A time dependent gene expression level of C3, C3aR and CTSL in CD4+ T cells

Abstract The T-cell receptor (TCR) zeta chain is a master sensor and regulator of lymphocyte responses which plays a critical role in TCR-mediated signal transduction. The abnormal expression of TCR zeta gene was found in some malignancies and immune disease. Cellular immune deficiency is the common feature in patients with chronic myeloid leukemia (CML). Our previous studies had showed that 60% of CML patients displayed an abnormally low expression of TCR zeta chain in peripheral blood mononuclear cells (PBMCs). To further estimate the changes of zeta chain expression of T-lymphocyte subsets in CML, TCR zeta chain gene expression level in purified CD4+ and CD8+ T cells sorted by MACS were detected by real-Time PCR with SYBR Green I technique, CD4+ and CD8+ T cells from both 10 cases with CML and 10 normal individuals were selected to analyze in the study. β2 -microglobulin was used as an endogenous reference. Relative changes in TCR zeta gene expression level were used by the 2−ΔCt×100% method. The relative mRNA expression level of TCR zeta gene in CD4+ and CD8+ T cells from CML was 4.06±4.82% and 3.82±4.25% respectively, whereas 7.75±2.52% and 5.19±1.25% were found in normal individuals. There was a wide range in the TCR zeta gene expression level of CD4+ T cells from CML patients (0.23–15.93%,) with a median value of 1.77%, whereas the a range of (4.74–12.81%) with a median of 7.93% was showed in normal controls (P=0.046). The expression level of TCR zeta gene in CD8+ T cells from controls ranged from 3.47% to 6.75% (median 5.52%) and in CML patients from 0.86% to 15.55% with a median of 2.51% (P=0.340). The results indicated that, compared with normal controls, zeta chain gene expression was obviously down-regulated in CD4+ T cells from patients with CML. However, there was not significant different in expression level of TCR zeta gene of CD8+ T cells between CML patients and normal individuals. Moreover, the expression level of TCR zeta gene is nonsignificant age-associated or gender-link in CD4+ T cells and CD8+ T cells from patients with CML. In conclusions, the results provide at first the difference of TCR-zeta gene expression pattern in CD4+ and CD8+ T cells from patients with CML, this may be reflective of signaling difference between the different T cell subtypes in immunodeficiency state of patients.

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Redox Imbalance in CD4+ T Cells of Relapsing-Remitting Multiple Sclerosis Patients

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/8860813 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Mohammad Javad Tavassolifar ◽

Abdorreza Naser Moghadasi ◽

Behnaz Esmaeili ◽

Omid Sadatpour ◽

Mohammad Vodjgani ◽

...

Keyword(s):

Gene Expression ◽

Multiple Sclerosis ◽

T Cells ◽

Cd4 T Cells ◽

Gene Expression Level ◽

Expression Level ◽

Related Factor ◽

Healthy Controls ◽

Relapsing Remitting ◽

Multiple Sclerosis Patients

As a prevalent autoimmune disease of the central nervous system in young adults, multiple sclerosis (MS) is mediated by T cells, particularly CD4+ subsets. Given the evidence that the perturbation in reactive oxygen species (ROS) production has a pivotal role in the onset and progression of MS, its regulation through the antioxidant molecules is too important. Here, we investigated the level of the redox system components in lymphocytes and CD4+ T cells of MS patients. The study was performed on relapsing-remitting MS (RRMS) patients ( n = 29 ) and age- and sex-matched healthy controls ( n = 15 ). Peripheral blood mononuclear cells (PBMCs) were cultured and stimulated by anti-CD3/CD28. The level of ROS, anion superoxide (O2-), and L-𝛾-glutamyl-Lcysteinylglycine (GSH) was measured by flow cytometry in lymphocytes/CD4+ T cells. The gene expression level of gp91phox, catalase, superoxide dismutase 1/2 (SOD), and nuclear factor-E2-related factor (Nrf2) was also measured by real-time PCR. We found that lymphocytes/CD4+ T cells of RRMS patients at the relapse phase significantly produced higher levels of ROS and O2- compared to patients at the remission phase ( P value < 0.001) and healthy controls ( P value < 0.001 and P value < 0.05, respectively). Interestingly, the gene expression level of gp91phox, known as the catalytic subunit of the NADPH oxidase, significantly increased in MS patients at the relapse phase ( P value < 0.05). Furthermore, the catalase expression augmented in patients at the acute phase ( P value < 0.05), while an increased expression of SOD1 and Nrf2 was found in RRMS patients at relapse and remission phases ( P value < 0.05). The increased production of ROS in CD4+ T cells of RRMS patients highlights the importance of amplifying antioxidant components as an efficient approach to ameliorate disease activity in MS patients.

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Expression Pattern of TCR-zeta Chain in Patients with Aplastic Anemia and Polycythemia Vera.

Blood ◽

10.1182/blood.v110.11.3763.3763 ◽

2007 ◽

Vol 110 (11) ◽

pp. 3763-3763

Author(s):

Lijian Yang ◽

Yangqiu Li ◽

Chuang Li ◽

Shaohua Chen ◽

Si Chen ◽

...

