Rapid assessment of antigen induced cytokine expression in memory T cells by flow cytometry

1998 ◽  
Vol 63 (1-2) ◽  
pp. 199-207 ◽  
Author(s):  
Vernon C Maino
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Memory T Cells ◽  
Rapid Assessment ◽  
Cytokine Expression

scholarly journals Oligoclonal Expansion and T-Cells Subpopulations in Patients with Aplastic Anemia at Different Stages of the Disease

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3864-3864
Author(s):  
Anastasia V. Abramova ◽  
Elena A. Mikhaylova ◽  
Zalina T. Fidarova ◽  
Yuliya O. Davydova ◽  
Nikolay M. Kapranov ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Aplastic Anemia ◽  
Memory T Cells ◽  
Clonal Expansion ◽  
Th Cells ◽  
Healthy Donors ◽  
Oligoclonal Expansion

Abstract Background. The main mechanism of the bone marrow (BM) failure in idiopathic aplastic anemia (AA) has an immunomediated character. Researching the T-cell clone's effect in the AA pathogenesis is very relevant at the present time. Oligoclonal expansion of T cells is frequent in AA and the identification of immunodominant T-cell clones can correlate with the disease activity and may possibly serve as response predictor to immunosuppressive therapy (IST). The aim. To identify T-cells subpopulations, expression of PD-1 and PD-L1 on T-cells and TCR-Vβ repertoires by flow cytometry in different groups of AA patients. Methods. Thirty AA patients (pts) with median age of 30.5 (19-71), m/f ratio 1:1,3 were divided in 3 groups: pts with newly diagnosed (ND) AA (n=13), pts with overall response to IST (OR) (n=10), non-response pts (NR) for 2 and more lines of IST (n=7). Flow cytometry was performed with BD FACS Canto II. We used commercial kit (IOTest® Beta Mark TCR Vb Repertoire) for evaluation of TCR-Vβ repertoire in the bone marrow (BM) of these patients. We performed analysis of BM samples from healthy donors as a control group (n=8). Due to low amount of donor samples the maximal value each of the 24 subclones (for CD4+ (T-helpers - Th) and CD8+ cells (T-cytotoxic cells - TCL)) was accepted as threshold. We concluded the presence of clonal expansion if TCR subclone exceeded this threshold. We identified different T-cell subpopulations in all 3 groups of AA and healthy donors by flow cytometry: double positive T-cells (CD3+CD4+CD8+), double negative T-cells (CD3+CD4- CD8-), Th (CD3+CD4+), TCL (CD3+CD8+), NK-T-cells (CD3+CD56+) out of CD3+ cells. Among Th and TCL cells was determined naive T-cells (CD28+CD95-), effector T-cells (CD28-CD95+), memory T-cells (CD28+CD95+), regulatory T-cells (CD4+CD127-CD25high) and subpopulations Th and TCL co-expressed PD-1 and PD-L1. Multiple comparisons were assessed by ANOVA or Kruskal Wallis test by GraphPad Prism software. Results. In our study all 30 AA patients had an immunodominant TCR-Vβ clones among Th and/or TCL cells. We identified the most common clonotypes in comparison with healthy donors - Vβ1, Vβ2, Vβ3 among the Th cells and Vβ3, Vβ9, Vβ13.1 among the TCL cells. In ND group Vβ1 was highly expanded in 5 (38.5%), Vβ3 - in 7 (53.8%) pts among Th, and Vβ3 - in 3 (23.1%) and Vβ9 - in 4 (30.8%) out of 13 pts among TCL. In OR group Vβ2 expansion was in 4 (40%) and Vβ3 - in 5 (50%) pts among Th; Vβ3 in 6 (60%) and Vβ9 in 6 (60%) out of 10 pts among TCL. In NR group the most frequent was Vβ13.1 clone in TCL - in 3 (42.9%) out of 7 pts. In NR group in overall clonal expansion was less frequent than in ND and OR groups. We also analyzed the previously mentioned subpopulations of T-cells in patients with AA in three groups (ND, OR, NR) compared to healthy donors (table 1). We obtained significant differences in the count of naive Th and TCL cells, memory T-cells in all three groups of AA patients compared to donors: proportion of naive Th and TCL cells was significantly higher and proportion of memory Th cells was lower in the donor group than in AA pts. The percent of TCL effectors was higher in ND AA pts compare to donors. We also found that cell count of activated Th (CD4+CD25+) was higher in the group of refractory pts. In OR pts proportion of PD-1-positive Th was higher than in donors. In NR pts Th and TCL co-expressed with PD-L1 were lower compare to donors (table 1). Conclusions. In our study we found immunodominant clonotypes in different AA pts and depletion of the pool of naive T cells. Dynamic observation of changes in the most common clonotypes in AA pts during treatment will provide suitable therapy tactics (allogenic bone marrow transplantation or IST). Disclosures No relevant conflicts of interest to declare.

