Publications
58 results found
Evans R, Flores-Borja F, Nassiri S, et al., 2019, Integrin-mediated macrophage adhesion promotes lymphovascular dissemination in breast cancer, Cell Reports, Vol: 27, Pages: 1967-1978.e4, ISSN: 2211-1247
Lymphatic vasculature is crucial for metastasis in triple-negative breast cancer (TNBC); however, cellular and molecular drivers controlling lymphovascular metastasis are poorly understood. We define a macrophage-dependent signaling cascade that facilitates metastasis through lymphovascular remodeling. TNBC cells instigate mRNA changes in macrophages, resulting in β4 integrin-dependent adhesion to the lymphovasculature. β4 integrin retains macrophages proximal to lymphatic endothelial cells (LECs), where release of TGF-β1 drives LEC contraction via RhoA activation. Macrophages promote gross architectural changes to lymphovasculature by increasing dilation, hyperpermeability, and disorganization. TGF-β1 drives β4 integrin clustering at the macrophage plasma membrane, further promoting macrophage adhesion and demonstrating the dual functionality of TGF-β1 signaling in this context. β4 integrin-expressing macrophages were identified in human breast tumors, and a combination of vascular-remodeling macrophage gene signature and TGF-β signaling scores correlates with metastasis. We postulate that future clinical strategies for patients with TNBC should target crosstalk between β4 integrin and TGF-β1.
Robertson FP, Yeung AC, Male V, et al., 2019, Urinary Neutrophil Gelatinase Associated Lipocalins (NGALs) predict acute kidney injury post liver transplant, HPB, Vol: 21, Pages: 473-481, ISSN: 1365-182X
BACKGROUND: Acute Kidney Injury, a common complication of liver transplant, is associated with a significant increase in the risk of morbidity, mortality and graft loss. Current diagnostic criteria leaves a delay in diagnosis allowing further potential irreversible damage. Early biomarkers of renal injury are of clinical importance and Neutrophil Gelatinase Associated Lipocalins (NGALs) and Syndecan-1 were investigated. METHODS: AKI was defined according to the Acute Kidney Injury Network criteria. Urine and blood samples were collected pre-operatively, immediately post-op and 24 h post reperfusion to allow measurement of NGAL and Syndecan-1 levels. RESULTS: 13 of 27 patients developed an AKI. Patients who developed AKI had significantly higher peak transaminases. Urinary NGAL, plasma NGAL and Syndecan-1 levels were significantly elevated in all patients post reperfusion. Urinary NGAL levels immediately post-op were significantly higher in patients who developed an AKI than those that didn't [1319 ng/ml vs 46.56 ng/ml, p ≤ 0.001]. ROC curves were performed and urinary NGAL levels immediately post-op were an excellent biomarker for AKI with an area under the curve of 0.948 (0.847-1.00). CONCLUSIONS: Urinary NGAL levels measured immediately post-op accurately predict the development of AKI and their incorporation into clinical practise could allow early protocols to be developed to treat post transplant AKI.
Cuff A, Perchet T, Dertschnig S, et al., 2019, Tbet promotes NK cell bone marrow egress via CXCR6 expression, Publisher: bioRxiv
Tbet-deficient mice have reduced NK cells in blood and spleen, but increased NK cells in bone marrow and lymph nodes, a phenotype that is thought to be due to a defect in S1PR5-mediated migration. Here, we revisit the role of Tbet in NK cell bone marrow egress. We definitively show that the accumulation of NK cells in the bone marrow of Tbet-deficient (Tbx21 −/− ) animals occurs because of a cell-intrinsic migration defect. We identify a profile of gene expression, co-ordinated by Tbet, which affects the localisation of NK cells in the bone marrow. The most underexpressed gene in the absence of Tbet was Cxcr6, and we confirmed that CXCR6 protein was also not expressed in Tbet-deficient NK cells. Cxcr6-expressing ILC progenitors and immature NK cells accumulate in the bone marrow of CXCR6-deficient mice. This suggests that CXCR6 is among the mediators of migration, controlled by Tbet, that work together to coordinate NK cell bone marrow egress. Understanding NK cell bone marrow egress will become increasingly important as we move into an era in which NK cell immunotherapies are being developed.
