79 results
Search Results
2. Paper electrophoresis of a purified specific antibody.
- Author
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TEKMAN S and UGUR A
- Subjects
- Humans, Antibodies, Antigens, Electrophoresis, Electrophoresis, Paper
- Published
- 1955
- Full Text
- View/download PDF
3. Detection of antigens as specific precipitates on paper electrophoresis strips
- Author
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S. Buttery
- Subjects
Multidisciplinary ,food.ingredient ,Chromatography ,Chemistry ,fungi ,food and beverages ,Paper electrophoresis ,humanities ,food ,Precipitin reaction ,Antigen ,Agar ,Humans ,Electrophoresis, Paper ,Antigens - Abstract
AGAR diffusion1 has been used for detecting antigens after paper electrophoresis. I have found that a precipitin reaction can be carried out directly on the paper.
- Published
- 1959
4. Paper electrophoresis of a purified specific antibody
- Author
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Ayten Ugur and S. Tekman
- Subjects
Electrophoresis ,Multidisciplinary ,biology ,Chemistry ,Paper electrophoresis ,biochemical phenomena, metabolism, and nutrition ,Gel electrophoresis of proteins ,Immune sera ,Antibodies ,Specific antibody ,Biochemistry ,biology.protein ,Humans ,Electrophoresis, Paper ,Antibody ,Antigens - Abstract
RECENTLY, several workers1 have tried to fractionate gamma-globulins and immune sera by electrophoresis. Such experiments have only given the mean properties of all gamma-globulin proteins, since non-antibody as well as antibody proteins occur in immune sera. We have now carried out paper electrophoresis of a purified specific antibody.
- Published
- 1955
5. The duration of antigen receptor signalling determines CD4+ versus CD8+ T-cell lineage fate.
- Author
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Yasutomo, Koji and Doyle, Carolyn
- Subjects
CD antigens ,ANTIGENS ,CD4 antigen ,T cell receptors - Abstract
Shows hat the CD4 versus CD8 lineage fate of immature thymocytes is controlled by the coreceptor-influenced duration of initial T-cell receptor-dependent signalling. Role of co-receptor in CD4/CD8 lineage choice; Thymocyte stimulation and preparation of CD69hi cells; Thymic reaggregate culture.
- Published
- 2000
- Full Text
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6. Atomic structure of anthrax protective antigen pore elucidates toxin translocation.
- Author
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Jiang, Jiansen, Pentelute, Bradley L., Collier, R. John, and Zhou, Z. Hong
- Subjects
ATOMIC structure ,ANTHRAX ,ANTIGENS ,CHROMOSOMAL translocation ,BACILLUS anthracis ,OLIGOMERS ,ELECTROPHYSIOLOGY ,CONFORMATIONAL analysis - Abstract
Anthrax toxin, comprising protective antigen, lethal factor, and oedema factor, is the major virulence factor of Bacillus anthracis, an agent that causes high mortality in humans and animals. Protective antigen forms oligomeric prepores that undergo conversion to membrane-spanning pores by endosomal acidification, and these pores translocate the enzymes lethal factor and oedema factor into the cytosol of target cells. Protective antigen is not only a vaccine component and therapeutic target for anthrax infections but also an excellent model system for understanding the mechanism of protein translocation. On the basis of biochemical and electrophysiological results, researchers have proposed that a phi (Φ)-clamp composed of phenylalanine (Phe)427 residues of protective antigen catalyses protein translocation via a charge-state-dependent Brownian ratchet. Although atomic structures of protective antigen prepores are available, how protective antigen senses low pH, converts to active pore, and translocates lethal factor and oedema factor are not well defined without an atomic model of its pore. Here, by cryo-electron microscopy with direct electron counting, we determine the protective antigen pore structure at 2.9-Å resolution. The structure reveals the long-sought-after catalytic Φ-clamp and the membrane-spanning translocation channel, and supports the Brownian ratchet model for protein translocation. Comparisons of four structures reveal conformational changes in prepore to pore conversion that support a multi-step mechanism by which low pH is sensed and the membrane-spanning channel is formed. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
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7. PD-1 blockade induces responses by inhibiting adaptive immune resistance.
- Author
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Tumeh, Paul C., Chmielowski, Bartosz, Glaspy, John A., Elashoff, David A., Ribas, Antoni, Harview, Christina L., Shintaku, I. Peter, Taylor, Emma J. M., Robert, Lidia, Spasic, Marko, Henry, Gina, Ciobanu, Voicu, West, Alisha N., Carmona, Manuel, Kivork, Christine, Seja, Elizabeth, Cherry, Grace, Gutierrez, Antonio J., Grogan, Tristan R., and Yearley, Jennifer H.
- Subjects
CANCER patients ,IMMUNE response ,ANTIGENS ,T cells ,MELANOMA - Abstract
Therapies that target the programmed death-1 (PD-1) receptor have shown unprecedented rates of durable clinical responses in patients with various cancer types. One mechanism by which cancer tissues limit the host immune response is via upregulation of PD-1 ligand (PD-L1) and its ligation to PD-1 on antigen-specific CD8
+ T cells (termed adaptive immune resistance). Here we show that pre-existing CD8+ T cells distinctly located at the invasive tumour margin are associated with expression of the PD-1/PD-L1 immune inhibitory axis and may predict response to therapy. We analysed samples from 46 patients with metastatic melanoma obtained before and during anti-PD-1 therapy (pembrolizumab) using quantitative immunohistochemistry, quantitative multiplex immunofluorescence, and next-generation sequencing for T-cell antigen receptors (TCRs). In serially sampled tumours, patients responding to treatment showed proliferation of intratumoral CD8+ T cells that directly correlated with radiographic reduction in tumour size. Pre-treatment samples obtained from responding patients showed higher numbers of CD8-, PD-1- and PD-L1-expressing cells at the invasive tumour margin and inside tumours, with close proximity between PD-1 and PD-L1, and a more clonal TCR repertoire. Using multivariate analysis, we established a predictive model based on CD8 expression at the invasive margin and validated the model in an independent cohort of 15 patients. Our findings indicate that tumour regression after therapeutic PD-1 blockade requires pre-existing CD8+ T cells that are negatively regulated by PD-1/PD-L1-mediated adaptive immune resistance. [ABSTRACT FROM AUTHOR]- Published
- 2014
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8. MPDL3280A (anti-PD-L1) treatment leads to clinical activity in metastatic bladder cancer.
- Author
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Powles, Thomas, Eder, Joseph Paul, Petrylak, Daniel P., Fine, Gregg D., Teng, Siew-leng, Shen, Xiaodong, Boyd, Zachary, Hegde, Priti S., Chen, Daniel S., Braiteh, Fadi S., Loriot, Yohann, Cruz, Cristina, Bellmunt, Joaquim, Burris, Howard A., and Vogelzang, Nicholas J.
- Subjects
METASTASIS ,BLADDER cancer treatment ,CANCER chemotherapy ,SOMATIC mutation ,ANTIGENS - Abstract
There have been no major advances for the treatment of metastatic urothelial bladder cancer (UBC) in the last 30 years. Chemotherapy is still the standard of care. Patient outcomes, especially for those in whom chemotherapy is not effective or is poorly tolerated, remain poor. One hallmark of UBC is the presence of high rates of somatic mutations. These alterations may enhance the ability of the host immune system to recognize tumour cells as foreign owing to an increased number of antigens. However, these cancers may also elude immune surveillance and eradication through the expression of programmed death-ligand 1 (PD-L1; also called CD274 or B7-H1) in the tumour microenvironment. Therefore, we examined the anti-PD-L1 antibody MPDL3280A, a systemic cancer immunotherapy, for the treatment of metastatic UBC. MPDL3280A is a high-affinity engineered human anti-PD-L1 monoclonal immunoglobulin-G1 antibody that inhibits the interaction of PD-L1 with PD-1 (PDCD1) and B7.1 (CD80). Because PD-L1 is expressed on activated T cells, MPDL3280A was engineered with a modification in the Fc domain that eliminates antibody-dependent cellular cytotoxicity at clinically relevant doses to prevent the depletion of T cells expressing PD-L1. Here we show that MPDL3280A has noteworthy activity in metastatic UBC. Responses were often rapid, with many occurring at the time of the first response assessment (6 weeks) and nearly all were ongoing at the data cutoff. This phase I expansion study, with an adaptive design that allowed for biomarker-positive enriched cohorts, demonstrated that tumours expressing PD-L1-positive tumour-infiltrating immune cells had particularly high response rates. Moreover, owing to the favourable toxicity profile, including a lack of renal toxicity, patients with UBC, who are often older and have a higher incidence of renal impairment, may be better able to tolerate MPDL3280A versus chemotherapy. These results suggest that MPDL3280A may have an important role in treating UBC-the drug received breakthrough designation status by the US Food and Drug Administration (FDA) in June 2014. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
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9. T-cell activation by transitory neo-antigens derived from distinct microbial pathways.
