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functional plasticity and robustness are essential characteristics of biological systems: lessons learned from klrg1-deficient mice.killer cell lectin-like receptor g1 (klrg1) receptor is considered to be a marker of terminally differentiated nk and t cells and is strongly induced by viral and other infections. klrg1 is a c-type lectin-like inhibitory receptor, which interacts with members of the cadherin family of molecules leading to the inhibition of t- and nk-cell function. a study in this issue of the european journal of immunology addresses the role of klrg1 in the maturation and differentiation of nk and t cells in vi ...201020373518
an improved tet-on system for gene expression in neurons delivered by a single lentiviral vector.in most cases, the successful application of gene therapy requires the development of vectors that can provide regulated control of therapeutic gene expression. we have reconstituted the tet-on (tetracycline-regulated transgene expression) two-component system in a single lentiviral vector with insertion of a chicken chromatin insulator (chs4) element between the two expression cassettes. optimization of this vector included an improved reverse tetracycline-dependent trans-activator (rtta) seque ...200920377365
cd4+cd25+ regulatory t cells suppress the immune responses of mouse embryo fibroblasts to murine cytomegalovirus infection.cytomegaloviruses (cmvs) cause common viral infectious diseases and are difficult for the host immune system to eliminate, which leads to persistent or chronic infection. to investigate the t cell immune response stimulated by murine cytomegalovirus (mcmv) infection and the role of cd4(+)cd25(+)foxp3(+) t regulatory cells (tregs) in this process, t cells containing various proportions of tregs were co-cultured with mcmv-infected mouse embryo fibroblasts (mefs). mcmv infection stimulated prolifer ...201020381532
immunoglobulin to zona pellucida 3 mediates ovarian damage and infertility after contraceptive vaccination in mice.antibodies reactive with the ovarian glycoprotein zona pellucida (zp) have been linked with human female infertility. anti-fertility vaccines that target zp antigens have been utilized to restrict pest animal populations and their efficacy is associated with ovary-specific antibody induction. however, the necessity for zona pellucida-specific antibody in mediating infertility has not been examined in vivo. a recombinant mouse cytomegalovirus vaccine encoding murine zona pellucida 3 that induces ...201020382503
catalytic m1gs rna as an antiviral agent in animals.the use of rnase p ribozyme (m1gs catalytic rna) for inhibition of murine cytomegalovirus (mcmv) propagation in mice is described in this chapter. general information about rnase p based technology is included and followed by detailed protocols focused on (1) construction and in vitro cleavage assay of the customized m1gs ribozyme, (2) stable expression of the m1gs rna and evaluation of its activity in inhibition of viral gene expression and growth in cultured cells, and (3) investigation of m1g ...201020387160
virus inhibition of rip3-dependent necrosis.viral infection activates cytokine expression and triggers cell death, the modulation of which is important for successful pathogenesis. necroptosis is a form of programmed necrosis dependent on two related rip homotypic interaction motif (rhim)-containing signaling adaptors, receptor-interacting protein kinases (rip) 1 and 3. we find that murine cytomegalovirus infection induces rip3-dependent necrosis. whereas rip3 kinase activity and rhim-dependent interactions control virus-associated necros ...201020413098
mhc class i d(k) expression in hematopoietic and nonhematopoietic cells confers natural killer cell resistance to murine cytomegalovirus.nk cell-mediated murine cytomegalovirus (mcmv) resistance (cmv(r)) is under h-2(k) control in ma/my mice, but the underlying gene(s) is unclear. prior genetic analysis mapped cmv(r) to the mhc class i (mhc-i) d(k) gene interval. because nk cell receptors are licensed by and responsive to mhc class i molecules, d(k) itself is a candidate gene. a 10-kb genomic d(k) fragment was subcloned and microinjected into mcmv-susceptible (cmv(s)) (ma/my.l-h2(b) x c57l)f(1) or (b6 x dba/2)f(2) embryos. transg ...201020421478
flt3 permits survival during infection by rendering dendritic cells competent to activate nk cells.a previously unappreciated signal necessary for dendritic cell (dc)-mediated activation of natural killer (nk) cells during viral infection was revealed by a recessive n-ethyl-n-nitrosourea-induced mutation called warmflash (wmfl). wmfl homozygotes displayed increased susceptibility to mouse cytomegalovirus (mcmv) infection. in response to mcmv infection in vivo, delayed nk cell activation was observed, but no intrinsic defects in nk cell activation or function were identified. rather, coculture ...201020457904
a spread-deficient cytomegalovirus for assessment of first-target cells in vaccination.human cytomegalovirus (hcmv) is a human pathogen that causes severe disease primarily in the immunocompromised or immunologically immature individual. to date, no vaccine is available. we describe use of a spread-deficient murine cmv (mcmv) as a novel approach for betaherpesvirus vaccination. to generate a spread-deficient mcmv, the conserved, essential gene m94 was deleted. immunization with mcmv-deltam94 is apathogenic and protective against wild-type challenge even in highly susceptible ifnal ...201020463067
mutations in the m112/m113-coding region facilitate murine cytomegalovirus replication in human cells.cytomegaloviruses, representatives of the betaherpesvirinae, cause opportunistic infections in immunocompromised hosts. they infect various cells and tissues in their natural host but are highly species specific. for instance, human cytomegalovirus (hcmv) does not replicate in mouse cells, and human cells are not permissive for murine cytomegalovirus (mcmv) infection. however, the underlying molecular mechanisms are so far poorly understood. in the present study we isolated and characterized a s ...201020519391
intact nkg2d-independent function of nk cells chronically stimulated with the nkg2d ligand rae-1.human tumors frequently express membrane-bound or soluble nk group 2, member d (nkg2d) ligands. this results in chronic engagement of nkg2d on the surfaces of nk and cd8(+) t cells and rapid internalization of the receptor. although it is well appreciated that this phenomenon impairs nkg2d-dependent function, careful analysis of nkg2d-independent functions in cells chronically stimulated through nkg2d is lacking. using a mouse model of chronic nkg2d ligand expression, we show that constant expos ...201020530257
dominant negative mutants of the murine cytomegalovirus m53 gene block nuclear egress and inhibit capsid maturation.the alphaherpesvirus proteins ul31 and ul34 and their homologues in other herpesvirus subfamilies cooperate at the nuclear membrane in the export of nascent herpesvirus capsids. we studied the respective betaherpesvirus proteins m53 and m50 in mouse cytomegalovirus (mcmv). recently, we established a random approach to identify dominant negative (dn) mutants of essential viral genes and isolated dn mutants of m50 (b. rupp, z. ruzsics, c. buser, b. adler, p. walther and u. h. koszinowski, j. virol ...201020610730