Kostrzewski T, Borg AJ, Meng Y, et al., 2018, Multiple levels of control determine how E4bp4/Nfil3 regulates NK cell development, Journal of Immunology, Vol: 200, Pages: 1370-1381, ISSN: 0022-1767
The transcription factor E4bp4/Nfil3 has been shown to have a critical role in the development of all innate lymphoid cell types including NK cells. In this study, we show that posttranslational modifications of E4bp4 by either SUMOylation or phosphorylation have profound effects on both E4bp4 function and NK cell development. We examined the activity of E4bp4 mutants lacking posttranslational modifications and found that Notch1 was a novel E4bp4 target gene. We observed that abrogation of Notch signaling impeded NK cell production and the total lack of NK cell development from E4bp4−/− progenitors was completely rescued by short exposure to Notch peptide ligands. This work reveals both novel mechanisms in NK cell development by a transcriptional network including E4bp4 with Notch, and that E4bp4 is a central hub to process extrinsic stimuli.
Male V, Stegmann KA, Easom NJ, et al., 2017, Natural killer cells in liver disease, Seminars in Liver Disease, Vol: 37, Pages: 198-209, ISSN: 0272-8087
Natural killer (NK) cells comprise one of the most abundant immune cell populations in human liver and the nature and functions of these cells have been a focus of recent interest. Here, we consider the possible roles of NK cells in diverse liver diseases, concentrating on data from patient studies. NK cells can be protective, killing virally infected and cancerous cells in the liver and limiting fibrosis by eliminating hepatic stellate cells. However, they can also be deleterious, contributing to pathology in viral hepatitis by killing hepatocytes and downregulating virus-specific T-cell responses. It has recently emerged that a large fraction of hepatic NK cells constitute a distinct liver-resident subset and we highlight the need to distinguish between circulating and liver-resident NK cells in future studies. There is also a need for further investigation into how NK cells are influenced by the liver microenvironment and what scope there is to harness their immunotherapeutic potential.
Cuff AO, Male V, 2017, Conventional NK cells and ILC1 are partially ablated in the livers of Ncr1 iCreTbx21 fl/fl mice [version 2; peer review: 2 approved], Wellcome Open Research, Vol: 2, ISSN: 2398-502X
Mouse liver contains both Eomes-dependent conventional natural killer (cNK) cells and Tbet-dependent liver-resident type I innate lymphoid cells (ILC1). In order to better understand the role of ILC1, we attempted to generate mice that would lack liver ILC1, while retaining cNK, by conditional deletion of Tbet in NKp46+ cells. Here we report that the Ncr1 iCreTbx21 fl/fl mouse has a roughly equivalent reduction in both the cNK and ILC1 compartments of the liver, limiting its utility for investigating the relative contributions of these two cell types in disease models. We also describe the phenotype of these mice with respect to NK cells, ILC1 and NKp46 + ILC3 in the spleen and small intestine lamina propria.
Cuff A, Male V, 2017, Conventional NK cells and ILC1 are partially ablated in the livers of Ncr1iCreTbx21fl/fl mice [version 1; peer review: 2 approved with reservations], Wellcome Open Research, Vol: 2, Pages: 1-12, ISSN: 2398-502X
Mouse liver contains both Eomes-dependent conventional natural killer (cNK) cells and Tbet-dependent liver-resident type I innate lymphoid cells (ILC1). In order to better understand the role of ILC1, we attempted to generate mice that would lack liver ILC1, while retaining cNK, by conditional deletion of Tbet in NKp46+ cells. Here we report that the Ncr1iCreTbx21fl/fl mouse has a roughly equivalent reduction in both the cNK and ILC1 compartments of the liver, limiting its utility for investigating the relative contributions of these two cell types in disease models. We also describe the phenotype of these mice with respect to NK cells, ILC1 and NKp46+ ILC3 in the spleen and small intestine lamina propria.
