Publications by authors named "Rene A W van Lier"

159 Publications

Hobit identifies tissue-resident memory T cell precursors that are regulated by Eomes.

Sci Immunol 2021 Aug;6(62)

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.

Tissue-resident memory CD8 T cells (T) constitute a noncirculating memory T cell subset that provides early protection against reinfection. However, how T arise from antigen-triggered T cells has remained unclear. Exploiting the T-restricted expression of Hobit, we used T reporter/deleter mice to study T differentiation. We found that Hobit was up-regulated in a subset of LCMV-specific CD8 T cells located within peripheral tissues during the effector phase of the immune response. These Hobit effector T cells were identified as T precursors, given that their depletion substantially decreased T development but not the formation of circulating memory T cells. Adoptive transfer experiments of Hobit effector T cells corroborated their biased contribution to the T lineage. Transcriptional profiling of Hobit effector T cells underlined the early establishment of T properties including down-regulation of tissue exit receptors and up-regulation of T-associated molecules. We identified Eomes as a key factor instructing the early bifurcation of circulating and resident lineages. These findings establish that commitment of T occurs early in antigen-driven T cell differentiation and reveal the molecular mechanisms underlying this differentiation pathway.
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http://dx.doi.org/10.1126/sciimmunol.abg3533DOI Listing
August 2021

How to Reliably Define Human CD8 T-Cell Subsets: Markers Playing Tricks.

Cold Spring Harb Perspect Biol 2021 Mar 29. Epub 2021 Mar 29.

Adaptive Immunity Laboratory and Landsteiner Laboratory of the AUMC at Sanquin Blood Supply Foundation, Amsterdam 1066 CX, The Netherlands.

In recent years, our understanding about the functional complexity of CD8 T-cell populations has increased tremendously. The immunology field is now facing challenges to translate these insights into phenotypic definitions that correlate reliably with distinct functional traits. This is key to adequately monitor and understand compound immune responses including vaccination and immunotherapy regimens. Here we will summarize our understanding of the current state in the human CD8 T-cell subset characterization field. We will address how reliably the currently used cell surface markers are connected to differentiation status and function of particular subsets. By restricting ourselves to CD8 αβ T cells, we will focus mostly on major histocompatibility complex (MHC) class I-restricted virus- and tumor-specific T cells. This comes with a major advantage as fluorescently labeled peptide-loaded MHC class I multimers have been widely used to identify and characterize these cells.
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http://dx.doi.org/10.1101/cshperspect.a037747DOI Listing
March 2021

CD8 and CD4 T Cell Populations in Human Kidneys.

Cells 2021 02 1;10(2). Epub 2021 Feb 1.

Department of Experimental Immunology, Amsterdam institute for Infection & Immunity, Amsterdam UMC, University of Amsterdam, 1105AZ Amsterdam, The Netherlands.

Background: At border sites, and in internal organs, tissue resident memory T cells (T contribute to the immune barrier against pathogens like viruses, bacteria, fungi, and cancer. However, information on the presence and function of these cells in the human kidney is scant. In order to better understand the T cell-mediated immunological defense in this organ, we aimed to determine phenotypic and functional aspects of CD8 and CD4 T cells present in healthy and allograft kidney tissue.

Methods: Using multichannel flow cytometry, we assessed the phenotype and function of T cells in healthy renal tissue samples ( = 5) and kidney allograft tissue ( = 7) and compared these aspects to T cells in peripheral blood from healthy controls ( = 13).

Results: Kidney tissue samples contained substantial amounts of CD8 and CD4 T cells. In contrast to the circulating cells, kidney T cells frequently expressed CD69 and CD103, and were more often actively cycling. Furthermore, nearly all kidney T cells expressed CXCR3, and often expressed CXCR6 compared to T cells in the circulation. Markedly, kidney T cells produced greater quantities of IFNγ than circulating cells and were frequently polyfunctional.

Conclusion: Functional T cells with the characteristic traits of T reside in human kidney tissues. These cells are more often actively cycling and frequently express CXCR3 and CXCR6.
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http://dx.doi.org/10.3390/cells10020288DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7912772PMC
February 2021

Low SARS-CoV-2 seroprevalence in blood donors in the early COVID-19 epidemic in the Netherlands.

Nat Commun 2020 11 12;11(1):5744. Epub 2020 Nov 12.

Department of Blood-borne Infections, Sanquin Research, Amsterdam, The Netherlands.

