Characterization of Circulating IL-10-Producing Cells in Septic Shock Patients: A Proof of Concept Study

Sepsis is a worldwide health priority characterized by the occurrence of severe immunosuppression associated with increased risk of death and secondary infections. Interleukin 10 (IL-10) is a potent immunosuppressive cytokine which plasma concentration is increased in septic patients in association with deleterious outcomes. Despite studies evaluating IL-10 production in specific subpopulations of purified cells, the concomitant description of IL-10 production in monocytes and lymphocytes in septic patients’ whole blood has never been performed. In this pilot study, we characterized IL-10 producing leukocytes in septic shock patients through whole blood intracellular staining by flow cytometry. Twelve adult septic shock patients and 9 healthy volunteers were included. Intracellular tumor necrosis factor-α (TNFα) and IL-10 productions after lipopolysaccharide stimulation by monocytes and IL-10 production after PMA/Ionomycine stimulation by lymphocytes were evaluated. Standard immunomonitoring (HLA-DR expression on monocytes, CD4+ T lymphocyte count) of patients was also performed. TNFα expression by stimulated monocytes was reduced in patients compared with controls while IL-10 production was increased. This was correlated with a reduced monocyte HLA-DR expression. B cells, CD4+, and CD4- T lymphocytes were the three circulating IL-10 producing lymphocyte subsets in both patients and controls. No difference in IL-10 production between patients and controls was observed for B and CD4- T cells. However, IL-10 production by CD4+ T lymphocytes significantly increased in patients in parallel with reduced CD4+ T cells number. Parameters reflecting altered monocyte (increased IL-10 production, decreased HLA-DR expression and decreased TNFα synthesis) and CD4+ T lymphocyte (increased IL-10 production, decreased circulating number) responses were correlated. Using a novel technique for intracellular cytokine measurement in whole blood, our results identify monocytes and CD4+ T cells as the main IL-10 producers in septic patients’ whole blood and illustrate the development of a global immunosuppressive profile in septic shock. Overall, these preliminary results add to our understanding of the global increase in IL-10 production induced by septic shock. Further research is mandatory to determine the pathophysiological mechanisms leading to such increased IL-10 production in monocytes and CD4+ T cells.

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Cytotoxic CD8+ T Cells Promote Granzyme B-Dependent Adverse Post-Ischemic Cardiac Remodeling

10.21203/rs.3.rs-51324/v1 ◽

2020 ◽

Author(s):

Icia Santos Zas ◽

Jeremie Lemarie ◽

Ivana Zlatanova ◽

Marine Cachanado ◽

Jean-Christophe Seghezzi ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Ventricular Remodeling ◽

Heart Function ◽

Ischemia Reperfusion ◽

Granzyme B ◽

Acute Ischemia ◽

Risk Of Death ◽

Increased Risk ◽

Acute Mi

Abstract Acute myocardial infarction (MI) is a common condition responsible for heart failure and sudden death. Here, we show that following acute MI in mice, CD8+ T lymphocytes are recruited and activated in the ischemic heart tissue, and release Granzyme B leading to cardiomyocyte apoptosis, adverse ventricular remodeling and deterioration of myocardial function. Depletion of CD8+ T lymphocytes decreases apoptosis within the ischemic myocardium, hampers inflammatory response, limits myocardial injury and improves heart function. These effects are recapitulated in mice with Granzyme B-deficient CD8+ T cells. The protective effect of CD8 depletion on heart function was confirmed by using a model of ischemia/reperfusion in pigs. Finally, we reveal that elevated circulating levels of Granzyme B in patients with acute MI predict increased risk of death at 1-year follow-up. Our work unravels an unsuspected deleterious role of CD8+ T lymphocytes following acute ischemia, and identifies novel therapeutic strategy targeting pathogenic CD8+ T lymphocytes in the setting of acute MI.