Pallett LJ, Davies J, Colbeck EJ, et al., 2017, IL-2high tissue-resident T cells in the human liver: Sentinels for hepatotropic infection, Journal of Experimental Medicine, Vol: 214, Pages: 1567-1580, ISSN: 0022-1007
The liver provides a tolerogenic immune niche exploited by several highly prevalent pathogens as well as by primary and metastatic tumors. We have sampled healthy and hepatitis B virus (HBV)-infected human livers to probe for a subset of T cells specialized to overcome local constraints and mediate immunity. We characterize a population of T-betloEomesloBlimp-1hiHobitlo T cells found within the intrahepatic but not the circulating memory CD8 T cell pool expressing liver-homing/retention markers (CD69+CD103+ CXCR6+CXCR3+). These tissue-resident memory T cells (TRM) are preferentially expanded in patients with partial immune control of HBV infection and can remain in the liver after the resolution of infection, including compartmentalized responses against epitopes within all major HBV proteins. Sequential IL-15 or antigen exposure followed by TGFβ induces liver-adapted TRM, including their signature high expression of exhaustion markers PD-1 and CD39. We suggest that these inhibitory molecules, together with paradoxically robust, rapid, cell-autonomous IL-2 and IFNγ production, equip liver CD8 TRM to survive while exerting local noncytolytic hepatic immunosurveillance.
Male V, Brady HJM, 2017, Murine thymic NK cells: a case of identity, European Journal of Immunology, Vol: 47, Pages: 797-799, ISSN: 0014-2980
Just over a decade ago, it was established that NK cells in the thymus do not follow precisely the same developmental pathway as conventional NK cells that develop in the bone marrow. Subsequently, it has emerged that NK cells are one branch of a family of innate lymphoid cells (ILCs). ILC1s and thymic NK cells have, however, sufficient similarities such that questions have been raised about how distinctive each cell type is from the other. In this issue of European Journal of Immunology, Gabrielli et al. [Eur. J. Immunol. 2017. 47: 800–805] make a detailed study of the transcription factor requirements of murine thymic NK cells. They provide a valuable insight into the distinctive identity of thymic NK cells with regard to Tbet, Nfil3, Id2, and Ets1. In addition, they clarify the nature of DX5 expression on NK cells and ILC‐like cells in the murine thymus.
Male V, 2017, Liver-resident NK cells: the human factor, Trends in Immunology, Vol: 38, Pages: 307-309, ISSN: 1471-4981
Mouse liver contains two natural killer (NK) cell populations, one of which recirculates while the other is tissue resident. Following this discovery, several groups have sought to identify liver-resident NK (lrNK) cells in humans. Here, I present an overview of recent advances in the field.
Irshad S, Flores-Borja F, Lawler K, et al., 2017, ROR gamma t(+) innate lymphoid cells promote lymph node metastasis of breast cancers, Cancer Research, Vol: 77, Pages: 1083-1096, ISSN: 0008-5472
Cancer cells tend to metastasize first to tumor-draining lymph nodes, but the mechanisms mediating cancer cell invasion into the lymphatic vasculature remain little understood. Here, we show that in the human breast tumor microenvironment (TME), the presence of increased numbers of RORγt+ group 3 innate lymphoid cells (ILC3) correlates with an increased likelihood of lymph node metastasis. In a preclinical mouse model of breast cancer, CCL21-mediated recruitment of ILC3 to tumors stimulated the production of the CXCL13 by TME stromal cells, which in turn promoted ILC3–stromal interactions and production of the cancer cell motile factor RANKL. Depleting ILC3 or neutralizing CCL21, CXCL13, or RANKL was sufficient to decrease lymph node metastasis. Our findings establish a role for RORγt+ILC3 in promoting lymphatic metastasis by modulating the local chemokine milieu of cancer cells in the TME. Cancer Res; 77(5); 1083–96.
