Analysis of the T-Cell Receptor Vα Gene Repertoire and Clonal Expansion in the Benzene-Exposed Group.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3874-3874
Author(s):  
Bo Li ◽  
Yangqiu Li ◽  
Shaohua Chen ◽  
Lijian Yang ◽  
Jiayu Chen ◽  
...  
Keyword(s):  
T Cells ◽  
Mononuclear Cells ◽  
Clonal Expansion ◽  
Cell Receptor ◽  
Exposed Group ◽  
Skewed Distribution ◽  
Gene Repertoire ◽  
Peripheral Blood Mononuclear ◽  
Pcr Products ◽  
Expression Rate

Abstract Benzene is a potent human leukemogen, but its mechanism of hematotoxicity is uncertain. It is well know that benzene inhibit T cells proliferation. Several reports revealed that clonal expansion TCR Vβ T cells could be found in workers exposed to benzene. In this study we observe the distribution of TCR Vα gene repertoire and clonal expansion in peripheral blood mononuclear cells from 9 donors and 16 workers exposed to benzene. Complementarity determining region 3 (CDR3) of TCR Vα subfamily genes were amplified using RT-PCR. The PCR products were further analyzed by genescan to evaluating clonality of T cells. 29 Vα subfamily could be detected in 9 donors.1~11 Vα subfamilies were identified in all but one of the workers studied. The most frequently expressed Vα subfamily were Vα3, Vα12 and Vα19 (68.8%), Vα14(56.3%), with a lower expression rate found in Vα5,Vα15, Vα16, Vα 22, Vα 23 and Vα 24 (6.3%). Clonal expansion T cells in one or more Vα subfamily were found in 12 out of all workers studied, including oligoclonal, oligocolonal trend and bioclonal patterns. The frequency of clonal expansion T cells in Vα12, Vα14 and Vα19 subfamilies were higher than others. In conclusion, skewed distribution and clonal expansion of TCR Vα subfamily T cells could be found in workers exposed to benzene. Vα12, Vα14 and Vα19 subfamilies may be highly sensitive to benzene exposed. This is the first report of clonal expansion TCR Vα T cells in the benzene-exposed group. The bias pattern of TCR Vα T cells may be due to the immune cytotocicity from benzene. However, whether the oligoclonality in some Vα subfamilies reflect the phenomenon of clone absense or may be a response clone to benzene-related impairment during exposed to benzene, remains an open question.

Oligoclonal Expansion of TCR V beta Subfamily T Cells in Patients with B-ALL.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3840-3840
Author(s):  
Yangqiu Li ◽  
Meijuan Huang ◽  
Yubing Zhou ◽  
Shaohua Chen ◽  
Lijian Yang
Keyword(s):  
T Cells ◽  
Peripheral Blood ◽  
Cell Clone ◽  
Clonal Expansion ◽  
Gene Repertoire ◽  
Peripheral Blood Mononuclear ◽  
Tumor Associated Antigen ◽  
Specific Cytotoxicity ◽  
Pcr Products

Abstract Clonal expansion of TCR V beta subfamily T cells were identified in peripheral blood and tumor infiltrated tissues lymphocytes from some solid tumors and leukemias, which were thought to be relative to tumor associated antigen and be benefited for design of the strategy of specific immunotherapy. The aim of this study is to investigate the situation of TCR V beta gene repertoire and clonal expansion in peripheral blood T cells from patients with B-ALL. The complementarity determining region 3 (CDR3) of TCR V beta 24 subfamily genes in peripheral blood mononuclear cell from 13 cases with untreated B-ALL were amplified using RT-PCR. The PCR products were further labelled by fluorescein and then analyzed by genescan technique for detection of the CDR3 size, to determine the clonality of T cells. The results showed that only 2–18 V beta subfamilies were identified in 13 B-ALL cases respectively. The frequency expression of V beta subfamilies was V beat 13 (84.6%) and V beta 21 (76.9%). No positive products were detected in V beta 4, 6, 8, 18, 20 and V beta 24 subfamilies. Clonal expansive T cells in one or more V beta subfamilies were found in all 13 cases. The display frequency of clonal expansive T cells in V beta 21 (46.1%) and V beta 23 (30.8%) subfamilies were higher than others. In conclusions, the results indicated that the dominant utilization of TCR V beta repertoire could be found in peripheral blood T cells from patients with B-ALL, meanwhile clonal expansive T cells were existed. It may be a feature of the host immune response for leukemia associated antigen. The clonal expansive tendency of T cells in V beta 21 and V beta 23 subfamilies was rather obvious, which maybe correlated with certain malignant B-cell clone. Isolation and amplification of the clonal expanded V beta T cells in vitro may be necessary for identification of the specific cytotoxicity for leukemia cells.