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Cytotoxic CD8+ T cells promote granzyme B-dependent adverse post-ischemic cardiac remodeling

Nature Communications ◽

10.1038/s41467-021-21737-9 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Icia Santos-Zas ◽

Jeremie Lemarié ◽

Ivana Zlatanova ◽

Marine Cachanado ◽

Jean-Christophe Seghezzi ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

T Cells ◽

T Lymphocytes ◽

Heart Function ◽

Ischemia Reperfusion ◽

Granzyme B ◽

Acute Ischemia ◽

Risk Of Death ◽

Increased Risk

AbstractAcute myocardial infarction is a common condition responsible for heart failure and sudden death. Here, we show that following acute myocardial infarction in mice, CD8+ T lymphocytes are recruited and activated in the ischemic heart tissue and release Granzyme B, leading to cardiomyocyte apoptosis, adverse ventricular remodeling and deterioration of myocardial function. Depletion of CD8+ T lymphocytes decreases apoptosis within the ischemic myocardium, hampers inflammatory response, limits myocardial injury and improves heart function. These effects are recapitulated in mice with Granzyme B-deficient CD8+ T cells. The protective effect of CD8 depletion on heart function is confirmed by using a model of ischemia/reperfusion in pigs. Finally, we reveal that elevated circulating levels of GRANZYME B in patients with acute myocardial infarction predict increased risk of death at 1-year follow-up. Our work unravels a deleterious role of CD8+ T lymphocytes following acute ischemia, and suggests potential therapeutic strategies targeting pathogenic CD8+ T lymphocytes in the setting of acute myocardial infarction.

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Flow Cytometric Analysis of T Cell Functions in CML Patients Under Imatinib Therapy

Blood ◽

10.1182/blood.v112.11.2633.2633 ◽

2008 ◽

Vol 112 (11) ◽

pp. 2633-2633

Author(s):

Deniz Goren Sahin ◽

Klara Dalva ◽

Sema Meric ◽

Gunhan Gurman ◽

Muhit Ozcan ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocytes ◽

T Lymphocyte ◽

Cd4 Cells ◽

Cell Functions ◽

Control Subjects ◽

Group 4 ◽

Hla Dr ◽

Healthy Control

Abstract Imatinib is currently the most chosen agent in the treatment of CML patients. There are published articles reporting that imatinib has suppressive effects on T lymphocytes. However there is little information avaliable about the effects of imatinib on the immune regulation after allogeneic stem cell transplantation and effect of graft versus leukemia. In light of these observations, in our study, primary objective was in vivo analysis of T lymphocyte functions by flow cytometry and the secondary objective was to evaluate the possible functional changes that might occur under imatinib therapy in CML patients. A total of 29 patients and 9 healthy control subjects were enrolled in this cross sectional, clinical-laboratory study. CML patients were divided into three groups as newly diagnosed patients having no treatment (group 1), patients receiving imatinib for 1 year (group 2) and patients receiving imatinib more than 1 year (group 3), respectively. Healthy control subjects were regarded as group 4. To evaluate T lymphocyte functions, cells were induced by phorbol myristate acetate (PMA) and ionomisin then CD4+ T-cells were selected and IL-4 and IFN-γ expression on these cells; how much percentage of CD3+ T cells were activated (CD3+CD69+); CD8+ T lymphocytes and the ratio and grade of expression of HLA-ABC and HLA-DR on those cells were evaluated, respectively. In our study, there was no significant difference in terms of mean number of CD4+ cells between the groups (p=0.125). However, there was a tendency toward higher CD4+ cells in group 4. Cytokine expression analyses (IL-4 and IFNgamma) were found not to be statistically significant between the groups. (p values 0.112 and 0.165 respectively). It was striking that group 4 has lower IL-4 and IFNgamma expression values but just failed to reach a statistical significant level. On the other hand, mean number of CD4+ cells, which did not express IL-4 and IFNgamma, were statistically higher in group 4 when compared to other groups. There were no significant differences in terms of CD3+ cells and CD69+ expression, which was an early activation marker of T cells. However, it was found differences in % activation values (p=0.002) and % activation of subjects in group 4 was found to be decreased when compared to that of other groups (Figure 1). CD8+ cell ratio was found to be statistically lower in all CML patient groups when compared to that of healthy control subjects (p=0.001). The expression of HLA-ABC and HLA-DR on CD8+ cells was similar between the groups. We could not show any inhibitory effect of imatinib on T cell functions that could support clinical observations. It will be a research area how this interaction will be in new and more potent 2nd and 3rd generation tyrosine kinase inhibitors. Figure 1. Graph showing distribution of mean percent activity between the groups. Note that group 4 shows significant lower values of mean percent activity. Figure 1. Graph showing distribution of mean percent activity between the groups. Note that group 4 shows significant lower values of mean percent activity.