Robertson F, Male V, Wright G, et al., 2017, Recruitment of inflammatory monocytes after liver transplantation and correlation with clinical outcome, Spring Meeting on Clinician Scientists in Training, Publisher: Elsevier, Pages: 84-84, ISSN: 0140-6736
Cuff AO, Robertson FP, Stegmann KA, et al., 2016, Eomeshi NK cells in human liver are long-lived and do not recirculate but can be replenished from the circulation, Journal of Immunology, Vol: 197, Pages: 4283-4291, ISSN: 1550-6606
Human liver contains an Eomeshi population of NK cells that is not present in the blood. In this study, we show that these cells are characterized by a molecular signature that mediates their retention in the liver. By examining liver transplants where donors and recipients are HLA mismatched, we distinguish between donor liver-derived and recipient-derived leukocytes to show that Eomeslo NK cells circulate freely whereas Eomeshi NK cells are unable to leave the liver. Furthermore, Eomeshi NK cells are retained in the liver for up to 13 y. Therefore, Eomeshi NK cells are long-lived liver-resident cells. We go on to show that Eomeshi NK cells can be recruited from the circulation during adult life and that circulating Eomeslo NK cells are able to upregulate Eomes and molecules mediating liver retention under cytokine conditions similar to those in the liver. This suggests that circulating NK cells are a precursor of their liver-resident counterparts.
Stegmann KA, Robertson F, Hansi N, et al., 2016, CXCR6 marks a novel subset of T-bet(lo)Eomes(hi) natural killer cells residing in human liver, Scientific Reports, Vol: 6, ISSN: 2045-2322
Natural killer cells (NK) are highly enriched in the human liver, where they can regulate immunity and immunopathology. We probed them for a liver-resident subset, distinct from conventional bone-marrow-derived NK. CXCR6+ NK were strikingly enriched in healthy and diseased liver compared to blood (p < 0.0001). Human hepatic CXCR6+ NK had an immature phenotype (predominantly CD56(bright)CD16-CD57-), and expressed the tissue-residency marker CD69. CXCR6+ NK produced fewer cytotoxic mediators and pro-inflammatory cytokines than the non-liver-specific CXCR6- fraction. Instead CXCR6+ NK could upregulate TRAIL, a key death ligand in hepatitis pathogenesis. CXCR6 demarcated liver NK into two transcriptionally distinct populations: T-bet(hi)Eomes(lo)(CXCR6-) and T-bet(lo)Eomes(hi)(CXCR6+); the latter was virtually absent in the periphery. The small circulating CXCR6+ subset was predominantly T-bet(hi)Eomes(lo), suggesting its lineage was closer to CXCR6- peripheral than CXCR6+ liver NK. These data reveal a large subset of human liver-resident T-bet(lo)Eomes(hi) NK, distinguished by their surface expression of CXCR6, adapted for hepatic tolerance and inducible anti-viral immunity.
Male V, Nisoli I, Kostrzewski T, et al., 2014, The transcription factor E4bp4/Nfil3 controls commitment to the NK lineage and directly regulates Eomes and Id2 expression, Journal of Experimental Medicine, Vol: 211, Pages: 635-642, ISSN: 1540-9538
The transcription factor E4bp4 (Nfil3) is essential for natural killer (NK) cell production. Here, we show that E4bp4 is required at the NK lineage commitment point when NK progenitors develop from common lymphoid progenitors (CLPs) and that E4bp4 must be expressed at the CLP stage for differentiation toward the NK lineage to occur. To elucidate the mechanism by which E4bp4 promotes NK development, we identified a central core of transcription factors that can rescue NK production from E4bp4−/− progenitors, suggesting that they act downstream of E4bp4. Among these were Eomes and Id2, which are expressed later in development than E4bp4. E4bp4 binds directly to the regulatory regions of both Eomes and Id2, promoting their transcription. We propose that E4bp4 is required for commitment to the NK lineage and promotes NK development by directly regulating the expression of the downstream transcription factors Eomes and Id2.
