Regulation of non-responsiveness and death in cytotoxic T cells by the agonistic potency of MHC : Peptide ligands

CD8+ T lymphocytes are important for immunological control of infections and tumors. The key interaction required to initiate the process of T cell activation is the engagement of the T cell receptor (TCR) with a major histocompatibility complex (MHC) class I/peptide complex on an antigen presenting cell (APC). Depending on the affinity of the interaction between the TCR and MHC class I molecule, different arrays of signaling pathways can be activated in the T cell. Molecular alterations in the peptide bound to the MHC class I can lead to a lower affinity of the MHCTCRinteraction resulting in incomplete or qualitatively different T-cell responses. Altered peptide ligands (APLs) exhibiting such activity are referred to as partial agonists and often occur naturally through genetic instability, which affects T cell epitopes derived from rapidly mutating viruses or tumor-associated cellular antigens. We studied the molecular basis of partial agonism using MHC class I/peptide tetramer complexes. By using tetramers assembled with a fully agonist peptide or its synthetic variant we could study the relationship between tetramer staining, cytokine production and different pathways of activation induced cell death (AICD). We found that positive tetramer staining correlated with at least two different activation programs in CD8+ T lymphocytes: full scale activation associated with Fas-dependent AICD and an incomplete activation followed by Fasindependent AICD. Further, we used raft-disrupting agents to assess the role of lipid rafts in determining the agonistic potency of different peptide ligands. We showed that overall binding of specific tetramers to CTLs was reduced upon raft disruption, although the half-life of tetramer:TCR complexes formed under these conditions was not affected. These findings suggest that different TCR complexes on the surface of CTLs may have different requirements for cholesterol and CTLs may be heterogeneous in their raft structure. In addition we analyzed programs of negative regulation of CD8+ T lymphocytes and the capacity of APLs to activate and modulate such programs. Upon specific triggering CD8+ T-cells become refractory to a secondary stimulus; a condition referred to Activation induced Non-Responsiveness (AINR). We have shown that TCRtriggering results in a novel degradation pathway of Lck, a kinase which plays a critical role in the initiation of T cell activation. Down-regulation of Lck through degradation correlated with AINR in CTLs. By blocking Lck degradation we could prevent the development of AINR. We further investigated how activation of CTLs with APLs affected Lck expression. The capacity of different peptide variants to induce Lck degradation correlated with their agonistic potency. Inefficient recognition of APLs by specific T lymphocytes is believed to contribute to the failure of the immune system to control certain tumor types and progressive viral diseases. To better understand the regulation of APL, activity by immunologic help, we analyzed the capacity of exogenous IL-2 and IL-15 to influence different aspects of activation triggered in CTLs by either fully or partially agonistic peptide ligands. We showed that signals induced by the lymphokines synergize with weak TCR signaling induced by partially agonistic APL, converting many of these peptides from inhibitory to stimulatory ligands. We also demonstrated that IL-2 and IL-15 suppress induction of a death receptor-independent apoptotic program triggered by partially agonistic APL. In conclusion, we have analyzed the molecular basis of partial agonism in CTL recognition of peptide epitopes and characterized molecular changes associated with death and AINR in specific CTLs. We have shown that structural changes in the sequence of CTL peptide epitopes may decrease the affinity of MHC/TCR interactions and generate APLs, which not only trigger incomplete activation programs but also induce and modulate negative regulation programs in CTLs. This APL induced signaling of suppressive nature appears to be more prominent in the absence of immunological help, suggesting that under conditions of immune deregulation APLs may actively suppress CTL responses against infectious agents or tumors

Methylseleninic Acid Sensitizes Ovarian Cancer Cells to T-Cell Mediated Killing by Decreasing PDL1 and VEGF Levels

Redox active selenium (Se) compounds at sub toxic doses act as pro-oxidants with cytotoxic effects on tumor cells and are promising future chemotherapeutic agents. However, little is known about how Se compounds affect immune cells in the tumor microenvironment. We demonstrate that the inorganic Se compound selenite and the organic methylseleninic acid (MSA) do not, despite their pro-oxidant function, influence the viability of immune cells, at doses that gives cytotoxic effects in ovarian cancer cell lines. Treatment of the ovarian cancer cell line A2780 with selenite and MSA increases NK cell mediated lysis, and enhances the cytolytic activity of T cells. Increased T cell function was observed after incubation of T cells in preconditioned media from tumor cells treated with MSA, an effect that was coupled to decreased levels of PDL1, HIF-1α, and VEGF. In conclusion, redox active selenium compounds do not kill or inactivate immune cells at doses required for anti-cancer treatment, and we demonstrate that MSA enhances T cell-mediated tumor cell killing via PDL1 and VEGF inhibition

Individualization of Hematopoietic Stem Cell Transplantation Using Alpha/Beta T-Cell Depletion

Allogeneic hematopoietic stem cell transplantation (HSCT) is associated with several potentially lethal complications. Higher levels of CD3+ T-cells in the graft have been associated with increased risk of graft-versus-host disease (GVHD), but also beneficial graft-versus-leukemia effect and reduced infections. To tackle post-transplant complications, donor lymphocyte infusions have been used but with an increased risk of GVHD. To reduce this risk, we performed depletion of αβ T-cells and treated 12 patients post-HSCT suffering from infections and/or poor immune reconstitution. The αβ T-cell depleted cell products were characterized by flow cytometry. The median log depletion of αβ T-cells was −4.3 and the median yield of γδ T-cells was 73.5%. The median CD34+ cell dose was 4.4 × 106/kg. All 12 patients were alive 3 months after infusion and after 1 year, two patients had died. No infusion-related side effects were reported and no severe acute GVHD (grade III-IV) developed in any patient post-infusion. Overall, 3 months after infusion 11 out of 12 patients had increased levels of platelets and/or granulocytes. In conclusion, we describe the use of αβ T-cell depleted products as stem cell boosters with encouraging results