The world is combating an ongoing COVID-19 pandemic with health-care systems, society and economies impacted in an unprecedented way. It is unclear how many people have contracted the causative coronavirus (SARS-CoV-2) unknowingly and are asymptomatic. Therefore, reported COVID-19 cases do not reflect the true scale of outbreak. Here we present the prevalence and distribution of antibodies to SARS-CoV-2 in a healthy adult population of the Netherlands, which is a highly affected country, using a high-performance immunoassay. Our results indicate that one month into the outbreak (i) the seroprevalence in the Netherlands was 2.7% with substantial regional variation, (ii) the hardest-hit areas showed a seroprevalence of up to 9.5%, (iii) the seroprevalence was sex-independent throughout age groups (18-72 years), and (iv) antibodies were significantly more often present in younger people (18-30 years). Our study provides vital information on the extent of exposure to SARS-CoV-2 in a country where social distancing is in place.
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http://dx.doi.org/10.1038/s41467-020-19481-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7665189PMC
November 2020

Divergent SARS-CoV-2-specific T- and B-cell responses in severe but not mild COVID-19 patients.

Eur J Immunol 2020 Dec 16;50(12):1998-2012. Epub 2020 Nov 16.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the current coronavirus disease 2019 (COVID-19) pandemic. Understanding the immune response that provides specific immunity but may also lead to immunopathology is crucial for the design of potential preventive and therapeutic strategies. Here, we characterized and quantified SARS-CoV-2-specific immune responses in patients with different clinical courses. Compared to individuals with a mild clinical presentation, CD4 T-cell responses were qualitatively impaired in critically ill patients. Strikingly, however, in these patients the specific IgG antibody response was remarkably strong. Furthermore, in these critically ill patients, a massive influx of circulating T cells into the lungs was observed, overwhelming the local T-cell compartment, and indicative of vascular leakage. The observed disparate T- and B-cell responses could be indicative of a deregulated immune response in critically ill COVID-19 patients.
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http://dx.doi.org/10.1002/eji.202048908DOI Listing
December 2020

Tissue-resident memory CD8 T cells shape local and systemic secondary T cell responses.

Nat Immunol 2020 09 13;21(9):1070-1081. Epub 2020 Jul 13.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands.

Tissue-resident memory CD8 T cells (T cells) are crucial in protecting against reinvading pathogens, but the impact of reinfection on their tissue confinement and contribution to recall responses is unclear. We developed a unique lineage tracer mouse model exploiting the T-defining transcription factor homolog of Blimp-1 in T cells (Hobit) to fate map the T progeny in secondary responses. After reinfection, a sizeable fraction of secondary memory T cells in the circulation developed downstream of T cells. These tissue-experienced ex-T cells shared phenotypic properties with the effector memory T cell population but were transcriptionally and functionally distinct from other secondary effector memory T cell cells. Adoptive transfer experiments of T cells corroborated their potential to form circulating effector and memory cells during recall responses. Moreover, specific ablation of primary T cell populations substantially impaired the secondary T cell response, both locally and systemically. Thus, T cells retain developmental plasticity and shape both local and systemic T cell responses on reinfection.
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http://dx.doi.org/10.1038/s41590-020-0723-4DOI Listing
September 2020

Clinical consequences of primary CMV infection after renal transplantation: a case-control study.

Transpl Int 2020 09 4;33(9):1116-1127. Epub 2020 Jul 4.

Renal Transplant Unit, Department of Internal Medicine, Academic Medical Center, Amsterdam, The Netherlands.

The impact of primary cytomegalovirus infection (pCMV) on renal allograft function and histology is controversial. We evaluated the influence on incidence of acute rejection, allograft loss, allograft function and interstitial fibrosis/tubular atrophy (IF/TA). Retrospective case-control study, recipients transplanted between 2000 and 2014. Risk of acute rejection and allograft loss for those who experienced pCMV infection compared with those who did not, within an exposure period of two months after transplantation. Besides, its influence on allograft function and histology at one to three years after transplantation. Of 113 recipients experienced pCMV infection, 306 remained CMV seronegative. pCMV infection in the exposure period could not be proven as increasing the risk for acute rejection [HR = 2.18 (95% CI 0.80-5.97) P = 0.13] or allograft loss [HR = 1.11 (95%CI 0.33-3.72) P = 0.87]. Combination of pCMV infection and acute rejection posed higher hazard for allograft loss than acute rejection alone [HR = 3.69 (95% CI 1.21-11.29) P = 0.02]. eGFR(MDRD) values did not significantly differ at years one [46 vs. 50], two [46 vs. 51] and three [46 vs. 52]. No association between pCMV infection and IF/TA could be demonstrated [OR = 2.15 (95%CI 0.73-6.29) P = 0.16]. pCMV infection was not proven to increase the risk for acute rejection or allograft loss. However, it increased the risk for rejection-associated allograft loss. In remaining functioning allografts, it was not significantly associated with decline in function nor with presence of IF/TA.
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http://dx.doi.org/10.1111/tri.13667DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7540315PMC
September 2020

Murine iNKT cells are depleted by liver damage via activation of P2RX7.