- Author
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Corbett, Alexandra J., Eckle, Sidonia B. G., Birkinshaw, Richard W., Liu, Ligong, Patel, Onisha, Mahony, Jennifer, Chen, Zhenjun, Reantragoon, Rangsima, Meehan, Bronwyn, Cao, Hanwei, Williamson, Nicholas A., Strugnell, Richard A., Van Sinderen, Douwe, Mak, Jeffrey Y. W., Fairlie, David P., Kjer-Nielsen, Lars, Rossjohn, Jamie, and McCluskey, James
- Subjects
T cell receptors ,VITAMIN B2 ,MICROBIAL cells ,ANTIGENS ,PEPTIDES ,BACTERIOLOGY ,CELL populations - Abstract
T cells discriminate between foreign and host molecules by recognizing distinct microbial molecules, predominantly peptides and lipids. Riboflavin precursors found in many bacteria and yeast also selectively activate mucosal-associated invariant T (MAIT) cells, an abundant population of innate-like T cells in humans. However, the genesis of these small organic molecules and their mode of presentation to MAIT cells by the major histocompatibility complex (MHC)-related protein MR1 (ref. 8) are not well understood. Here we show that MAIT-cell activation requires key genes encoding enzymes that form 5-amino-6-d-ribitylaminouracil (5-A-RU), an early intermediate in bacterial riboflavin synthesis. Although 5-A-RU does not bind MR1 or activate MAIT cells directly, it does form potent MAIT-activating antigens via non-enzymatic reactions with small molecules, such as glyoxal and methylglyoxal, which are derived from other metabolic pathways. The MAIT antigens formed by the reactions between 5-A-RU and glyoxal/methylglyoxal were simple adducts, 5-(2-oxoethylideneamino)-6-d-ribitylaminouracil (5-OE-RU) and 5-(2-oxopropylideneamino)-6-d-ribitylaminouracil (5-OP-RU), respectively, which bound to MR1 as shown by crystal structures of MAIT TCR ternary complexes. Although 5-OP-RU and 5-OE-RU are unstable intermediates, they became trapped by MR1 as reversible covalent Schiff base complexes. Mass spectra supported the capture by MR1 of 5-OP-RU and 5-OE-RU from bacterial cultures that activate MAIT cells, but not from non-activating bacteria, indicating that these MAIT antigens are present in a range of microbes. Thus, MR1 is able to capture, stabilize and present chemically unstable pyrimidine intermediates, which otherwise convert to lumazines, as potent antigens to MAIT cells. These pyrimidine adducts are microbial signatures for MAIT-cell immunosurveillance. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
10. Computational design of ligand-binding proteins with high affinity and selectivity.
- Author
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Tinberg, Christine E., Khare, Sagar D., Dou, Jiayi, Doyle, Lindsey, Nelson, Jorgen W., Schena, Alberto, Jankowski, Wojciech, Kalodimos, Charalampos G., Johnsson, Kai, Stoddard, Barry L., and Baker, David
- Subjects
CARRIER proteins ,LIGANDS (Biochemistry) ,DIGOXIGENIN ,BINDING sites ,ANTIGENS ,MOLECULAR recognition ,BIOSENSOR research - Abstract
The ability to design proteins with high affinity and selectivity for any given small molecule is a rigorous test of our understanding of the physiochemical principles that govern molecular recognition. Attempts to rationally design ligand-binding proteins have met with little success, however, and the computational design of protein-small-molecule interfaces remains an unsolved problem. Current approaches for designing ligand-binding proteins for medical and biotechnological uses rely on raising antibodies against a target antigen in immunized animals and/or performing laboratory-directed evolution of proteins with an existing low affinity for the desired ligand, neither of which allows complete control over the interactions involved in binding. Here we describe a general computational method for designing pre-organized and shape complementary small-molecule-binding sites, and use it to generate protein binders to the steroid digoxigenin (DIG). Of seventeen experimentally characterized designs, two bind DIG; the model of the higher affinity binder has the most energetically favourable and pre-organized interface in the design set. A comprehensive binding-fitness landscape of this design, generated by library selections and deep sequencing, was used to optimize its binding affinity to a picomolar level, and X-ray co-crystal structures of two variants show atomic-level agreement with the corresponding computational models. The optimized binder is selective for DIG over the related steroids digitoxigenin, progesterone and β-oestradiol, and this steroid binding preference can be reprogrammed by manipulation of explicitly designed hydrogen-bonding interactions. The computational design method presented here should enable the development of a new generation of biosensors, therapeutics and diagnostics. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
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11. Thymus-derived regulatory T cells contribute to tolerance to commensal microbiota.
- Author
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Cebula, Anna, Seweryn, Michal, Rempala, Grzegorz A., Pabla, Simarjot Singh, McIndoe, Richard A., Denning, Timothy L., Bry, Lynn, Kraj, Piotr, Kisielow, Pawel, and Ignatowicz, Leszek
- Subjects
THYMUS ,AUTOIMMUNITY ,ANTIGENS ,T cell receptors ,CD4 antigen ,NUCLEOTIDE sequence - Abstract
Peripheral mechanisms preventing autoimmunity and maintaining tolerance to commensal microbiota involve CD4
+ Foxp3+ regulatory T (Treg ) cells generated in the thymus or extrathymically by induction of naive CD4+ Foxp3− T cells. Previous studies suggested that the T-cell receptor repertoires of thymic Treg cells and induced Treg cells are biased towards self and non-self antigens, respectively, but their relative contribution in controlling immunopathology, such as colitis and other untoward inflammatory responses triggered by different types of antigens, remains unresolved. The intestine, and especially the colon, is a particularly suitable organ to study this question, given the variety of self-, microbiota- and food-derived antigens to which Treg cells and other T-cell populations are exposed. Intestinal environments can enhance conversion to a regulatory lineage and favour tolerogenic presentation of antigens to naive CD4+ T cells, suggesting that intestinal homeostasis depends on microbiota-specific induced Treg cells. Here, to identify the origin and antigen-specificity of intestinal Treg cells, we performed single-cell and high-throughput sequencing of the T-cell receptor repertoires of CD4+ Foxp3+ and CD4+ Foxp3− T cells, and analysed their reactivity against specific commensal species. We show that thymus-derived Treg cells constitute most Treg cells in all lymphoid and intestinal organs, including the colon, where their repertoire is heavily influenced by the composition of the microbiota. Our results suggest that thymic Treg cells, and not induced Treg cells, dominantly mediate tolerance to antigens produced by intestinal commensals. [ABSTRACT FROM AUTHOR]- Published
- 2013
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12. MR1 presents microbial vitamin B metabolites to MAIT cells.
- Author
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Kjer-Nielsen, Lars, Patel, Onisha, Corbett, Alexandra J., Le Nours, Jérôme, Meehan, Bronwyn, Liu, Ligong, Bhati, Mugdha, Chen, Zhenjun, Kostenko, Lyudmila, Reantragoon, Rangsima, Williamson, Nicholas A., Purcell, Anthony W., Dudek, Nadine L., McConville, Malcolm J., O'Hair, Richard A. J., Khairallah, George N., Godfrey, Dale I., Fairlie, David P., Rossjohn, Jamie, and McCluskey, James
- Subjects
ANTIGENS ,METABOLITES ,HLA histocompatibility antigens ,MAJOR histocompatibility complex ,VITAMIN B2 ,FLAVINS - Abstract
Antigen-presenting molecules, encoded by the major histocompatibility complex (MHC) and CD1 family, bind peptide- and lipid-based antigens, respectively, for recognition by T cells. Mucosal-associated invariant T (MAIT) cells are an abundant population of innate-like T cells in humans that are activated by an antigen(s) bound to the MHC class I-like molecule MR1. Although the identity of MR1-restricted antigen(s) is unknown, it is present in numerous bacteria and yeast. Here we show that the structure and chemistry within the antigen-binding cleft of MR1 is distinct from the MHC and CD1 families. MR1 is ideally suited to bind ligands originating from vitamin metabolites. The structure of MR1 in complex with 6-formyl pterin, a folic acid (vitamin B9) metabolite, shows the pterin ring sequestered within MR1. Furthermore, we characterize related MR1-restricted vitamin derivatives, originating from the bacterial riboflavin (vitamin B2) biosynthetic pathway, which specifically and potently activate MAIT cells. Accordingly, we show that metabolites of vitamin B represent a class of antigen that are presented by MR1 for MAIT-cell immunosurveillance. As many vitamin biosynthetic pathways are unique to bacteria and yeast, our data suggest that MAIT cells use these metabolites to detect microbial infection. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
13. Melanomas resist T-cell therapy through inflammation-induced reversible dedifferentiation.
- Author
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Landsberg, Jennifer, Kohlmeyer, Judith, Renn, Marcel, Bald, Tobias, Rogava, Meri, Cron, Mira, Fatho, Martina, Lennerz, Volker, Wölfel, Thomas, Hölzel, Michael, and Tüting, Thomas
- Subjects
MELANOMA treatment ,T cells ,CYTOKINES ,TUMOR necrosis factors ,ANTIGENS ,CANCER relapse ,DISEASE remission ,SPONTANEOUS cancer regression ,THERAPEUTICS - Abstract
Adoptive cell transfer therapies (ACTs) with cytotoxic T cells that target melanocytic antigens can achieve remissions in patients with metastatic melanomas, but tumours frequently relapse. Hypotheses explaining the acquired resistance to ACTs include the selection of antigen-deficient tumour cell variants and the induction of T-cell tolerance. However, the lack of appropriate experimental melanoma models has so far impeded clear insights into the underlying mechanisms. Here we establish an effective ACT protocol in a genetically engineered mouse melanoma model that recapitulates tumour regression, remission and relapse as seen in patients. We report the unexpected observation that melanomas acquire ACT resistance through an inflammation-induced reversible loss of melanocytic antigens. In serial transplantation experiments, melanoma cells switch between a differentiated and a dedifferentiated phenotype in response to T-cell-driven inflammatory stimuli. We identified the proinflammatory cytokine tumour necrosis factor (TNF)-? as a crucial factor that directly caused reversible dedifferentiation of mouse and human melanoma cells. Tumour cells exposed to TNF-? were poorly recognized by T cells specific for melanocytic antigens, whereas recognition by T cells specific for non-melanocytic antigens was unaffected or even increased. Our results demonstrate that the phenotypic plasticity of melanoma cells in an inflammatory microenvironment contributes to tumour relapse after initially successful T-cell immunotherapy. On the basis of our work, we propose that future ACT protocols should simultaneously target melanocytic and non-melanocytic antigens to ensure broad recognition of both differentiated and dedifferentiated melanoma cells, and include strategies to sustain T-cell effector functions by blocking immune-inhibitory mechanisms in the tumour microenvironment. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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14. Pregnancy imprints regulatory memory that sustains anergy to fetal antigen.
- Author
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Rowe, Jared H., Ertelt, James M., Xin, Lijun, and Way, Sing Sing
- Subjects
PREGNANCY complications ,ANERGY ,ANTIGENS ,IMMUNOSUPPRESSION ,T cells - Abstract
Pregnancy is an intricately orchestrated process where immune effector cells with fetal specificity are selectively silenced. This requires the sustained expansion of immune-suppressive maternal FOXP3
+ regulatory T cells (Treg cells), because even transient partial ablation triggers fetal-specific effector T-cell activation and pregnancy loss. In turn, many idiopathic pregnancy complications proposed to originate from disrupted fetal tolerance are associated with blunted maternal Treg expansion. Importantly, however, the antigen specificity and cellular origin of maternal Treg cells that accumulate during gestation remain incompletely defined. Here we show that pregnancy selectively stimulates the accumulation of maternal FOXP3+ CD4 cells with fetal specificity using tetramer-based enrichment that allows the identification of rare endogenous T cells. Interestingly, after delivery, fetal-specific Treg cells persist at elevated levels, maintain tolerance to pre-existing fetal antigen, and rapidly re-accumulate during subsequent pregnancy. The accelerated expansion of Treg cells during secondary pregnancy was driven almost exclusively by proliferation of fetal-specific FOXP3+ cells retained from prior pregnancy, whereas induced FOXP3 expression and proliferation of pre-existing FOXP3+ cells each contribute to Treg expansion during primary pregnancy. Furthermore, fetal resorption in secondary compared with primary pregnancy becomes more resilient to partial maternal FOXP3+ cell ablation. Thus, pregnancy imprints FOXP3+ CD4 cells that sustain protective regulatory memory to fetal antigen. We anticipate that these findings will spark further investigation on maternal regulatory T-cell specificity that unlocks new strategies for improving pregnancy outcomes and novel approaches for therapeutically exploiting Treg cell memory. [ABSTRACT FROM AUTHOR]- Published
- 2012
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15. Expression of tumour-specific antigens underlies cancer immunoediting.