scholarly journals Differential effects of IL-21 and IL-15 on perforin expression, lysosomal degranulation, and proliferation in CD8 T cells of patients with human immunodeficiency virus-1 (HIV)

Blood ◽  
2006 ◽  
Vol 109 (9) ◽  
pp. 3873-3880 ◽  
Author(s):  
Lesley White ◽  
Subramaniam Krishnan ◽  
Natasa Strbo ◽  
Huanliang Liu ◽  
Michael A. Kolber ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cd8 T Cells ◽  
Highly Active Antiretroviral Therapy ◽  
Cell Activation ◽  
Mononuclear Cells ◽  
Cell Receptor ◽  
Peripheral Blood Mononuclear ◽  
Perforin Expression

Abstract An urgent need exists to devise strategies to augment antiviral immune responses in patients with HIV who are virologically well controlled and immunologically stable on highly active antiretroviral therapy (HAART). The objective of this study was to compare the immunomodulatory effects of the cytokines interleukin (IL)–21 with IL-15 on CD8 T cells in patients with HIV RNA of less than 50 copies/mL and CD4 counts greater than 200 cells/mm.3 Patient CD8 T cells displayed skewed maturation and decreased perforin expression compared with healthy controls. Culture of freshly isolated patient peripheral-blood mononuclear cells (PBMCs) for 5 hours to 5 days with IL-21 resulted in up-regulation of perforin in CD8 T cells, including memory and effector subsets and virus-specific T cells. IL-21 did not induce T-cell activation or proliferation, nor did it augment T-cell receptor (TCR)–induced degranulation. Treatment of patient PBMCs with IL-15 resulted in induction of perforin in association with lymphocyte proliferation and augmentation of TCR-induced degranulation. Patient CD8 T cells were more responsive to cytokine effects than the cells of healthy volunteers. We conclude that CD8 T cells of patients with HIV can be modulated by IL-21 to increase perforin expression without undergoing overt cellular activation. IL-21 could potentially be useful for its perforin-enhancing properties in anti-HIV immunotherapy.

CD3-Zeta Gene Expression in Workers Benzene-Exposed and Benzene- Poisoned Workers

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4925-4925
Author(s):  
Bo Li ◽  
Yangqiu Li ◽  
Shaohua Chen ◽  
Lijian Yang ◽  
Wei Yu ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Mononuclear Cells ◽  
Skewed Distribution ◽  
Gene Expression Level ◽  
Expression Level ◽  
Peripheral Blood Mononuclear ◽  
Invariant Structure ◽  
Normal Individuals ◽  
Exposed Workers