murine cytomegalovirus major immediate-early protein 3 interacts with cellular and viral proteins in viral dna replication compartments and is important for early gene activation.murine cytomegalovirus (mcmv) immediate-early protein 3 (ie3) is essential for successful viral infection. this study developed mcmvs with an egfp-fused ie3 gene in order to study ie3 gene expression, subnuclear distribution and biological function, as well as to examine the interaction of ie3 with cellular and viral proteins. the generated viruses included mcmvie3gfp, in which ie1 was completely removed by the in-frame fusion of exons 3 and 5 and the c terminus of ie3 was tagged with egfp, and ...201020631086
in vivo imaging assay for the convenient evaluation of antiviral compounds against cytomegalovirus in mice.evaluation of newly identified antiviral compounds against cytomegalovirus (cmv) in vivo still requires laborious and time-consuming experiments using a large number of animals. in this study, we examined an in vivo imaging assay for the evaluation of antiviral compounds using a recombinant murine cmv expressing egfp (mcmv-gfp). we found the followings: (1) fluorescent signals were detectable from 1 day after subcutaneous inoculation of the viruses into the backs of mice, and reached to the peak ...201020633578
excess neutrophil infiltration during cytomegalovirus brain infection of interleukin-10-deficient mice.wild-type mice control murine cytomegalovirus (mcmv) brain infection, but identical infection is lethal to animals deficient in interleukin (il)-10. here, we report that mcmv-infected il-10 knockout (ko) mice displayed a marked increase in neutrophil infiltration into the infected, il-10-deficient brain when compared to wild-type animals. enhanced microglial cell activation, determined by mhc class ii up-regulation, overexpression of cxcl2, and elevated p-selectin mrna levels were observed. in v ...201020655600
glycosylation contributes to variability in expression of murine cytomegalovirus m157 and enhances stability of interaction with the nk-cell receptor ly49h.nk cell-mediated resistance to murine cytomegalovirus (mcmv) is controlled by allelic ly49 receptors, including activating ly49h (c57bl/6 strain) and inhibitory ly49i (129 strain), which specifically recognize mcmv m157, a glycosylphosphatidylinositol-linked protein with homology to mhc class i. although the ly49 receptors retain significant homology to classic carbohydrate-binding lectins, the role of glycosylation in ligand binding is unclear. herein, we show that m157 is expressed in multiple ...201020662096
human herpesvirus 7 in allogeneic hemopoietic stem cell transplant recipients in the central clinical hospital in warsaw: a three-year survey.human herpesvirus 7 (hhv-7) is spread worldwide and has been described as a potential pathogen in immunosuppressed patients. different clinical manifestations have been described including fever and skin rash; hhv-7 may also be a possible cofactor for cytomegalovirus disease in transplant recipients.201120689313
il-10 restricts memory t cell inflation during cytomegalovirus infection.the beta-herpesvirus cmv induces a substantial and progressive expansion of virus-specific memory cd8 t cells, which protect the host against viral reactivation from latency. in this paper, we report that this expansion, or "inflation," of memory t cells is amplified dramatically during mouse cmv infection of il-10 knockout (il-10(-/-)) mice. t cells from il-10(-/-) mice were oligoclonal, exhibited a highly activated phenotype, expressed antiviral cytokines, and degranulated in response to cogna ...201020713884
murine cytomegalovirus perturbs endosomal trafficking of major histocompatibility complex class i molecules in the early phase of infection.murine cytomegalovirus (mcmv) functions interfere with protein trafficking in the secretory pathway. in this report we used δm138-mcmv, a recombinant virus with a deleted viral fc receptor, to demonstrate that mcmv also perturbs endosomal trafficking in the early phase of infection. this perturbation had a striking impact on cell surface-resident major histocompatibility complex class i (mhc-i) molecules due to the complementary effect of mcmv immunoevasins, which block their egress from the sec ...201020719942
biphasic role of 4-1bb in the regulation of mouse cytomegalovirus-specific cd8(+) t cells.the initial requirement for the emergence of cmv-specific cd8(+) t cells is poorly understood. mice deficient in the cosignaling tnf superfamily member, 4-1bb, surprisingly developed exaggerated early cd8(+) t-cell responses to mouse cmv (mcmv). cd8(+) t cells directed against acute mcmv epitopes were enhanced, demonstrating that 4-1bb naturally antagonizes these primary populations. paradoxically, 4-1bb-deficient mice displayed reduced accumulation of memory cd8(+) t cells that expand during ch ...201020722077
the impact of cd4+foxp3+ treg on immunity to murine cytomegalovirus after bone marrow transplantation depends on the peripheral or thymic source of t cell regeneration.the adoptive transfer of regulatory t cells (treg) in murine models has been shown to ameliorate graft-versus-host disease while it may preserve the graft-versus-leukemia effect. however, the impact of treg on infectious immunity after bone marrow transplantation (bmt) is still unclear. immunocompetence against opportunistic viral infections depends on the kinetics of t cell recovery after bmt through two distinctive processes, i.e. lymphopenia-induced proliferation (lip) of mature t cells and g ...201020801217
expression and regulation of the chemokine cxcl16 in crohn's disease and models of intestinal inflammation.cxcl16 mediates adhesion and phagocytosis of both gram-negative and gram-positive bacteria and is a strong chemoattractant for cxcr6+ t cells. in this study, we determined the so far unknown expression and signal transduction of the novel cxcl16-cxcr6 chemokine-ligand receptor system in intestinal inflammation in vivo and in vitro.201020848509
iκbζ is essential for natural killer cell activation in response to il-12 and il-18.iκbζ, encoded by nfibiz, is a nuclear iκb-like protein harboring ankyrin repeats. iκbζ has been shown to regulate il-6 production in macrophages and th17 development in t cells. however, the role of iκbζ in natural killer (nk) cells has not be understood. in the present study, we found that the expression of iκbζ was rapidly induced in response to il-18 in nk cells, but not in t cells. analysis of nfkbiz(-/-) mice revealed that iκbζ was essential for the production of ifn-γ production and cytoto ...201020876105
interleukin-10 repletion suppresses pro-inflammatory cytokines and decreases liver pathology without altering viral replication in murine cytomegalovirus (mcmv)-infected il-10 knockout mice.to determine the role of interleukin-10 (il-10) in protecting against the deleterious pro-inflammatory cytokine response to murine cytomegalovirus (mcmv), we studied the impact of il-10 repletion in mcmv-infected il-10 knockout (ko) mice.201020922456