- Author
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DuPage, Michel, Mazumdar, Claire, Schmidt, Leah M., Cheung, Ann F., and Jacks, Tyler
- Subjects
ANTIGENS ,TUMOR antigens ,CARCINOGENS ,CANCER genetics ,SARCOMA ,LABORATORY mice ,DIAGNOSIS - Abstract
Cancer immunoediting is a process by which immune cells, particularly lymphocytes of the adaptive immune system, protect the host from the development of cancer and alter tumour progression by driving the outgrowth of tumour cells with decreased sensitivity to immune attack. Carcinogen-induced mouse models of cancer have shown that primary tumour susceptibility is thereby enhanced in immune-compromised mice, whereas the capacity for such tumours to grow after transplantation into wild-type mice is reduced. However, many questions about the process of cancer immunoediting remain unanswered, in part because of the known antigenic complexity and heterogeneity of carcinogen-induced tumours. Here we adapted a genetically engineered, autochthonous mouse model of sarcomagenesis to investigate the process of cancer immunoediting. This system allows us to monitor the onset and growth of immunogenic and non-immunogenic tumours induced in situ that harbour identical genetic and histopathological characteristics. By comparing the development of such tumours in immune-competent mice with their development in mice with broad immunodeficiency or specific antigenic tolerance, we show that recognition of tumour-specific antigens by lymphocytes is critical for immunoediting against sarcomas. Furthermore, primary sarcomas were edited to become less immunogenic through the selective outgrowth of cells that were able to escape T lymphocyte attack. Loss of tumour antigen expression or presentation on major histocompatibility complex I was necessary and sufficient for this immunoediting process to occur. These results highlight the importance of tumour-specific-antigen expression in immune surveillance, and potentially, immunotherapy. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
16. Antibody-based protection against HIV infection by vectored immunoprophylaxis.
- Author
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Balazs, Alejandro B., Chen, Joyce, Hong, Christin M., Rao, Dinesh S., Yang, Lili, and Baltimore, David
- Subjects
IMMUNOGLOBULINS ,HIV prevention ,ANTIBODY diversity ,ANTIGENS ,EPITOPES ,GENETIC transformation - Abstract
Despite tremendous efforts, development of an effective vaccine against human immunodeficiency virus (HIV) has proved an elusive goal. Recently, however, numerous antibodies have been identified that are capable of neutralizing most circulating HIV strains. These antibodies all exhibit an unusually high level of somatic mutation, presumably owing to extensive affinity maturation over the course of continuous exposure to an evolving antigen. Although substantial effort has focused on the design of immunogens capable of eliciting antibodies de novo that would target similar epitopes, it remains uncertain whether a conventional vaccine will be able to elicit analogues of the existing broadly neutralizing antibodies. As an alternative to immunization, vector-mediated gene transfer could be used to engineer secretion of the existing broadly neutralizing antibodies into the circulation. Here we describe a practical implementation of this approach, which we call vectored immunoprophylaxis (VIP), which in mice induces lifelong expression of these monoclonal antibodies at high concentrations from a single intramuscular injection. This is achieved using a specialized adeno-associated virus vector optimized for the production of full-length antibody from muscle tissue. We show that humanized mice receiving VIP appear to be fully protected from HIV infection, even when challenged intravenously with very high doses of replication-competent virus. Our results suggest that successful translation of this approach to humans may produce effective prophylaxis against HIV. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
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17. Basigin is a receptor essential for erythrocyte invasion by Plasmodium falciparum.
- Author
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Crosnier, Cécile, Bustamante, Leyla Y., Bartholdson, S. Josefin, Bei, Amy K., Theron, Michel, Uchikawa, Makoto, Mboup, Souleymane, Ndir, Omar, Kwiatkowski, Dominic P., Duraisingh, Manoj T., Rayner, Julian C., and Wright, Gavin J.
- Subjects
ERYTHROCYTES ,PLASMODIUM falciparum ,ANTIGENS ,IMMUNOGLOBULINS ,PROTEIN-protein interactions - Abstract
Erythrocyte invasion by Plasmodium falciparum is central to the pathogenesis of malaria. Invasion requires a series of extracellular recognition events between erythrocyte receptors and ligands on the merozoite, the invasive form of the parasite. None of the few known receptor-ligand interactions involved are required in all parasite strains, indicating that the parasite is able to access multiple redundant invasion pathways. Here, we show that we have identified a receptor-ligand pair that is essential for erythrocyte invasion in all tested P. falciparum strains. By systematically screening a library of erythrocyte proteins, we have found that the Ok blood group antigen, basigin, is a receptor for PfRh5, a parasite ligand that is essential for blood stage growth. Erythrocyte invasion was potently inhibited by soluble basigin or by basigin knockdown, and invasion could be completely blocked using low concentrations of anti-basigin antibodies; importantly, these effects were observed across all laboratory-adapted and field strains tested. Furthermore, Ok
a? erythrocytes, which express a basigin variant that has a weaker binding affinity for PfRh5, had reduced invasion efficiencies. Our discovery of a cross-strain dependency on a single extracellular receptor-ligand pair for erythrocyte invasion by P. falciparum provides a focus for new anti-malarial therapies. [ABSTRACT FROM AUTHOR]- Published
- 2011
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18. Commensal microbiota and myelin autoantigen cooperate to trigger autoimmune demyelination.
- Author
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Berer, Kerstin, Mues, Marsilius, Koutrolos, Michail, Rasbi, Zakeya Al, Boziki, Marina, Johner, Caroline, Wekerle, Hartmut, and Krishnamoorthy, Gurumoorthy
- Subjects
COMMENSALISM ,ANTIGENS ,AUTOIMMUNITY ,DEMYELINATION ,T cells ,B cells ,INFECTION - Abstract
Active multiple sclerosis lesions show inflammatory changes suggestive of a combined attack by autoreactive T and B lymphocytes against brain white matter. These pathogenic immune cells derive from progenitors that are normal, innocuous components of the healthy immune repertoire but become autoaggressive upon pathological activation. The stimuli triggering this autoimmune conversion have been commonly attributed to environmental factors, in particular microbial infection. However, using the relapsing-remitting mouse model of spontaneously developing experimental autoimmune encephalomyelitis, here we show that the commensal gut flora-in the absence of pathogenic agents-is essential in triggering immune processes, leading to a relapsing-remitting autoimmune disease driven by myelin-specific CD4
+ T cells. We show further that recruitment and activation of autoantibody-producing B cells from the endogenous immune repertoire depends on availability of the target autoantigen, myelin oligodendrocyte glycoprotein (MOG), and commensal microbiota. Our observations identify a sequence of events triggering organ-specific autoimmune disease and these processes may offer novel therapeutic targets. [ABSTRACT FROM AUTHOR]- Published
- 2011
- Full Text
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19. Cross-dressed dendritic cells drive memory CD8+ T-cell activation after viral infection.
- Author
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Wakim, Linda M. and Bevan, Michael J.
- Subjects
T cells ,LYMPHOCYTES ,ANTIGENS ,ANTIGEN-antibody reactions ,IMMUNE response ,DENDRITIC cells ,VIRUS diseases - Abstract
After an infection, cytotoxic T lymphocyte precursors proliferate and become effector cells by recognizing foreign peptides in the groove of major histocompatibility complex (MHC) class I molecules expressed by antigen-presenting cells (APCs). Professional APCs specialized for T-cell activation acquire viral antigen either by becoming infected themselves (direct presentation) or by phagocytosis of infected cells, followed by transfer of antigen to the cytosol, processing and MHC class I loading in a process referred to as cross-presentation. An alternative way, referred to as 'cross-dressing', by which an uninfected APC could present antigen was postulated to be by the transfer of preformed peptide-MHC complexes from the surface of an infected cell to the APC without the need of further processing. Here we show that this mechanism exists and boosts the antiviral response of mouse memory CD8
+ T cells. A number of publications have demonstrated sharing of peptide-loaded MHC molecules in vitro. Our in vitro experiments demonstrate that cross-dressing APCs do not acquire peptide-MHC complexes in the form of exosomes released by donor cells. Rather, the APCs and donor cells have to contact each other for the transfer to occur. After a viral infection, we could isolate cross-dressed APCs able to present viral antigen in vitro. Furthermore, using the diphtheria toxin system to selectively eliminate APCs that could only acquire viral peptide-MHC complexes by cross-dressing, we show that such presentation can promote the expansion of resting memory T cells. Notably, naive T cells were excluded from taking part in the response. Cross-dressing is a mechanism of antigen presentation used by dendritic cells that may have a significant role in activating previously primed CD8+ T cells. [ABSTRACT FROM AUTHOR]- Published
- 2011
- Full Text
- View/download PDF
20. Co-adjuvant effects of retinoic acid and IL-15 induce inflammatory immunity to dietary antigens.
- Author
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DePaolo, R. W., Abadie, V., Tang, F., Fehlner-Peach, H., Hall, J. A., Wang, W., Marietta, E. V., Kasarda, D. D., Waldmann, T. A., Murray, J. A., Semrad, C., Kupfer, S. S., Belkaid, Y., Guandalini, S., and Jabri, B.