scholarly journals Pharmacokinetic and Pharmacodynamic Study of NKTR-255, a Polymer-Conjugated Human IL-15, in Cynomolgus Monkey

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2952-2952
Author(s):  
Takahiro Miyazaki ◽  
Peiwen Kuo ◽  
Mekhala Maiti ◽  
Palakshi Obalapur ◽  
Murali Addepalli ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Nk Cells ◽  
Cd8 T Cells ◽  
Cynomolgus Monkey ◽  
Memory T Cells ◽  
Granzyme B ◽  
Employment Equity ◽  
Equity Ownership ◽  
Dose Dependent

Abstract Introduction IL-15 is a common gamma chain cytokine that activates and provides a survival benefit to T-cells and NK cells and has long been recognized as having potential as an immunotherapeutic agent for the treatment of cancer. Therapeutic use of native IL-15 has been challenging due to, for example, its unfavorable pharmacokinetic and safety properties. NKTR-255 is a polymer-conjugated human IL-15 that retains binding affinity to the alpha subunit of IL-15 receptor and exhibits reduced clearance to thereby provide a sustained pharmacodynamics response. Here we investigate the biological effects of NKTR-255 in naïve cynomolgus monkey. Methods In vitro monkey whole blood was treated with NKTR255 and the percentage of pSTAT5 positive populations in each NK, CD4 T and CD8 T cells was determined by flow cytometry. In an PK/PD study, monkeys received single IV doses of 0.001, 0.003, 0.01, 0.03, or 0.1 mg/kg NKTR-255. Blood samples were collected to determine the plasma concentrations of NKTR-255 and to assess the effects of NKTR-255 on NK and CD8 T cells at multiple time points; flow cytometry was used to measure STAT5 phosphorylation, Ki-67 expression and frequency of cell populations. Granzyme B expression was assessed in NK and CD8 T cells by flow cytometry. Results NKTR-255 induced dose-dependent phosphorylation of STAT5 in monkey whole blood (EC50 values NK cells: 6.9 ng/ml, CD8 T cells: 39 ng/ml, CD4 T cells: 53 ng/ml). The half-life and clearance of NKTR-255 were 26x longer and 38x lower, respectively, than IL-15. NKTR-255 engaged the IL-15 signaling pathway, in vivo, demonstrating both robust and sustained STAT5 phosphorylation in lymphocytes. NKTR-255 drove the proliferation of total CD8 T cells and NK cells in a dose-dependent manner, with dramatic and durable increases observed in Ki67 positive population and absolute cell numbers (NK cells: 6.1 fold; CD8 T cells: 7.8 fold from baseline on day 5 at 0.1 mg/kg). These effects were strongly biased towards CD8 T cells and NK cells, with substantially less induction of CD4 T cells. The Ki67 response analyses of the T cell subpopulation revealed a higher response of memory populations than for naive T cells. Among memory T cells, effector memory T cells showed the highest response over stem cell memory T cells and central memory T cells. Finally, NKTR-255 also increased the expression of Granzyme B in both NK and CD8 T cells, concomitant with an enhancement in target cell lysis. Conclusions Nektar has generated a novel and potent molecule in NKTR-255 that not only preserves the relevant biology of IL-15, but additionally provides enhanced PK and PD properties relative to the native IL-15 cytokine. NKTR-255 is being developed as an immune-stimulatory agent to target NK and CD8 T cell biology for the treatment of cancer. Disclosures Miyazaki: Nektar Therapeutics: Employment, Equity Ownership. Kuo:Nektar Therapeutics: Employment, Equity Ownership. Maiti:Nektar Therapeutics: Employment, Equity Ownership. Obalapur:Nektar Therapeutics: Employment, Equity Ownership. Addepalli:Nektar Therapeutics: Employment, Equity Ownership. Rubas:Nektar Therapeutics: Employment, Equity Ownership. Sims:Nektar Therapeutics: Employment, Equity Ownership. Zhang:Nektar Therapeutics: Employment, Equity Ownership. Madakamutil:Nektar Therapeutics: Employment, Equity Ownership. Zalevsky:Nektar Therapeutics: Employment, Equity Ownership.

