Search

Helper T (TT‑Helfer(TH)-Lymphozyten spielen eine entscheidende Rolle bei der Auslösung und Aufrechterhaltung chronischer rheumatischer Entzündungen. Über die Ausschüttung von entzündungsfördernden Botenstoffen und die Expression kostimulatorischer Oberflächenmoleküle koordinieren sie die Rekrutierung und Aktivierung von Effektorzellen, die letztendlich die Immunpathologie und Gewebezerstörung auslösen. Allerdings waren Therapieansätze, die auf die Eliminierung von TH-Zellen abzielten, aufgrund ihrer mangelnden Selektivität nicht erfolgreich. Am Deutschen Rheuma-Forschungszentrum (DRFZ) wollen wir deshalb TH-Zellen , die an chronischen Entzündungsreaktionen beteiligt sind, besser verstehen. Unser Ziel ist es, basierend darauf neue therapeutische Strategien zu entwickeln, die pathogene TH-Zellen bei rheumatischen Entzündungen gezielt und selektiv bekämpfen. Die Erkenntnisse des DRFZ zu diesem Thema fasst der vorliegende Beitrag zusammen.

The cumulation of cultural codes forms and structures the integral matrix of information capacity - the cognitive thesaurus (CT) of the psyche. The dissociation of the parameters of the current balance of somatic/mental homeostasis initiates a compensatory need. The "need" extracts from the CT (actualizes) the "image" of the cultural code - a representable informational equivalent of the "need" ("dominant"). Being the content of consciousness at the time of relevance, "dominant" acquires the properties of a resonant operator, a dominant focus, forming control mental constructs. Actualization of the "dominant" initiates the activity of the "action result acceptor." For the actual dominant, the "action result acceptor" is an indicator/corrector, a frequency matching censor (resonance): perceptual dynamics of operational images of external presentation ("code key") to the parameters of the "dominant" (code). Decoding the physical parameters of the "code" provides the potential to regulate the resonant impact on the arsenal of "cultural codes" of CT through moderation of the "code key." The CT, through the "action result acceptor" (the extra-conscious regulator of the actual autonoetic forms of mental activity), programs and formats the somatic/psychic continuum of the individual. The integral matrix of the CT, having a location outside the conscious sector of the psyche, including the imperative corrector of the perceived sector of the psyche associated with the emotional register, determining/correcting the main vectors of the regulation of the life continuum, is meaningfully close to the philosophical/psychological criteria of the main cognitive regulator of the person activity, defined as the soul.

The unification of cultural codes forms the basic set of code symbols (image+sense), which are the basis for recognition, orientation, social cooperation, and response to external appeals. The result of unification is a stable system of deterministic goal-setting and behavioral equivalents. Cultural codes, as a derivative of upbringing, education, and cultural landscape, are the result of spatiotemporal transformation (from fields of reception to displacement from consciousness and compression) of organized information constructs - "dominants" encoded in the form of electro/magnetic patterns. "Dominant" in this message is a generalized definition of organized, hierarchical constructs (packages) of information carriers, forced out of the conscious levels of the psyche and continuing to circulate in closed, reverberant neural circuits. The "dominants" forced out of consciousness circulate through recurrent neural circuits, the activity of which is reduced to several stable states, performing the functions of information retention. The electrical and magnetic parameters of the repressed "dominants" are the basic potential of frequency resonance upon presentation of a perceptual/cognitive construct that is close in frequency parameters - a "code key" at the entrance to the psyche system. The basic principle of information exchange is the formation of the initial resonance potential: the arsenal of "dominants," the concentration of information in frequency electro-magnetic patterns. The interaction of the "information universe" with the arsenal of "dominants" is a resonant process of frequency coincidence of the patterns of the "code key" and the "dominant" displaced from consciousness, which results in actualization, the appearance of an object of resonance in the field of voluntary attention, in consciousness. The updated "dominant," as a "resonant operator," determines the dynamics of the system as a whole for the period of relevance. The phase transition in the socio/cultural environment is the result of changes in the hidden meanings of social frames. The dynamics of the perception of the transformation of frames under changing (appearing new) hidden meanings ["habituation">"acceptability">"acceptability”] determines the growth of the frustration potential associated with acquired needs. The transformation of basic cultural codes rebuilds the mechanism of reproduction of the ethnocultural matrix and congruent social structures.

