lymphocytes.

Integrin αEβ7 is involved in the intestinal barrier injury of sepsis.

Background: IL-9-producing CD4(+) T (Th9) cell was related to acute intestinal barrier injury in sepsis. Integrin αEβ7 was an important lymphocyte homing receptor on the surface of intestinal Th9 cells. However, the roles of αEβ7 in the intestinal injury caused by Th9 cells were not clear in sepsis.Methods: To investigate the roles of αEβ7 in the intestinal injury caused by Th9 cells in sepsis model, the Th9 cells percentages, αEβ7, E-cadherin, IL-9, and D-lactate levels in both serum and intestinal tissue were measured. The intestinal histopathology, epithelium apoptosis, and mucosal permeability measurement were also performed. The survival rate of septic rats was recorded daily for 14 days.Results: Rats were assigned to four cohorts: control cohort, sepsis cohort, sepsis+αEβ7i (αEβ7 inhibition) cohort, and sepsis+αEβ7e (αEβ7 overexpression) cohort. The Th9 cells percentages, αEβ7, IL-9, and D-lactate levels of the sepsis cohort were significantly higher than those of the control cohort. The levels of these variables were also elevated progressively in the sepsis+αEβ7i cohort, sepsis cohort, and sepsis+αEβ7e cohort. The E-cadherin levels were decreased progressively in the control cohort, sepsis+αEβ7i cohort, sepsis cohort, and sepsis+αEβ7e cohort. Moreover, αEβ7 overexpression could decrease the 14-day survival rate. The findings of histopathology staining, apoptosis detection, and intestinal permeability test also confirmed that the barrier injury was deteriorated or relieved by elevating or decreasing the αEβ7 expression levels, respectively.Conclusions: Integrin αEβ7 was closely associated with the intestinal barrier injury caused by Th9 lymphocytes in sepsis.

Integrin αE(CD103)β7 in Epithelial Cancer.

Simple SummaryThe immune system in cancer is a central focus of research and clinical developments alike. We delineate our current view on αE(CD103)β7 integrin (CD103) expressing tumor infiltrating T lymphocytes in epithelial tumors. CD103 binds to E-cadherin within epithelial tissues and can be induced by TGFβ. It appears to play a role in the formation and function of the immunological synapse between cytotoxic T cell and tumor cell. Infiltration of CD103-expressing T cells in epithelial tumors is often associated with a better prognosis for patients. An exception seems to be epithelial skin tumors, where CD103 expression is not associated with better prognosis. We also show new data that CD103 is significantly more highly expressed in squamous cell carcinomas of the skin than in basal cell carcinomas or some other skin tumors, although the overall expression pattern is heterogeneous. A better understanding of αE(CD103)β7 integrin may facilitate some immunological antitumor therapies.Interactions of both the innate and the adaptive immune system with tumors are complex and often influence courses and therapeutic treatments in unanticipated ways. Based on the concept that CD8+T cells can mediate important antitumor effects, several therapies now aim to amplify their specific activity. A subpopulation of CD8+ tissue-resident T lymphocytes that express the αE(CD103)β7 integrin has raised particular interest. This receptor presumably contributes to the recruitment and retention of tumor-infiltrating immune cells through interaction with its ligand, E-cadherin. It appears to have regulatory functions and is thought to be a component of some immunological synapses. In TGF-rich environments, the αE(CD103)β7/E-cadherin-interaction enhances the binding strength between tumor cells and infiltrating T lymphocytes. This activity facilitates the release of lytic granule contents and cytokines as well as further immune responses and the killing of target cells. Expression of αE(CD103)β7 in some tumors is associated with a rather favorable prognosis, perhaps with the notable exception of squamous cell carcinoma of the skin. Although epithelial skin tumors are by far the most common tumors of fair-skinned people, there have been very few studies on the distribution of αE(CD103)β7 expressing cells in these neoplasms. Given this background, we describe here that αE(CD103)β7 is scarcely present in basal cell carcinomas, but much more abundant in squamous cell carcinomas with heterogeneous distribution. Notwithstanding a substantial number of studies, the role of αE(CD103)β7 in the tumor context is still far from clear. Here, we summarize the essential current knowledge on αE(CD103)β7 and outline that it is worthwhile to further explore this intriguing receptor with regard to the pathophysiology, therapy, and prognosis of solid tumors.

Regulation and Role of αE Integrin and Gut Homing Integrins in Migration and Retention of Intestinal Lymphocytes during Inflammatory Bowel Disease.