Eur J Immunol 2020 10 27;50(10):1515-1524. Epub 2020 May 27.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

Invariant natural killer T cells (iNKT) constitute up to 50% of liver lymphocytes and contribute to immunosurveillance as well as pathogenesis of the liver. Systemic activation of iNKT cells induces acute immune-mediated liver injury. However, how tissue damage events regulate iNKT cell function and homeostasis remains unclear. We found that specifically tissue-resident iNKT cells in liver and spleen express the tissue-damage receptor P2RX7 and the P2RX7-activating ectoenzyme ARTC2. P2RX7 expression restricted formation of iNKT cells in the liver suggesting that liver iNKT cells are actively restrained under homeostatic conditions. Deliberate activation of P2RX7 in vivo by exogenous NAD resulted in a nearly complete iNKT cell ablation in liver and spleen in a P2RX7-dependent manner. Tissue damage generated by acetaminophen-induced liver injury reduced the number of iNKT cells in the liver. The tissue-damage-induced iNKT cell depletion was driven by P2RX7 and localized to the site of injury, as iNKT cells in the spleen remained intact. The depleted liver iNKT cells reconstituted only slowly compared to other lymphocytes such as regulatory T cells. These findings suggest that tissue-damage-mediated depletion of iNKT cells acts as a feedback mechanism to limit iNKT cell-induced pathology resulting in the establishment of a tolerogenic environment.
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http://dx.doi.org/10.1002/eji.201948509DOI Listing
October 2020

GITR shapes humoral immunity by controlling the balance between follicular T helper cells and regulatory T follicular cells.

Immunol Lett 2020 06 4;222:73-79. Epub 2020 Apr 4.

Department of Hematopoiesis, Sanquin Research, Amsterdam, the Netherlands; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, the Netherlands; Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands. Electronic address:

Follicular helper CD4 T-cells (Tfh) control humoral immunity by driving affinity maturation and isotype-switching of activated B-cells. Tfh localize within B-cell follicles and, upon encounter with cognate antigen, drive B-cell selection in germinal centers (GCs) as GC-Tfh. Tfh functionality is controlled by Foxp3-expressing Tfh, which are known as regulatory T follicular cells (Tfr). Thus far, it remains unclear which factors determine the balance between these functionally opposing follicular T-cell subsets. Here, we demonstrate in human and mouse that Tfh and GC-Tfh, as well as their regulatory counterparts, express glucocorticoid-induced TNF receptor related protein (GITR) on their surface. This costimulatory molecule not only helps to identify follicular T-cell subsets, but also increases the ratio of Tfh vs. Tfr, both within and outside the GC. Correspondingly, GITR triggering increases the number of IL-21 producing CD4 T-cells, which also produce more IFN-γ and IL-10. The latter are known switch factors for IgG2c and IgG1, respectively, which corresponds to a concomitant increase in IgG2c and IgG1 production upon GITR-mediated costimulation. These results demonstrate that GITR can skew the functional balance between Tfh and Tfr, which offers new therapeutic possibilities in steering humoral immunity.
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http://dx.doi.org/10.1016/j.imlet.2020.03.008DOI Listing
June 2020

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition).

Eur J Immunol 2019 Oct;49(10):1457-1973

Flow Cytometry Laboratory, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München, German Research Center for Environmental Health, München, Germany.

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.
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http://dx.doi.org/10.1002/eji.201970107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7350392PMC
October 2019

Blimp-1 Rather Than Hobit Drives the Formation of Tissue-Resident Memory CD8 T Cells in the Lungs.

Front Immunol 2019 7;10:400. Epub 2019 Mar 7.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.

