Identification and Characterization of a Master Transcription Factor of Th1 Cells, T-bet, Within Flounder (Paralichthys olivaceus)

T-bet, a T-box family member, is a transcription factor essential for the differentiation of naive CD4+ T cells into Th1 cells that are involved in both innate and adaptive immune responses. In this study, the transcription factor T-bet of flounder (Paralichthys olivaceus) was cloned and characterized, and its expression profile after infection was analyzed. T-bet+ cells were identified in flounder, and the expression and localization of T-bet in T lymphocyte subsets and B lymphocytes were investigated. Finally, the proliferation of T-bet+ cells, T lymphocyte subsets, and B lymphocytes were studied after stimulation with IFN-γ, IL-2, and IL-6, respectively, and the variations of some transcription factors and cytokines in CD4+ T lymphocyte subsets were detected. The results showed that T-bet in flounder consists of 619 aa with a conserved T-box DNA binding domain. T-bet was abundantly expressed in the spleen, head kidney, and heart, and it was significantly upregulated after infection with Vibrio anguillarum, Edwardsiella tarda, and Hirame rhabdovirus, especially in the group of Edwardsiella tarda. A polyclonal antibody against recombinant protein of T-bet was prepared, which specifically recognized the natural T-bet molecule in flounder. T-bet+ cells were found to be distributed in the lymphocytes of peripheral blood, spleen, and head kidney, with the highest proportion in spleen, and the positive signals of T-bet occurred in the cell nucleus. T-bet was also detected in the sorted CD4-1+, CD4-2+, CD8+ T lymphocytes, and IgM+ B lymphocytes. In addition, T-bet+ cells, coordinated with CD4-1+ and CD4-2+ T lymphocytes, were proliferated after stimulation with IFN-γ, IL-2, and IL-6. Especially in sorted CD4-1+ and CD4-2+ T lymphocytes, IFN-γ and IL-2 were able to upregulate the expression of T-bet, forming a positive feedback loop in Th1-type cytokine secretion. These results suggest that T-bet may act as a master transcription factor regulating flounder CD4+ T lymphocytes involved in a Th1-type immune response.

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Kinetics of T lymphocyte subsets and B lymphocytes in response to immunostimulants in flounder (Paralichthys olivaceus): implications for CD4+ T lymphocyte differentiation

Scientific Reports ◽

10.1038/s41598-020-69542-6 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Jing Xing ◽

Hong-fei Tian ◽

Xiao-qian Tang ◽

Xiu-zhen Sheng ◽

Wen-bin Zhan

Keyword(s):

B Lymphocytes ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

Paralichthys Olivaceus ◽

T Lymphocyte Subsets ◽

Lymphocyte Differentiation ◽

Kinetics Of

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Flow cytometric analysis of T lymphocytes and cytokines in aqueous humor of patients with varicella zoster virus-mediated acute retinal necrosis

BMC Ophthalmology ◽

10.1186/s12886-021-01951-1 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Hao Kang ◽

Yunbo Wei ◽

Ming Liu ◽

Di Yu ◽

Yong Tao

Keyword(s):