Targeting interactions between α4β7 integrin and endothelial adhesion molecule MAdCAM-1 to inhibit lymphocyte migration to the gastrointestinal tract is an effective therapy in inflammatory bowel disease (IBD). Following lymphocyte entry into the mucosa, a subset of these cells expresses αEβ7 integrin, which is expressed on proinflammatory lymphocytes, to increase cell retention. The factors governing lymphocyte migration into the intestinal mucosa and αE integrin expression in healthy subjects and IBD patients remain incompletely understood. We evaluated changes in factors involved in lymphocyte migration and differentiation within tissues. Both ileal and colonic tissue from active IBD patients showed upregulation of ICAM-1, VCAM-1, and MAdCAM-1 at the gene and protein levels compared with healthy subjects and/or inactive IBD patients. β1 and β7 integrin expression on circulating lymphocytes was similar across groups. TGF-β1 treatment induced expression of αE on both β7+ and β7- T cells, suggesting that cells entering the mucosa independently of MAdCAM-1/α4β7 can become αEβ7+ ITGAE gene polymorphisms did not alter protein induction following TGF-β1 stimulation. Increased phospho-SMAD3, which is directly downstream of TGF-β, and increased TGF-β-responsive gene expression were observed in the colonic mucosa of IBD patients. Finally, in vitro stimulation experiments showed that baseline β7 expression had little effect on cytokine, chemokine, transcription factor, and effector molecule gene expression in αE+ and αE- T cells. These findings suggest cell migration to the gut mucosa may be altered in IBD and α4β7-, and α4β7+ T cells may upregulate αEβ7 in response to TGF-β once within the gut mucosa.

Integrin αEβ7+ T cells direct intestinal stem cell fate decisions via adhesion signaling.

Intestinal stem cell (ISC) differentiation is regulated precisely by a niche in the crypt, where lymphocytes may interact with stem and transient amplifying (TA) cells. However, whether and how lymphocyte-stem/TA cell contact affects ISC differentiation is largely unknown. Here, we uncover a novel role of T cell-stem/TA cell contact in ISC fate decisions. We show that intestinal lymphocyte depletion results in skewed ISC differentiation in mice, which can be rescued by T cell transfer. Mechanistically, integrin αEβ7 expressed on T cells binds to E-cadherin on ISCs and TA cells, triggering E-cadherin endocytosis and the consequent Wnt and Notch signaling alterations. Blocking αEβ7-E-cadherin adhesion suppresses Wnt signaling and promotes Notch signaling in ISCs and TA cells, leading to defective ISC differentiation. Thus, αEβ7+ T cells regulate ISC differentiation at single-cell level through cell-cell contact-mediated αEβ7-E-cadherin adhesion signaling, highlighting a critical role of the T cell-stem/TA cell contact in maintaining intestinal homeostasis.

Anti-Integrins for the Treatment of Inflammatory Bowel Disease: Current Evidence and Perspectives.

Leukocyte trafficking to the gastrointestinal tract is recognized to play a role in the pathogenesis of inflammatory bowel disease (IBD). Integrins are expressed on immune cells and interact with cell adhesion molecules (CAM) to mediate leukocyte trafficking. Blockade of the gut-tropic integrin α4β7 and its subunits has been exploited as a therapeutic target in IBD. Natalizumab (anti-α4) is approved for moderate to severe Crohn’s disease (CD), but its use is limited due to potential risk of progressive multifocal leukoencephalopathy. Vedolizumab (anti-α4β7) is approved for the treatment of ulcerative colitis (UC) and CD. It is the most widely used anti-integrin therapy in IBD and has been shown to be effective in both induction and maintenance therapy, with a favorable safety profile. Several models incorporating clinical, genetic, immune, gut microbial, and vitamin D markers to predict response to vedolizumab in IBD have been developed. Etrolizumab (anti-β7) blocks leukocyte trafficking via α4β7 and cell adhesion via αEβ7 integrins. Large phase 3 clinical trials evaluating efficacy of etrolizumab in the induction and maintenance of patients with IBD are underway. Other investigational anti-integrin therapies include abrilumab (anti-α4β7 IgG2), PN-943 (orally administered and gut-restricted α4β7 antagonist peptide), AJM300 (orally active small molecule inhibitor of α4), and ontamalimab (anti-MAdCAM-1 IgG).