in vivo impact of cytomegalovirus evasion of cd8 t-cell immunity: facts and thoughts based on murine models.cytomegaloviruses (cmvs) co-exist with their respective host species and have evolved to avoid their elimination by the hosts' immune effector mechanisms and to persist in a non-replicative state, known as viral latency. there is evidence to suggest that latency is nevertheless a highly dynamic condition during which episodes of viral gene desilencing, which can be viewed as incomplete reactivations, cause intermittent antigenic activity that stimulates cd8 memory-effector t cells and drives the ...201020933556
inhibition of programmed cell death by cytomegaloviruses.the elimination of infected cells by programmed cell death (pcd) is one of the most ancestral defense mechanisms against infectious agents. this mechanism should be most effective against intracellular parasites, such as viruses, which depend on the host cell for their replication. however, even large and slowly replicating viruses like the cytomegaloviruses (cmvs) can prevail and persist in face of cellular suicide programs and other innate defense mechanisms. during evolution, these viruses ha ...201020969904
cathepsin c limits acute viral infection independently of nk cell and cd8(+) t-cell cytolytic function.destruction of target cells by cytotoxic t lymphocytes (ctls) or natural killer (nk) cells requires the coordinated action of the pore forming protein perforin (pfp) and the granzyme (gzm) family of serine proteases. the activation of a number of serine proteases, including gzma and b, is predominately mediated by cathepsin c (catc). deficiencies in catc-null mice were therefore expected to replicate the defects observed in gzmab-deficient mice. we have previously determined that gzmab-deficient ...201020975734
[role of allitridin in the blocking of murine cytomegalovirus induced regulatory t cells amplification in vitro].to investigate the effect of allitridin on murine cytomegalovirus (mcmv) induced regulatory t cells (treg) amplification in vitro.201020979868
cytomegalovirus microrna expression is tissue specific and is associated with persistence.micrornas (mirnas) are a class of small noncoding rnas involved in posttranscriptional regulation. mirnas are utilized in organisms ranging from plants to higher mammals, and data have shown that dna viruses also use this method for host and viral gene regulation. here, we report the sequencing of the small rnas in rat cytomegalovirus (rcmv)-infected fibroblasts and persistently infected salivary glands. we identified 24 unique mirnas that mapped to hairpin structures found within the viral geno ...201020980502
b7-mediated costimulation of cd4 t cells constrains cytomegalovirus persistence.cytomegalovirus (cmv) utilizes multiple strategies to modulate immunity and promote lifelong, persistent/latent infection, including suppressing t cell activation pathways. here we examined the role of b7 costimulatory ligands in establishing immune détente from both the host and virus perspectives. mice lacking both b7.1 and b7.2 showed reduced early expansion of cmv-specific cd4 t cells, consequently allowing for enhanced levels of persistent virus replication. in turn, a cmv mutant lacking ex ...201020980516
cytomegalovirus immunoevasin reveals the physiological role of "missing self" recognition in natural killer cell dependent virus control in vivo.cytomegaloviruses (cmvs) are renowned for interfering with the immune system of their hosts. to sidestep antigen presentation and destruction by cd8(+) t cells, these viruses reduce expression of major histocompatibility complex class i (mhc i) molecules. however, this process sensitizes the virus-infected cells to natural killer (nk) cell-mediated killing via the "missing self" axis. mouse cytomegalovirus (mcmv) uses m152 and m06 encoded proteins to inhibit surface expression of mhc i molecules ...201021078887
can we build it better? using bac genetics to engineer more effective cytomegalovirus vaccines.the magnitude and durability of immunity to human cytomegalovirus (hcmv) following natural infection is compromised by the presence of immune modulation genes that appear to promote evasion of host clearance mechanisms. since immunity to hcmv offers limited protection, rational design of effective vaccines has been challenging. in this issue of the jci, slavuljica and colleagues employ techniques to genetically modify the highly related mouse cmv (mcmv), in the process generating a virus that wa ...201021099107
recombinant mouse cytomegalovirus expressing a ligand for the nkg2d receptor is attenuated and has improved vaccine properties.human cmv (hcmv) is a major cause of morbidity and mortality in both congenitally infected and immunocompromised individuals. development of an effective hcmv vaccine would help protect these vulnerable groups. nk group 2, member d (nkg2d) is a potent activating receptor expressed by cells of the innate and adaptive immune systems. its importance in hcmv immune surveillance is indicated by the elaborative evasion mechanisms evolved by the virus to avoid nkg2d. in order to study this signaling pa ...201021099111
the activator protein 1 binding motifs within the human cytomegalovirus major immediate-early enhancer are functionally redundant and act in a cooperative manner with the nf-{kappa}b sites during acute infection.human cytomegalovirus (hcmv) infection causes a rapid induction of c-fos and c-jun, the major subunits of activator protein 1 (ap-1), which in turn have been postulated to activate the viral immediate-early (ie) genes. accordingly, the major ie promoter (miep) enhancer, a critical control region for initiating lytic hcmv infection and reactivation from the latent state, contains one well-characterized ap-1 site and a second candidate interaction site. in this study we explored the role of these ...201021106746
plasmacytoid dendritic cell ablation impacts early interferon responses and antiviral nk and cd8(+) t cell accrual.plasmacytoid dendritic cells (pdcs) mediate type i interferon (ifn-i) responses to viruses that are recognized through the toll-like receptor 7 (tlr7) or tlr9 signaling pathway. however, it is unclear how pdcs regulate the antiviral responses via innate and adaptive immune cells. we generated diphtheria toxin receptor transgenic mice to selectively deplete pdcs by administration of diphtheria toxin. pdc-depleted mice were challenged with viruses known to activate pdcs. in murine cytomegalovirus ...201021130004
the role of decay accelerating factor in the immunopathogenesis of cytomegalovirus infection.a wide variety of the host immune elements play an influential role in the defence against cytomegalovirus (cmv) infection. however, the role of complement in the clearance of cmv infection is less well studied. decay accelerating factor (daf/cd55) is a membrane-bound complement regulatory protein that inhibits the formation and accelerates the decay of c3-convertase. here we hypothesize that murine cmv (mcmv) utilizes daf as an immunoevasive strategy through down-regulation of host adaptive res ...201021166665