scholarly journals Dissociation of CD154 and Cytokine Expression Patterns in CD38+ CD4+ Memory T Cells in Chronic HIV-1 Infection

2010 ◽  
Vol 55 (4) ◽  
pp. 439-445 ◽  
Author(s):  
Enrique Espinosa ◽  
Christopher E Ormsby ◽  
Gustavo Reyes-Terán ◽  
Robert Asaad ◽  
Scott F Sieg ◽  
...  
Keyword(s):  
T Cells ◽  
Memory T Cells ◽  
Expression Patterns ◽  
Cytokine Expression ◽  
Hiv 1

scholarly journals OP0042 BLOCKING OF CD103+ TISSUE RESIDENT MEMORY T CELLS (TRM) AS A THERAPEUTIC STRATEGY IN SJOGREN’S SYNDROME

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 22.3-22
Author(s):  
D. Mauro ◽  
X. Lin ◽  
G. Guggino ◽  
D. Chong ◽  
S. Raimondo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Salivary Glands ◽  
Memory T Cells ◽  
Granzyme B ◽  
Down Regulation ◽  
Therapeutic Targeting ◽  
Post Immunization ◽  
Resident Memory T Cells ◽  
The Impact

Background:Tissue-resident memory T cells (TRM), are a recently identified T cells population featuring tissue localization and expression of markers of tissue homing, CD69 and CD103. Recently, the expansion of CD8+ TRMs and their involvement in the sialadenitis was described in a murine model of SS. However, CD4+ and CD8+ TRM’s functional relevance in pSS is still not fully understood, and the TRM therapeutic targeting unexplored.Objectives:The study aimed to address the role of CD4+ and CD8+ TRMs in the pathogenesis of pSS and to explore the therapeutic targeting of the tissue residency marker of TRM CD103.Methods:An animal model of experimental (ESS) obtained by immunization of female C57BL/6 mice (n=10) with salivary glands (SG) protein extract and Freund’s complete adjuvant used to investigate the dynamic of infiltration of SG by CD4+ and CD8+ TRMs, their frequency, and the impact of CD103 blockade. For the therapeutic intervention, at 10-weeks post-immunization, the salivary gland was cannulated via Wharton’s duct, and an anti-CD103 neutralizing antibody or vehicle-injected. The mice’s saliva flow rate was assessed, and SGs were analyzed by Flow-cytometry and immunohistochemistry (IHC).The frequency and localization of TRMs was analyzed in minor SG of sicca syndrome (nSS) and pSS patients (n=39) by flow cytometry and IHC. The expression of genes involved in the tissue retention of TRMs was assessed in SG by RT-PCR.Results:Upon the ESS progression, a significant progressive increase in CD45+CD103+ cells frequency was observed from 5wk to 20wk post-immunization (p<0.001), where the CD8+ were the most abundant, followed by CD4+. Consistently, CD103+CD8+ T cells were detected within the lymphocytic infiltration of SG from ESS mice. Sorted purified SG CD10+CD3+CD8+ T cells showed higher Granzyme B, TNF-alpha expression compared to CD103-CD3+CD8+ at both mRNA and protein levels. Notably, ESS mice treated with anti-CD103 showed improvement in salivary function (p<0.05) and reduced lymphocytic infiltrations measured as focus score (FS) (p<0.01) and area-fraction (p<0.01). Consistently, anti-CD103 treatment consistently reduced CD103+ cells and IFN-gamma+, Granzyme B+, and TNFa+ CD8+ cells. We next performed phenotypic analysis of CD45+CD103+ immune cells in the SG of pSS patients observing an increase in both with CD8+CD103+CD69+ and CD4+CD103+CD69+ (p<0.05). Finally, IHC showed that the expansion of TRMs in pSS salivary glands was accompanied by a down-regulation of E-cadherin glandular expression and their migration outside the epithelium in the context of inflammatory infiltrates. SG of patients with pSS showed a significant up-regulation of BLIMP1, KFL-2, and S1PR1 and down-regulation of ITGB2. CXCL9 and CXCL10, and IL-15 involved in the tissue recruitment and long-term survival of TRMs were significantly modulated in pSS salivary glands.Conclusion:TRM are expanded and activated in the SG of pSS and ESS, participating in the organization of tissue inflammation. Although the mechanisms behind this expansion are still not fully understood, CD103 could be a valuable novel therapeutic target to prevent lymphocytic infiltrations and glandular destruction in Sjogren syndrome.Disclosure of Interests:None declared

scholarly journals lncRNA028466 regulates Th1/Th2 cytokine expression and associates with Echinococcus granulosus antigen P29 immunity