Recent advances in inflammatory bowel disease therapy

Inflammatory bowel disease (IBD) is a group of chronic, relapsing disorders of the gastrointestinal (GI) tract that significantly affect patient's quality of life. The main goals of IBD treatment are long-lasting clinical remission without serious adverse events. The lack of fully effective treatment urges researchers to seek for new therapeutic strategies and design of novel anti-inflammatory compounds.In this review, we focus on the latest advances in the IBD therapy. We characterize the clinical efficacy and mechanism of action of stem cell-, antibody- and small molecule-based methods of treatment that already reached clinic or are currently evaluated in clinical studies. The scope of this article is on agents targeting interleukin 12 and 23, integrins α4β7 and αEβ7, mucosal vascular addressin cell adhesion molecule, janus kinases, sphingosine-1-phosphate and toll-like receptor 9. We also describe recent advances in the discovery of biomarkers in IBD.

Combined blockade of α4β7 and αEβ7 integrins leads to a greater reduction of gut proinflammatory T lymphocytes over either pathway alone

Abstract Therapeutic approaches to treat inflammatory bowel diseases (IBD) include targeting integrins that mediate adhesion and migration of lymphocytes to the gastrointestinal (GI) tract. Etrolizumab is a humanized monoclonal antibody that selectively binds the β7 subunit of the heterodimeric integrins of α4β7 and αEβ7. The relative contribution of α4β7 and αEβ7 integrin family members to lymphocyte migration and retention in GI tractremains to be characterized. Here we applied single-cell technologies to healthy and IBD patient biopsies and found that αEβ7 is highly expressed in a subset of proinflammatory CD4+T cells and majority of cytotoxic intraepithelial CD8+T cells. In order to understand the role of αEβ7 in gut T cell accumulation, we used a photo-convertible reporter system to determine that concurrent inhibition of α4β7 and αEβ7 together led to a greater reduction of polyclonal or antigen specific T cell accumulation in the intestinal mucosa and epithelium compared to single blockade of either α4β7 or αEβ7. Further intra-vital two-photon microscopy and photo-specific labeling experiments revealed that blockade of αEβ7 reduces T cell interactions with basolateral epithelial surface, increases the migratory speed of activated T cells in the intestinal mucosa, and facilitates effector T cell egress from the intestinal mucosal through lymphatic vessels. Our data suggests co-blockade of α4β7 and αEβ7 together offers a superior inhibition of T cell accumulation in gastrointestinal tissues through a stepwise inhibition of T cell migration and subsequent tissue retention.

Abstract P3-08-13: Prognostic significance of intraepithelial CD103-positive tumor-infiltrating lymphocytes in breast cancer

Abstract Cluster of differentiation 103 (CD103), also known as αEβ7 integrin, is expressed on subsets of CD8+ T lymphocytes and a biomarker for tissue resident memory CD8+ T cells (TRM). CD103 expression is upregulated upon antigen recognition concurrent with transforming growth factor β (TGFβ) stimulation. CD103 is primarily expressed in the intra-epithelial tumor-infiltrating lymphocytes (iTILs) compared with that in stromal TILs (sTILs). Here, we investigated the prognostic significance of CD103+ iTILs in invasive breast cancer (IBC). Immunohistochemistry was performed for CD103, TGFβ isoforms (1, 2, and 3), and TGFβ receptors on tissue microarrays of 1173 IBC samples. CD103 expression was analyzed using Aperio ScanScope slide scanner. An area with the highest density of CD103+ iTILs was captured at a magnification of × 200, and CD103+ iTILs per unit area (mm2) were enumerated. The threshold for CD103+ iTILs (high vs low) was determined based on the mean number. Immunohistochemical staining for TGFβ isoforms (1, 2, and 3) and their receptors was quantitated using H-score and calculated as: staining intensity (0-3) × the percentage of positive tumor cells (0-100%). For statistical analyses, we categorized cases as positive or negative based on the mean value of H-score. The number of CD103+ iTILs were compared with clinicopathological characteristics, and tumor cell expression of TGFβ and its receptors. Further, the relationship between CD103+ iTILs density and prognosis was investigated according to the molecular subtypes. CD103+ iTILs were observed in 713 (60.8%) cases with a mean ± standard deviation of 34.2 ± 94.6. Increased abundance of CD103+ iTILs was found to be significantly associated with histological grade 3, high Ki-67 labelling index, non-luminal subtypes, high sTIL density, and positive expression of TGFβ3 and its receptor (all, P< 0.001). Furthermore, high CD103+ iTIL density was significantly associated with prolonged overall survival (OS) (P = 0.02) and disease-free survival (DFS) (P = 0.001). Subgroup analysis according to molecular subtypes showed that CD103+ iTILs was prognostic only for triple-negative breast cancer (OS, P = 0.003; DFS, P = 0.005). Multivariate analysis of CD103+ iTILs with clinicopathological prognostic factors revealed that CD103+ iTILs was an independent prognostic factor for OS (P = 0.005) and DFS (P = 0.006) for the triple-negative subtype. Our study revealed that breast cancer cell expression of TGFβ3 may be involved in recruitment of CD103+ iTILs. In breast cancer, CD103+ iTILs indicates good prognosis which is an independent favorable prognostic factor in patients with triple negative subtype and more reliable than sTIL density. Citation Format: Young Kyung Bae, Min Hui Park, Jung Eun Choi, Su Hwan Kang, Gyungyub Gong. Prognostic significance of intraepithelial CD103-positive tumor-infiltrating lymphocytes in breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-08-13.