Background & aimsStudies investigating the gut-liver axis have largely focused on bacteria, whereas little is known about commensal fungi. We characterized fecal fungi in patients with non-alcoholic fatty liver disease (NAFLD) and investigated their role in a fecal microbiome-humanized mouse model of Western diet-induced steatohepatitis.MethodsWe performed fungal internal transcribed spacer 2 sequencing using fecal samples from 78 patients with NAFLD, 16 controls and 73 patients with alcohol use disorder. Anti-Candida albicans (C.albicans) IgG was measured in blood samples from 17 controls and 79 patients with NAFLD. Songbird, a novel multinominal regression tool, was used to investigate mycobiome changes. Germ-free mice were colonized with feces from patients with non-alcoholic steatohepatitis (NASH), fed a Western diet for 20 weeks and treated with the antifungal amphotericin B.ResultsThe presence of non-obese NASH or F2-F4 fibrosis was associated with a distinct fecal mycobiome signature. Changes were characterized by an increased log-ratio for Mucor sp./Saccharomyces cerevisiae (S. cerevisiae) in patients with NASH and F2-F4 fibrosis. The C.albicans/S. cerevisiae log-ratio was significantly higher in non-obese patients with NASH when compared with non-obese patients with NAFL or controls. We observed a different fecal mycobiome composition in patients with NAFLD and advanced fibrosis compared to those with alcohol use disorder and advanced fibrosis. Plasma anti-C.albicans IgG was increased in patients with NAFLD and advanced fibrosis. Gnotobiotic mice, colonized with human NASH feces and treated with amphotericin B were protected from Western diet-induced steatohepatitis.ConclusionsNon-obese patients with NAFLD and more advanced disease have a different fecal mycobiome composition to those with mild disease. Antifungal treatment ameliorates diet-induced steatohepatitis in mice. Intestinal fungi could be an attractive target to attenuate NASH.Lay summaryNon-alcoholic fatty liver disease is one of the most common chronic liver diseases and is associated with changes in the fecal bacterial microbiome. We show that patients with non-alcoholic fatty liver disease and more severe disease stages have a specific composition of fecal fungi and an increased systemic immune response to Candida albicans. In a fecal microbiome-humanized mouse model of Western diet-induced steatohepatitis, we show that treatment with antifungals reduces liver damage.

Bone marrow plasma cells (BMPC) emerge as a consequence of immune reactions and are considered the source of antibodies that protect against recurrent infectious diseases throughout life. Despite their importance, it remains unclear if these cells reflect different activation environments or the differentiation/maturation stages of their precursors. Here we track the recruitment of plasma cells, generated in primary and secondary immune reactions to SARS-CoV-2 spike protein vaccines, to the human bone marrow. Trajectories based on single cell transcriptomes and antigen-receptor clonotypes of antibody-secreting cells exiting the immune reaction and of those residing in the bone marrow, allow to follow the evolution of the immune response to these vaccines, leading to sequential colonization of these cells to different compartments (clans) of BMPC, and their establishment as long-lived (memory) plasma cells. In primary immune reactions, both CD19low (clans 1 and 4) and CD19high (clan 0) BMPC are generated. In secondary immune reactions, mostly CD19high BMPC of the largest compartment (clan 0) are generated, resulting from the reactivation of memory B lymphocytes. The latter is also observed in vaccinated convalescent individuals and upon recall vaccination against diphtheria/tetanus/pertussis (DTP). Thus, humoral immunological memory, i.e. serum antibodies secreted by long-lived memory BMPC, is generated already in the primary immune response, more so in the secondary, and it represents the evolution of the immune response.

SARS-CoV-2 is a single-stranded RNA virus that causes coronavirus disease 2019 (COVID-19). Given its acute and often self-limiting course, components of the innate immune system are likely central in controlling virus replication thereby determining clinical outcome. Natural killer (NK) cells are innate lymphocytes with notable activity against a broad range of viruses, including RNA viruses1,2. NK cell function may be altered during COVID-19 despite increased representation of NK cells with an activated and ‘adaptive’ phenotype3,4. Here we show that viral load decline in COVID-19 correlates with NK cell status and that NK cells can control SARS-CoV-2 replication by recognizing infected target cells. In severe COVID-19, NK cells show remarkable defects in virus control, cytokine production and cell-mediated cytotoxicity despite high expression of cytotoxic effector molecules. Single-cell RNA-sequencing (scRNA-seq) of NK cells along the time course of the entire COVID-19 disease spectrum reveals a unique gene expression signature. Transcriptional networks of interferon-driven NK cell activation are superimposed by a dominant TGFβ response signature with reduced expression of genes related to cell-cell adhesion, granule exocytosis and cell-mediated cytotoxicity. In severe COVID-19, serum levels of TGFβ peak during the first 2 weeks of infection, and serum obtained from these patients profoundly inhibits NK cell function in a TGFβ-dependent manner. Our data reveal that untimely production of TGFβ is a hallmark of severe COVID-19 and may inhibit NK cell function and early virus control.