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Chan Wang ◽  
Song-Hao Yang ◽  
Nan Niu ◽  
Jia Tao ◽  
Xian-Cai Du ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
T Cells ◽  
Echinococcus Granulosus ◽  
Noncoding Rna ◽  
Cytokine Expression ◽  
Qrt Pcr ◽  
Th2 Cytokine ◽  
Ifn Γ ◽  
Th1 And Th2

Abstract Background Cystic echinococcosis (CE) is a parasitic disease that is caused by Echinococcus granulosus (Eg). The recombinant Echinococcus granulosus antigen P29 (rEg.P29) was shown to confer effective immunity to sheep and mice during E. granulosus secondary infection in our previous study. In this study, we sought to investigate the ability of long noncoding RNA 028466 (lncRNA028466) as a regulator for the protective immunity mediated by rEg.P29 vaccination and to study the effects of lncRNA028466 on CD4+T cell differentiation in mice spleen. Methods Female BALB/c mice were divided into two groups and were vaccinated subcutaneously with rEg.P29 antigen and PBS as a control (12 mice each group). Following prime-boost vaccination, CD4+T, CD8+T, and B cells from the spleen were isolated by flow cytometry. Quantitative real-time PCR (qRT-PCR) was performed to measure the expression of lncRNA028466 in these three kinds of cells. Then, lncRNA028466 was overexpressed and knocked down in naive CD4+T cells, and Th1 and Th2 cytokine expression was detected. qRT-PCR, western blot, and ELISA were performed to evaluate the production of IFN-γ, IL-2, IL-4, and IL-10, and flow cytometry was performed to detect the differentiation of Th1 and Th2 subgroups. Results lncRNA028466 was significantly decreased after the second week of immunization with rEg.P29 antigen. The proportion of CD4+ T cells was increased after rEg.P29 immunization. Overexpression of lncRNA028466 facilitated the production of IL-4, IL-10 and suppressed the production of IFN-γ, IL-2. Furthermore, after transfection with siRNA028466, IL-2 production was facilitated and IL-10 production was suppressed in naive CD4+ T cells. Conclusions Immunization with rEg.P29 downregulated the expression of lncRNA028466, which was related to a higher Th1 immune response and a lower Th2 immune response. Our results suggest that lncRNA028466 may be involved in rEg.P29-mediated immune response by regulating cytokine expression of Th1 and Th2.

838 Phenotypic and functional signatures of peripheral and tumor-resident γδ T cells are informative for outcome of checkpoint blockade in melanoma

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A890-A890
Author(s):  
Graham Pawelec ◽  
Kilian Wistuba-Hamprecht ◽  
Kilian Wistuba-Hamprecht
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Replicative Senescence ◽  
Γδ T Cells ◽  
Cytokine Expression ◽  
Checkpoint Blockade ◽  
Phenotypic Characterization ◽  
List Type ◽  
Gammadelta T Cells