- Subjects
LYMPHOID tissue ,TRETINOIN ,ANTIGENS ,CELIAC disease ,IMMUNOLOGICAL adjuvants ,CELLULAR immunity - Abstract
Under physiological conditions the gut-associated lymphoid tissues not only prevent the induction of a local inflammatory immune response, but also induce systemic tolerance to fed antigens. A notable exception is coeliac disease, where genetically susceptible individuals expressing human leukocyte antigen (HLA) HLA-DQ2 or HLA-DQ8 molecules develop inflammatory T-cell and antibody responses against dietary gluten, a protein present in wheat. The mechanisms underlying this dysregulated mucosal immune response to a soluble antigen have not been identified. Retinoic acid, a metabolite of vitamin A, has been shown to have a critical role in the induction of intestinal regulatory responses. Here we find in mice that in conjunction with IL-15, a cytokine greatly upregulated in the gut of coeliac disease patients, retinoic acid rapidly activates dendritic cells to induce JNK (also known as MAPK8) phosphorylation and release the proinflammatory cytokines IL-12p70 and IL-23. As a result, in a stressed intestinal environment, retinoic acid acted as an adjuvant that promoted rather than prevented inflammatory cellular and humoral responses to fed antigen. Altogether, these findings reveal an unexpected role for retinoic acid and IL-15 in the abrogation of tolerance to dietary antigens. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
21. The structural basis for autonomous dimerization of the pre-T-cell antigen receptor.
- Author
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Siew Siew Pang, Berry, Richard, Zhenjun Chen, Lars Kjer-Nielsen, Matthew A. Perugini, King, Glenn F., Christina Wang, Sock Hui Chew, La Gruta, Nicole L., Williams, Neal K., Beddoe, Travis, Tiganis, Tony, Cowieson, Nathan P., Godfrey, Dale I., Purcell, Anthony W., Wilce, Matthew C. J., McCluskey, James, and Rossjohn, Jamie
- Subjects
T-cell receptor genes ,CELL membranes ,IMMUNOGLOBULINS ,B cells ,ANTIGENS ,GENE expression - Abstract
The pre-T-cell antigen receptor (pre-TCR), expressed by immature thymocytes, has a pivotal role in early T-cell development, including TCR β-selection, survival and proliferation of CD4
− CD8− double-negative thymocytes, and subsequent αβ T-cell lineage differentiation. Whereas αβTCR ligation by the peptide-loaded major histocompatibility complex initiates T-cell signalling, pre-TCR-induced signalling occurs by means of a ligand-independent dimerization event. The pre-TCR comprises an invariant α-chain (pre-Tα) that pairs with any TCR β-chain (TCRβ) following successful TCR β-gene rearrangement. Here we provide the basis of pre-Tα-TCRβ assembly and pre-TCR dimerization. The pre-Tα chain comprised a single immunoglobulin-like domain that is structurally distinct from the constant (C) domain of the TCR α-chain; nevertheless, the mode of association between pre-Tα and TCRβ mirrored that mediated by the Cα-Cβ domains of the αβTCR. The pre-TCR had a propensity to dimerize in solution, and the molecular envelope of the pre-TCR dimer correlated well with the observed head-to-tail pre-TCR dimer. This mode of pre-TCR dimerization enabled the pre-Tα domain to interact with the variable (V) β domain through residues that are highly conserved across the Vβ and joining (J) β gene families, thus mimicking the interactions at the core of the αβTCR's Vα-Vβ interface. Disruption of this pre-Tα-Vβ dimer interface abrogated pre-TCR dimerization in solution and impaired pre-TCR expression on the cell surface. Accordingly, we provide a mechanism of pre-TCR self-association that allows the pre-Tα chain to simultaneously 'sample' the correct folding of both the V and C domains of any TCR β-chain, regardless of its ultimate specificity, which represents a critical checkpoint in T-cell development. This unusual dual-chaperone-like sensing function of pre-Tα represents a unique mechanism in nature whereby developmental quality control regulates the expression and signalling of an integral membrane receptor complex. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
22. Human melanoma-initiating cells express neural crest nerve growth factor receptor CD271.
- Author
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Boiko, Alexander D., Razorenova, Olga V., van de Rijn, Matt, Swetter, Susan M., Johnson, Denise L., Ly, Daphne P., Butler, Paris D., Yang, George P., Joshua, Benzion, Kaplan, Michael J., Longaker, Michael T., and Weissman, Irving L.
- Subjects
MELANOMA ,NERVE growth factor ,CELL transplantation ,LABORATORY mice ,METASTASIS ,STEM cells ,T-cell lymphoma ,ANTIGENS ,TUMOR growth - Abstract
The question of whether tumorigenic cancer stem cells exist in human melanomas has arisen in the last few years. Here we show that in melanomas, tumour stem cells (MTSCs, for melanoma tumour stem cells) can be isolated prospectively as a highly enriched CD271
+ MTSC population using a process that maximizes viable cell transplantation. The tumours sampled in this study were taken from a broad spectrum of sites and stages. High-viability cells isolated by fluorescence-activated cell sorting and re-suspended in a matrigel vehicle were implanted into T-, B- and natural-killer-deficient Rag2−/− γc−/− mice. The CD271+ subset of cells was the tumour-initiating population in 90% (nine out of ten) of melanomas tested. Transplantation of isolated CD271+ melanoma cells into engrafted human skin or bone in Rag2−/− γc−/− mice resulted in melanoma; however, melanoma did not develop after transplantation of isolated CD271− cells. We also show that in mice, tumours derived from transplanted human CD271+ melanoma cells were capable of metastatsis in vivo. CD271+ melanoma cells lacked expression of TYR, MART1 and MAGE in 86%, 69% and 68% of melanoma patients, respectively, which helps to explain why T-cell therapies directed at these antigens usually result in only temporary tumour shrinkage. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
23. Induction of tumour immunity by targeted inhibition of nonsense-mediated mRNA decay.
- Author
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Pastor, Fernando, Kolonias, Despina, Giangrande, Paloma H., and Gilboa, Eli
- Subjects
TUMORS ,IMMUNE system ,PATHOGENIC microorganisms ,VACCINATION ,IMMUNE response ,RNA ,ANTIGENS ,OLIGONUCLEOTIDES ,CELLS - Abstract
The main reason why tumours are not controlled by the immune system is that, unlike pathogens, they do not express potent tumour rejection antigens (TRAs). Tumour vaccination aims at stimulating a systemic immune response targeted to, mostly weak, antigens expressed in the disseminated tumour lesions. Main challenges in developing effective vaccination protocols are the identification of potent and broadly expressed TRAs and effective adjuvants to stimulate a robust and durable immune response. Here we describe an alternative approach in which the expression of new, and thereby potent, antigens are induced in tumour cells by inhibiting nonsense-mediated messenger RNA decay (NMD). Small interfering RNA (siRNA)-mediated inhibition of NMD in tumour cells led to the expression of new antigenic determinants and their immune-mediated rejection. In subcutaneous and metastatic tumour models, tumour-targeted delivery of NMD factor-specific siRNAs conjugated to oligonucleotide aptamer ligands led to significant inhibition of tumour growth that was superior to that of vaccination with granulocyte–macrophage colony-stimulating factor (GM-CSF)-expressing irradiated tumour cells, and could be further enhanced by co-stimulation. Tumour-targeted NMD inhibition forms the basis of a simple, broadly useful, and clinically feasible approach to enhance the antigenicity of disseminated tumours leading to their immune recognition and rejection. The cell-free chemically synthesized oligonucleotide backbone of aptamer–siRNAs reduces the risk of immunogenicity and enhances the feasibility of generating reagents suitable for clinical use. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
24. The kinetics of two-dimensional TCR and pMHC interactions determine T-cell responsiveness.
- Author
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Jun Huang, Zarnitsyna, Veronika I., Liu, Baoyu, Edwards, Lindsay J., Ning Jiang, Evavold, Brian D., and Cheng Zhu
- Subjects
T-cell receptor genes ,MAJOR histocompatibility complex ,PEPTIDE drugs ,PATHOGENIC microorganisms ,ANTIGENS ,BIOLOGICAL membranes ,IMMUNE complexes ,CHEMICAL agonists ,DYNAMICS - Abstract
The T-cell receptor (TCR) interacts with peptide-major histocompatibility complexes (pMHC) to discriminate pathogens from self-antigens and trigger adaptive immune responses. Direct physical contact is required between the T cell and the antigen-presenting cell for cross-junctional binding where the TCR and pMHC are anchored on two-dimensional (2D) membranes of the apposing cells. Despite their 2D nature, TCR–pMHC binding kinetics have only been analysed three-dimensionally (3D) with a varying degree of correlation with the T-cell responsiveness. Here we use two mechanical assays to show high 2D affinities between a TCR and its antigenic pMHC driven by rapid on-rates. Compared to their 3D counterparts, 2D affinities and on-rates of the TCR for a panel of pMHC ligands possess far broader dynamic ranges that match that of their corresponding T-cell responses. The best 3D predictor of response is the off-rate, with agonist pMHC dissociating the slowest. In contrast, 2D off-rates are up to 8,300-fold faster, with the agonist pMHC dissociating the fastest. Our 2D data suggest rapid antigen sampling by T cells and serial engagement of a few agonist pMHCs by TCRs in a large self pMHC background. Thus, the cellular environment amplifies the intrinsic TCR–pMHC binding to generate broad affinities and rapid kinetics that determine T-cell responsiveness. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
25. TCR–peptide–MHC interactions in situ show accelerated kinetics and increased affinity.
- Author
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Huppa, Johannes B., Axmann, Markus, Mörtelmaier, Manuel A., Lillemeier, Björn F., Newell, Evan W., Brameshuber, Mario, Klein, Lawrence O., Schütz, Gerhard J., and Davis, Mark M.