Creativity, the integrative function of the psyche of Homo sapiens (HS), which arose about 50,000 years ago, allowed HS to project the image of the goal (IG), transformed into a “symbol,” into the external environment. The projection constructs of the psyche have become autonomous fragments of the environment in the HS perception, not being its derivatives. Objective reality, perceived by HS, has acquired non-inherent properties: a mental product of the psyche that integrates the virtual and real components of the environment. In other words, the role of HS has evolved over the last 10,000 years, since the beginning of the agrarian revolution: from (1) a dependent subject controlled by external forces in an animated world to (2) “the crown of God's creation” in “theism” and, (3) to the status "Higher power" during the period, in which "God is Dead." Initially, HS exists in an incompletely real environment, with an increasing component of virtuality. With symbolic virtual content, HS supplemented or duplicated the entire surrounding world, creating a two-component habitat (virtual and real). The emergence and development of conceptual thinking (ConceptT) led to a partial "devirtualization" of the environment, the removal of restrictions on scientific knowledge, the rapid growth of technology and social dynamics. The result of technological development was, in the recent past, a temporary resolution of the primary frustration: the establishment of the current equilibrium in the relationships with the regulatory "dissociated symbol"—the virtual "information universe” (IU). The IU, defining as the interference of "media" with the "information body" of the Internet, we consider as a unified information space, integrated with reality and in total constituting the HS habitat. Clip thinking (ClipT), qualitatively different from ConceptT, is a new operating system of the psyche, a moderator of adaptation to a new, virtualized environment. A technological derivative of mental activity HS—the IU—without the participation of conscious forms of mental activity, transforms the algorithms of thinking, i.e. formats the psyche as a whole with adaptation to qualitative and quantitative changes in the virtual component of the environment, and to the perspectives on the development of technologies during the singularity.

The perception of environmental images that are not in causal relationships and are not interconnected sequences forms what is called clip thinking (ClipT). The concomitant decrease in the analytical-synthetic psychic functions is the result of “images” from the environment (not constructs of the psyche of the subject) being crowded out by a sequence of other images, and so are not kept in the field of voluntary attention for sufficient time for fixation. As a result, along with analytical and synthetic functions, from the sequential series of operational functions of conceptual thinking (ConceptT), come the closing steps of the perception phase—the default mode network (DMN) activity. The result is changes in the reciprocal relationship between the perception and processing of incoming information. The deficiency changes in DMN cause an imbalance of structures that provide "focus and defocus," difficulty fixing attention and filtering information, interruption of social relations, and other disruptions. ClipT forms the ability to perceive a larger (compared to ConceptT) amount of short volumes of information per unit of time, with an increase in the total number and range of heterogeneous units of incoming information, increasing the multitasking of thinking and allowing the simultaneous execution of several types of activities. The prospective dynamics in the development of ClipT forms global unified communicative codes with the potency of the functional association of subject groups that do not have stratification and quantitative restrictions. The distribution of information in these associations, which have almost identical characteristics of operational symbols (OSs), and the high speed of distribution of content on an unlimited scale of social networks, is similar to infection mechanisms during epidemics. ClipT is at the initial stages of development, adapting the psyche to accelerating changes in the environmental information parameters. We believe that ClipT and ConceptT are various interfering processes of the transforming psyche of Homo sapiens. The development of ClipT creates the conditions for the psyche to transition to a new qualitative level, with a change in structure and a transition to a new class of systems. In the conditions of increasing density, speed, randomness of the information flow and increasing degree of uncertainty, we consider the new phase transition as the attractor of the described changes in thinking (the psyche as a whole) of HS—the singularity period determining the changes (transformation of thinking is the most accessible integral research feature of significant, but less noticeable changes in the psyche as a whole) that take place with the future final state of a unified information system, including external devices and adaptive integral changes in the properties of the psyche as a whole.