comparative analysis of the lambda-interferons il-28a and il-29 regarding their transcriptome and their antiviral properties against hepatitis c virus.specific differences in signaling and antiviral properties between the different lambda-interferons, a novel group of interferons composed of il-28a, il-28b and il-29, are currently unknown. this is the first study comparatively investigating the transcriptome and the antiviral properties of the lambda-interferons il-28a and il-29.201021170333
functional consequences of natural sequence variation of murine cytomegalovirus m157 for ly49 receptor specificity and nk cell activation.the ly49h activating receptor on c57bl/6 (b6) nk cells plays a key role in early resistance to murine cytomegalovirus (mcmv) infection through specific recognition of the mcmv-encoded mhc class i-like molecule m157 expressed on infected cells. the m157 molecule is also recognized by the ly49i inhibitory receptor from the 129/j mouse strain. the m157 gene is highly sequence variable among mcmv isolates, with many m157 variants unable to bind ly49h(b6). in this study, we have sought to define if m ...201021187440
impact of β2 integrin deficiency on mouse natural killer cell development and function.natural killer (nk) cells are innate immune cells that express members of the leukocyte β2 integrin family in humans and mice. these cd11/cd18 heterodimers play critical roles in leukocyte trafficking, immune synapse formation, and costimulation. the cell-surface expression of one of these integrins, cd11b/cd18, is also recognized as a major marker of mouse nk-cell maturation, but its function on nk cells has been largely ignored. using n-ethyl-n-nitrosourea (enu) mutagenesis, we generated a mou ...201121239699
differential requirements for cd45 in nk-cell function reveal distinct roles for syk-family kinases.the protein tyrosine phosphatase cd45 is an important regulator of src-family kinase activity. we found that in the absence of cd45, natural killer (nk) cells are defective in protecting the host from mouse cytomegalovirus infection. we show that although cd45 is necessary for all immunoreceptor tyrosine-based activation motif (itam)-specific nk-cell functions and processes such as degranulation, cytokine production, and expansion during viral infection, the impact of cd45 deficiency on itam sig ...201121245479
murine cytomegalovirus infection of neural stem cells alters neurogenesis in the developing brain.congenital cytomegalovirus (cmv) brain infection causes serious neuro-developmental sequelae including: mental retardation, cerebral palsy, and sensorineural hearing loss. but, the mechanisms of injury and pathogenesis to the fetal brain are not completely understood. the present study addresses potential pathogenic mechanisms by which this virus injures the cns using a neonatal mouse model that mirrors congenital brain infection. this investigation focused on, analysis of cell types infected wi ...201121249143
salivary gland nk cells are phenotypically and functionally unique.natural killer (nk) cells and cd8(+) t cells play vital roles in containing and eliminating systemic cytomegalovirus (cmv). however, cmv has a tropism for the salivary gland acinar epithelial cells and persists in this organ for several weeks after primary infection. here we characterize a distinct nk cell population that resides in the salivary gland, uncommon to any described to date, expressing both mature and immature nk cell markers. using rorγt reporter mice and nude mice, we also show tha ...201121249177
activation receptor-induced tolerance of mature nk cells in vivo requires signaling through the receptor and is reversible.nk cell responses are determined by signals received through activating and inhibitory cell surface receptors. ly49h is an nk cell-specific activating receptor that accounts for the genetic resistance to murine cmv (mcmv). the ly49h receptor has been shown to interact with two adaptor proteins (dap12 and dap10). in the context of mcmv infection, interaction of m157 (the mcmv-encoded ligand for ly49h) with ly49h results in activation of ly49h-expressing nk cells. chronic exposure of ly49h with m1 ...201121263069
exploitation of herpesviral transactivation allows quantitative reporter gene-based assessment of virus entry and neutralization.herpesviral entry is a highly elaborated process requiring many proteins to act in precise conjunction. neutralizing antibodies interfere with this process to abrogate viral infection. based on promoter transactivation of a reporter gene we established a novel method to quantify herpesvirus entry and neutralization by antibodies. following infection with mouse and human cytomegalovirus and herpes simplex virus 1 we observed promoter transactivation resulting in substantial luciferase expression ...201121264213
genetic labeling reveals altered turnover and stability of innate lymphocytes in latent mouse cytomegalovirus infection.mouse cmv (mcmv) infection rapidly induces the proliferation of nk cells, which correlates with immunological protection. whether nk cells primed during acute response against mcmv are maintained for the long term is not known. in this study, we used tcrdh2begfp mice in which maturing nk cells are genetically labeled with a pulse of very stable histone-2b-egfp. in this system, we found that the reporter protein was diluted out upon nk cell division during acute mcmv infection. at the same time, ...201121270406
a dominant role for the immunoproteasome in cd8+ t cell responses to murine cytomegalovirus.murine cytomegalovirus (mcmv) is an important animal model of human cytomegalovirus (hcmv), a ß-herpesvirus that infects the majority of the world's population and causes disease in neonates and immunocompromised adults. cd8(+) t cells are a major part of the immune response to mcmv and hcmv. processing of peptides for presentation to cd8(+) t cells may be critically dependent on the immunoproteasome, expression of which is affected by mcmv. however, the overall importance of the immunoproteasom ...201121304910
salivary glands act as mucosal inductive sites via the formation of ectopic germinal centers after site-restricted mcmv infection.we investigated the hypothesis that salivary gland inoculation stimulates formation of ectopic germinal centers (gcs), transforming the gland into a mucosal inductive site. intraglandular infection of mice with murine cytomegalovirus (mcmv; control: uv-inactivated mcmv) induces salivary gland ectopic follicles comprising cognate interactions between cd4(+) and b220(+) lymphocytes, igm(+) and isotype-switched igg(+) and iga(+) b cells, antigen presenting cells, and follicular dendritic cells. b c ...201121307334
lack of tnf-alpha promotes caspase-3-independent apoptosis during murine cytomegalovirus retinitis.both caspase-dependent and caspase-independent apoptosis contribute to retinal damage during murine cytomegalovirus (mcmv) retinitis, and tnf-a is among the inducers of apoptosis. the aim of this study was to determine the contribution of tnf-a by studying virus replication and apoptosis in immunosuppressed (is) tnf-a(-/-) mice.201121310911