Abstract Benzene is an industrial chemical and component of cigarette smoke, gasoline, and automobile emissions. Benzene’s toxic effects on the blood and bone marrow can induce aplastic anemia and leukemia. Benzene is known to be highly toxic to a variety of cell types including lymphocytes. Our previous study showed that skewed distribution and clonal expansion of TCR Vα subfamily T cells had been found in benzene-exposed workers, indicating that the disorder of T cell immune function might relate to benzene-exposed. The TCR expressed on the surface of T cells is associated with an invariant structure-CD3 and composed the TCR/CD3 complex. The CD3ζ chain plays an important role in the complex which involved in signal transduction. Little is known about the feature of CD3 ζ chain expression in benzene-exposed workers. To further identify the expression level of CD3 ζ gene in benzene-exposed workers, real-time PCR with SYBR Green±technique was used for detecting CD3 ζ gene expression level in peripheral blood mononuclear cells from 29 benzene-exposed workers, 42 benzene-poisoned workers and 20 normal individuals. β2- microglobulin gene (β2M) was used as an endogenous reference. Relative changes in CD3 ζ gene expression level was used by the 2−ΔCt×100% method (ΔCt=Ct(ζ) −Ct(β2M) ). CD3ζ gene could be detected in all of normal individuals, however, only 21 out of 29 benzeneexposed workers could be detected the CD3ζ gene with a mean expression level of 15.0 ±24.9, and in 33 of all 45 benzene-poisoned workers with mean value of 19.8 ±29.7. The detectable CD3 ζ gene expression level in both benzene-exposed and benzene-poisoned groups increased significantly compared with that in normal individuals (3.00±2.11, P< 0.05). This is, to our knowledge, the first description of the effect of benzene-exposed on the expression of the CD3 ζ gene. The abnormality expression of CD3 ζ gene might lead to immune dysfunction in benzene-exposed and benzene-poisoned workers. In addition, CD3ζ gene could not be detected in a part of samples, whether the absence of CD3ζ gene might related to dysfunction of T cells in workers with benzene-exposed and benzenepoisoned, it remains an open question.

Cytolytic activity and regulatory functions of inhibitory NK cell receptor–expressing T cells expanded from granulocyte colony-stimulating factor–mobilized peripheral blood mononuclear cells

Blood ◽  
2004 ◽  
Vol 104 (3) ◽  
pp. 768-774 ◽  
Author(s):  
Junji Tanaka ◽  
Tomomi Toubai ◽  
Yutaka Tsutsumi ◽  
Yoko Miura ◽  
Naoko Kato ◽  
...  
Keyword(s):  
T Cells ◽  
Mononuclear Cells ◽  
Hla Class I ◽  
Cell Receptor ◽  
Cytolytic Activity ◽  
Leukemic Cells ◽  
Peripheral Blood Mononuclear ◽  
Hla Class I Molecules ◽  
Blood Mononuclear Cells ◽  
Mobilized Peripheral Blood

AbstractInhibitory natural killer cell receptor (NKR)–expressing cells may induce a graft-versus-leukemia/tumor (GVL/T) effect against leukemic cells and tumor cells that have mismatched or decreased expression of HLA class I molecules and may not cause graft-versus-host disease (GVHD) against host cells that have normal expression of HLA class I molecules. In our study, we were able to expand inhibitory NKR (CD94/NKG2A)–expressing CD8+ T cells from granulocyte colonystimulating factor (G-CSF)–mobilized peripheral blood mononuclear cells (G-PBMCs) by more than 500-fold using stimulation by an anti-CD3 monoclonal antibody with interleukin 15 (IL-15). These expanded and purified CD94-expressing cells attacked various malignant cell lines, including solid cancer cell lines, as well as the patients' leukemic cells but not autologous and allogeneic phytohemagglutinin (PHA) blasts in vitro. Also, these CD94-expressing cells prevented the growth of K562 leukemic cells and CW2 colon cancer cells in NOD/SCID mice in vivo. On the other hand, the CD94-expressing cells have low responsiveness to alloantigen in mixed lymphocyte culture (MLC) and have high transforming growth factor (TGF)–β1– but low IL-2– producing capacity. Therefore, CD94-expressing cells with cytolytic activity against the recipient's leukemic and tumor cells without enhancement of alloresponse might be able to be expanded from donor G-PBMCs.