Tissue-resident memory CD8 T (T) cells that develop in the epithelia at portals of pathogen entry are important for improved protection against re-infection. CD8 T cells within the skin and the small intestine are long-lived and maintained independently of circulating memory CD8 T cells. In contrast to CD8 T cells at these sites, CD8 T cells that arise after influenza virus infection within the lungs display high turnover and require constant recruitment from the circulating memory pool for long-term persistence. The distinct characteristics of CD8 T cell maintenance within the lungs may suggest a unique program of transcriptional regulation of influenza-specific CD8 T cells. We have previously demonstrated that the transcription factors Hobit and Blimp-1 are essential for the formation of CD8 T cells across several tissues, including skin, liver, kidneys, and the small intestine. Here, we addressed the roles of Hobit and Blimp-1 in CD8 T cell differentiation in the lungs after influenza infection using mice deficient for these transcription factors. Hobit was not required for the formation of influenza-specific CD8 T cells in the lungs. In contrast, Blimp-1 was essential for the differentiation of lung CD8 T cells and inhibited the differentiation of central memory CD8 T (T) cells. We conclude that Blimp-1 rather than Hobit mediates the formation of CD8 T cells in the lungs, potentially through control of the lineage choice between T and T cells during the differentiation of influenza-specific CD8 T cells.
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http://dx.doi.org/10.3389/fimmu.2019.00400DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6416215PMC
July 2020

Expression of IL-7Rα and KLRG1 defines functionally distinct CD8 T-cell populations in humans.

Eur J Immunol 2019 05 25;49(5):694-708. Epub 2019 Mar 25.

Department of Experimental Immunology, Amsterdam Infection and Immunity Institute, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

During acute viral infections in mice, IL-7Rα and KLRG1 together are used to distinguish the short-lived effector cells (SLEC; IL-7Rα KLRG ) from the precursors of persisting memory cells (MPEC; IL-7Rα KLRG1 ). We here show that these markers can be used to define distinct subsets in the circulation and lymph nodes during the acute phase and in "steady state" in humans. In contrast to the T cells in the circulation, T cells derived from lymph nodes hardly contain any KLRG1-expressing cells. The four populations defined by IL-7Rα and KLRG1 differ markedly in transcription factor, granzyme and chemokine receptor expression. When studying renal transplant recipients experiencing a primary hCMV and EBV infection, we also found that after viral control, during latency, Ki-67-negative SLEC can be found in the peripheral blood in considerable numbers. Thus, combined analyses of IL-7Rα and KLRG1 expression on human herpes virus-specific CD8 T cells can be used to separate functionally distinct subsets in humans. As a noncycling IL-7Rα KLRG1 population is abundant in healthy humans, we conclude that this combination of markers not only defines short-lived effector cells during the acute response but also stable effector cells that are formed and remain present during latent herpes infections.
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http://dx.doi.org/10.1002/eji.201847897DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6593687PMC
May 2019

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Sci Immunol 2018 12;3(30)

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.

Tissue-resident memory T cells (T) are noncirculating immune cells that contribute to the first line of local defense against reinfections. Their location at hotspots of pathogen encounter frequently exposes T to tissue damage. This history of danger-signal exposure is an important aspect of T-mediated immunity that has been overlooked so far. RNA profiling revealed that T from liver and small intestine express P2RX7, a damage/danger-associated molecular pattern (DAMP) receptor that is triggered by extracellular nucleotides (ATP, NAD). We confirmed that P2RX7 protein was expressed in CD8 T but not in circulating T cells (T) across different infection models. Tissue damage induced during routine isolation of liver lymphocytes led to P2RX7 activation and resulted in selective cell death of T P2RX7 activation in vivo by exogenous NAD led to a specific depletion of T while retaining T The effect was absent in P2RX7-deficient mice and after P2RX7 blockade. TCR triggering down-regulated P2RX7 expression and made T resistant to NAD-induced cell death. Physiological triggering of P2RX7 by sterile tissue damage during acetaminophen-induced liver injury led to a loss of previously acquired pathogen-specific local T in wild-type but not in P2RX7 KO T cells. Our results highlight P2RX7-mediated signaling as a critical pathway for the regulation of T maintenance. Extracellular nucleotides released during infection and tissue damage could deplete T locally and free niches for new and infection-relevant specificities. This suggests that the recognition of tissue damage promotes persistence of antigen-specific over bystander T in the tissue niche.
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http://dx.doi.org/10.1126/sciimmunol.aau1022DOI Listing
December 2018

Functional Heterogeneity of CD4 Tumor-Infiltrating Lymphocytes With a Resident Memory Phenotype in NSCLC.

Front Immunol 2018 16;9:2654. Epub 2018 Nov 16.

Sanquin Research, Department of Hematopoiesis, and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.