Synchronization of the curves of central and peripheral hemodynamics, according to the data obtained during catheterization, showed the intersection points of the graphical pressure curves, which were shown in previous works. The three-chamber block of the ventricles (with leading role of LV) in the isometric contraction phase (D(Q)) generates three wave impulses: 1) retrograde impulse of CS directed to the zone of the integral combination (hemodynamics, metabolites of metabolic zones, hormones) and partial interference of venous flows coming to RA (end section of SC); 2) anterograde impulse of RV, which heads to the exchange zone of the lung (initial section of PC) and outrunning of the “venous bolus”; 3) anterograde impulse of LV, ahead of the “arterial bolus” (initial section of SC), leaving along Ao into the vascular bed to the exchange zones of peripheral organs. We have interpreted intersections of hemodynamic curves as “zone of temporal equalization of pressure” (ZTEP), both in the heart and in remote topographic zones. Based on the results of previous works, we built separate ZTEP plots for the high- and low-energy phases of the cardiac cycle (CC), relative to CMIP. The genesis and development of key points of the low-energy phase are considered. We believe that the distributed ZTEP matrix created in the vascular bed during each CC, consisting of short-term synchronous equal-sized pressor structures interfering with the Ao wave impulse, has a regulatory effect on the CC phase sequence, the hemodynamics of organs (including peripheral resistance), and metabolism. The systemic distributed ZTEP association—a formed CMIP—is a synchronized (between ZTEPs, with the phases of CC, and ECG), high-speed, information-regulatory structure of the interaction of hemodynamics and metabolism of both central and peripheral organs, which is involved in the control and regulation of homeostasis as a whole.

The commensal microflora is a source for multiple antigens that may induce cross-reactive antibodies against host proteins and pathogens. However, whether commensal bacteria can induce cross-reactive antibodies against SARS-CoV-2 remains unknown. Here we report that several commensal bacteria contribute to the generation of cross-reactive IgA antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein. We identified SARS-CoV-2 unexposed individuals with RBD-binding IgA antibodies at their mucosal surfaces. Conversely, neutralising monoclonal anti-RBD antibodies recognised distinct commensal bacterial species. Some of these bacteria, such as Streptococcus salivarius, induced a cross-reactive anti-RBD antibodies upon supplementation in mice. Conversely, severely ill COVID-19 patients showed reduction of Streptococcus and Veillonella in their oropharynx and feces and a reduction of anti-RBD IgA at mucosal surfaces. Altogether, distinct microbial species of the human microbiota can induce secretory IgA antibodies cross-reactive for the RBD of SARS-CoV-2.

By catheterization, the integral indicators of synchronization and interaction of blood flows, designated as “venous and arterial boluses,” were obtained, studied and analyzed in healthy people on the pathway: right heart–lung–left heart. It has been confirmed that the complete CC of the BB from RA to the ejection from the LV has a length equal to two completed heart contraction cycles. Interaction of venous and arterial boluses, with differentiated external myocardial exposure, along the path “venous block of the heart–lung–arterial block of the heart,” forms averaged (compensated by the flexible septum) variable pressure values between the stages of intracardiac routes of BBs (unidirectional, synchronous, but spread in the space). The complex of these pressure values creates an intracardiac pressure balance at the border of high- and low-energy processes of the heart. We defined the sequential dynamics of these values as CMIP. Our mathematical and graphical data demonstrate the presence of direct and inversecardio-cerebral wave connections, where the waveguides are the vessels of entry and exit from the skull. We believe that CMIP is a universal, central rhythmic process, a regulator that determines the sequence and intensity of the CC phases, HR, and synchronous nervous and wave effects on brain structures. The modulating effect of CMIP on brain structures, providing some sensory-motor reactions, behavioral functions and forms of behavior, occurs outside the realm of consciousness. Our data suggest that the modulating effect of CMIP on the brain is carried out not only along the neural pathways, but also by the vascular wave structures that combine the heart and brain into a single hydrodynamic structure with phase-varying volume and configuration, as well as variable patterns of regulatory impulses.