Specific Cytotoxicity and Clonal Expansion of TCR Vβ Subfamily T Cells Induced by PML-RARα Peptide.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3904-3904
Author(s):  
Yangqiu Li ◽  
Ji Tang ◽  
Lijian Yang ◽  
Shaohua Chen ◽  
Yubing Zhou
Keyword(s):  
T Cells ◽  
Cord Blood ◽  
Mononuclear Cells ◽  
Statistical Significance ◽  
Clonal Expansion ◽  
Cell Receptor ◽  
Control Group ◽  
Effector Cells ◽  
Specific Cytotoxicity

Abstract The analysis of T cell receptor (TCR) Vβ repertoire is one of the sensitive methods to identify the clonal expansion T cells which response to tumor associated antigens. Understanding the clonality and restricted usage of TCR Vβ repertoire of expanded T-cells induced by PML-RARα peptide may be useful in helping design the new immunotherapeutic strategy specifically for acute promyelocytic leukemia (APL).The aim of the present study was to investigate the specific cytotoxicity and clonality of TCR Vβ repertoire in cord blood T cells induced by PML-RARα peptide (LSSCITQGKAIETQSSSSEE) in vitro. Cord blood mononuclear cells were amplified by IL-2, anti-CD3 and anti-CD28 antibody with different concentration (16.7μg /ml, 33.3μg /ml or 50μg /ml respectively) of synthetic PML-RARα peptide. The induced T cells were collected at different time points after culture (3, 6, 9, 10, 12 or 15 days). The expression and clonality of TCR Vβ subfamilies within induced T cells were analyzed by using RT-PCR and genescan technique. The cytotoxicity of induced T cells was detected by LDH release assay. The results showed that the best condition for T cells induction and amplication was at a concentration with 16.7μg /ml of PML-RARα peptide and at a culture duration with 10 to 15 day. TCR Vβ repertoire analysis showed that restricted expression and cloanl expansion of TCR Vβ subfamily cord blood T cells could be identified after induction by PML-RARα peptide. Clonal expanded T cells were found in Vβ13, Vβ14 and Vβ16 subfamlies respectively. The induced T cells were showed to have the specific cytotoxicity for NB4 cell line (effector cells: tagerted cells=20:1), the cytotoxicity rates were 49.65±6.7% (p<0.05) at day 10th and 73.13±8.42% (p<0.01) at day 15th after culture, which show statistical significance in compare to the control group (without PML-RARα induction). In conclusions, the PML-RARα peptide could induce the clonal expansion T cells from cord blood in vitro, which may have specific cytotoxicity for PML-RARα+ cells.

scholarly journals Neutralization of Interleukin-10 from CD14+ Monocytes Enhances Gamma Interferon Production in Peripheral Blood Mononuclear Cells from Mycobacterium avium subsp. paratuberculosis-Infected Goats

2009 ◽  
Vol 16 (7) ◽  
pp. 1003-1011 ◽  
Author(s):  
Kari R. Lybeck ◽  
Anne K. Storset ◽  
Ingrid Olsen
Keyword(s):  
T Cells ◽  
Mycobacterium Avium ◽  
Mononuclear Cells ◽  
Cell Receptor ◽  
Interleukin 10 ◽  
Gamma Interferon ◽  
Regulatory Mechanisms ◽  
Interferon Production ◽  
Peripheral Blood Mononuclear

ABSTRACT The gamma interferon assay is used to identify Mycobacterium avium subsp. paratuberculosis-infected animals. It has been suggested that regulatory mechanisms could influence the sensitivity of the test when it is performed with cells from cattle and that the neutralization of interleukin-10 (IL-10) in vitro would increase the gamma interferon responses. To investigate the regulatory mechanisms affecting the gamma interferon assay with cells from goats, blood was collected from M. avium subsp. paratuberculosis-infected, M. avium subsp. paratuberculosis-exposed, and noninfected goats. Neutralization of IL-10 by a monoclonal antibody resulted in increased levels of gamma interferon production in M. avium subsp. paratuberculosis purified protein derivative (PPDj)-stimulated samples from both infected and exposed goats. However, the levels of gamma interferon release were also increased in unstimulated cells and in PPDj-stimulated cells from some noninfected animals following neutralization. Depletion of putative regulatory CD25high T cells had no clear effect on the number of gamma-interferon-producing cells. The IL-10-producing cells were identified to be mainly CD14+ major histocompatibility complex class II-positive monocytes in both PPDj-stimulated and control cultures and not regulatory T cells. However, possible regulatory CD4+ CD25+ T cells produced IL-10 in response to concanavalin A stimulation. The numbers of CD4+, CD8+, and CD8+ γδT-cell receptor-positive cells producing gamma interferon increased following IL-10 neutralization. These results provide insight into the source and the role of IL-10 in gamma interferon assays with cells from goats and suggest that IL-10 from monocytes can regulate both innate and adaptive gamma interferon production from several cell types. Although IL-10 neutralization increased the sensitivity of the gamma interferon assay, the specificity of the test could be compromised.

