European heart health survey 2019.

BACKGROUND: Rising life expectancy in the western population is increasing the prevalence of heart valve diseases (HVD). HYPOTHESIS: The level of awareness and initial screening for HVD should be sufficient. The potential impact of HVD on the daily activities of the elderly population in Europe might affect our society. METHODS AND RESULTS: A survey was conducted, including a total of 12 832 people aged ≥ 60 years in 11 European countries. Of all the people surveyed, 5.6% could correctly describe aortic valve stenosis. Most participants (75.0%) claimed they regularly do activities like sports or social activities, 29.2% provide care for a family member, friend or acquaintance. The majority (69.2%) would be prevented from doing these activities by symptoms such as chest pain, fatigue or shortness of breath. Having chest pain (76.5%) and shortness of breath (57.8%) were reasons for most people to arrange an appointment with their GP, whereas only 26.2% would visit a GP for fatigue. 67.6% of respondents claimed to be checked with a stethoscope by their GP occasionally, never, or only when they ask. The preferred treatment option for HVD is a keyhole procedure (45.8%), whereas open heart surgery would only be preferred by 7.0%. CONCLUSION: Knowledge about HVD is still low. Neither appointments with a GP driven by symptoms nor regular use of a stethoscope are a reliable guarantee for early diagnosis. With the over 60s in Europe playing an active role in social life, awareness campaigns and regular heart health checks may guarantee early diagnosis and treatment of HVD.

Dendritic cell type determines the mechanism of bystander suppression by adaptive T regulatory cells specific for the minor antigen HA-1.

One hallmark of acquired tolerance is bystander suppression, a process whereby Ag-specific (adaptive) T regulatory cells (TR) inhibit the T effector cell response both to specific Ag and to a colocalized third-party Ag. Using peripheral blood T cells from recipients of HLA-identical kidney transplants as responders in the trans vivo-delayed type hypersensitivity assay, we found that dendritic cells (DC), but not monocyte APCs, could mediate bystander suppression of EBV-specific recall response. When HA-1(H) peptide was added to mixtures of plasmacytoid DC (pDC) and T cells, bystander suppression of the response to a colocalized recall Ag occurred primarily via indolamine-2,3-dioxygenase (IDO) production. Similarly, addition of HA-1(H) peptide to cocultures of T cells and pDC, but not myeloid DC (mDC), induced IDO activity in vitro. When mDC presented HA-1(H) peptide to Ag-specific CD8+ TR, cytokine release (TGF-beta, IL-10, or both) was the primary mode of bystander suppression. Bystander suppression via mDC was reversed not only by Ab to TGF-beta and its receptor on T cells, but also by Ab to thrombospondin-1. EBV addition did not induce IDO or thrombospondin-1 in T-DC cocultures, suggesting that these DC products are not induced by T effector cells, but only by TR cells. These results shed light upon the mechanism of bystander suppression by donor Ag-specific TR in patients with organ transplant tolerance and underscores the distinct and critical roles of mDC and pDCs in this phenomenon.

Novel role for the N-terminus domain of the a2 isoform of vacuolar ATPase in interleukin-1beta production.

Interleukin-1beta (IL-1beta) is a mediator cytokine that is released by macrophages and epithelial cells in pregnancy and tumorigenesis before antigen recognition. a2V-ATPase is a protein expressed during pregnancy and tumorigenesis and has a novel role in immune regulation. It is expressed as a 70 kDa molecule in intracellular vesicles. Upon cell stimulation it migrates to the surface followed by the cleavage of a 20 kDa portion (a2 N-terminus domain, a2NTD). This study aimed to determine whether a2NTD could induce IL-1beta production in immune cells. Peripheral blood mononuclear cells (PMBC) were stimulated with a2NTD and analyzed for cytokine gene expression by gene arrays. Supernatants were analyzed for IL-1beta by enzyme-linked immunosorbent assay, and cells were analyzed for intracellular expression of IL-1alpha, IL-1beta, and TNF-alpha by flow cytometry. When PBMC were cultured with a2NTD, there was a 2.5-fold increase in IL1A and IL1B gene expression and no induction of TNF gene expression. There was a 72-fold increase in IL-1beta in supernatants of PBMC cultured with a2NTD. Finally, there was a 204-fold increase in intracellular expression of IL-1beta in monocytes incubated with a2NTD. These results indicate a regulatory role for a2NTD in IL-1 cytokine production and suggest a unique role for this molecule in inflammation.

In vitro parameters of donor-antigen-specific tolerance.

Donor-antigen-specific tolerance to the allograft is increasingly considered a reachable goal in the field of transplantation. As our understanding of the processes that govern donor-specific tolerance increases, so must our understanding of ways to detect this state of affairs. Unfortunately, this is not a straightforward procedure, as the mechanisms which govern tolerance are multiple and varied. Previously, the mixed lymphocyte reaction was used as standard to detect unresponsiveness. This approach is not valid for detecting tolerance because it only measures both direct pathway, naïve and memory responses, whereas the indirect pathway and 'pure' memory responses are more informative parameters for detecting tolerance. Techniques, such as the trans vivo delayed-type hypersensitivity assay, ELISPOT and antigen-specific HLA tetramer analysis address this problem, and the numbers of cell subsets, such as dendritic cells and NKT cells, can also aid us in detecting donor-antigen-specific tolerance.