Resident memory T cells (T) inhabit peripheral tissues and are critical for protection against localized infections. Recently, it has become evident that CD103 T are not only important in combating secondary infections, but also for the elimination of tumor cells. In several solid cancers, intratumoral CD103CD8 tumor infiltrating lymphocytes (TILs), with T properties, are a positive prognostic marker. To better understand the role of T in tumors, we performed a detailed characterization of CD8 and CD4 TIL phenotype and functional properties in non-small cell lung cancer (NSCLC). Frequencies of CD8 and CD4 T cell infiltrates in tumors were comparable, but we observed a sharp contrast in T ratios compared to surrounding lung tissue. The majority of both CD4 and CD8 TILs expressed CD69 and a subset also expressed CD103, both hallmarks of T. While CD103CD8 T cells were enriched in tumors, CD103CD4 T cell frequencies were decreased compared to surrounding lung tissue. Furthermore, CD103CD4 and CD103CD8 TILs showed multiple characteristics of T, such as elevated expression of CXCR6 and CD49a, and decreased expression of T-bet and Eomes. In line with the immunomodulatory role of the tumor microenvironment, CD8 and CD4 TILs expressed high levels of inhibitory receptors 2B4, CTLA-4, and PD-1, with the highest levels found on CD103 TILs. Strikingly, CD103CD4 TILs were the most potent producers of TNF-α and IFN-γ, while other TIL subsets lacked such cytokine production. Whereas, CD103CD4PD-1 TILs produced the most effector cytokines, CD103CD4PD-1 and CD69CD4PD-1 TILs produced CXCL13. Furthermore, a large proportion of TILs expressed co-stimulatory receptors CD27 and CD28, unlike lung T, suggesting a less differentiated phenotype. Agonistic triggering of these receptors improved cytokine production of CD103CD4 and CD69CD8 TILs. Our findings thus provide a rationale to target CD103CD4 TILs and add co-stimulation to current therapies to improve the efficacy of immunotherapies and cancer vaccines.
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http://dx.doi.org/10.3389/fimmu.2018.02654DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6250821PMC
October 2019

Tissue-resident memory T cells populate the human brain.

Nat Commun 2018 11 2;9(1):4593. Epub 2018 Nov 2.

Department of Neuroimmunology, Netherlands Institute for Neuroscience, Meibergdreef 47, 1105BA, Amsterdam, The Netherlands.

Most tissues are populated by tissue-resident memory T cells (T cells), which are adapted to their niche and appear to be indispensable for local protection against pathogens. Here we show that human white matter-derived brain CD8 T cells can be subsetted into CD103CD69 and CD103CD69 T cells both with a phenotypic and transcription factor profile consistent with T cells. Specifically, CD103 expression in brain CD8 T cells correlates with reduced expression of differentiation markers, increased expression of tissue-homing chemokine receptors, intermediate and low expression of the transcription factors T-bet and eomes, increased expression of PD-1 and CTLA-4, and low expression of cytolytic enzymes with preserved polyfunctionality upon activation. Brain CD4 T cells also display T cell-associated markers but have low CD103 expression. We conclude that the human brain is surveilled by T cells, providing protection against neurotropic virus reactivation, whilst being under tight control of key immune checkpoint molecules.
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http://dx.doi.org/10.1038/s41467-018-07053-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6214977PMC
November 2018

Blimp-1 induces and Hobit maintains the cytotoxic mediator granzyme B in CD8 T cells.

Eur J Immunol 2018 10 16;48(10):1644-1662. Epub 2018 Aug 16.

Dept of Hematopoiesis, Sanquin Research and Landsteiner Laboratory Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.

CD8 T cells acquire cytotoxic molecules including granzyme B during effector differentiation. Both tissue-resident memory CD8 T cells (Trm) and circulating CD45RA effector-type T cells (Temra) cells have the ability to retain granzyme B protein expression into the memory phase, but it is unclear how this persistence of cytolytic activity is regulated during steady state. Previously, we have described that the transcriptional regulators Hobit and Blimp-1 have overlapping target genes that include granzyme B, but their impact on the regulation of cytotoxicity in Trm and Temra cells during homeostasis has remained unclear. We examined the expression regulation of Hobit and Blimp-1 in murine and human CD8 T-cells to determine their timeframe of activity. While Blimp-1 mRNA was expressed throughout effector and memory T cells, Blimp-1 protein, was only transiently expressed during the effector stage. In contrast, Hobit mRNA and protein expression was stably maintained during quiescence, but downregulated after activation. Notably, Blimp-1 was required for expression of granzyme B in murine effector T cells and Trm, while Hobit specifically regulated granzyme B in murine Trm during the memory phase. These findings suggest that Blimp-1 initiates cytotoxic effector function and that Hobit maintains cytotoxicity in a deployment-ready modus in Trm.
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http://dx.doi.org/10.1002/eji.201847771DOI Listing
October 2018

Neutrophils Kill Antibody-Opsonized Cancer Cells by Trogoptosis.