- Subjects
ANTIGENS ,IMMUNOGLOBULINS ,T cells ,LYMPHOCYTES ,IMMUNE response ,IMMUNOLOGY ,CELLULAR immunity ,T cell receptors ,MAJOR histocompatibility complex - Abstract
The recognition of foreign antigens by T lymphocytes is essential to most adaptive immune responses. It is driven by specific T-cell antigen receptors (TCRs) binding to antigenic peptide–major histocompatibility complex (pMHC) molecules on other cells. If productive, these interactions promote the formation of an immunological synapse. Here we show that synaptic TCR–pMHC binding dynamics differ significantly from TCR–pMHC binding in solution. We used single-molecule microscopy and fluorescence resonance energy transfer (FRET) between fluorescently tagged TCRs and their cognate pMHC ligands to measure the kinetics of TCR–pMHC binding in situ. When compared with solution measurements, the dissociation of this complex was increased significantly (4–12-fold). Disruption of actin polymers reversed this effect, indicating that cytoskeletal dynamics destabilize this interaction directly or indirectly. Nevertheless, TCR affinity for pMHC was significantly elevated as the result of a large (about 100-fold) increase in the association rate, a likely consequence of complementary molecular orientation and clustering. In helper T cells, the CD4 molecule has been proposed to bind cooperatively with the TCR to the same pMHC complex. However, CD4 blockade had no effect on the synaptic TCR affinity, nor did it destabilize TCR–pMHC complexes, indicating that the TCR binds pMHC independently of CD4. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
26. Enhancing CD8 T-cell memory by modulating fatty acid metabolism.
- Author
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Pearce, Erika L., Walsh, Matthew C., Cejas, Pedro J., Harms, Gretchen M., Hao Shen, Li-San Wang, Jones, Russell G., and Yongwon Choi
- Subjects
T cells ,LYMPHOCYTES ,CANCER ,ANTIGENS ,IMMUNITY ,CANCER treatment ,FATTY acids - Abstract
CD8 T cells, which have a crucial role in immunity to infection and cancer, are maintained in constant numbers, but on antigen stimulation undergo a developmental program characterized by distinct phases encompassing the expansion and then contraction of antigen-specific effector (T
E ) populations, followed by the persistence of long-lived memory (TM ) cells. Although this predictable pattern of CD8 T-cell responses is well established, the underlying cellular mechanisms regulating the transition to TM cells remain undefined. Here we show that tumour necrosis factor (TNF) receptor-associated factor 6 (TRAF6), an adaptor protein in the TNF-receptor and interleukin-1R/Toll-like receptor superfamily, regulates CD8 TM -cell development after infection by modulating fatty acid metabolism. We show that mice with a T-cell-specific deletion of TRAF6 mount robust CD8 TE -cell responses, but have a profound defect in their ability to generate TM cells that is characterized by the disappearance of antigen-specific cells in the weeks after primary immunization. Microarray analyses revealed that TRAF6-deficient CD8 T cells exhibit altered expression of genes that regulate fatty acid metabolism. Consistent with this, activated CD8 T cells lacking TRAF6 display defective AMP-activated kinase activation and mitochondrial fatty acid oxidation (FAO) in response to growth factor withdrawal. Administration of the anti-diabetic drug metformin restored FAO and CD8 TM -cell generation in the absence of TRAF6. This treatment also increased CD8 TM cells in wild-type mice, and consequently was able to considerably improve the efficacy of an experimental anti-cancer vaccine. [ABSTRACT FROM AUTHOR]- Published
- 2009
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- View/download PDF
27. A yeast-endonuclease-generated DNA break induces antigenic switching in Trypanosoma brucei.
- Author
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Boothroyd, Catharine E., Dreesen, Oliver, Leonova, Tatyana, Ly, K. Ina, Figueiredo, Luisa M., Cross, George A. M., and Papavasiliou, F. Nina
- Subjects
TRYPANOSOMA brucei ,ENDONUCLEASES ,YEAST research ,ANTIGENS ,AFRICAN trypanosomiasis ,GLYCOPROTEINS ,GENE conversion ,GENETIC recombination - Abstract
Trypanosoma brucei is the causative agent of African sleeping sickness in humans and one of the causes of nagana in cattle. This protozoan parasite evades the host immune system by antigenic variation, a periodic switching of its variant surface glycoprotein (VSG) coat. VSG switching is spontaneous and occurs at a rate of about 10
-2 –10-3 per population doubling in recent isolates from nature, but at a markedly reduced rate (10-5 –10-6 ) in laboratory-adapted strains. VSG switching is thought to occur predominantly through gene conversion, a form of homologous recombination initiated by a DNA lesion that is used by other pathogens (for example, Candida albicans, Borrelia sp. and Neisseria gonorrhoeae) to generate surface protein diversity, and by B lymphocytes of the vertebrate immune system to generate antibody diversity. Very little is known about the molecular mechanism of VSG switching in T. brucei. Here we demonstrate that the introduction of a DNA double-stranded break (DSB) adjacent to the ∼70-base-pair (bp) repeats upstream of the transcribed VSG gene increases switching in vitro ∼250-fold, producing switched clones with a frequency and features similar to those generated early in an infection. We were also able to detect spontaneous DSBs within the 70-bp repeats upstream of the actively transcribed VSG gene, indicating that a DSB is a natural intermediate of VSG gene conversion and that VSG switching is the result of the resolution of this DSB by break-induced replication. [ABSTRACT FROM AUTHOR]- Published
- 2009
- Full Text
- View/download PDF
28. Adaptation of HIV-1 to human leukocyte antigen class I.
- Author
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Kawashima, Yuka, Pfafferott, Katja, Frater, John, Matthews, Philippa, Payne, Rebecca, Addo, Marylyn, Gatanaga, Hiroyuki, Fujiwara, Mamoru, Hachiya, Atsuko, Koizumi, Hirokazu, Kuse, Nozomi, Oka, Shinichi, Duda, Anna, Prendergast, Andrew, Crawford, Hayley, Leslie, Alasdair, Brumme, Zabrina, Brumme, Chanson, Allen, Todd, and Brander, Christian
- Subjects
LETTERS to the editor ,ANTIGENS - Abstract
The rapid and extensive spread of the human immunodeficiency virus (HIV) epidemic provides a rare opportunity to witness host–pathogen co-evolution involving humans. A focal point is the interaction between genes encoding human leukocyte antigen (HLA) and those encoding HIV proteins. HLA molecules present fragments (epitopes) of HIV proteins on the surface of infected cells to enable immune recognition and killing by CD8
+ T cells; particular HLA molecules, such as HLA-B*57, HLA-B*27 and HLA-B*51, are more likely to mediate successful control of HIV infection. Mutation within these epitopes can allow viral escape from CD8+ T-cell recognition. Here we analysed viral sequences and HLA alleles from >2,800 subjects, drawn from 9 distinct study cohorts spanning 5 continents. Initial analysis of the HLA-B*51-restricted epitope, TAFTIPSI (reverse transcriptase residues 128–135), showed a strong correlation between the frequency of the escape mutation I135X and HLA-B*51 prevalence in the 9 study cohorts (P = 0.0001). Extending these analyses to incorporate other well-defined CD8+ T-cell epitopes, including those restricted by HLA-B*57 and HLA-B*27, showed that the frequency of these epitope variants (n = 14) was consistently correlated with the prevalence of the restricting HLA allele in the different cohorts (together, P < 0.0001), demonstrating strong evidence of HIV adaptation to HLA at a population level. This process of viral adaptation may dismantle the well-established HLA associations with control of HIV infection that are linked to the availability of key epitopes, and highlights the challenge for a vaccine to keep pace with the changing immunological landscape presented by HIV. [ABSTRACT FROM AUTHOR]- Published
- 2009
- Full Text
- View/download PDF
29. SUMOylation regulates Rad18-mediated template switch.
- Author
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Branzei, Dana, Vanoli, Fabio, and Foiani, Marco
- Subjects
SACCHAROMYCES cerevisiae ,DNA replication ,MESSENGER RNA ,DNA damage ,ANTIGENS ,SISTER chromatid exchange ,GENETIC recombination - Abstract
Replication by template switch is thought to mediate DNA damage-bypass and fillings of gaps. Gap-filling repair requires homologous recombination as well as Rad18- and Rad5-mediated proliferating cell nuclear antigen (PCNA) polyubiquitylation. However, it is unclear whether these processes are coordinated, and the physical evidence for Rad18–Rad5-dependent template switch at replication forks is still elusive. Here we show, using genetic and physical approaches, that in budding yeast (Saccharomyces cerevisiae) Rad18 is required for the formation of X-shaped sister chromatid junctions (SCJs) at damaged replication forks through a process involving PCNA polyubiquitylation and the ubiquitin-conjugating enzymes Mms2 and Ubc13. The Rad18–Mms2-mediated damage-bypass through SCJs requires the small ubiquitin-like modifier (SUMO)-conjugating enzyme Ubc9 and SUMOylated PCNA, and is coordinated with Rad51-dependent recombination events. We propose that the Rad18–Rad5–Mms2-dependent SCJs represent template switch events. Altogether, our results unmask a role for PCNA ubiquitylation and SUMOylation pathways in promoting transient damage-induced replication-coupled recombination events involving sister chromatids at replication forks. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
30. Generation of a prostate from a single adult stem cell.
- Author
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Leong, Kevin G., Wang, Bu-Er, Johnson, Leisa, and Gao, Wei-Qiang
- Subjects
STEM cell research ,ANIMAL models of prostate cancer ,ANTIANDROGENS ,ANTIGENS ,PHENOTYPES ,CARCINOGENESIS ,CANCER immunology - Abstract
The existence of prostate stem cells (PSCs) was first postulated from the observation that normal prostate regeneration can occur after repeated cycles of androgen deprivation and replacement in rodents. Given the critical role of PSCs in maintaining prostate tissue integrity and their potential involvement in prostate tumorigenesis, it is important to define specific markers for normal PSCs. Several cell-surface markers have been reported to identify candidate PSCs, including stem cell antigen-1 (Sca-1, also known as Ly6a), CD133 (Prom1) and CD44 (refs 3—10). However, many non-PSCs in the mouse prostate also express these markers and thus identification of a more defined PSC population remains elusive. Here we identify CD117 (c-kit, stem cell factor receptor) as a new marker of a rare adult mouse PSC population, and demonstrate that a single stem cell defined by the phenotype Lin
- Sca-1+ CD133+ CD44+ CD117+ can generate a prostate after transplantation in vivo. CD117 expression is predominantly localized to the region of the mouse prostate proximal to the urethra and is upregulated after castration-induced prostate involution—two characteristics consistent with that of a PSC marker. CD117+ PSCs can generate functional, secretion-producing prostates when transplanted in vivo. Moreover, CD117+ PSCs have long-term self-renewal capacity, as evidenced by serial isolation and transplantation in vivo. Our data establish that single cells in the adult mouse prostate with multipotent, self-renewal capacity are defined by a Lin- Sca-1+ CD133+ CD44+ CD117+ phenotype. [ABSTRACT FROM AUTHOR]- Published