murine model for congenital cmv infection and hearing impairment.congenital cytomegalovirus (cmv) infection is the leading cause of sensorineural hearing loss (snhl), and snhl is the most frequent sequela of congenital cmv infection. but the pathogenic mechanism remains unknown, and there is no ideal cmv intrauterine infection animal model to study the mechanisms by which snhl develops.201121320351
single cell detection of latent cytomegalovirus reactivation in host tissue.the molecular mechanisms leading to reactivation of latent cytomegalovirus are not well understood. to study reactivation, the few cells in an organ tissue that give rise to reactivated virus need to be identified, ideally at the earliest possible time point in the process. to this end, mouse cytomegalovirus (mcmv) reporter mutants were designed to simultaneously express the red fluorescent protein mcherry and the secreted gaussia luciferase (gluc). whereas gluc can serve to assess infection at ...201121325477
differential b7-cd28 costimulatory requirements for stable and inflationary mouse cytomegalovirus-specific memory cd8 t cell populations.cmv establishes a lifelong persistent infection, and viral immune-modulating strategies are important in facilitating this. a particularly diverse cd8 t cell response develops as a result of this host-virus détente, with the cmv-specific memory t cell pool displaying unique functions and phenotypes. to gain insight into the factors that regulate cmv-specific cd8 t cell responses, we examined the influence of the b7-cd28 costimulatory pathway on magnitude, kinetics, and phenotype. initial expansi ...201121357256
expression and iron-dependent regulation of succinate receptor gpr91 in retinal pigment epithelium.gpr91, a succinate receptor, is expressed in retinal ganglion cells and induces vascular endothelial growth factor (vegf) expression. rpe also expresses vegf, but whether this cell expresses gpr91 is not known. excessive iron is also proangiogenic, and hemochromatosis is associated with iron overload. therefore, we examined the expression and iron-dependent regulation of gpr91 in the rpe.201121357408
transmission of murine cytomegalovirus in breast milk: a model of natural infection in neonates.vertical transmission of viruses in breast milk can expose neonates to infectious pathogens at a time when the capacity of their immune system to control infections is limited. we developed a mouse model to study the outcomes of acquisition of murine cytomegalovirus (mcmv) when neonates are breastfed by mothers with acute or latent infection. breast milk leukocytes collected from lactating mice were examined for the presence of mcmv ie-1 mrna by reverse transcription-pcr (rt-pcr) with southern a ...201121367905
mouse embryonic stem cells inhibit murine cytomegalovirus infection through a multi-step process.in humans, cytomegalovirus (cmv) is the most significant infectious cause of intrauterine infections that cause congenital anomalies of the central nervous system. currently, it is not known how this process is affected by the timing of infection and the susceptibility of early-gestational-period cells. embryonic stem (es) cells are more resistant to cmv than most other cell types, although the mechanism responsible for this resistance is not well understood. using a plaque assay and evaluation ...201121407806
host defense against viral infection involves interferon mediated down-regulation of sterol biosynthesis.little is known about the protective role of inflammatory processes in modulating lipid metabolism in infection. here we report an intimate link between the innate immune response to infection and regulation of the sterol metabolic network characterized by down-regulation of sterol biosynthesis by an interferon regulatory loop mechanism. in time-series experiments profiling genome-wide lipid-associated gene expression of macrophages, we show a selective and coordinated negative regulation of the ...201121408089
the mouse cytomegalovirus glycoprotein m155 inhibits cd40 expression and restricts cd4 t cell responses.cytomegaloviruses (cmv) utilize a variety of immunomodulatory strategies to facilitate the establishment of lifelong persistence in their infected hosts. we show that the mouse cmv (mcmv) m155 open reading frame (orf) is required for the posttranscriptional inhibition of cd40 expression in infected antigen-presenting cells. consistent with the known importance of cd40-mediated costimulation of t cells, a m155-deficient virus induces enhanced mcmv epitope-specific cd4 t cell responses.201121411536
cytomegalovirus-induced sensorineural hearing loss with persistent cochlear inflammation in neonatal mice.congenital cytomegalovirus (cmv) infection is the leading cause of sensorineural hearing loss (snhl) in children. during murine (m)cmv-induced encephalitis, the immune response is important for both the control of viral dissemination and the clearance of virus from the brain. while the importance of cmv-induced snhl has been described, the mechanisms surrounding its pathogenesis and the role of inflammatory responses remain unclear. this study presents a neonatal mouse model of profound snhl in ...201121416394
self mhc class i-licensed nk cells enhance adaptive cd8 t-cell viral immunity.mhc class i (mhc i) is essential to nk- and t-cell effector and surveillance functions. however, it is unknown whether mhc i polymorphism influences adaptive immunity through nk cells. previously, we found that mhc i d(k), a cognate ligand for the ly49g2 inhibitory receptor, was essential to nk control of murine (m)cmv infection. here we assessed the significance of nk inhibitory receptor recognition of mcmv on cd8 t cells in genetically defined mhc i d(k) disparate mice. we observed that d(k)-l ...201121436069
p-selectin mediates the microvascular dysfunction associated with persistent cytomegalovirus infection in normocholesterolemic and hypercholesterolemic mice.please cite this paper as: senchenkov, khoretonenko, leskov, ostanin, and stokes (2011). p-selectin mediates the microvascular dysfunction associated with persistent cytomegalovirus infection in normocholesterolemic and hypercholesterolemic mice. microcirculation 18(6), 452-462. abstract: objective: cytomegalovirus has been implicated in cardiovascular disease, possibly through the induction of inflammatory processes. p-selectin and l-selectin are adhesion molecules that mediate early microvasc ...201121457388
[a murine model with murine cytomegalovirus infection resulting in colon inflammation after allogenetic skin transplantation].to provide a reliable animal model for study of human cmv disease in gastrointestinal track, we tried to infect with murine cytomegalovirus (mcmv) in mice that were received allogenetic skin transplantation under immunosuppression. (1) skin transplantation was performed between 18 donor c57bl/6 mice and 72 recipient balb/c mice. (2) all recipient mice were then given cyclosporine at 12 mg/kg daily for 2 weeks by intraperitoneal injection. mice were randomly divided into 3 groups. two experimenta ...201121462500