Crotta S, Gkioka A, Male V, et al., 2014, The Transcription Factor E4BP4 Is Not Required for Extramedullary Pathways of NK Cell Development, JOURNAL OF IMMUNOLOGY, Vol: 192, Pages: 2677-2688, ISSN: 0022-1767
NK cells contribute to antitumor and antiviral immunosurveillance. Their development in the bone marrow (BM) requires the transcription factor E4BP4/NFIL3, but requirements in other organs are less well defined. In this study, we show that CD3−NK1.1+NKp46+CD122+ NK cells of immature phenotype and expressing low eomesodermin levels are found in thymus, spleen, and liver of E4BP4-deficient mice, whereas numbers of mature, eomesoderminhigh conventional NK cells are drastically reduced. E4BP4-deficient CD44+CD25− double-negative 1 thymocytes efficiently develop in vitro into NK cells with kinetics, phenotype, and functionality similar to wild-type controls, whereas no NK cells develop from E4BP4-deficient BM precursors. In E4BP4/Rag-1 double-deficient (DKO) mice, NK cells resembling those in Rag-1–deficient controls are found in similar numbers in the thymus and liver. However, NK precursors are reduced in DKO BM, and no NK cells develop from DKO BM progenitors in vitro. DKO thymocyte precursors readily develop into NK cells, but DKO BM transfers into nude recipients and NK cells in E4BP4/Rag-1/IL-7 triple-KO mice indicated thymus-independent NK cell development. In the presence of T cells or E4BP4-sufficient NK cells, DKO NK cells have a selective disadvantage, and thymic and hepatic DKO NK cells show reduced survival when adoptively transferred into lymphopenic hosts. This correlates with higher apoptosis rates and lower responsiveness to IL-15 in vitro. In conclusion, we demonstrate E4BP4-independent development of NK cells of immature phenotype, reduced fitness, short t1/2, and potential extramedullary origin. Our data identify E4BP4-independent NK cell developmental pathways and a role for E4BP4 in NK cell homeostasis.
Male V, Brady HJM, 2014, Transcriptional Control of NK Cell Differentiation and Function, TRANSCRIPTIONAL CONTROL OF LINEAGE DIFFERENTIATION IN IMMUNE CELLS, Vol: 381, Pages: 173-187, ISSN: 0070-217X
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- Citations: 10
Wilkens J, Male V, Ghazal P, et al., 2013, Uterine NK Cells Regulate Endometrial Bleeding in Women and Are Suppressed by the Progesterone Receptor Modulator Asoprisnil, JOURNAL OF IMMUNOLOGY, Vol: 191, Pages: 2226-2235, ISSN: 0022-1767
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- Citations: 62
Male V, Gardner L, Moffett A, 2012, Isolation of cells from the feto-maternal interface., Curr Protoc Immunol, Vol: Chapter 7
The mucosal lining of the human uterus is host to a specialized population of leukocytes, which, during pregnancy, interact with invading placental cells (trophoblast) of fetal origin. Of particular interest are uterine natural killer cells, which account for around 70% of the leukocytes at this site during the first trimester of pregnancy, and seem to be specially adapted to recognize invading trophoblast cells. In order to understand the interactions between mucosal immune cells and trophoblast, and those among the immune cells themselves, it is useful to be able to isolate and culture these cells. Here, we describe protocols for the isolation of leukocytes, stromal cells, and trophoblast cells from the feto-maternal interface.
Male V, Nisoli I, Gascoyne DM, et al., 2012, E4BP4: an unexpected player in the immune response, TRENDS IN IMMUNOLOGY, Vol: 33, Pages: 98-102, ISSN: 1471-4906
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- Citations: 74
Male V, Sharkey A, Masters L, et al., 2011, The effect of pregnancy on the uterine NK cell KIR repertoire, European Journal of Immunology, Vol: 41, Pages: 3017-3027, ISSN: 0014-2980
The major leukocyte population in the decidua during the first trimester of pregnancy consists of NK cells that express receptors capable of recognizing MHC class I molecules expressed by placental trophoblast. These include members of the killer immunoglobulin‐like receptor (KIR) family, the two‐domain KIR (KIR2D), which recognize HLA‐C. Interactions between decidual NK (dNK) cell KIR2D and placental HLA‐C contribute to the success of pregnancy and dNK cells express KIR2D at higher frequency than peripheral NK (pNK) cells. Thus, they are biased toward recognizing HLA‐C. In order to investigate when this unusual KIR repertoire appears, we compared the phenotype of NK cells isolated from non‐pregnant (endometrium) and pregnant (decidua) human uterine mucosa. Endometrial NK (eNK) cells did not express KIR2D at a higher level than matched pNK cells, so the bias toward HLA‐C recognition occurs as a response to pregnancy. Furthermore, HLA‐C expression was upregulated on uterine stromal cells as the mucosa transformed from endometrium to decidua at the onset of pregnancy. As uterine NK (uNK) cells can mature from NK precursors and acquire KIR expression in utero, the pregnancy‐specific bias of uNK cells toward HLA‐C recognition could arise as developing uNK cells interact with uterine stromal cells, which express higher levels of HLA‐C during pregnancy.