Hemodynamic indices of healthy people obtained by catheterization in such vascular areas as the chambers of the heart (both ventricles, both atria, coronary sinus), IVC, SVC, RHV, right renal vein, sigmoid sinus, and aorta were analyzed. Using the mean values of hemodynamic parameters, we constructed graphs of the curves of the central, arterial, and venous pressure, synchronized with each other, with an ECG, and with the peripheral pulse wave. In the present work, to generalize the results obtained, we constructed integral curves of the dynamics of the blood substrate (“bolus,” BB) vector from the entrance (the fusion of venous flows: SVC, IVC, RHV, and coronary sinus) to the exit from the heart-lung-heart system in the form of LVEF. It was revealed that the complete CC consists of two BBs, simultaneously passing the hemodynamic pathway: heart-lung-heart. One BB completes the transformation, leaving PC after contact with the external gaseous medium in the lung and the formation of the pulse wave spheroid in LV, going into SC in the form of LVEF; another BB, after the formation of the pulse wave spheroid in RV, goes into PC, followed by hemodynamic, metabolic and gas transformation. CMIP is a universal indicator of the quality and stability of the relationship of all hemodynamic structures (flows in vessels and heart chambers) included in integral hemodynamic flows (IC-1, IC-2) throughout the complete CC. Changes in the configuration or level of CMIP is an indicator of imbalance and restructuring of the integral indicators of blood flow (therefore, components of hemodynamic flows), indicating a deviation in the parameters of stability of homeostasis as a whole.

Bone marrow (BM) stromal cells are important in the development and maintenance of cells of the immune system. Using single cell RNA sequencing, we here explore the functional and phenotypic heterogeneity of individual transcriptomes of 1167 murine BM mesenchymal stromal cells. These cells exhibit a tremendous heterogeneity of gene expression, which precludes the identification of defined subpopulations. However, according to the expression of 108 genes involved in the communication of stromal cells with hematopoietic cells, we have identified 14 non‐overlapping subpopulations, with distinct cytokine or chemokine gene expression signatures. With respect to the maintenance of subsets of immune memory cells by stromal cells, we identified distinct subpopulations expressing Il7, Il15 and Tnfsf13b. Together, this study provides a comprehensive dissection of the BM stromal heterogeneity at the single cell transcriptome level and provides a basis to understand their lifestyle and their role as organizers of niches for the long‐term maintenance of immune cells.

Summary In order to ascertain the significance of transmembrane tumor necrosis factor (tmTNF) reverse signaling in vivo, we generated a triple transgenic mouse model (3TG, TNFR1−/−, TNFR2−/−, and tmTNFKI/KI) in which all canonical tumor necrosis factor (TNF) signaling was abolished. In bone-marrow-derived macrophages harvested from these mice, various anti-TNF biologics induced the expression of genes characteristic of alternative macrophages and also inhibited the expression of pro-inflammatory cytokines mainly through the upregulation of arginase-1. Injections of TNF inhibitors during arthritis increased pro-resolutive markers in bone marrow precursors and joint cells leading to a decrease in arthritis score. These results demonstrate that the binding of anti-TNF biologics to tmTNF results in decreased arthritis severity. Collectively, our data provide evidence for the significance of tmTNF reverse signaling in the modulation of arthritis. They suggest a complementary interpretation of anti-TNF biologics effects in the treatment of inflammatory diseases and pave the way to studies focused on new arginase-1-dependent therapeutic targets.

The pathogenesis of severe COVID-19 reflects an inefficient immune reaction to SARS-CoV-2. Here we analyze, at the single cell level, plasmablasts egressed into the blood to study the dynamics of adaptive immune response in COVID-19 patients requiring intensive care. Before seroconversion in response to SARS-CoV-2 spike protein, peripheral plasmablasts display a type 1 interferon-induced gene expression signature; however, following seroconversion, plasmablasts lose this signature, express instead gene signatures induced by IL-21 and TGF-β, and produce mostly IgG1 and IgA1. In the sustained immune reaction from COVID-19 patients, plasmablasts shift to the expression of IgA2, thereby reflecting an instruction by TGF-β. Despite their continued presence in the blood, plasmablasts are not found in the lungs of deceased COVID-19 patients, nor does patient IgA2 binds to the dominant antigens of SARS-CoV-2. Our results thus suggest that, in severe COVID-19, SARS-CoV-2 triggers a chronic immune reaction that is instructed by TGF-β, and is distracted from itself., Our understanding on the humoral immunity induced by SARS-CoV-2 is still lacking. Here the authors analyze B cell responses at the single cell level to find that, in severe COVID-19 patients, plasmablasts shift from IFN to TGFβ instruction to produce IgA antibodies that are not specific to dominant SARS-CoV-2 antigens.