T Lymphocytes ◽

Varicella Zoster Virus ◽

Aqueous Humor ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

Flow Cytometric Analysis ◽

T Lymphocyte Subsets ◽

Varicella Zoster ◽

Flow Cytometric ◽

Cd8 T Lymphocytes

Abstract Background The purpose of this study is to investigate the aqueous humor (AH) T lymphocyte subsets and cytokines of acute retinal necrosis (ARN) to elucidate the immunologic inflammatory features of this disorder. Methods Three patients with ARN infected with varicella zoster virus (VZV) who underwent multiple intravitreal injections of ganciclovir were enrolled in this study. The control group consisted of four non-infectious patients with acute anterior uveitis (AAU). Flow cytometric analysis was performed on the lymphocyte subsets from the AH and peripheral blood (PB) samples during the active phase of intraocular inflammation. Five inflammatory cytokines were measured in each AH sample and various clinical characteristics were also assessed. Results VZV deoxyribonucleic acid (DNA) was detected by real-time polymerase chain reaction (PCR) in AH from all the ARN patients, who showed higher CD8+ T lymphocytes population in AH than the AAU patients (p = 0.006). CD4/CD8 ratios of T lymphocytes and the percentage of CD8 + CD25+ T lymphocytes in AH were significantly lower in ARN than in AAU (p = 0.006; p = 0.012). In the ARN patients, the percentages of CD4+ and CD8+ T lymphocytes in AH were higher than those found in PB. The percentage of CD4 + CD25+ T lymphocytes in AH was significantly higher than the proportion in PB in the AAU patients (p = 0.001). Immunoregulatory cytokine Interleukin-10 in AH was significantly elevated in the ARN patients in comparison with the case of the AAU patients (p = 0.036). In ARN, the copy number of VZV DNA in AH positively correlated with the percentage of CD8+ T lymphocytes in AH and negatively correlated with the CD4/CD8 ratio in AH during the course of disease treatment (p = 0.009, r = 0.92; p = 0.039, r = − 0.834). Conclusion The ARN patients caused by VZV had different intraocular T lymphocyte subsets and cytokines profile than those of the non-infectious patients. High percentages of CD8+ T lymphocytes and low CD4/CD8 T cell ratios may be a potential biomarker for diagnosis of viral-infectious uveitis. T lymphocytes examination at the inflammatory sites has the potential to become a useful research tool for differentiating viral and non-viral uveitis.

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Flow Cytometric Analysis of T Lymphocytes and Cytokines in Aqueous Humor of Patients with Varicella Zoster Virus-mediated Acute Retinal Necrosis

10.21203/rs.3.rs-56692/v1 ◽

2020 ◽

Author(s):

Hao Kang ◽

Yunbo Wei ◽

Ming Liu ◽

Di Yu ◽

Yong Tao

Keyword(s):

T Lymphocytes ◽

Varicella Zoster Virus ◽

Aqueous Humor ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

Flow Cytometric Analysis ◽

T Lymphocyte Subsets ◽

Varicella Zoster ◽

Flow Cytometric ◽

Cd8 T Lymphocytes

Abstract Background: The purpose of this study is to investigate the aqueous humor (AH) T lymphocyte subsets and cytokines of acute retinal necrosis (ARN) to elucidate the immunologic inflammatory features of this disorder.Methods: Three patients with ARN infected with varicella zoster virus (VZV) who underwent multiple intravitreal injections of ganciclovir were enrolled in this study. The control group consisted of four non-viral infectious patients with acute anterior uveitis (AAU). Flow cytometric analysis was performed on the lymphocyte subsets from the AH and peripheral blood (PB) samples during the active phase of intraocular inflammation. Five inflammatory cytokines were measured in each AH sample and various clinical characteristics were also assessed.Results: VZV DNA was detected by real-time polymerase chain reaction (PCR) in AH from all the ARN patients, who showed higher CD8+ T lymphocytes population in AH than the AAU patients (p=0.006). CD4/CD8 ratios of T lymphocytes and the percentage of CD8+CD25+ T lymphocytes in AH were significantly lower in ARN than in AAU (p=0.006; p=0.012). In the ARN patients, the percentages of CD4+ and CD8+ T lymphocytes in AH were higher than those found in PB. The percentage of CD4+CD25+ T lymphocytes in AH was significantly higher than the proportion in PB in the AAU patients (p=0.001). Immunoregulatory cytokine Interleukin-10 in AH was significantly elevated in the ARN patients in comparison with the case of the AAU patients (p=0.036). In ARN, the copy number of VZV DNA in AH positively correlated with the percentage of CD8+ T lymphocytes in AH and negatively correlated with the CD4/CD8 ratio in AH during the course of disease treatment (p=0.009, r=0.92; p=0.039, r=-0.834).Conclusion: The ARN patients caused by VZV had different intraocular T lymphocyte subsets and cytokines profile than those of the non-viral infectious patients. High percentages of CD8+ T lymphocytes and low CD4/CD8 T cell ratios may be a potential biomarker for diagnosis of viral-infectious uveitis. T lymphocytes examination at the inflammatory sites has the potential to become a useful research tool for differentiating viral and non-viral uveitis.