Cell Rep 2018 06;23(13):3946-3959.e6

Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands; Department of Molecular Cell Biology and Immunology, VU Medical Center, Amsterdam, the Netherlands. Electronic address:

Destruction of cancer cells by therapeutic antibodies occurs, at least in part, through antibody-dependent cellular cytotoxicity (ADCC), and this can be mediated by various Fc-receptor-expressing immune cells, including neutrophils. However, the mechanism(s) by which neutrophils kill antibody-opsonized cancer cells has not been established. Here, we demonstrate that neutrophils can exert a mode of destruction of cancer cells, which involves antibody-mediated trogocytosis by neutrophils. Intimately associated with this is an active mechanical disruption of the cancer cell plasma membrane, leading to a lytic (i.e., necrotic) type of cancer cell death. Furthermore, this mode of destruction of antibody-opsonized cancer cells by neutrophils is potentiated by CD47-SIRPα checkpoint blockade. Collectively, these findings show that neutrophil ADCC toward cancer cells occurs by a mechanism of cytotoxicity called trogoptosis, which can be further improved by targeting CD47-SIRPα interactions.
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http://dx.doi.org/10.1016/j.celrep.2018.05.082DOI Listing
June 2018

Tissue-resident memory T cells at the center of immunity to solid tumors.

Nat Immunol 2018 06 18;19(6):538-546. Epub 2018 May 18.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

Immune responses in tissues are constrained by the physiological properties of the tissue involved. Tissue-resident memory T cells (T cells) are a recently discovered lineage of T cells specialized for life and function within tissues. Emerging evidence has shown that T cells have a special role in the control of solid tumors. A high frequency of T cells in tumors correlates with favorable disease progression in patients with cancer, and studies of mice have shown that T cells are necessary for optimal immunological control of solid tumors. Here we describe what defines T cells as a separate lineage and how these cells are generated. Furthermore, we discuss the properties that allow T cells to operate in normal and transformed tissues, as well as implications for the treatment of patients with cancer.
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http://dx.doi.org/10.1038/s41590-018-0114-2DOI Listing
June 2018

Proteomic Analyses of Human Regulatory T Cells Reveal Adaptations in Signaling Pathways that Protect Cellular Identity.

Immunity 2018 05 8;48(5):1046-1059.e6. Epub 2018 May 8.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands. Electronic address:

To obtain a molecular definition of regulatory T (Treg) cell identity, we performed proteomics and transcriptomics on various populations of human regulatory and conventional CD4 T (Tconv) cells. A protein expression signature was identified that defines all Treg cells, and another signature that defines effector Treg cells. These signatures could not be extrapolated from transcriptome data. Unique cell-biological and metabolic features in Treg cells were defined, as well as specific adaptations in cytokine, TCR, and costimulatory receptor signaling pathways. One such adaptation-selective STAT4 deficiency-prevented destabilization of Treg cell identity and function by inflammatory cytokines, while these signals could still induce critical transcription factors and homing receptors via other pathways. Furthermore, our study revealed surface markers that identify FOXP3CD4 T cells with distinct functional properties. Our findings suggest that adaptation in signaling pathways protect Treg cell identity and present a resource for further research into Treg cell biology.
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http://dx.doi.org/10.1016/j.immuni.2018.04.008DOI Listing
May 2018

The Contribution of Cytomegalovirus Infection to Immune Senescence Is Set by the Infectious Dose.

Front Immunol 2017 10;8:1953. Epub 2018 Jan 10.

Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, Netherlands.

The relationship between human cytomegalovirus (HCMV) infections and accelerated immune senescence is controversial. Whereas some studies reported a CMV-associated impaired capacity to control heterologous infections at old age, other studies could not confirm this. We hypothesized that these discrepancies might relate to the variability in the infectious dose of CMV occurring in real life. Here, we investigated the influence of persistent CMV infection on immune perturbations and specifically addressed the role of the infectious dose on the contribution of CMV to accelerated immune senescence. We show in experimental mouse models that the degree of mouse CMV (MCMV)-specific memory CD8 T cell accumulation and the phenotypic T cell profile are directly influenced by the infectious dose, and data on HCMV-specific T cells indicate a similar connection. Detailed cluster analysis of the memory CD8 T cell development showed that high-dose infection causes a differentiation pathway that progresses faster throughout the life span of the host, suggesting a virus-host balance that is influenced by aging and infectious dose. Importantly, short-term MCMV infection in adult mice is not disadvantageous for heterologous superinfection with lymphocytic choriomeningitis virus (LCMV). However, following long-term CMV infection the strength of the CD8 T cell immunity to LCMV superinfection was affected by the initial CMV infectious dose, wherein a high infectious dose was found to be a prerequisite for impaired heterologous immunity. Altogether our results underscore the importance of stratification based on the size and differentiation of the CMV-specific memory T cell pools for the impact on immune senescence, and indicate that reduction of the latent/lytic viral load can be beneficial to diminish CMV-associated immune senescence.
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http://dx.doi.org/10.3389/fimmu.2017.01953DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5768196PMC
January 2018