BackgroundImmune checkpoint blockade (ICB) set a milestone in cancer immunotherapy, but still only a fraction of patients responds. Thus, there is an urgent need for biomarkers predicting outcome, and also for understanding the responsible mechanisms. γδ T cells constitute a numerically minor subset of 1-10% of the peripheral T cell compartment in healthy people and have a major role in defense against multiple microbial and non-microbial challenges. Unlike the majority of T cells, γδ T cells bind their ligands in an MHC-independent manner. We previously studied γδ T cells, that also express checkpoint molecules, in patients in the pre-checkpoint blockade era and thereafter, and identified correlations between subset frequencies of these unconventional T cells and patients‘ overall survival (OS). Here, we present a detailed phenotyping and functional investigation of tumor-resident as well as peripheral γδ T cells.MethodsPhenotyping was performed in stage IV melanoma patients before and under PD-1+/-CTLA-4 blockade using as basis our published OMIP-20 protocol.1 Cytokine expression patterns and proliferative capacities were determined as described according to our established protocols.2 Primary flow cytometry data analysis was performed using FlowJo (BD) and correlations with clinical meta data were determined using Prism (GraphPad) and SPSS (IBM).ResultsWe found previously that low frequencies of peripheral Vδ1 γδ T cells were associated with prolonged OS. Here, we investigated functional aspects and abundance of γδ T cells within the tumor as well as in the blood. The peripheral Vδ1 but not the Vδ2 differentiation signature revealed significantly lower proportions of naive and effector cells as well as an accumulation of late differentiated cells in patients with high Vδ1 frequencies. The cytokine expression pattern (IFNγ, TNF and IL-17) and the degranulation marker CD107a were different in patients with high versus low peripheral Vδ1 frequencies. The proliferative capabilities of Vδ1 cells in melanoma were limited in comparison to healthy subjects. Both Vδ1 and Vδ2 cells were found in tumor tissues, and these analyses are ongoing, including analyses of replicative senescence through CD57 expression.ConclusionsOur data provide novel insights into the role of γδ T cells in cancer rejection. The previously found negative correlation of Vδ1 T cells with OS is likely due to an accumulation of mal-functioning, probably exhausted Vδ1 T cells in patients with poor outcome of ICB. Thus, we suggest that Vδ1 T cells are promising candidates for future exploitation in novel ICB-approaches.Ethics ApprovalThis study was approved by K. Wistuba-Hamprecht´s Ethics Committee (approval nos. 490/2014BO1 and 792/2016BO2).ReferencesWistuba-Hamprecht K, Pawelec G, Derhovanessian E. OMIP-020: Phenotypic characterization of human gammadelta T-cells by multicolor flow cytometry. Cytometry A 2014;85:522–524. doi:10.1002/cyto.a.22470Beucke N, et al. Pitfalls in the characterization of circulating and tissue-resident human gammadelta T cells. J Leukoc Biol 2020. doi:10.1002/JLB.5MA1219-296R

PS02.124: NOVEL APPROACHES AND CONSIDERATIONS FOR IMMUNOTHERAPY IN OBESITY-ASSOCIATED OESOPHAGEAL ADENOCARCINOMA

2018 ◽  
Vol 31 (Supplement_1) ◽  
pp. 156-156
Author(s):  
Melissa Conroy ◽  
Karen Galvin ◽  
Margaret Dunne ◽  
John Reynolds ◽  
Joanne Lysaght
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Chemokine Receptor ◽  
Cytokine Expression ◽  
Receptor Expression ◽  
Oesophageal Adenocarcinoma ◽  
Cell Trafficking ◽  
Tumour Immunity ◽  
T Cell Trafficking

Abstract Background Immunotherapies are transforming cancer treatment for inoperable and advanced disease. However, the incidence of obesity-associated cancers, including oesophageal adenocarcinoma (OAC) continue to increase. Treatment with immune-based therapies present a unique challenge for immunologists, as they need to enhance anti-tumour immunity without exacerbating pre-existing carcinogenic inflammation. Therefore, we examined the effect of PD-1 blockade in omentum and liver of OAC patients, sites of activated inflammatory T cells, which play a key role in obesity-associated inflammation. In addition, we examined novel ways to reduce the infiltration of inflammatory T cells to these sites in a bid to reduce adipose tissue inflammation. Methods Blood, omentum and liver samples were obtained from consenting OAC patients and treated with anti-PD-1 antibody. CD4+ and CD8+ T cell activation, cytokine expression and cytotoxic potential were assessed by flow cytometry. To identify potential chemokine pathways to alter T cell trafficking to the omentum and liver, chemokine receptor expression was examined, along with levels of secreted chemokines using flow cytometry and ELISAs. Pre-treatment of T cells with chemokine receptor antagonists was performed prior to chemotaxis assays using an in vitro transwell system. Results In addition to OAC tumour, omentum and liver were found to be enriched with substantial populations of PD-1 expressing T cells. Treatment of omental and hepatic T cells with anti-PD-1 did not enhance inflammatory cytokine expression or proliferation, but transiently increased CD107a expression by CD8+ T cells. MIP-1α, MIP-1β and IP-10 mediate T cell trafficking to the omentum and liver in OAC. OAC-derived T cells preferentially migrate to the adipose and liver tissue conditioned media of obese OAC patients and this can be significantly reduced using a MIP-1α receptor antagonist. Conclusion This study provides evidence that anti-PD-1 treatment is unlikely to exacerbate pre-existing T cell-mediated inflammation outside the tumour in obesity-associated cancers. Furthermore, novel CCR1 antagonists may be used to attenuate pathological inflammation in obesity. This dual targeting approach aims to enhance anti-tumour immunity through the use of immune checkpoint inhibitors, and has identified a novel immunotherapeutic approach to reduce obesity-associated inflammation by targeting key chemokine pathways. In vivo testing is required to determine the efficacy of this approach. Disclosure All authors have declared no conflicts of interest.