CD8αα+ T cells show amoeboid shape and frequent morphological change in vitro, and localize to small intestinal intraepithelial region in vivo

Intraepithelial lymphocytes (IELs) are very unique in the intestinal immune system. They include γδT cells and CD4–CD8–TCRαβ+T cells (double negative: DNT), both of which are specific for the intestine, in addition to CD4+ and CD8+ T cells. IELs exist within the monolayer of the intestinal epithelial cells and dynamically move between lamina propria (LP) and intraepithelial (IE) region. The localization and movement patterns of IEL subsets and the regulatory factors have been unknown. Here, we developed a novel in vitro live imaging system and quantified the motility and morphological changes among subsets of IELs. We identified CD8αα as the key regulatory factor. IELs, especially γδ and DNT cells, showed amoeboid shape and frequent morphological change, while most T cells in MLN or SP showed round shape in vitro. TCR signal, IL-15, gut microbes, CCL25, and integrin αEβ7 expression were non-essential for IEL movement in vitro. CD8αα+ cells showed higher motility and larger morphological changes than CD8αα– cells. Adoptive transferred CD8αα+CD4-IELs localized to IE region of recipient NSG mice, while CD8αα–CD4-IELs localized to the LP. Our results showed that the CD8αα/TL signal is essential for the localization of IELs to IE region in vivo. CD8αα/TL may be an effective target to increase the number of IELs, which protects against intestinal infection, allergy, tumorigenesis or inflammation.

αEβ7 Expression Increases With the Extent of Cutaneous Involvement in Mycosis Fungoides.

Cell adhesion molecules are essential to lymphocyte migration in neoplastic and inflammatory skin diseases. Our aim was to investigate possible differences in cell adhesion molecule expression between mycosis fungoides (MF) and inflammatory skin diseases (drug reactions and allergic contact dermatitis). We selected 33 biopsies from patients with MF and 10 biopsies of patients with inflammatory skin diseases from Department of Pathology-Universidade Federal de São Paulo (UNIFESP) from January 1997 to December 2013. Expression of α4β1 integrin and αEβ7 integrin was assessed by immunohistochemistry in intraepidermal lymphocytes by counting 4 microscopic epidermal fields (×400) and comparing those between the 2 groups. We observed increased expression of integrin αEβ7 in intraepidermal lymphocytes in advanced stages of MF (T3 and T4). αEβ7 expression was detected in intraepidermal dendritic cells of MF and inflammatory diseases samples. The expression of E-cadherin in epidermal cells in MF outlined Pautrier microabscesses, whereas in inflammatory diseases, spongiosis reduced its expression in keratinocytes. The findings presented here support the idea that the lymphocyte migratory mechanism observed in neoplasms is similar to that of inflammatory processes of the skin.

Function of Human Tumor-Infiltrating Lymphocytes in Early-Stage Non-Small Cell Lung Cancer.

Cancer progression is marked by dysfunctional tumor-infiltrating lymphocytes (TIL) with high inhibitory receptor (IR) expression. Because IR blockade has led to clinical responses in some patients with non-small cell lung cancer (NSCLC), we investigated how IRs influenced CD8+ TIL function from freshly digested early-stage NSCLC tissues using a killing assay and intracellular cytokine staining after in vitro T-cell restimulation. Early-stage lung cancer TIL function was heterogeneous with only about one third of patients showing decrements in cytokine production and lytic function. TIL hypofunction did not correlate with clinical factors, coexisting immune cells (macrophages, neutrophils, or CD4+ T regulatory cells), nor with PD-1, TIGIT, TIM-3, CD39, or CTLA-4 expression. Instead, we found that the presence of the integrin αeβ7 (CD103), characteristic of tissue-resident memory cells (TRM), was positively associated with cytokine production, whereas expression of the transcription factor Eomesodermin (Eomes) was negatively associated with TIL function. These data suggest that the functionality of CD8+ TILs from early-stage NSCLCs may be influenced by competition between an antitumor CD103+ TRM program and an exhaustion program marked by Eomes expression. Understanding the mechanisms of T-cell function in the progression of lung cancer may have clinical implications for immunotherapy.