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Flow Cytometry Study of Lymphocyte Subsets in Malnourished and Well-Nourished Children with Bacterial Infections

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.11.3.577-580.2004 ◽

2004 ◽

Vol 11 (3) ◽

pp. 577-580 ◽

Cited By ~ 45

Author(s):

Oralia Nájera ◽

Cristina González ◽

Guadalupe Toledo ◽

Laura López ◽

Rocío Ortiz

Keyword(s):

Flow Cytometry ◽

B Lymphocytes ◽

T Lymphocyte ◽

Bacterial Infections ◽

B Lymphocyte ◽

Lymphocyte Subsets ◽

Protein Energy Malnutrition ◽

T Lymphocyte Subsets ◽

Malnourished Children ◽

Protein Energy

ABSTRACT Protein-energy malnutrition is the primary cause of immune deficiency in children across the world. It has been related to changes in peripheral T-lymphocyte subsets. The aim of the present study was to evaluate the effects of infection and malnutrition on the proportion of peripheral-lymphocyte subsets in well-nourished non-bacterium-infected (WN), well-nourished bacterium-infected (WNI), and malnourished bacterium-infected (MNI) children by flow cytometry. A prospectively monitored cohort of 15 MNI, 12 WNI, and 17 WN children was studied. All the children were 3 years old or younger and had only bacterial infections. Results showed a significant decrease in the proportion of T CD3+ (P < 0.05 for relative and P < 0.03 for absolute values), CD4+ (P < 0.01 for relative and absolute values), and CD8+ (P < 0.05 for relative values) lymphocyte subsets in WNI children compared to the results seen with WN children. Additionally, B lymphocytes in MNI children showed significant lower values (CD20+ P < 0.02 for relative and P < 0.05 for absolute values) in relation to the results seen with WNI children. These results suggest that the decreased proportions of T-lymphocyte subsets observed in WNI children were associated with infection diseases and that the incapacity to increase the proportion of B lymphocyte was associated with malnutrition. This low proportion of B lymphocytes may be associated with the mechanisms involved in the immunodeficiency of malnourished children.

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Methrotexate Treatment Inmunomodulates Abnormal Cytokine Expression by T CD4 Lymphocytes Present in DMARD-Naïve Rheumatoid Arthritis Patients

International Journal of Molecular Sciences ◽

10.3390/ijms21186847 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6847

Author(s):

Jorge Monserrat Sanz ◽

Cristina Bohórquez ◽

Ana Maria Gómez ◽

Atusa Movasat ◽

Ana Pérez ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

T Lymphocytes ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

Cytokine Expression ◽

Effector Memory ◽

T Lymphocyte Subsets ◽

Relevant Role ◽

Circulating Levels ◽

New Onset

CD4+T-lymphocytes are relevant in the pathogenesis of rheumatoid arthritis (RA), however, their potential involvement in early RA remains elusive. Methotrexate (MTX) is a commonly used disease-modifying antirheumatic drug (DMARD), but its mechanism has not been fully established. In 47 new-onset DMARD-naïve RA patients, we investigated the pattern of IFNγ, IL-4 and IL-17A expression by naïve (TN), central (TCM), effector memory (TEM) and effector (TE) CD4+ subsets; their STAT-1, STAT-6 and STAT-3 transcription factors phosphorylation, and the circulating levels of IFNγ, IL-4 and IL-17. We also studied the RA patients after 3 and 6 months of MTX treatment and according their clinical response. CD4+T-lymphocyte subsets and cytokine expression were measured using flow cytometry. New-onset DMARD-naïve RA patients showed a significant expansion of IL-17A+, IFNγ+ and IL-17A+IFNγ+ CD4+T-lymphocyte subsets and increased intracellular STAT-1 and STAT-3 phosphorylation. Under basal conditions, nonresponder patients showed increased numbers of circulating IL-17A producing TN and TMC CD4+T-lymphocytes and IFNγ producing TN, TCM, TEM CD4+T-lymphocytes with respect to responders. After 6 months, the numbers of CD4+IL-17A+TN remained significantly increased in nonresponders. In conclusion, CD4+T-lymphocytes in new-onset DMARD-naïve RA patients show IL-17A and IFNγ abnormalities in TN, indicating their relevant role in early disease pathogenesis. Different patterns of CD4+ modulation are identified in MTX responders and nonresponders.