cytomegalovirus e127 protein interacts with the inhibitory cd200 receptor.cd200 is a cell surface glycoprotein that binds an inhibitory receptor (cd200r) on myeloid cells. cd200 orthologues are present in many species of virus, and we show that the rat cytomegalovirus cd200 orthologue (e127) is expressed at the cell surface on infected cells. it binds the host cd200r with the same affinity as that of the host protein, and thus this protein acts as a close mimic of the host protein and has the potential to downregulate immune responses to the virus.201121471232
temporal profiling of the coding and noncoding murine cytomegalovirus transcriptomes.the global transcriptional program of murine cytomegalovirus (mcmv), involving coding, noncoding, and antisense transcription, remains unknown. here we report an oligonucleotide custom microarray platform capable of measuring both coding and noncoding transcription on a genome-wide scale. by profiling mcmv wild-type and immediate-early mutant strains in fibroblasts, we found rapid activation of the transcriptome by 6.5 h postinfection, with absolute dependency on ie3, but not ie1 or ie2, for gen ...201121471238
partial functional complementation between human and mouse cytomegalovirus chemokine receptor homologues.the human cytomegalovirus (cmv) proteins us28 and ul33 are homologous to chemokine receptors (ckrs). knockout of the mouse cmv m33 protein (ul33 homologue) results in substantial attenuation of salivary gland infection/replication and reduced efficiency of reactivation from tissue explants. m33-mediated g protein-coupled signaling is critical for the salivary gland phenotype. in this report, we demonstrate that us28 and (to a lesser degree) ul33 restore reactivation from tissue explants and part ...201121490099
virus-induced differential expression of nuclear receptors and coregulators in dendritic cells: implication to interferon production.we investigated mrna expression of 49 nuclear hormone receptors (nrs) and 35 transcriptional coregulators in mouse bone marrow-derived dendritic cells (dcs) upon infection with newcastle disease virus (ndv) or murine cytomegalovirus (mcmv). these viruses regulated mrna expression of some nrs among which nor1 and lxrα were highly induced at mrna and protein levels. exogenous expression of the latter nrs repressed irf3- or irf7-induced transactivation of the interferon β promoter and ndv infection ...201121492741
distinct mhc class i-dependent nk cell-activating receptors control cytomegalovirus infection in different mouse strains.recognition of mouse cytomegalovirus (mcmv)-infected cells by activating nk cell receptors was first described in the context of ly49h, which confers resistance to c57bl/6 mice. we investigated the ability of other activating ly49 receptors to recognize mcmv-infected cells in mice from various h-2 backgrounds. we observed that ly49p1 from nod/ltj mice, ly49l from balb mice, and ly49d2 from pwk/pas mice respond to mcmv-infected cells in the context of h-2d(k) and the viral protein m04/gp34. recog ...201121518798
nk cell receptor/h2-d-dependent host resistance to viral infection is quantitatively modulated by h2 inhibitory signals.the cytomegalovirus resistance locus cmv3 has been linked to an epistatic interaction between two loci: a natural killer (nk) cell receptor gene and the major histocompatibility complex class i (mhc-i) locus. to demonstrate the interaction between cmv3 and h2(k), we generated double congenic mice between ma/my and balb.k mice and an f(2) cross between fvb/n (h-2(q)) and balb.k (h2(k)) mice, two strains susceptible to mouse cytomegalovirus (mcmv). only mice expressing h2(k) in conjunction with cm ...201121533075
t cell responses in experimental viral retinitis: mechanisms, peculiarities and implications for gene therapy with viral vectors.t lymphocytes play a decisive role in the course and clinical outcome of viral retinal infection. this review focuses on aspects of the adaptive cellular immune response against viral pathogens in the retina. two distinct models to study adaptive cell mediated immune responses in viral retinitis are presented: (i) experimental retinitis induced by murine cytomegalovirus (mcmv), where the immune system prevents necrotizing damage to the retina and (ii) retinitis induced by the non-cytopathic lymp ...201121550416
functional interaction of nuclear domain 10 and its components with cytomegalovirus after infections: cross-species host cells versus native cells.species-specificity is one of the major characteristics of cytomegaloviruses (cmvs) and is the primary reason for the lack of a mouse model for the direct infection of human cmv (hcmv). it has been determined that cmv cross-species infections are blocked at the post-entry level by intrinsic cellular defense mechanisms, but few details are known. it is important to explore how cmvs interact with the subnuclear structure of the cross-species host cell. in our present study, we discovered that nucl ...201121552525
small molecule inhibitors of the host cell cox/areg/egfr/erk pathway attenuate cytomegalovirus-induced pathogenesis.as with other herpesviruses, human cytomegalovirus (hcmv) has the ability to establish lifelong persistence and latent infection following primary exposure, salivary glands (smgs) being the primary site of both. in the immunocompromised patient, hcmv is a common cause of opportunistic infections, and subsequent morbidity and mortality. elucidating the molecular pathogenesis of cmv-induced disease is critical to the development of more effective and safer drug therapies. in the present study, we ...201121565184
t cell help permits memory cd8(+) t cell inflation during cytomegalovirus latency.cd4(+) t cells are implied to sustain cd8(+) t cell responses during persistent infections. as cd4(+) t cells are often themselves antiviral effectors, they might shape cd8(+) t cell responses via help or via controlling antigen load. we used persistent mouse cytomegalovirus (mcmv) infection to dissect the impact of cd4(+) t cells on virus-specific cd8(+) t cells, distinguishing between increased viral load in absence of cd4(+) t cells and cd4(+) t cell-mediated helper mechanisms. absence of t h ...201121590767
effects of allitridin on acute and chronic mouse cytomegalovirus infection.this study investigated the effects of allitridin on acute and chronic mouse cytomegalovirus (mcmv) infections in vivo. the results demonstrated that allitridin reduced the titers of mcmv in salivary glands, and reductions in viral loads were confirmed by determining viral dna and rna levels in susceptible organs during the acute infection phase. although allitridin did not eliminate mcmv, treatment reduced viral levels and facilitated healing of pathologic lesions in organs, particularly during ...201121604182
batf3 transcription factor-dependent dc subsets in murine cmv infection: differential impact on t cell priming and memory inflation.priming of cd8(+) t cells specific for viruses that interfere with the mhc class i presentation pathway is a challenge for the immune system and is believed to rely on cross-presentation. cytomegalovirus (cmv) infection induces vigorous cd8(+) t cell responses despite its potent immune evasion strategies. furthermore, cd8(+) t cells specific for a subset of viral epitopes accumulate and are maintained at high levels exhibiting an activated phenotype - referred to as 'inflationary t cells'. takin ...201121604258