scholarly journals HIV disease progression correlates with the generation of dysfunctional naive CD8low T cells

Blood ◽  
2011 ◽  
Vol 117 (7) ◽  
pp. 2189-2199 ◽  
Author(s):  
David Favre ◽  
Cheryl A. Stoddart ◽  
Brinda Emu ◽  
Rebecca Hoh ◽  
Jeffrey N. Martin ◽  
...  
Keyword(s):  
T Cells ◽  
Hiv Infection ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Cell Receptor ◽  
Interferon Α ◽  
Hiv Disease ◽  
Peripheral Blood Mononuclear ◽  
Class I Mhc ◽  
Mhc I

Abstract HIV infection can result in depletion of total CD4+ T cells and naive CD8+ T cells, and in the generation of dysfunctional effector CD8+ T cells. In this study, we show that naive CD8+ T cells in subjects with progressive HIV disease express low levels of CD8α and CD8β chains. Such naive CD8low T cells display broad signaling defects across the T-cell receptor complex, and their appearance correlates with generalized up-regulation of major histocompatibility complex class I (MHC-I) antigens on peripheral blood mononuclear cells (PBMCs). To explore a causal link between increased MHC-I up-regulation and the generation of naive CD8low T cells, we used the humanized SCID-hu Thy/Liv mouse model to show that HIV infection of the thymus and interferon α (IFNα) treatment alone result in MHC-I up-regulation and in the generation of dysfunctional CD3highCD8+CD4− single-positive 8 (SP8) thymocytes with low expression of CD8. We suggest that dysfunctional naive CD8low T cells are generated as a result of IFNα-mediated up-regulation of MHC-I on stromal cells in the thymus and antigen-presenting cells in the periphery, and that dysfunction in this naive compartment contributes to the immunodeficiency of HIV disease. This study is registered at www.clinicaltrials.gov as NCT00187512.

Oligoclonal Vβ21 with Different Vα Partner in T Cells Associated with CML Cell Antigens

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4236-4236
Author(s):  
Yangqiu Li ◽  
Xianfeng Zha ◽  
Shaohua Chen ◽  
Lijian Yang ◽  
Yuhong Lu ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Mononuclear Cells ◽  
Target Therapy ◽  
Treatment Strategies ◽  
Clinical Problem ◽  
Improvement Strategy ◽  
Peripheral Blood Mononuclear ◽  
Gene Mutant ◽  
Pcr Products

Abstract Chronic myeloid leukemia (CML) is one of most common leukemia, although target therapy with imatinib is significant improved the survival rate, disease relapse and drug resistant with abl gene mutant are still the major clinical problem, improvement strategy to reverse the outcome of CML is the key point for the researchers. T-cell immunodeficiency was suggested in tumor patients for many years, understanding T cell immune status in leukemia patients might suggest treatment strategies to enhance immune competence in all. In order to investigate the clonal expansion and the feature of CDR3 sequence of T cells in patients with CML, the CDR3 of TCR Vα 29 and TCR Vβ 24 subfamily genes were amplified in peripheral blood mononuclear cells (PBMCs) from two case with CML using RT-PCR, the PCR products were further analyzed by genescan technique to evaluate clonality of the detectable TCR Vα and Vβ subfamilies. The oligoclonal PCR products were analyzed by sequencing to define the sequence of CDR3. Oligoclonal Vβ21 T cells could be identified in both CML cases, which might partner with different oligoclonal Vα subfamilies, including Vα7, Vα13 or Vα18. The CDR3 sequences from two Vβ21 and three Vα oligoclonal TCR genes were identified in Vβ21NDβNJβ1.1, Vβ21NDβNJβ2.7, Vα7NJα23, Vα13NJα49, Vα18NJα5 respectively. The length and motifs of CDR3 seem different in all five TCR genes, however, the motifs in both Vβ21 genes (GAV or LRV) share the valine as last residue of CDR3 motif, and all of three Vα gene (Vα7:G; Vα13:GDEAD or Vα18:GG ) share the Glycine as first residue in the CDR3 motif. All of five TCR sequences were already submitted to the Genbank (Accession No: EU395806, EU379940, FJ009444, EU589347 or EU770971 respectively). The results indicated that oligoclonal Vβ21 genes with different Vα partners which share part of CDR3 motifs might recognize the similar CML associated antigen epitope in CML patients. Farther investigation will be constructed the recombination vector containing the antigen specific TCRs for gene transfer to establish TCR modified-T cells, which was expected to employ for specific immunotherapy in CML.