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Characterization of flounder (Paralichthys olivaceus) CD4+ T lymphocyte subsets in response to Th-type antigens

Fish & Shellfish Immunology ◽

10.1016/j.fsi.2019.04.270 ◽

2019 ◽

Vol 91 ◽

pp. 464

Author(s):

Hongfei Tian ◽

Jing Xing ◽

Xiaoqian Tang ◽

Xiuzhen Sheng ◽

Wenbin Zhan

Keyword(s):

T Lymphocyte ◽

Lymphocyte Subsets ◽

Paralichthys Olivaceus ◽

T Lymphocyte Subsets

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Establishment of Normal Reference Intervals for CD3+, CD4+, CD8+, and CD4+to CD8+Ratio of T Lymphocytes in HIV Negative Adults from University of Gondar Hospital, North West Ethiopia

AIDS Research and Treatment ◽

10.1155/2014/267450 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 2

Author(s):

Addisu Gize ◽

Biniam Mathewos ◽

Beyene Moges ◽

Meseret Workineh ◽

Lealem Gedefaw

Keyword(s):

T Lymphocytes ◽

T Lymphocyte ◽

Lymphocyte Subsets ◽

Reference Interval ◽

Reference Intervals ◽

University Hospital ◽

T Lymphocyte Subsets ◽

Cross Sectional ◽

North West ◽

Normal Reference

Background.Reference values for the CD3+, CD4+, CD8+, and CD4+to CD8+ratio T lymphocyte subsets are adopted from textbooks. But for appropriate diagnosis, treatment, and follow-up of patients, correct interpretations of the laboratory results from normal reference interval are mandatory. This study was, therefore, planned to establish normal reference interval for T lymphocytes subset count and CD4+to CD8+ratio.Methods.A cross-sectional study was conducted on apparently healthy adult individuals who visited voluntary counseling and HIV testing clinic Gondar University Hospital from April to May, 2013. Whole blood was analyzed using fluorescence-activated cell sorting (BD FACS, San Jose, CA) machine to enumerate the T-cell subpopulations.Results.Out of the total 320 study participants, 161 (50.3%) were men and 159 (49.7%) were women. The normal reference intervals were (655–2,823 cells/μL), (321–1,389 cells/μL), and (220–1,664 cells/μL) for CD3+, CD4+, and CD8+T lymphocyte subsets, respectively, and CD4+to CD8+ratio was 0.5–2.5.Conclusion.The overall CD3+T lymphocytes reference interval in the current study was wide; low CD4+T lymphocytes, CD4 to CD8 ratio, and high CD8+T lymphocytes values were observed.

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The Levels of T Lymphocyte Subsets in Immune Thrombocytopenia Associated with Anti-GPIIb/IIIa- and/or Anti-GPIbα-Mediated Responses Are Differentially Sensitive to Dexamethasone

Acta Haematologica ◽

10.1159/000491977 ◽

2018 ◽

Vol 140 (1) ◽

pp. 60-66 ◽

Cited By ~ 5

Author(s):

Yang Chen ◽

Yanyan Xie ◽

Min Ruan ◽

Jinning Shi

Keyword(s):

T Lymphocytes ◽

T Lymphocyte ◽

Immune Thrombocytopenia ◽

Lymphocyte Subsets ◽

Response Rate ◽

Control Group ◽

Dexamethasone Treatment ◽

T Lymphocyte Subsets ◽

Cd8 T Lymphocytes ◽

Significant Difference

Objective: The aim of this work was to investigate the influence of T lymphocyte subsets and platelet-specific autoantibodies on immune thrombocytopenia (ITP) with dexamethasone therapy. Methods: The samples were obtained from patients before therapy. T lymphocyte subsets were measured by flow cytometry, and platelet-specific autoantibodies were evaluated by modified monoclonal antibody immobilization of platelet antigen assay. Results: A total of 50 ITP patients were involved in the study. Twenty-three were anti-GPIbα antibody positive and were treated with dexamethasone, with a response rate of 47.8%. Twenty-seven cases were anti-GPIbα antibody negative, with a response rate of 77.8%. A significant difference was detected (p < 0.05). The level of CD4+ T lymphocytes in ITP patients was lower compared with the control group (p < 0.05). The level of CD8+ T lymphocytes was higher than that in the normal controls (p < 0.05). Additionally, the patients with a higher level of CD8+ T lymphocytes and lower level of CD4+ T lymphocytes were more likely to respond to dexamethasone treatment. Moreover, we observed that ITP patients associated with anti-GPIIb/IIIa antibodies had lower levels of CD4+ T lymphocytes and higher CD8+ T lymphocyte levels. Conclusions: There was insensitivity to dexamethasone treatment in ITP patients who were anti-GPIbα antibody positive. The detection of T lymphocyte subsets is useful in ITP patients for forecasting the outcome of dexamethasone treatment. There were some relationships between the different antibodies and the levels of T lymphocyte subsets.

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