comparative genome and virion structurethe two major lineages in the betaherpesvirinae are the cytomegaloviruses (the cytomegalovirus and muromegalovirus genera, plus a number of other viruses whose taxonomy is only partially defined) and the roseolovirus genus (see chapter 1). the best characterized members of these lineages are hcmv (the prototype of the subfamily) and hhv-6, respectively. cytomegaloviruses are present in a wide range of mammalian species, and have been termed “salivary gland viruses” because of the ...200721348083
[fluvastatin's effect on atherogenesis in apolipoprotein-e knockout mice infected by cytomegalovirus].the goal of this study was to investigate whether murine cytomegalovirus (mcmv) is able to exacerbate the atherosclerotic process in apolipoprotein e knockout (apoe -/-) mice, and the effect of fluvastatin on the atherogenesis.201021604568
the role of interleukin-17 in murine cytomegalovirus interstitial pneumonia in mice with skin transplants.we hypothesized that the t helper (th)17 response plays an important role in murine cytomegalovirus (mcmv) interstitial pneumonia. balb/c mice with skin grafts from c57/bj6 mice were intranasally inoculated with 1.0 × 10(5) pfu mcmv. lung tissues and skin grafts were histologically evaluated and expression of interleukins (il)-17, -6 and -8, monocyte chemotactic protein (mcp)-1 and interferon (ifn)-γ in serum and bronchoalveolar lavage (bal) fluid, intracellular il-4, -17, and ifn-γ, in spleen l ...201121623953
antigen-presenting cells of hematopoietic origin prime cytomegalovirus-specific cd8 t cells but are not sufficient for driving memory inflation during viral latency.expansion of the cd8 t cell memory pool, also known as 'memory inflation', for certain but not all viral epitopes in latently infected host tissues is a special feature of the immune response to cytomegalovirus. the ld-presented murine cytomegalovirus (mcmv) ie1 peptide is the prototype of an epitope that is associated with memory inflation. based on the detection of ie1 transcripts in latently infected lungs it was previously proposed that episodes of viral gene expression and antigenic activit ...201121632567
h2b homology region of major immediate-early (mie) protein 1 (ie1) is essential for murine cytomegalovirus to disrupt nuclear domain 10 (nd10), but not important for viral replication in cell culture.abstract: cytomegalovirus (cmv) major immediate-early protein 1 (ie1) has multiple functions and is important for efficient viral infection. as does its counterpart in human cmv, murine cmv (mcmv) ie1 also functions as a disruptor of mouse cell nd10 (nuclear domain 10), where many different gene-regulation proteins congregate. it still remains unclear how mcmv ie1 disperses nd10 and whether this dispersion could have any effect on viral replication. mcmv ie1 has 595 amino acids and multiple func ...201121632568
suppression of tlr9 immunostimulatory motifs in the genome of a gammaherpesvirus.multiple receptors within the innate immune system have evolved to recognize nucleic acids as signatures of viral infection. it is believed that this specificity is essential for viral detection, as viruses often lack other invariant features that can serve as suitable targets for innate receptors. one such innate receptor, tlr9, has been implicated in the detection of many dsdna viruses. in this study, we investigate the detection of murine gammaherpesvirus 68 (mhv68) by tlr9. we find that the ...201121666062
virus progeny of mcmv bacterial artificial chromosome psm3fr shows reduced growth in salivary glands due to a fixed mutation of mck-2.mcmv (murine cytomegalovirus) smith strain has been cloned as a bacterial artificial chromosome (bac) named psm3fr and has since then been used for analysis of virus gene functions in vitro and in vivo. when sequencing the complete bac genome, we identified a frameshift mutation within the orf encoding mcmv chemokine homolog mck-2. this mutation would result in a truncated mck-2 protein. when mice were infected with psm3fr-derived virus, we observed a reduced virus production in salivary glands ...201121813614
reversible inhibition of mcmv replication by ifn{gamma} in primary macrophages involves a primed type i ifn signaling sub-network for full establishment of an immediate-early antiviral-state.activated macrophages play a central role in controlling inflammatory responses to infection and are tightly regulated to rapidly mount responses to infectious challenge. type-i (ifna/ß) and type-ii (ifn?) interferon play a crucial role in activating macrophages and subsequently restricting viral infections. both types of ifns signal through related but distinct signaling pathways, inducing a vast number of interferon stimulated genes, overlapping but distinguishable. the exact mechanism by whic ...201121775459
m94 is essential for the secondary envelopment of murine cytomegalovirus.the gene m94 of murine cytomegalovirus (mcmv) as well as its homologues ul16 in alphaherpesviruses is involved in viral morphogenesis. for a better understanding of its role in the viral life cycle, a library of random m94 mutants was generated by modified transposon-based linker scanning mutagenesis. a comprehensive set of m94 mutants was reinserted into the mcmv genome and tested for their capacity to complement the m94 null mutant. thereby, 34 loss-of-function mutants of m94 were identified, ...201121715482
Natural Killer Cell Licensing During Viral Infection.Natural Killer (NK) cell functionality is controlled by inhibitory receptors that recognize self-MHC class I. NK cells that do not interact with self-MHC class I are hypo-responsive to many stimuli and fail to reject MHC class I-deficient cells. Thus, although the mechanisms are unknown, interactions with MHC class I "licensed" NK cells respond efficiently. Surprisingly, these licensed NK cells fail to control viral infection. During mouse cytomegalovirus (MCMV) infection, SHP-1 signaling downst ...201221842363
dissecting the requirements for maintenance of the cmv-specific memory t-cell pool.cytomegalovirus (cmv) infection promotes a broad t-cell response, with the resulting memory cells displaying diverse phenotypes. cmv establishes lifelong persistence/latency, and it is thought that viral antigens expressed during this period may regulate the expansion and/or maintenance of "inflationary" cd8 t-memory populations that display an effector memory phenotype. we show here that mouse cmv (mcmv)-specific inflationary memory t cells do not decrease in number after thymectomy, indicating ...201121721929
comparing the kinetics of nk cells, cd4, and cd8 t cells in murine cytomegalovirus infection.nk cells recognize virus-infected cells with germline-encoded activating and inhibitory receptors that do not undergo genetic recombination or mutation. accordingly, nk cells are often considered part of the innate immune response. the innate response comprises rapid early defenders that do not form immune memory. however, there is increasing evidence that experienced nk cells provide increased protection to secondary infection, a hallmark of the adaptive response. in this study, we compare the ...201121697462