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Proteome Analysis of CD4+ T Cells Reveals Differentially Expressed Proteins in Infertile Polycystic Ovary Syndrome Patients

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530320666201119152323 ◽

2020 ◽

Vol 20 ◽

Author(s):

Fatemeh Nasri ◽

Maryam Zare ◽

Mehrnoosh Doroudchi ◽

Behrouz Gharesi-Fard

Keyword(s):

Mass Spectrometry ◽

T Cells ◽

T Lymphocytes ◽

Gel Electrophoresis ◽

Cd4 T Cells ◽

Polycystic Ovary ◽

Proteome Analysis ◽

Two Dimensional ◽

Ovary Syndrome ◽

Two Dimensional Gel Electrophoresis

Background: Polycystic ovary syndrome (PCOS) is the most frequent endocrine disorder affecting 6–7% of premenopausal women. Recent studies revealed that the immune system especially CD4+ T helper cells are important in the context PCOS. Proteome analysis of CD4+ T lymphocytes can provide valuable information regarding the biology of these cells in the context of PCOS. Objective: To investigate immune dysregulation in CD4+ T lymphocytes at the protein level in the context of PCOS using two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: In the present study, we applied two-dimensional gel electrophoresis / mass spectrometry to identify proteins differentially expressed by peripheral blood CD4+ T cells in ten PCOS women compared with ten healthy women. Western blot technique was used to confirm the identified proteins. Results: Despite the overall proteome similarities, there were significant differences in the expression of seven spots between two groups (P <0.05). Three proteins, namely phosphatidylethanolamine-binding protein 1, proteasome activator complex subunit 1 and triosephosphate isomerase 1 were successfully identified by Mass technique and confirmed by western blot. All characterized proteins were over-expressed in CD4+ T cells from patients compared to CD4+ T cells from controls (P <0.05). In-silico analysis suggested that the over-expressed proteins interact with other proteins involved in cellular metabolism especially glycolysis and ferroptosis pathway. Conclusion: These findings suggest that metabolic adjustments in CD4+ T lymphocytes, which is in favor of increased glycolysis and Th2 differentiation are important in the context of PCOS.

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Development of Autologous, Oligoclonal, Poorly Functioning T Lymphocytes in a Patient With Autosomal Recessive Severe Combined Immunodeficiency Caused by Defects of the Jak3 Tyrosine Kinase

Blood ◽

10.1182/blood.v91.3.949 ◽

1998 ◽

Vol 91 (3) ◽

pp. 949-955 ◽

Cited By ~ 32

Author(s):

Duilio Brugnoni ◽

Luigi D. Notarangelo ◽

Alessandra Sottini ◽

Paolo Airò ◽

Marta Pennacchio ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Tyrosine Kinase ◽

Cd4 T Cells ◽

Severe Combined Immunodeficiency ◽

Combined Immunodeficiency ◽

T Helper ◽

T Helper 1 ◽

In Vitro Susceptibility ◽

And Function

Abstract Defects of the common gamma chain subunit of the cytokine receptors (γc) or of Jak3, a tyrosine kinase required for γc signal transduction, result in T−B+ severe combined immunodeficiency (SCID). However, atypical cases, characterized by progressive development of T lymphocytes, have been also reported. We describe a child with SCID caused by Jak3 gene defects, which strongly but not completely affect Jak3 protein expression and function, who developed a substantial number (>3,000/μL) of autologous CD3+CD4+ T cells. These cells showed a primed/activated phenotype (CD45R0+ Fas+HLA-DR+ CD62Llo), defective secretion of T-helper 1 and T-helper 2 cytokines, reduced proliferation to mitogens, and a high in vitro susceptibility to spontaneous (caused by downregulation of bcl-2 expression) as well as activation-induced cell death. A restricted T-cell receptor repertoire was observed, with oligoclonal expansion within each of the dominant segments. These features resemble those observed in γc-/y and in Jak3−/−mice, in which a population of activated, anergic T cells (predominantly CD4+) also develops with age. These results suggest that residual Jak3 expression and function or other Jak3-independent signals may also permit the generation of CD4+ T cells that undergo in vivo clonal expansion in humans; however, these mechanisms do not allow development of CD8+ T cells, nor do they fully restore the functional properties of CD4+ T lymphocytes.