- 2008
- Full Text
- View/download PDF
31. Autophagy in thymic epithelium shapes the T-cell repertoire and is essential for tolerance.
- Author
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Nedjic, Jelena, Aichinger, Martin, Emmerich, Jan, Mizushima, Noboru, and Klein, Ludger
- Subjects
EPITOPES ,ANTIGENS ,IMMUNOSPECIFICITY ,CELLS ,EPITHELIAL cells ,EPITHELIUM ,EXFOLIATIVE cytology ,POLYPEPTIDES ,PEPTIDES - Abstract
Recognition of self-antigen-derived epitopes presented by major histocompatibility complex class II (MHC II) molecules on thymic epithelial cells (TECs) is critical for the generation of a functional and self-tolerant CD4 T-cell repertoire. Whereas haematopoietic antigen-presenting cells generate MHC-II–peptide complexes predominantly through the processing of endocytosed polypeptides, it remains unknown if and how TECs use unconventional pathways of antigen presentation. Here we address the role of macroautophagy, a process that has recently been shown to allow for endogenous MHC II loading, in T-cell repertoire selection in the mouse thymus. In contrast to most other tissues, TECs had a high constitutive level of autophagy. Genetic interference with autophagy specifically in TECs led to altered selection of certain MHC-II-restricted T-cell specificities and resulted in severe colitis and multi-organ inflammation. Our findings indicate that autophagy focuses the MHC-II–peptide repertoire of TECs on their intracellular milieu, which notably comprises a wide array of otherwise strictly ‘tissue-specific’ self antigens. In doing so, it contributes to T-cell selection and is essential for the generation of a self-tolerant T-cell repertoire. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
32. The structural basis for an essential subunit interaction in influenza virus RNA polymerase.
- Author
-
Obayashi, Eiji, Yoshida, Hisashi, Kawai, Fumihiro, Shibayama, Naoya, Kawaguchi, Atsushi, Nagata, Kyosuke, Tame, Jeremy R. H., and Sam-Yong Park
- Subjects
INFLUENZA A virus ,INFLUENZA viruses ,PATHOGENIC microorganisms ,VIRAL replication ,RNA polymerases ,BINDING sites ,GENETIC transcription ,ANTIGENS ,VIRUSES - Abstract
Influenza A virus is a major human and animal pathogen with the potential to cause catastrophic loss of life. The virus reproduces rapidly, mutates frequently and occasionally crosses species barriers. The recent emergence in Asia of avian influenza related to highly pathogenic forms of the human virus has highlighted the urgent need for new effective treatments. Here we demonstrate the importance to viral replication of a subunit interface in the viral RNA polymerase, thereby providing a new set of potential drug binding sites entirely independent of surface antigen type. No current medication targets this heterotrimeric polymerase complex. All three subunits, PB1, PB2 and PA, are required for both transcription and replication. PB1 carries the polymerase active site, PB2 includes the capped-RNA recognition domain, and PA is involved in assembly of the functional complex, but so far very little structural information has been reported for any of them. We describe the crystal structure of a large fragment of one subunit (PA) of influenza A RNA polymerase bound to a fragment of another subunit (PB1). The carboxy-terminal domain of PA forms a novel fold, and forms a deep, highly hydrophobic groove into which the amino-terminal residues of PB1 can fit by forming a 3
10 helix. [ABSTRACT FROM AUTHOR]- Published
- 2008
- Full Text
- View/download PDF
33. Memory CD4 T cells emerge from effector T-cell progenitors.
- Author
-
Harrington, Laurie E., Janowski, Karen M., Oliver, James R., Zajac, Allan J., and Weaver, Casey T.
- Subjects
CELLULAR immunity ,T cells ,CELL differentiation ,MEMORY ,ANTIGENS ,VACCINATION ,PATHOGENIC microorganisms ,THERAPEUTICS ,INFECTION - Abstract
A hallmark of adaptive immunity is the generation of memory T cells that confer long-lived, antigen-specific protection against repeat challenges by pathogens. Understanding the mechanisms by which memory T cells arise is important for rational vaccination strategies and improved therapeutic interventions for chronic infections and autoimmune disorders. The large clonal expansion of CD8 T cells in response to some infections has made the development of CD8 T-cell memory more amenable to study, giving rise to a model of memory cell differentiation in which a fraction of fully competent effector T cells transition into long-lived memory T cells. Delineation of CD4 T-cell memory development has proved more difficult as a result of limitations on tracking the smaller populations of CD4 effector T cells generated during a pathogenic challenge, complicating efforts to determine whether CD4 memory T cells are direct descendants of effector T cells or whether they develop by alternative pathways. Here, using two complementary cytokine reporter mouse models to identify interferon (IFN)-γ-positive effector T cells and track their fate, we show that the lineage relationship between effector and memory CD4 T cells resembles that for CD8 T cells responding to the same pathogen. We find that, in parallel with effector CD8 T cells, IFN-γ-positive effector CD4 T cells give rise to long-lived memory T cells capable of anamnestic responses to antigenic rechallenge. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
34. TANK-binding kinase-1 delineates innate and adaptive immune responses to DNA vaccines.
- Author
-
Ishii, Ken J., Kawagoe, Tatsukata, Koyama, Shohei, Matsui, Kosuke, Kumar, Himanshu, Kawai, Taro, Uematsu, Satoshi, Takeuchi, Osamu, Takeshita, Fumihiko, Coban, Cevayir, and Akira, Shizuo
- Subjects
VACCINES ,ANTIGENS ,DNA ,IMMUNOLOGICAL adjuvants ,T-cell receptor genes ,B cells ,PLASMIDS ,INTERFERONS ,CARRIER proteins - Abstract
Successful vaccines contain not only protective antigen(s) but also an adjuvant component that triggers innate immune activation and is necessary for their optimal immunogenicity. In the case of DNA vaccines, this consists of plasmid DNA; however, the adjuvant element(s) as well as its intra- and inter-cellular innate immune signalling pathway(s) leading to the encoded antigen-specific T- and B-cell responses remain unclear. Here we demonstrate in vivo that TANK-binding kinase 1 (TBK1), a non-canonical IκB kinase, mediates the adjuvant effect of DNA vaccines and is essential for its immunogenicity in mice. Plasmid-DNA-activated, TBK1-dependent signalling and the resultant type-I interferon receptor-mediated signalling was required for induction of antigen-specific B and T cells, which occurred even in the absence of innate immune signalling through a well known CpG DNA sensor—Toll-like receptor 9 (TLR9) or Z-DNA binding protein 1 (ZBP1, also known as DAI, which was recently reported as a potential B-form DNA sensor). Moreover, bone-marrow-transfer experiments revealed that TBK1-mediated signalling in haematopoietic cells was critical for the induction of antigen-specific B and CD4
+ T cells, whereas in non-haematopoietic cells TBK1 was required for CD8+ T-cell induction. These data suggest that TBK1 is a key signalling molecule for DNA-vaccine-induced immunogenicity, by differentially controlling DNA-activated innate immune signalling through haematopoietic and non-haematopoietic cells. [ABSTRACT FROM AUTHOR]- Published
- 2008
- Full Text
- View/download PDF
35. Roquin represses autoimmunity by limiting inducible T-cell co-stimulator messenger RNA.
- Author
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Di Yu, Andy Hee-Meng Tan, Xin Hu, Athanasopoulos, Vicki, Simpson, Nicholas, Silva, Diego G., Hutloff, Andreas, Giles, Keith M., Leedman, Peter J., Kong Peng Lam, Goodnow, Christopher C., and Vinuesa, Carola G.
- Subjects
AUTOIMMUNITY ,T cells ,MESSENGER RNA ,IMMUNE response ,IMMUNOLOGY ,PATHOGENIC microorganisms ,ANTIGENS ,LYMPHOCYTES ,AUTOIMMUNE diseases - Abstract
Immune responses are normally targeted against microbial pathogens and not self-antigens by mechanisms that are only partly understood. Here we define a newly discovered pathway that prevents autoimmunity by limiting the levels on T lymphocytes of a co-stimulatory receptor, the inducible T-cell co-stimulator (ICOS). In sanroque mice homozygous for an M199R mutation in the ROQ domain of Roquin (also known as Rc3h1), increased Icos expression on T cells causes the accumulation of lymphocytes that is associated with a lupus-like autoimmune syndrome. Roquin normally limits Icos expression by promoting the degradation of Icos messenger RNA. A conserved segment in the unusually long ICOS 3′ untranslated mRNA is essential for regulation by Roquin. This segment comprises a 47-base-pair minimal region complementary to T-cell-expressed microRNAs including miR-101, the repressive activity of which is disrupted by base-pair inversions predicted to abrogate miR-101 binding. These findings illuminate a critical post-transcriptional pathway within T cells that regulates lymphocyte accumulation and autoimmunity, and highlights the therapeutic potential of partially antagonising the ICOS pathway. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
36. Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and present them to antiviral B cells.
- Author
-
Junt, Tobias, Moseman, E. Ashley, Iannacone, Matteo, Massberg, Steffen, Lang, Philipp A., Boes, Marianne, Fink, Katja, Henrickson, Sarah E., Shayakhmetov, Dmitry M., Di Paolo, Nelson C., van Rooijen, Nico, Mempel, Thorsten R., Whelan, Sean P., and von Andrian, Ulrich H.
- Subjects
IMMUNITY ,PATHOGENIC microorganisms ,IMMUNE response ,MACROPHAGES ,LYMPH nodes ,B cells ,ANTIGENS ,ANTIGEN-antibody reactions ,ANTIGEN presenting cells ,IMMUNOLOGY - Abstract
Lymph nodes prevent the systemic dissemination of pathogens such as viruses that infect peripheral tissues after penetrating the body’s surface barriers. They are also the staging ground of adaptive immune responses to pathogen-derived antigens. It is unclear how virus particles are cleared from afferent lymph and presented to cognate B cells to induce antibody responses. Here we identify a population of CD11b
+ CD169+ MHCII+ macrophages on the floor of the subcapsular sinus (SCS) and in the medulla of lymph nodes that capture viral particles within minutes after subcutaneous injection. Macrophages in the SCS translocated surface-bound viral particles across the SCS floor and presented them to migrating B cells in the underlying follicles. Selective depletion of these macrophages compromised local viral retention, exacerbated viraemia of the host, and impaired local B-cell activation. These findings indicate that CD169+ macrophages have a dual physiological function. They act as innate ‘flypaper’ by preventing the systemic spread of lymph-borne pathogens and as critical gatekeepers at the lymph–tissue interface that facilitate the recognition of particulate antigens by B cells and initiate humoral immune responses. [ABSTRACT FROM AUTHOR]- Published
- 2007
- Full Text
- View/download PDF
37. IgH class switching and translocations use a robust non-classical end-joining pathway.
- Author
-
Yan, Catherine T., Boboila, Cristian, Souza, Ellen Kris, Franco, Sonia, Hickernell, Thomas R., Murphy, Michael, Gumaste, Sunil, Geyer, Mark, Zarrin, Ali A., Manis, John P., Rajewsky, Klaus, and Alt, Frederick W.