absence of vasoactive intestinal peptide expression in hematopoietic cells enhances th1 polarization and antiviral immunity in mice.vasoactive intestinal peptide (vip) induces regulatory dendritic cells (dc) in vitro that inhibit cellular immune responses. we tested the role of physiological levels of vip on immune responses to murine cmv (mcmv) using vip-knockout (vip-ko) mice and radiation chimeras engrafted with syngenic vip-ko hematopoietic cells. vip-ko mice had less weight loss and better survival following mcmv infection compared with wild-type (wt) littermates. mcmv-infected vip-ko mice had lower viral loads, faster ...201121677142
Expression of the RAE-1 family of stimulatory NK-cell ligands requires activation of the PI3K pathway during viral infection and transformation.Natural killer (NK) cells are lymphocytes that play a major role in the elimination of virally-infected cells and tumor cells. NK cells recognize and target abnormal cells through activation of stimulatory receptors such as NKG2D. NKG2D ligands are self-proteins, which are absent or expressed at low levels on healthy cells but are induced upon cellular stress, transformation, or viral infection. The exact molecular mechanisms driving expression of these ligands remain poorly understood. Here we ...201121966273
Flagellin, a TLR5 agonist, reduces graft-versus-host disease in allogeneic hematopoietic stem cell transplantation recipients while enhancing antiviral immunity.Graft-versus-host disease (GVHD) is a major cause of morbidity and mortality in patients treated with allogeneic hematopoietic stem cell transplantation (HSCT). Posttransplant immunosuppressive drugs incompletely control GVHD and increase susceptibility to opportunistic infections. In this study, we used flagellin, a TLR5 agonist protein (~50 kDa) extracted from bacterial flagella, as a novel experimental treatment strategy to reduce both acute and chronic GVHD in allogeneic HSCT recipients. On ...201122013117
il-10 restricts activation-induced death of nk cells during acute murine cytomegalovirus infection.il-10 is an immunomodulatory cytokine that acts to antagonize t cell responses elicited during acute and chronic infections. thus, the il-10r signaling pathway provides a potential therapeutic target in strategies aimed at combating infectious diseases. in this study, we set out to investigate whether il-10 expression had an effect on nk cells. murine cmv infection provides the best characterized in vivo system to evaluate the nk cell response, with nk cells being critical in the early control o ...201121849677
Squalamine as a broad-spectrum systemic antiviral agent with therapeutic potential.Antiviral compounds that increase the resistance of host tissues represent an attractive class of therapeutic. Here, we show that squalamine, a compound previously isolated from the tissues of the dogfish shark (Squalus acanthias) and the sea lamprey (Petromyzon marinus), exhibits broad-spectrum antiviral activity against human pathogens, which were studied in vitro as well as in vivo. Both RNA- and DNA-enveloped viruses are shown to be susceptible. The proposed mechanism involves the capacity o ...201121930925
ganciclovir transiently attenuates murine cytomegalovirus-associated renal allograft inflammation.prophylactic ganciclovir (gcv) is used in high-risk renal transplant patients to prevent acute cytomegalovirus (cmv) disease, but its impact on inflammation within the allograft itself remains undefined.201121878840
[influence of allitridin on transcription, expression and function of il-12 genes in mice infected by murine cytomegalovirus].to investigate whether allitridin could interfere with the effects of murine cytomegalovirus (mcmv) infection on the transcription, expression and function of il-12 genes in order to further explore the mechanism of allitridin against mcmv.201122007556
absence of cross-presenting cells in the salivary gland and viral immune evasion confine cytomegalovirus immune control to effector cd4 t cells.horizontal transmission of cytomegaloviruses (cmv) occurs via prolonged excretion from mucosal surfaces. we used murine cmv (mcmv) infection to investigate the mechanisms of immune control in secretory organs. cd4 t cells were crucial to cease mcmv replication in the salivary gland (sg) via direct secretion of ifnγ that initiated antiviral signaling on non-hematopoietic cells. in contrast, cd4 t cell helper functions for cd8 t cells or b cells were dispensable. despite sg-resident mcmv-specific ...201121901102
virally-induced upregulation of heparan sulfate on b cells via the action of type i ifn.cell surface heparan sulfate (hs) is an important coreceptor for many cytokines, chemokines, and growth factors. in this study, we report that splenic murine b cells express very little hs and that upon infection with either gammaherpesvirus (murine gammaherpesvirus 68) or betaherpesvirus (murine cytomegalovirus), hs is rapidly upregulated at the surface of b cells. hs upregulation was not observed in mice deficient for the type i ifn (ifn-i) receptor. additionally, treatment of wild-type mice w ...201122048770
polarized distribution of heme transporters in retinal pigment epithelium and their regulation in the iron-overload disease hemochromatosis.flvcr, bcrp, and pcft/hcp-1 represent the three heme transporters identified thus far in mammalian cells, but there is very little known about their expression and regulation in the retina. in this study, the expression of these transporters in mouse retina and retinal pigment epithelium (rpe) and their regulation in the iron-overload disease hemochromatosis were examined.201122058337
A replicating cytomegalovirus-based vaccine encoding a single Ebola virus nucleoprotein CTL epitope confers protection against Ebola virus.Human outbreaks of Ebola virus (EBOV) are a serious human health concern in Central Africa. Great apes (gorillas/chimpanzees) are an important source of EBOV transmission to humans due to increased hunting of wildlife including the 'bush-meat' trade. Cytomegalovirus (CMV) is an highly immunogenic virus that has shown recent utility as a vaccine platform. CMV-based vaccines also have the unique potential to re-infect and disseminate through target populations regardless of prior CMV immunity, whi ...201121858240
tuning of natural killer cell reactivity by nkp46 and helios calibrates t cell responses.natural killer (nk) cells are lymphocytes involved in antimicrobial and antitumoral immune responses. using n-ethyl-n-nitrosourea mutagenesis in mice, we identified a mutant with increased resistance to viral infections because of the presence of hyperresponsive nk cells. whole-genome sequencing and functional analysis revealed a loss-of-function mutation in the ncr1 gene encoding the activating receptor nkp46. the down-regulation of nk cell activity by nkp46 was associated with the silencing of ...201222267813
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