Apps R, Sharkey A, Gardner L, et al., 2011, <i>Ex vivo</i> functional responses to HLA-G differ between blood and decidual NK cells, MOLECULAR HUMAN REPRODUCTION, Vol: 17, Pages: 577-586
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- Citations: 33
Apps R, Sharkey A, Gardner L, et al., 2011, Genome-wide expression profile of first trimester villous and extravillous human trophoblast cells, PLACENTA, Vol: 32, Pages: 33-43, ISSN: 0143-4004
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- Citations: 100
Male V, Hughes T, McClory S, et al., 2010, Immature NK Cells, Capable of Producing IL-22, Are Present in Human Uterine Mucosa, JOURNAL OF IMMUNOLOGY, Vol: 185, Pages: 3913-3918, ISSN: 0022-1767
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- Citations: 140
Moffett A, Male VH, 2010, NK cells and reproduction, NATURAL KILLER CELLS: BASIC SCIENCE AND CLINICAL APPLICATION, Editors: Lotze, Thomson, Publisher: ELSEVIER SCIENCE BV, Pages: 403-416, ISBN: 978-0-12-370454-2
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- Citations: 1
Male V, Trundley A, Gardner L, et al., 2010, Natural Killer Cells in Human Pregnancy, NATURAL KILLER CELL PROTOCOLS, Vol: 612, Pages: 447-463, ISSN: 1064-3745
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- Citations: 32
Thomas R, Apps R, Qi Y, et al., 2009, HLA-C cell surface expression and control of HIV/AIDS correlate with a variant upstream of <i>HLA</i>-<i>C</i>, NATURE GENETICS, Vol: 41, Pages: 1290-U46, ISSN: 1061-4036
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- Citations: 237
Apps R, Gardner L, Traherne J, et al., 2008, Natural-killer cell ligands at the maternal-fetal interface: UL-16 binding proteins, MHC class-I chain related molecules, HLA-F and CD48, Human Reproduction, Vol: 23, Pages: 2535-2548, ISSN: 1460-2350
BACKGROUNDIn the early stages of human placentation, the decidua is invaded by fetal extravillous trophoblast (EVT) cells. Interactions between EVT cells and local decidual leukocytes are likely to contribute to immunological accommodation of the semi-allogeneic fetus.METHODS AND RESULTSNatural-killer group 2 member D (NKG2D) and 2B4 (CD244) are receptors ubiquitously expressed by the distinctive population of CD56 bright, uterine natural-killer cells, which dominate the decidua at the time of implantation. Here, we investigate the UL-16 binding protein (ULBP) and MHC class-I chain related molecule (MIC) ligands of NKG2D, the CD48 ligand of 2B4 and the non-classical HLA class-I molecule, HLA-F, at the maternal–fetal interface of normal pregnancies. For many of these molecules, significant mRNA expression was detected by RT-PCR in decidual and placental tissue throughout gestation. Flow cytometry of isolated cells or immunohistological staining of implantation site sections was then performed. No protein expression of NKG2D ligands or HLA-F could be detected in decidual leukocytes or fetal trophoblast cells from the first trimester. An NKG2D-Fc fusion protein identified no novel ligands for this promiscuous receptor at the maternal–fetal interface. Strong surface protein expression of CD48 by decidual leukocytes but not by trophoblast cells was detected by flow cytometry. Histological staining showed a clear aggregation of CD48+ cells around transformed spiral arteries of the implantation site.CONCLUSIONSWe conclude that the role of NKG2D and 2B4 is not focussed on trophoblast recognition in normal pregnancy, but is more likely involved in cross-talk among maternal cells of the placental bed.
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