- Subjects
IMMUNOGLOBULINS ,EXONS (Genetics) ,B cells ,GENETIC recombination ,ANTIGENS ,DOUBLE-stranded RNA ,DNA repair ,HOMOLOGY (Biology) ,IMMUNOGLOBULIN G - Abstract
Immunoglobulin variable region exons are assembled in developing B cells by V(D)J recombination. Once mature, these cells undergo class-switch recombination (CSR) when activated by antigen. CSR changes the heavy chain constant region exons (Ch) expressed with a given variable region exon from Cμ to a downstream Ch (for example, Cγ, Cε or Cα), thereby switching expression from IgM to IgG, IgE or IgA. Both V(D)J recombination and CSR involve the introduction of DNA double-strand breaks and their repair by means of end joining. For CSR, double-strand breaks are introduced into switch regions that flank Cμ and a downstream Ch, followed by fusion of the broken switch regions. In mammalian cells, the ‘classical’ non-homologous end joining (C-NHEJ) pathway repairs both general DNA double-strand breaks and programmed double-strand breaks generated by V(D)J recombination. C-NHEJ, as observed during V(D)J recombination, joins ends that lack homology to form ‘direct’ joins, and also joins ends with several base-pair homologies to form microhomology joins. CSR joins also display direct and microhomology joins, and CSR has been suggested to use C-NHEJ. Xrcc4 and DNA ligase IV (Lig4), which cooperatively catalyse the ligation step of C-NHEJ, are the most specific C-NHEJ factors; they are absolutely required for V(D)J recombination and have no known functions other than C-NHEJ. Here we assess whether C-NHEJ is also critical for CSR by assaying CSR in Xrcc4- or Lig4-deficient mouse B cells. C-NHEJ indeed catalyses CSR joins, because C-NHEJ-deficient B cells had decreased CSR and substantial levels of IgH locus (immunoglobulin heavy chain, encoded by Igh) chromosomal breaks. However, an alternative end-joining pathway, which is markedly biased towards microhomology joins, supports CSR at unexpectedly robust levels in C-NHEJ-deficient B cells. In the absence of C-NHEJ, this alternative end-joining pathway also frequently joins Igh locus breaks to other chromosomes to generate translocations. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
38. Structural basis of Dscam isoform specificity.
- Author
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Meijers, Rob, Puettmann-Holgado, Roland, Skiniotis, Georgios, Jin-Huan Liu, Walz, Thomas, Jia-Huai Wang, and Schmucker, Dietmar
- Subjects
IMMUNOGLOBULIN genes ,IMMUNOGLOBULIN D ,DROSOPHILA melanogaster ,CELL receptors ,IMMUNE response ,NEURAL circuitry ,GENES ,PEPTIDES ,ANTIGENS ,NEURAL receptors - Abstract
The Dscam gene gives rise to thousands of diverse cell surface receptors thought to provide homophilic and heterophilic recognition specificity for neuronal wiring and immune responses. Mutually exclusive splicing allows for the generation of sequence variability in three immunoglobulin ecto-domains, D2, D3 and D7. We report X-ray structures of the amino-terminal four immunoglobulin domains (D1–D4) of two distinct Dscam isoforms. The structures reveal a horseshoe configuration, with variable residues of D2 and D3 constituting two independent surface epitopes on either side of the receptor. Both isoforms engage in homo-dimerization coupling variable domain D2 with D2, and D3 with D3. These interactions involve symmetric, antiparallel pairing of identical peptide segments from epitope I that are unique to each isoform. Structure-guided mutagenesis and swapping of peptide segments confirm that epitope I, but not epitope II, confers homophilic binding specificity of full-length Dscam receptors. Phylogenetic analysis shows strong selection of matching peptide sequences only for epitope I. We propose that peptide complementarity of variable residues in epitope I of Dscam is essential for homophilic binding specificity. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
39. Toll-dependent selection of microbial antigens for presentation by dendritic cells.
- Author
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Blander, J. Magarian and Medzhitov, Ruslan
- Subjects
ANTIGENS ,DENDRITIC cells ,PATHOGENIC microorganisms ,INFECTION ,CELL receptors ,IMMUNITY - Abstract
Dendritic cells constitutively sample the tissue microenvironment and phagocytose both microbial and host apoptotic cells. This leads to the induction of immunity against invading pathogens or tolerance to peripheral self antigens, respectively. The outcome of antigen presentation by dendritic cells depends on their activation status, such that Toll-like receptor (TLR)-induced dendritic cell activation makes them immunogenic, whereas steady-state presentation of self antigens leads to tolerance. TLR-inducible expression of co-stimulatory signals is one of the mechanisms of self/non-self discrimination. However, it is unclear whether or how the inducible expression of co-stimulatory signals would distinguish between self antigens and microbial antigens when both are encountered by dendritic cells during infection. Here we describe a new mechanism of antigen selection in dendritic cells for presentation by major histocompatibility complex class II molecules (MHC II) that is based on the origin of the antigen. We show that the efficiency of presenting antigens from phagocytosed cargo is dependent on the presence of TLR ligands within the cargo. Furthermore, we show that the generation of peptide–MHC class II complexes is controlled by TLRs in a strictly phagosome-autonomous manner. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
40. Apolipoprotein-mediated pathways of lipid antigen presentation.
- Author
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van den Elzen, Peter, Garg, Salil, León, Luis, Brigl, Manfred, Leadbetter, Elizabeth A., Gumperz, Jenny E., Dascher, Chris C., Tan-Yun Cheng, Sacks, Frank M., Illarionov, Petr A., Besra, Gurdyal S., Kent, Sally C., Moody, D. Branch, and Brenner, Michael B.
- Subjects
APOLIPOPROTEINS ,LIPIDS ,ANTIGENS ,MOLECULES ,T cells ,CELLS - Abstract
Peptide antigens are presented to T cells by major histocompatibility complex (MHC) molecules, with endogenous peptides presented by MHC class I and exogenous peptides presented by MHC class II. In contrast to the MHC system, CD1 molecules bind lipid antigens that are presented at the antigen-presenting cell (APC) surface to lipid antigen-reactive T cells. Because CD1 molecules survey endocytic compartments, it is self-evident that they encounter antigens from extracellular sources. However, the mechanisms of exogenous lipid antigen delivery to CD1-antigen-loading compartments are not known. Serum apolipoproteins are mediators of extracellular lipid transport for metabolic needs. Here we define the pathways mediating markedly efficient exogenous lipid antigen delivery by apolipoproteins to achieve T-cell activation. Apolipoprotein E binds lipid antigens and delivers them by receptor-mediated uptake into endosomal compartments containing CD1 in APCs. Apolipoprotein E mediates the presentation of serum-borne lipid antigens and can be secreted by APCs as a mechanism to survey the local environment to capture antigens or to transfer microbial lipids from infected cells to bystander APCs. Thus, the immune system has co-opted a component of lipid metabolism to develop immunological responses to lipid antigens. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
41. Sodium channel mutation leading to saxitoxin resistance in clams increases risk of PSP.
- Author
-
Bricelj, V. Monica, Connell, Laurie, Konoki, Keiichi, MacQuarrie, Scott P., Scheuer, Todd, Catterall, William A., and Trainer, Vera L.
- Subjects
PARALYTIC shellfish poisoning ,PARALYSIS ,SEAFOOD poisoning ,RED tide ,ANTITOXINS ,ANTIGENS - Abstract
Bivalve molluscs, the primary vectors of paralytic shellfish poisoning (PSP) in humans, show marked inter-species variation in their capacity to accumulate PSP toxins (PSTs) which has a neural basis. PSTs cause human fatalities by blocking sodium conductance in nerve fibres. Here we identify a molecular basis for inter-population variation in PSP resistance within a species, consistent with genetic adaptation to PSTs. Softshell clams (Mya arenaria) from areas exposed to‘red tides’are more resistant to PSTs, as demonstrated by whole-nerve assays, and accumulate toxins at greater rates than sensitive clams from unexposed areas. PSTs lead to selective mortality of sensitive clams. Resistance is caused by natural mutation of a single amino acid residue, which causes a 1,000-fold decrease in affinity at the saxitoxin-binding site in the sodium channel pore of resistant, but not sensitive, clams. Thus PSTs might act as potent natural selection agents, leading to greater toxin resistance in clam populations and increased risk of PSP in humans. Furthermore, global expansion of PSP to previously unaffected coastal areas might result in long-term changes to communities and ecosystems. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
42. Recognition of bacterial glycosphingolipids by natural killer T cells.
- Author
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Kinjo, Yuki, Wu, Douglass, Kim, Gisen, Xing, Guo-Wen, Poles, Michael A., Ho, David D., Tsuji, Moriya, Kawahara, Kazuyoshi, Wong, Chi-Huey, and Kronenberg, Mitchell
- Subjects
GLYCOSPHINGOLIPIDS ,KILLER cells ,T cells ,BACTERIA ,SPHINGOLIPIDS ,ANTIGENS ,IMMUNE response - Abstract
Natural killer T (NKT) cells constitute a highly conserved T lymphocyte subpopulation that has the potential to regulate many types of immune responses through the rapid secretion of cytokines. NKT cells recognize glycolipids presented by CD1d, a class I-like antigen-presenting molecule. They have an invariant T-cell antigen receptor (TCR)a-chain, but whether this invariant TCR recognizes microbial antigens is still controversial. Here we show that most mouse and human NKT cells recognize glycosphingolipids from Sphingomonas, Gram-negative bacteria that do not contain lipopolysaccharide. NKT cells are activated in vivo after exposure to these bacterial antigens or bacteria, and mice that lack NKT cells have a marked defect in the clearance of Sphingomonas from the liver. These data suggest that NKT cells are T lymphocytes that provide an innate-type immune response to certain microorganisms through recognition by their antigen receptor, and that they might be useful in providing protection from bacteria that cannot be detected by pattern recognition receptors such as Toll-like receptor 4. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