Here we have analyzed the dynamics of the adaptive immune response triggered by SARS-CoV-2 in severely affected COVID-19 patients, as reflected by activated B cells egressing into the blood, at the single cell level. Early on, before seroconversion in response to SARS-CoV-2 spike protein, activated peripheral B cells displayed a type 1 interferon-induced gene expression signature. After seroconversion, activated B cells lost this signature, expressed IL-21- and TGF-β-induced gene expression signatures, and mostly IgG1 and IgA1. In the sustained immune reaction of the COVID-19 patients, until day 59, activated peripheral B cells shifted to expression of IgA2, reflecting instruction by TGF-β. Despite the continued generation of activated B cells, those cells were not found in the lungs of deceased COVID-19 patients, nor did the IgA2 bind to dominant antigens of SARS-CoV-2. In severe COVID-19, SARS-CoV-2 thus triggers a chronic immune reaction distracted from itself and instructed by TGF-β.

Summary In order to ascertain the significance of transmembrane tumor necrosis factor (tmTNF) reverse signaling in vivo, we generated a triple transgenic mouse model (3TG, TNFR1−/−, TNFR2−/−, and tmTNFKI/KI) in which all canonical tumor necrosis factor (TNF) signaling was abolished. In bone-marrow-derived macrophages harvested from these mice, various anti-TNF biologics induced the expression of genes characteristic of alternative macrophages and also inhibited the expression of pro-inflammatory cytokines mainly through the upregulation of arginase-1. Injections of TNF inhibitors during arthritis increased pro-resolutive markers in bone marrow precursors and joint cells leading to a decrease in arthritis score. These results demonstrate that the binding of anti-TNF biologics to tmTNF results in decreased arthritis severity. Collectively, our data provide evidence for the significance of tmTNF reverse signaling in the modulation of arthritis. They suggest a complementary interpretation of anti-TNF biologics effects in the treatment of inflammatory diseases and pave the way to studies focused on new arginase-1-dependent therapeutic targets., Graphical abstract, Highlights • In vivo demonstration of tmTNF reverses signaling existence • tmTNF reverse signaling induces anti-oxidative stress response • tmTNF reverse signaling induces an arginase-1-mediated anti-inflammatory response • Reverse signaling is a complementary mechanism to TNF neutralization by anti-TNF, Immunology; Cell Biology

Foxp3+ regulatory T cells (Treg cells) are crucial for the maintenance of immune homeostasis both in lymphoid tissues and in non-lymphoid tissues. Here we demonstrate that the ability of intestinal Treg cells to constrain microbiota-dependent interleukin (IL)-17-producing helper T cell (TH17 cell) and immunoglobulin A responses critically required expression of the transcription factor c-Maf. The terminal differentiation and function of several intestinal Treg cell populations, including RORγt+ Treg cells and follicular regulatory T cells, were c-Maf dependent. c-Maf controlled Treg cell-derived IL-10 production and prevented excessive signaling via the kinases PI(3)K (phosphatidylinositol-3-OH kinase) and Akt and the metabolic checkpoint kinase complex mTORC1 (mammalian target of rapamycin) and expression of inflammatory cytokines in intestinal Treg cells. c-Maf deficiency in Treg cells led to profound dysbiosis of the intestinal microbiota, which when transferred to germ-free mice was sufficient to induce exacerbated intestinal TH17 responses, even in a c-Maf-competent environment. Thus, c-Maf acts to preserve the identity and function of intestinal Treg cells, which is essential for the establishment of host-microbe symbiosis.

RORγt

Hemodynamic indices studied in healthy people were obtained by catheterization in various vascular areas: the chambers of the heart (ventricles, atria, coronary sinus), pulmonary trunk, sigmoid sinus, aorta, inferior vena cava, superior vena cava (SVC), and right hepatic vein (RHV). Using the mean values of the hemodynamic parameters, we constructed graphics of the curves of the central, arterial, and venous pressure, synchronized with each other, with an ECG, and with pulse wave. The complex of hemodynamic curves, supplemented by curves of RHV and SVC, revealed a coincidence in one zone of temporal equalization of pressure (ZTEP) for values of the left atrium, coronary sinus, right ventricle, sigmoid sinus, SVC, and RHV(wedged). We identified this ZTEP as a hemodynamic trigger (T-point), assuming that this point is the trigger point for the launch of the high-energy processes of the right heart, forming the following cardiac cycle: the beginning of the systole of right ventricle, the opening of pulmonary valve.