Combining Flow and Mass Cytometry in the Search for Biomarkers in Chronic Graft-versus-Host Disease

Chronic graft-versus-host disease (cGVHD) is a debilitating complication arising in around half of all patients treated with an allogeneic hematopoietic stem cell transplantation. Even though treatment of severe cGVHD has improved during recent years, it remains one of the main causes of morbidity and mortality in affected patients. Biomarkers in blood that could aid in the diagnosis and classification of cGVHD severity are needed for the development of novel treatment strategies that can alleviate symptoms and reduce the need for painful and sometimes complicated tissue biopsies. Methods that comprehensively profile complex biological systems such as the immune system can reveal unanticipated markers when used with the appropriate methods of data analysis. Here, we used mass cytometry, flow cytometry, enzyme-linked immunosorbent assay, and multiplex assays to systematically profile immune cell populations in 68 patients with varying grades of cGVHD. We identified multiple subpopulations across T, B, and NK-cell lineages that distinguished patients with cGVHD from those without cGVHD and which were associated in varying ways with severity of cGVHD. Specifically, initial flow cytometry demonstrated that patients with more severe cGVHD had lower mucosal-associated T cell frequencies, with a concomitant higher level of CD38 expression on T cells. Mass cytometry could identify unique subpopulations specific for cGVHD severity albeit with some seemingly conflicting results. For instance, patients with severe cGVHD had an increased frequency of activated B cells compared to patients with moderate cGVHD while activated B cells were found at a reduced frequency in patients with mild cGVHD compared to patients without cGVHD. Moreover, results indicate it may be possible to validate mass cytometry results with clinically viable, smaller flow cytometry panels. Finally, no differences in levels of blood soluble markers could be identified, with the exception for the semi-soluble combined marker B-cell activating factor/B cell ratio, which was increased in patients with mild cGVHD compared to patients without cGVHD. These findings suggest that interdependencies between such perturbed subpopulations of cells play a role in cGVHD pathogenesis and can serve as future diagnostic and therapeutic targets

State-society relations in contemporary Russia: New forms of political and social contention

Much existing analysis of Russian state–society relations focuses on public, active forms of contention such as the “opposition” and protest movements. There is need for a more holistic perspective which adds study of a range of overt, “co-opted”, and hidden forms of interaction to this focus on public contention. A theoretical and empirical basis for understanding state–society relations in today's Russia involves broadening the concept of “contentious politics” to include models of “consentful” as well as “dissentful” contention. A diffused model of contentious politics can situate claim-making along the axes of consentful and dissentful motivations, and compliant and contentious behaviours

Associations between depressive symptoms and disease progression in older patients with chronic kidney disease: results of the EQUAL study

Background Depressive symptoms are associated with adverse clinical outcomes in patients with end-stage kidney disease; however, few small studies have examined this association in patients with earlier phases of chronic kidney disease (CKD). We studied associations between baseline depressive symptoms and clinical outcomes in older patients with advanced CKD and examined whether these associations differed depending on sex. Methods CKD patients (>= 65 years; estimated glomerular filtration rate <= 20 mL/min/1.73 m(2)) were included from a European multicentre prospective cohort between 2012 and 2019. Depressive symptoms were measured by the five-item Mental Health Inventory (cut-off <= 70; 0-100 scale). Cox proportional hazard analysis was used to study associations between depressive symptoms and time to dialysis initiation, all-cause mortality and these outcomes combined. A joint model was used to study the association between depressive symptoms and kidney function over time. Analyses were adjusted for potential baseline confounders. Results Overall kidney function decline in 1326 patients was -0.12 mL/min/1.73 m(2)/month. A total of 515 patients showed depressive symptoms. No significant association was found between depressive symptoms and kidney function over time (P = 0.08). Unlike women, men with depressive symptoms had an increased mortality rate compared with those without symptoms [adjusted hazard ratio 1.41 (95% confidence interval 1.03-1.93)]. Depressive symptoms were not significantly associated with a higher hazard of dialysis initiation, or with the combined outcome (i.e. dialysis initiation and all-cause mortality). Conclusions There was no significant association between depressive symptoms at baseline and decline in kidney function over time in older patients with advanced CKD. Depressive symptoms at baseline were associated with a higher mortality rate in men

Lymphocytes in Placental Tissues: Immune Regulation and Translational Possibilities for Immunotherapy

Immune modulation at the fetomaternal interface is crucial to ensure that the fetal allograft is not rejected. In the present review, the focus is to describe basic functions of lymphocyte populations and how they may contribute to fetomaternal immune regulation, as well as determining what proportions and effector functions of these cells are reported to be present in placental tissues in humans. Also explored is the possibility that unique cell populations at the fetomaternal interface may be targets for adoptive cell therapy. Increasing the understanding of immune modulation during pregnancy can give valuable insight into other established fields such as allogeneic hematopoietic stem cell transplantation and solid organ transplantation. In these settings, lymphocytes are key components that contribute to inflammation and rejection of either patient or donor tissues following transplantation. In contrast, an allogeneic fetus eludes rejection by the maternal immune system