43. Exogenous and endogenous glycolipid antigens activate NKT cells during microbial infections.
- Author
-
Mattner, Jochen, DeBord, Kristin L., Ismail, Nahed, Goff, Randal D., Cantu, Carlos, Zhou, Dapeng, Saint-Mezard, Pierre, Wang, Vivien, Gao, Ying, Yin, Ning, Hoebe, Kasper, Schneewind, Olaf, Walker, David, Beutler, Bruce, Teyton, Luc, Savage, Paul B., and Bendelac, Albert
- Subjects
GLYCOLIPIDS ,ANTIGENS ,KILLER cells ,T cells ,LEUCOCYTES ,PATHOGENIC microorganisms ,CELL receptors - Abstract
CD1d-restricted natural killer T (NKT) cells are innate-like lymphocytes that express a conserved T-cell receptor and contribute to host defence against various microbial pathogens. However, their target lipid antigens have remained elusive. Here we report evidence for microbial, antigen-specific activation of NKT cells against Gram-negative, lipopolysaccharide (LPS)-negative alpha-Proteobacteria such as Ehrlichia muris and Sphingomonas capsulata. We have identified glycosylceramides from the cell wall of Sphingomonas that serve as direct targets for mouse and human NKT cells, controlling both septic shock reaction and bacterial clearance in infected mice. In contrast, Gram-negative, LPS-positive Salmonella typhimurium activates NKT cells through the recognition of an endogenous lysosomal glycosphingolipid, iGb3, presented by LPS-activated dendritic cells. These findings identify two novel antigenic targets of NKT cells in antimicrobial defence, and show that glycosylceramides are an alternative to LPS for innate recognition of the Gram-negative, LPS-negative bacterial cell wall. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
44. The immunoglobulin superfamily protein Izumo is required for sperm to fuse with eggs.
- Author
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Inoue, Naokazu, Ikawa, Masahito, Isotani, Ayako, and Okabe, Masaru
- Subjects
IMMUNOGLOBULINS ,PROTEINS ,MEMBRANE fusion ,SPERMATOZOA ,EGGS ,ANTIGENS ,MONOCLONAL antibodies ,MOLECULAR cloning - Abstract
Representing the 60 trillion cells that build a human body, a sperm and an egg meet, recognize each other, and fuse to form a new generation of life. The factors involved in this important membrane fusion event, fertilization, have been sought for a long time. Recently, CD9 on the egg membrane was found to be essential for fusion, but sperm-related fusion factors remain unknown. Here, by using a fusion-inhibiting monoclonal antibody and gene cloning, we identify a mouse sperm fusion-related antigen and show that the antigen is a novel immunoglobulin superfamily protein. We have termed the gene Izumo and produced a gene-disrupted mouse line. Izumo
-/- mice were healthy but males were sterile. They produced normal-looking sperm that bound to and penetrated the zona pellucida but were incapable of fusing with eggs. Human sperm also contain Izumo and addition of the antibody against human Izumo left the sperm unable to fuse with zona-free hamster eggs. [ABSTRACT FROM AUTHOR]- Published
- 2005
- Full Text
- View/download PDF
45. Somatic diversification of variable lymphocyte receptors in the agnathan sea lamprey.
- Author
-
Pancer, Zeev, Amemiya, Chris T., Ehrhardt, Götz R. A., Ceitlin, Jill, Gartland, G. Larry, and Cooper, Max D.
- Subjects
LYMPHOCYTE receptors ,IMMUNE response ,SEA lamprey ,ANTIGENS ,LEUCINE ,TRINUCLEOTIDE repeats - Abstract
Although jawless vertebrates are apparently capable of adaptive immune responses, they have not been found to possess the recombinatorial antigen receptors shared by all jawed vertebrates. Our search for the phylogenetic roots of adaptive immunity in the lamprey has instead identified a new type of variable lymphocyte receptors (VLRs) composed of highly diverse leucine-rich repeats (LRR) sandwiched between amino- and carboxy-terminal LRRs. An invariant stalk region tethers the VLRs to the cell surface by means of a glycosyl-phosphatidyl-inositol anchor. To generate rearranged VLR genes of the diversity necessary for an anticipatory immune system, the single lamprey VLR locus contains a large bank of diverse LRR cassettes, available for insertion into an incomplete germline VLR gene. Individual lymphocytes express a uniquely rearranged VLR gene in monoallelic fashion. Different evolutionary strategies were thus used to generate highly diverse lymphocyte receptors through rearrangement of LRR modules in agnathans (jawless fish) and of immunoglobulin gene segments in gnathostomes (jawed vertebrates). [ABSTRACT FROM AUTHOR]
- Published
- 2004
- Full Text
- View/download PDF
46. Detecting selection using a single genome sequence of M. tuberculosis and P. falciparum.
- Author
-
Plotkin, Joshua B., Dushoff, Jonathan, and Fraser, Hunter B.
- Subjects
MYCOBACTERIUM tuberculosis ,PLASMODIUM falciparum ,GENOMES ,ANTIGENS ,PROTEOMICS ,GENOMICS - Abstract
Selective pressures on proteins are usually measured by comparing nucleotide sequences. Here we introduce a method to detect selection on the basis of a single genome sequence. We catalogue the relative strength of selection on each gene in the entire genomes of Mycobacterium tuberculosis and Plasmodium falciparum. Our analysis confirms that most antigens are under strong selection for amino-acid substitutions, particularly the PE/PPE family of putative surface proteins in M. tuberculosis and the EMP1 family of cytoadhering surface proteins in P. falciparum. We also identify many uncharacterized proteins that are under strong selection in each pathogen. We provide a genome-wide analysis of natural selection acting on different stages of an organism's life cycle: genes expressed in the ring stage of P. falciparum are under stronger positive selection than those expressed in other stages of the parasite's life cycle. Our method of estimating selective pressures requires far fewer data than comparative sequence analysis, and it measures selection across an entire genome; the method can readily be applied to a large range of sequenced organisms. [ABSTRACT FROM AUTHOR]
- Published
- 2004
- Full Text
- View/download PDF
47. Immune recognition of a human renal cancer antigen through post-translational protein splicing.
- Author
-
Hanada, Ken-Ichi, Yewdell, Jonathan W., and Yang, James C.
- Subjects
IMMUNE recognition ,RENAL cancer ,PROTEIN engineering ,MOLECULAR oncology ,ANTIGENS ,T cells - Abstract
Cytotoxic T lymphocytes (CTLs) detect and destroy cells displaying class I molecules of the major histocompatibility complex (MHC) that present oligopeptides derived from aberrant self or foreign proteins. Most class I peptide ligands are created from proteins that are degraded by proteasomes and transported, by the transporter associated with antigen processing, from the cytosol into the endoplasmic reticulum, where peptides bind MHC class I molecules and are conveyed to the cell surface. C2 CTLs, cloned from human CTLs infiltrating a renal cell carcinoma, kill cancer cells overexpressing fibroblast growth factor-5 (FGF-5). Here we show that C2 cells recognize human leukocyte antigen-A3 MHC class I molecules presenting a nine-residue FGF-5 peptide generated by protein splicing. This process, previously described strictly in plants and unicellular organisms, entails post-translational excision of a polypeptide segment followed by ligation of the newly liberated carboxy-terminal and amino-terminal residues. The occurrence of protein splicing in vertebrates has important implications for the complexity of the vertebrate proteome and for the immune recognition of self and foreign peptides. [ABSTRACT FROM AUTHOR]
- Published
- 2004
- Full Text
- View/download PDF
48. Phagosomes are competent organelles for antigen cross-presentation.
- Author
-
Houde, Mathieu, Bertholet, Sylvie, Gagnon, Etienne, Brunet, Sylvain, Goyette, Guillame, Laplante, Annie, Princiotta, Michael F., Thibault, Pierre, Sacks, David, and Desjardins, Michel
- Subjects
PHAGOCYTOSIS ,ANTIGENS ,ENDOPLASMIC reticulum - Abstract
The ability to process microbial antigens and present them at the surface of cells is an important aspect of our innate ability to clear infections. It is generally accepted that antigens in the cytoplasm are loaded in the endoplasmic reticulum and presented at the cell surface on major histocompatibility complex (MHC) class I molecules, whereas peptides present in endo/phagocytic compartments are presented on MHC class II molecules. Despite the apparent segregation of the class I and class II pathways, antigens from intracellular pathogens including mycobacteria, Escherichia coli, Salmonella typhimurium, Brucella abortus and Leishmania, have been shown to elicit an MHC class-I-dependent CD8
+ T-cell response, a process referred to as cross-presentation. The cellular mechanisms allowing the cross-presentation pathway are poorly understood. Here we show that phagosomes display the elements and properties needed to be self-sufficient for the cross-presentation of exogenous antigens, a newly ascribed function linked to phagocytosis mediated by the endoplasmic reticulum. [ABSTRACT FROM AUTHOR]- Published
- 2003
- Full Text
- View/download PDF
49. Protein kinase Cδ controls self-antigen-induced B-cell tolerance.
- Author
-
MecklenbrÄuker, Ingrid, Saijo, Kaoru, Nai-Ying Zheng, Leitges, Michael, and Tarakhovsky, Alexander
- Subjects
B cells ,ANTIGENS ,PROTEIN kinases - Abstract
Interaction of a B cell expressing self-specific B-cell antigen receptor (BCR) with an auto-antigen results in either clonal deletion or functional inactivation[SUP1-3]. Both of these processes lead to B-cell tolerance and are essential for the prevention of auto-immune diseases. Whereas clonal deletion results in the death of developing autoreactive B cells, functional inactivation of self-reactive B lymphocytes leads to complex changes in the phenotype of peripheral B cells, described collectively as anergy[SUP1-3]. Here we demonstrate that deficiency in protein kinase Cλ (PKCλ) prevents B-cell tolerance, and allows maturation and terminal differentiation of self-reactive B cells in the presence of the tolerizing antigen. The importance of PKC-λ in B-cell tolerance is further underscored by the appearance of autoreactive antiDNA and anti-nuclear antibodies in the serum of PKC-λ-deficient mice. As deficiency of PKC-λ does not affect BCR-mediated B-cell activation in vitro and in vivo, our data suggest a selective and essential role of PKC-&lambda in tolerogenic, but not immunogenic, B-cell responses. [ABSTRACT FROM AUTHOR]
- Published
- 2002
50. H3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes.
- Author
-
Bouillet, Philippe, Purton, Jared F., Godfrey, Dale I., Li-Chen Zhang, Coultas, Leigh, Puthalakath, Hamsa, Pellegrini, Marc, Cory, Suzanne, Adams, Jerry M., and Strasser, Andreas
- Subjects
APOPTOSIS ,ANTIGENS - Abstract
Assesses the requirement for apoptosis on autoreactive thymocytes. Assembly of genes coding for antigen receptors by combinational linking of gene segments; Induction of autoimmune disease; Effects of bim deficiency to thymocyte.
- Published
- 2002
- Full Text
- View/download PDF
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