Abstract MP49: Single Cell Multiplex Immunophenotyping Using Mass Cytometry And CITE-Seq Reveals Decreases In Circulating PD-1 + CD8 + Memory T Cells With Features Of Exhaustion In Human Hypertension

Hypertension ◽  
2021 ◽  
Vol 78 (Suppl_1) ◽  
Author(s):  
Matthew R Alexander ◽  
Charles D Smart ◽  
Bethany L Dale ◽  
Fernando Elijovich ◽  
Cara Wogsland ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Single Cell ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
Memory T Cells ◽  
Gene Set Enrichment Analysis ◽  
Mass Cytometry ◽  
Human Hypertension ◽  
Adaptive Immune Cells

Emerging evidence from animal models has demonstrated the importance of multiple innate and adaptive immune cells in hypertension. We hypothesized that the abundance and phenotype of circulating immune cell subsets are altered in human hypertension. To test this, we performed high dimensional single cell profiling of human peripheral blood mononuclear cells using mass cytometry. Unsupervised computational analysis revealed a 40% decrease in CD8 + memory T cells in hypertensive individuals. Using Phenograph to identify subsets of these cells revealed a selective 60% decrease in PD-1 + CD8 + memory T cells in hypertension. This observation was confirmed in a validation cohort using flow cytometry in which PD-1 + CD8 + memory T cells were significantly decreased 44% in hypertensive compared to control individuals. To determine the phenotype of these PD-1 + CD8 + memory T cells, we performed Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq) on four control and four hypertensive individuals. Using antibodies to identify PD-1 + and PD-1 - CD8 + memory T cells, gene set enrichment analysis of the coordinate single cell transcriptomic data revealed that PD-1 + cells exhibit over-representation of features of both immunologically active effector T cells and hypofunctional exhausted T cells. Thus, clustering analysis of PD-1 + CD8 + memory T cells was performed which demonstrated 4 distinct subclusters. One of these subclusters was decreased in hypertension and exhibited selective expression of multiple inhibitory receptors characteristic of exhausted T cells. At the protein level, this subcluster was marked by expression of the inhibitory receptor LAG3 and low levels of CD57. Combining these markers to identify PD-1 + LAG3 + CD57 - CD8 + memory T cells permitted identification of exhausted cells which demonstrated a significant 35% decrease in hypertensive compared to control individuals using flow cytometry. Taken together, these results demonstrate novel and reproducible decreases in circulating PD-1 + CD8 + memory T cells with features of exhaustion in human hypertension. These findings provide new insights into the pathogenesis of human hypertension including loss and/or re-invigoration of exhausted T cells.

scholarly journals RESEARCH OF RESTORATION PROCESSES AFTER INDUCTION OF LYMPHOPENIA BY CYCLOPHOSPHANE IN AN EXPERIMENTAL MODEL ON MOUSE

2020 ◽  
Vol 2 (29(56)) ◽  
pp. 5-9
Author(s):  
E.K Grinko ◽  
S.N. Marzanova ◽  
A.D. Doneckova
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Experimental Model ◽  
Memory T Cells ◽  
Cell Recovery ◽  
Central Memory T Cells ◽  
Delayed Recovery ◽  
Spleen Lymphocytes

Objective: to evaluate the dynamics of T-cell recovery after exposure to cyclophosphamide (CP). CP was injected at a dose of 125 mg/kg, followed by killing of mice on days 0, 10, 20, 30, 60 and determination of the subpopulation composition by flow cytometry. It was shown that thymocytes are more vulnerable to the action of the CP than spleen lymphocytes. Cell restoration in the thymus begins earlier with delayed recovery of spleen lymphocytes. Due to CP, there is an increase in the conversion of the naive T-cells into central memory T-cells with a relative accumulation of the latter.

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