Detection of microchimerism by minor histocompatibility antigen HA-1 allele-specific nested polymerase chain reaction.

Minor histocompatibility antigens (mHags) can induce T-cell reactivities with important consequences for the graft-versus-leukemia effect and the development of graft-versus-host disease in HLA-matched stem cell transplantation settings. Recently, mHag-specific T cells were also demonstrated in multiparous woman and in solid organ transplant recipients. Microchimeric cells have been detected in the latter settings. To study whether microchimerism is instrumental in the induction and/or maintenance of mHag T cells, we developed an HA-1 allele-specific nested polymerase chain reaction. To optimize and validate the reliability of this method at different levels of microchimerism, serial dilutions of HA-1(H) cells titrated into HA-1(R) cells were tested. We demonstrated that the HA-1(H) allele can be reliably and consistently detected at concentrations as low as 1:10(5) without losing specificity. The developed HA-1-specific nested polymerase chain reaction is an important tool that facilitates the detection of HA-1 microchimerism in various clinical specimens and that promotes investigation of the effects of microchimerism on induction of mHag-specific T cells in the various settings of immunization.

Regeneration and tolerance factor prevents bystander T-cell death associated with human immunodeficiency virus infection.

Human immunodeficiency virus (HIV) infection is characterized by a depletion of T cells. This depletion is caused both by the virus-induced death of infected T cells and by the death of uninfected cells (bystander depletion) by a mechanism which is largely uncharacterized. Regeneration and tolerance factor (RTF) is a subunit of the vacuolar ATPase and a protein that is involved with activation and apoptosis. Anti-RTF antibodies mediate apoptosis in T lymphocytes. When anti-RTF was added to lymphocytes from an HIV-positive individual, they underwent larger amounts of apoptosis than cells taken from healthy controls. When lymphocytes were examined by Western blotting, those from HIV-positive individuals exhibited increased levels of expression of the 50-kDa protein (P < 0.001). A 70-kDa protein was the predominant form of RTF in uninfected control lymphocytes, being expressed in 100% of individuals studied. The expression of the 50-kDa protein in HIV-positive individuals correlated with decreased absolute CD4 counts with a sensitivity of 92% and a positive predictive value of 86%. When uninfected lymphocytes were stimulated with anti-CD3 and anti-CD28, no RTF was detected during early stimulation but a 50-kDa protein was expressed during late stimulation. When the susceptibilities of the lymphocytes to anti-RTF-induced apoptosis were measured, they correlated with the size of the RTF protein expressed. The cells were not susceptible to apoptosis when the 70-kDa RTF was present but were susceptible when the 50-kDa RTF was present. We propose that the increase in the levels of the 50-kDa RTF on cells from HIV-positive individuals is important in preventing the cell from undergoing apoptosis.

Regeneration and tolerance factor modulates the effect of adenosine triphosphate-induced interleukin 1 beta secretion in human macrophages.

These studies characterize a molecule known as regeneration and tolerance factor (RTF), which controls inflammation by regulating interleukin 1 beta (IL-1 beta) secretion. Recently, it has been demonstrated that the interaction of adenosine triphosphate (ATP) with the P2X7 purinoceptor induces the secretion of IL-1 beta and initiates the inflammatory response. In these experiments, that the addition of ATP to macrophages was found to induce P2X7 activation and secretion of IL-1 beta. This secretion is enhanced with anti-RTF antibody in combination with exogenous ATP (p< 0.005). RTF is also revealed to be able to influence surface ATPase activity and, increase PI incorporation, which is an indicator of P2X7 activation. We demonstrate that RTF has a role in controlling IL-1 beta secretion by regulating P2X7 activity.

Minor H antigen HA-1-specific regulator and effector CD8+ T cells, and HA-1 microchimerism, in allograft tolerance.

The role of the hematopoietic lineage-restricted minor histocompatibility (H) antigen HA-1 in renal allograft tolerance was explored. We obtained peripheral blood samples from three recipients of histocompatibility leukocyte antigen (HLA)-matched, HA-1-mismatched renal transplants, one of which had discontinued immunosuppression >30 yr ago while sustaining normal kidney function. Peripheral blood mononuclear cells (PBMCs) were injected into the footpads of severe combined immunodeficiency mice to measure human delayed type hypersensitivity (DTH) responses. All three patients manifested regulated DTH responses to HA-1H peptide. By differential tetramer staining intensities, we observed two distinct minor H antigen HA-1-specific CD8+ T cell subsets. The one that stained dimly had the characteristics of a T regulatory (TR) cell and produced interleukin (IL) 10 and/or transforming growth factor (TGF) beta. These HA-1-specific TR cells coexisted with bright tetramer-binding CD8+ T effector (TE) cells. The CD8+ TE cells mediated HA-1-specific DTH and produced interferon-gamma. Suppression of these TE functions by TR cells was TGFbeta, IL-10, and cytotoxic T lymphocyte-associated antigen 4 dependent. In addition, HA-1 microchimerism was detected in two recipients, primarily in the dendritic cell fraction of the PBMCs. This is the first demonstration of coexisting CD8+ memory TR and TE cells, both specific for the same HA-1 antigen, in the context of renal allograft tolerance.