scholarly journals Generation and first characterization of TRDC-knockout pigs lacking γδ T cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Bjoern Petersen ◽  
Robert Kammerer ◽  
Antje Frenzel ◽  
Petra Hassel ◽  
Tung Huy Dau ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Mononuclear Cells ◽  
Flow Cytometric Analysis ◽  
Γδ T Cells ◽  
Cell Receptor ◽  
Lymphoid Tissues ◽  
Γδ T Cell ◽  
Peripheral Blood Mononuclear

AbstractThe TRDC-locus encodes the T cell receptor delta constant region, one component of the γδ T cell receptor which is essential for development of γδ T cells. In contrast to peptide recognition by αβ T cells, antigens activating γδ T cells are mostly MHC independent and not well characterized. Therefore, the function of γδ T cells and their contribution to protection against infections is still unclear. Higher numbers of circulating γδ T cells compared to mice, render the pig a suitable animal model to study γδ T cells. Knocking-out the porcine TRDC-locus by intracytoplasmic microinjection and somatic cell nuclear transfer resulted in healthy living γδ T cell deficient offspring. Flow cytometric analysis revealed that TRDC-KO pigs lack γδ T cells in peripheral blood mononuclear cells (PBMC) and spleen cells. The composition of the remaining leucocyte subpopulations was not affected by the depletion of γδ T cells. Genome-wide transcriptome analyses in PBMC revealed a pattern of changes reflecting the impairment of known or expected γδ T cell dependent pathways. Histopathology did not reveal developmental abnormalities of secondary lymphoid tissues. However, in a vaccination experiment the KO pigs stayed healthy but had a significantly lower neutralizing antibody titer as the syngenic controls.

scholarly journals Single-cell immune profiling reveals distinct immune response in asymptomatic COVID-19 patients

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Xiang-Na Zhao ◽  
Yue You ◽  
Xiao-Ming Cui ◽  
Hui-Xia Gao ◽  
Guo-Lin Wang ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Nk Cells ◽  
Single Cell ◽  
Temporal Dynamics ◽  
Severe Disease ◽  
Clonal Expansion ◽  
Cell Receptor ◽  
Peripheral Blood Mononuclear ◽  
Asymptomatic Patients

AbstractWhile some individuals infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) present mild-to-severe disease, many SARS-CoV-2-infected individuals are asymptomatic. We sought to identify the distinction of immune response between asymptomatic and moderate patients. We performed single-cell transcriptome and T-cell/B-cell receptor (TCR/BCR) sequencing in 37 longitudinal collected peripheral blood mononuclear cell samples from asymptomatic, moderate, and severe patients with healthy controls. Asymptomatic patients displayed increased CD56briCD16− natural killer (NK) cells and upregulation of interferon-gamma in effector CD4+ and CD8+ T cells and NK cells. They showed more robust TCR clonal expansion, especially in effector CD4+ T cells, but lack strong BCR clonal expansion compared to moderate patients. Moreover, asymptomatic patients have lower interferon-stimulated genes (ISGs) expression in general but large interpatient variability, whereas moderate patients showed various magnitude and temporal dynamics of the ISGs expression across multiple cell populations but lower than a patient with severe disease. Our data provide evidence of different immune signatures to SARS-CoV-2 in asymptomatic infections.

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