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Macrophage-dependent Apoptosis of CD4+ T Lymphocytes from HIV-infected Individuals Is Mediated by FasL and Tumor Necrosis Factor

Journal of Experimental Medicine ◽

10.1084/jem.185.1.55 ◽

1997 ◽

Vol 185 (1) ◽

pp. 55-64 ◽

Cited By ~ 181

Author(s):

Andrew D. Badley ◽

David Dockrell ◽

Margaret Simpson ◽

Ron Schut ◽

David H. Lynch ◽

...

Keyword(s):

T Cells ◽

Tumor Necrosis Factor ◽

T Lymphocytes ◽

Cd4 T Cells ◽

Tumor Necrosis ◽

Human Macrophages ◽

Infected Cells ◽

Induced Apoptosis ◽

Antigen Presenting ◽

Necrosis Factor

Apoptosis of bystander uninfected CD4+ T lymphocytes by neighboring HIV-infected cells is observed in cell culture and in lymphoid tissue of HIV-infected individuals. This study addresses whether antigen-presenting cells such as human macrophages mediate apoptosis of CD4+ T cells from HIV-infected individuals. Uninfected human macrophages, and to a larger degree, HIV-infected macrophages mediate apoptosis of T cells from HIV-infected, but not from uninfected control individuals. This macrophage-dependent killing targets CD4+, but not CD8+ T lymphocytes from HIV-infected individuals, and direct contact between macrophages and lymphocytes is required. Additional analyses indicated that the apoptosis-inducing ligands, FasL and tumor necrosis factor (TNF), mediate this macrophage-induced apoptosis of CD4+ T cells. These results support a role for macrophage-associated FasL and TNF in the selective depletion of CD4+ T cells in HIV-infected individuals.

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B and T lymphocyte attenuator exhibits structural and expression polymorphisms and is highly induced in anergic CD4+ T cells

The Journal of Immunology ◽

10.4049/jimmunol.174.9.5884a ◽

2005 ◽

Vol 174 (9) ◽

pp. 5884a-5884 ◽

Cited By ~ 3

Author(s):

Michelle A. Hurchla ◽

John R. Sedy ◽

Maya Gavrielli ◽

Charles G. Drake ◽

Theresa L. Murphy ◽

...

Keyword(s):

T Cells ◽

T Lymphocyte ◽

Cd4 T Cells

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Interleukin-10 induces a long-term antigen-specific anergic state in human CD4+ T cells.

Journal of Experimental Medicine ◽

10.1084/jem.184.1.19 ◽

1996 ◽

Vol 184 (1) ◽

pp. 19-29 ◽

Cited By ~ 549

Author(s):

H Groux ◽

M Bigler ◽

J E de Vries ◽

M G Roncarolo

Keyword(s):

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Interleukin 10 ◽

Necrosis Factor Alpha ◽

T Cell Anergy ◽

Factor Alpha ◽

Cell Anergy ◽

Anergic T Cells ◽

Macrophage Colony

Human CD4+ T cells, activated by allogeneic monocytes in a primary mixed lymphocyte reaction in the presence of exogenous interleukin (IL) 10, specifically failed to proliferate after restimulation with the same alloantigens. A comparable state of T cell unresponsiveness could be induced by activation of CD4+ T cells by cross-linked anti-CD3 monoclonal antibodies (mAbs) in the presence of exogenous IL-10. The anergic T cells failed to produce IL-2, IL-5, IL-10, interferon gamma, tumor necrosis factor alpha, and granulocyte/macrophage colony-stimulating factor. The IL-10-induced anergic state was long-lasting. T cell anergy could not be reversed after restimulation of the cells with anti-CD3 and anti-CD28 mAbs, although CD3 and CD28 expression was normal. In addition, restimulation of anergized T cells with anti-CD3 mAbs induced normal Ca2+ fluxes and resulted in increased CD3, CD28, and class II major histocompatibility complex expression, indicating that calcineurin-mediated signaling occurs in these anergic cells. However, the expression of the IL-2 receptor alpha chain was not upregulated, which may account for the failure of exogenous IL-2 to reverse the anergic state. Interestingly, anergic T cells and their nonanergic counterparts showed comparable levels of proliferation and cytokine production after activation with phorbol myristate acetate and Ca2+ ionophore, indicating that a direct activation of a protein kinase C-dependent pathway can overcome the tolerizing effect of IL-10. Taken together, these data demonstrate that IL-10 induces T cell anergy and therefore may play an important role in the induction and maintenance of antigen-specific T cell tolerance.

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