Keyword(s):

Gene Expression ◽

T Cells ◽

Aplastic Anemia ◽

Polycythemia Vera ◽

Expression Pattern ◽

Gene Expression Pattern ◽

Gene Expression Level ◽

Expression Level ◽

Chain Gene ◽

Zeta Chain

Abstract TCR zeta chain of TCR-CD3 receptor complex is though to be critical for T cells development and T cell effector function. The absence of TCR zeta chain not only influences the TCR expression on the cell membrane, but also impairs the proliferative response and the level of activation of mature T cells. Recently, the abnormal expression of TCR zeta chain was found different disease, for example in hematological malignancy. In the present study, we analyzed the expression level of TCR zeta chain gene in T cells from patients with aplastic anemia (AA) and polycythemia vera (PV), thereby to estimate the feature of T cells activation status. Real-time PCR with SYBR Green I technique was used for detecting the expression level of TCR zeta chain gene in peripheral blood mononuclear cells of patients with AA (25 cases),PV (12 cases), and 30 normal individuals. β2-microglobulin gene was used as an endogeneous reference. Evaluation of TCR zeta chain gene expression level were used the 2−ΔΔCt method. Compared with the normal control, TCR zeta chain gene expression level could be divided into two groups: 10 patients with AA (40%) and 5 patients with PV (41.7%) were over expression, while 15 patients with AA (60%) and 7 patients with PV (58.3%) were down expression. The expression level of TCR zeta gene is nonsignificat age-associated in both AA and PV patients. Moreover, the expression level of TCR zeta gene is nonsignificant age-associated or gender-link in T cells from patients with AA. The signaling pathway of T cells activation might display some disorder in patients with AA or PV. The effect of the change of TCR zeta chain gene expression pattern in AA and PV patients should be further confirmed.

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

Blood ◽

10.1182/blood.v112.11.4925.4925 ◽

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.

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Single-Cell RNA Sequencing Unveils the Hallmarks of Populations at Different Stages of HTLV-1 Infection

Blood ◽

10.1182/blood-2021-152438 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 3323-3323

Author(s):

Yan Huang ◽

Peifang Jiang ◽

Jiazheng Li ◽

Yanxin Chen ◽

Zhengjun Wu ◽

...

Keyword(s):

Gene Expression ◽

T Cells ◽

T Cell ◽

Single Cell ◽

Cd4 T Cells ◽

Treg Cells ◽

Cd4 T Cell ◽

Expression Level ◽

Normal People ◽

Healthy Donors

Abstract Background Adult T-cell leukemia-lymphoma (ATL) is an aggressive mature T-cell neoplasm caused by human T -cell leukemia virus type 1 (HTLV-1). Up to 5% of infected individuals develop to ATL. HTLV-1 preferentially infects CD4 + T cells, and stimulates cell proliferation and prevents cell death by apoptosis. The viral oncogene-encoded proteins, Tax and HBZ, play important roles in viral infection and cell immortalization. However, the host factor of developing from carrier to patient is not clear. Results To characterize the heterogeneity of ATL patients, we performed single-cell RNA-sequencing (10x Genomics) analysis on single cell suspensions isolated from PBMCs of nine samples, including three ATL patients, three HTLV-1 asymptomatic carriers as well as three healthy donors (HD). We acquired 82977 high-quality cells with a median of 1718 genes detected per cell. Unsupervised clustering using Seurat followed by visualization in t-Stochastic Neighbor Embedding (t-SNE) identified 29 distinct cell clusters (Figure 1A). The single cell profiling of ATL patients were significantly different from that of carriers and healthy donors, while the latter two had little difference (Figure 1B). Based on singleR packages and marker genes of each cluster, 4 major cell populations (T cells, NK cells, B cells and myeloid cells) and other rare cell types were annotated, such as erythrocyte cluster and eosinophils cluster. We observed an enrichment of CD4 + T cell from patients in 4 cluster (Figure 1C), which proportion of cells was higher than that of carriers and healthy donors. According to cell type annotation, cells from cluster 11 were CD4 + CD25 + Foxp3 + Treg cells. Based on the increasing proportion of cluster 11 in healthy people, carriers and patients, without significant statistical differences, we assumed that Foxp3 + Treg cells were involved in the evolution of ATL tumor cells. That was identical with published literatures. To investigate the differences between tumor and normal CD4 + T cell, the gene expression was compared among 7 clusters of CD4 + T cell from three groups. Using a threshold of p value < 0.05 and | fold change| >2. Through integrated analysis, we identified 26 commonly upregulated genes (gene expression level: patients > carriers > HD) and 9 downregulated genes (gene expression level: patients < carriers < HD. To further analyze the biological function of the common DEGs, gene ontology (GO) analysis showed that these genes could be mainly categorized into plasma membrane and protein binding. Subsequently, we validated the mRNA expression level of upregulated common DEGs among three groups by qRT-PCR. The isolated CD4 + T cell using CD4 microbeads of a total of 6 patients, 3 carriers and 9 normal specimens were included. The result showed that the mRNA expression levels of gene CADM1 and RGS13 in patients were higher than those in carriers and healthy donors, although there was no statistical difference between patients and carriers, and the expression levels of carriers tended to be higher than those in normal people (Figure 1D and E). Previously, CADM1 has been revealed to be highly expressed in HTLV-1-infected CD4 + T cells. Our study confirmed this result by single-cell profiling. RGS13, a member of the regulators of G protein signaling (RGS) family, participates in cellular communication. The role of RGS13 in ATL needs to be investigated. Conclusions This study is the first time to analyze the single-cell RNA level of ATL patients, HTLV-1 virus carriers and normal people. The peripheral blood cell composition of the patient is significantly different from that of the carriers and healthy donors, while it is similar between carriers and normal people. CD4 + T cells are the main cell population of patients. The proportion of CD4 + CD25 + Foxp3 + Treg cells increased gradually in healthy people, carriers and patients. DEGs analysis showed that CADM1 and RGS13 were highly expressed in CD4 + T cells of patients, followed by carriers, validated by 18 clinical samples. However, the molecular mechanism of RGS13 in the process from HTLV-1 infection to ATL needs to be further studied. Figure 1 Figure 1. Disclosures Hu: Astellas Pharma, Inc.: Research Funding.

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