Tumor immunity requires border patrol to fight the enemy within.

Nat Immunol 2017 07;18(8):870-872

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory for Blood Cell Research, Amsterdam, the Netherlands.

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http://dx.doi.org/10.1038/ni.3792DOI Listing
July 2017

Cytomegalovirus (CMV) research in immune senescence comes of age: overview of the 6th International Workshop on CMV and Immunosenescence.

Geroscience 2017 06 17;39(3):245-249. Epub 2017 Jun 17.

Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

Cytomegalovirus (CMV) is one of the most complex and most ubiquitous latent persistent viruses, with a considerable ability to evade and manipulate the immune system. Following an early-life infection, most immunocompetent humans spend several decades living with CMV, and, because the virus in these hosts does not cause manifest disease, CMV can be considered part of normal aging for more than half of humanity. However, there is accumulating evidence that CMV carriage is not a null event and that both potentially harmful and potentially beneficial outcomes emanate from the interaction of CMV with its mammalian hosts. This article provides an overview of the 6th International Workshop on CMV and Immunosenescence, highlighting the advances in the field made in the past two years, as related to CMV epidemiology/geroscience, CMV virology with an accent on latency, and CMV immune evasion and immune recognition of the virus and its antigens.
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http://dx.doi.org/10.1007/s11357-017-9984-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5505886PMC
June 2017

Label-free Analysis of CD8 T Cell Subset Proteomes Supports a Progressive Differentiation Model of Human-Virus-Specific T Cells.

Cell Rep 2017 05;19(5):1068-1079

Sanquin Research and Landsteiner laboratory, 1066CX Amsterdam, the Netherlands.

Pathogens trigger T cells to express distinct sets of effector proteins. To better understand the molecular mechanisms that drive functional specification, we used high-resolution mass spectrometry and label-free protein quantification to measure proteomic differences between the seven largest circulating human CD8 T cell subsets. Hierarchical clustering of the proteomes placed naive and CD45RA-expressing effector-type T cells at the extremes of the spectrum, with central memory and other effector memory stages located in between. Prominent differences between the subsets included expression of specific granzymes, signaling proteins, and molecules involved in metabolic regulation and cell adhesion. Remarkably, whereas most of the proteomic relationships between the subsets occurred in linear variations, a small proportion of proteins was regulated only in discrete subsets. The data obtained from this proteome analysis correspond best to a progressive differentiation model in which specific stable traits are gradually acquired during development.
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http://dx.doi.org/10.1016/j.celrep.2017.04.014DOI Listing
May 2017

The Transcription Factor Hobit Identifies Human Cytotoxic CD4 T Cells.

Front Immunol 2017 24;8:325. Epub 2017 Mar 24.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory , Amsterdam , Netherlands.

The T cell lineage is commonly divided into CD4-expressing helper T cells that polarize immune responses through cytokine secretion and CD8-expressing cytotoxic T cells that eliminate infected target cells by virtue of the release of cytotoxic molecules. Recently, a population of CD4 T cells that conforms to the phenotype of cytotoxic CD8 T cells has received increased recognition. These cytotoxic CD4 T cells display constitutive expression of granzyme B and perforin at the protein level and mediate HLA class II-dependent killing of target cells. In humans, this cytotoxic profile is found within the human cytomegalovirus (hCMV)-specific, but not within the influenza- or Epstein-Barr virus-specific CD4 T cell populations, suggesting that, in particular, hCMV infection induces the formation of cytotoxic CD4 T cells. We have previously described that the transcription factor Homolog of Blimp-1 in T cells (Hobit) is specifically upregulated in CD45RA effector CD8 T cells that arise after hCMV infection. Here, we describe the expression pattern of Hobit in human CD4 T cells. We found Hobit expression in cytotoxic CD4 T cells and accumulation of Hobit CD4 T cells after primary hCMV infection. The Hobit CD4 T cells displayed highly overlapping characteristics with Hobit CD8 T cells, including the expression of cytotoxic molecules, T-bet, and CX3CR1. Interestingly, γδ T cells that arise after hCMV infection also upregulate Hobit expression and display a similar effector phenotype as cytotoxic CD4 and CD8 T cells. These findings suggest a shared differentiation pathway in CD4, CD8, and γδ T cells that may involve Hobit-driven acquisition of long-lived cytotoxic effector function.
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http://dx.doi.org/10.3389/fimmu.2017.00325DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5364140PMC
March 2017

The cellular immune system comes of age.

J Allergy Clin Immunol 2017 06 22;139(6):1793-1794. Epub 2017 Mar 22.

Department of Blood Cell Research, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands; Department of Pediatric Hematology, Immunology, and Infectious Disease, Emma Children's Hospital, Academic Medical Center, Amsterdam, The Netherlands.

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http://dx.doi.org/10.1016/j.jaci.2017.02.023DOI Listing
June 2017

Programs for the persistence, vigilance and control of human CD8 lung-resident memory T cells.

Nat Immunol 2016 Dec 24;17(12):1467-1478. Epub 2016 Oct 24.

Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, Amsterdam, the Netherlands.

Tissue-resident memory T cells (T cells) in the airways mediate protection against respiratory infection. We characterized T cells expressing integrin α (CD103) that reside within the epithelial barrier of human lungs. These cells had specialized profiles of chemokine receptors and adhesion molecules, consistent with their unique localization. Lung T cells were poised for rapid responsiveness by constitutive expression of deployment-ready mRNA encoding effector molecules, but they also expressed many inhibitory regulators, suggestive of programmed restraint. A distinct set of transcription factors was active in CD103 T cells, including Notch. Genetic and pharmacological experiments with mice revealed that Notch activity was required for the maintenance of CD103 T cells. We have thus identified specialized programs underlying the residence, persistence, vigilance and tight control of human lung T cells.
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http://dx.doi.org/10.1038/ni.3589DOI Listing
December 2016

Clinically Relevant Reactivation of Polyomavirus BK (BKPyV) in HLA-A02-Positive Renal Transplant Recipients Is Associated with Impaired Effector-Memory Differentiation of BKPyV-Specific CD8+ T Cells.

PLoS Pathog 2016 Oct 10;12(10):e1005903. Epub 2016 Oct 10.

Department of Experimental Immunology, Amsterdam, the Netherlands.

Polyomavirus BK (BKPyV) frequently reactivates in immunosuppressed renal transplant recipients (RTRs) and may lead to graft loss due to BKPyV-induced interstitial nephritis (BKVN). Little is known on the differentiation of CD8+ T cells targeting BKPyV in RTRs. Here we investigated whether BKPyV-specific CD8+ T cell differentiation differs in RTRs with varying degrees of BKPyV reactivation and/or BKVN. Using combinatorial encoding with tetramers carrying BKPyV major capsid protein (VP1) and large T antigen protein (LTAG) epitopes, we investigated CD8+ T cell responses to BKPyV in longitudinally obtained PBMC samples from 46 HLA-A02-positive RTRs and 20 healthy adults. We were also able to isolate BKPyV-specific CD8+ T cells from five renal allografts, two of which were affected by BKVN. Before transplantation, BKPyV-specific CD8+ T cells targeting VP1 and LTAG epitopes appeared predominantly as central-memory and CD27+/CD28+ effector-memory (TEM), and naïve-like PD-1-expressing cells, respectively. After viral reactivation, BKPyV-specific CD8+ T cells assumed CD28- TEM and TEMRA states in patients who were able to control BKPyV, whereas differentiation lagged behind in patients with severe viral reactivation or BKVN. Furthermore, VP1-specific CD69+/CD103+ tissue-resident memory (TRM) cells accumulated in BKVN-affected allografts but lacked signs of effector differentiation. In contrast, granzyme B-expressing effector cells were detected in allografts not affected by BKVN. In conclusion, effector-memory differentiation of BKPyV-specific CD8+ T cells in patients with high viral load or BKVN is impaired. Further characterization of the specific mechanisms behind this altered cellular differentiation is necessary to develop therapies that can prevent the emergence of BKVN.
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http://dx.doi.org/10.1371/journal.ppat.1005903DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5056763PMC
October 2016

Immune checkpoint blockade: Which switches to hit and how much?

Immunol Lett 2016 12 20;180:73-74. Epub 2016 Sep 20.

Sanquin Research, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. Electronic address:

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http://dx.doi.org/10.1016/j.imlet.2016.08.014DOI Listing
December 2016
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