Significance In spite of TNF involvement in the pathogenesis of multiple sclerosis (MS), systemic TNF neutralization in MS patients was not successful. One of the possible reasons is that TNF possesses both pathogenic and protective features that may be related to TNFR1 versus TNFR2 receptor engagement. This study uncovers one of such protective functions of TNF mediated by intrinsic TNFR2 signaling in Treg cells. In mice bearing humanized TNF and TNFR2 genetic loci, TNFR2 ablation restricted to Treg cells led to reduced capacity to control Th17 cell responses, exacerbated experimental autoimmune encephalomyelitis (EAE) development, and affected the maintenance of Treg cells. These findings provide support for the emerging role of TNFR2 signaling in autoimmunity, as demonstrated here in mice with conditional inactivation of TNFR2., TNF is a multifunctional cytokine involved in autoimmune disease pathogenesis that exerts its effects through two distinct TNF receptors, TNFR1 and TNFR2. While TNF- and TNFR1-deficient (but not TNFR2-deficient) mice show very similar phenotypes, the significance of TNFR2 signaling in health and disease remains incompletely understood. Recent studies implicated the importance of the TNF/TNFR2 axis in T regulatory (Treg) cell functions. To definitively ascertain the significance of TNFR2 signaling, we generated and validated doubly humanized TNF/TNFR2 mice, with the option of conditional inactivation of TNFR2. These mice carry a functional human TNF-TNFR2 (hTNF-hTNFR2) signaling module and provide a useful tool for comparative evaluation of TNF-directed biologics. Conditional inactivation of TNFR2 in FoxP3+ cells in doubly humanized TNF/TNFR2 mice down-regulated the expression of Treg signature molecules (such as FoxP3, CD25, CTLA-4, and GITR) and diminished Treg suppressive function in vitro. Consequently, Treg-restricted TNFR2 deficiency led to significant exacerbation of experimental autoimmune encephalomyelitis (EAE), accompanied by reduced capacity to control Th17-mediated immune responses. Our findings expose the intrinsic and beneficial effects of TNFR2 signaling in Treg cells that could translate into protective functions in vivo, including treatment of autoimmunity.

Hemodynamic indices studied in practically healthy people were obtained by catheterization in various vascular areas: the chambers of the heart (ventricles, atria, coronary sinus), pulmonary trunk, aorta, inferior vena cava, superior vena cava, right hepatic vein, and sigmoid sinus. Using the mean values of the hemodynamic parameters, we constructed graphics of the "curves" of the central, arterial, and venous pressure, synchronized with each other and an ECG, and with the radial pulse wave recorded by a non-invasive method. The obtained data, which demonstrate the projection coincidences of the characteristic points of peripheral pulse wave with the key indicators of the phases of cardiac cycle (CC), made it possible to transform the results obtained during the invasive examination into indicators of the non-invasive technique. This transformation became possible not only at the characteristic points of the deployed peripheral pulse wave, but in each anacrotic and dicrotic segment, which are understood as the projection areas of the synchronized hemodynamic and wave processes of the vascular bed. We believe it possible to catalog the forms of pulse waves, as well as their projection segments, to obtain accurate diagnostic information about the phases of CC and organ hemodynamics in humans in norm and with pathological conditions, using a non-invasive methood based on basic information obtained by invasive methods.

In humans and mice, mucosal immune responses are dominated by IgA antibodies and the cytokine TGF-β, suppressing unwanted immune reactions but also targeting Ig class switching to IgA. It had been suggested that eosinophils promote the genera- tion and maintenance of mucosal IgA-expressing plasma cells. Here, we demonstrate that not eosinophils, but specific bacteria determine mucosal IgA production. Co-housing of eosinophil-deficient mice with mice having high intestinal IgA levels, as well as the intentional microbiota transfer induces TGF-β expression in intestinal T follicular helper cells, thereby promoting IgA class switching in Peyer’s patches, enhancing IgA+ plasma cell numbers in the small intestinal lamina propria and levels of mucosal IgA. We show that bacteria highly enriched for the genus Anaeroplasma are sufficient to induce these changes and enhance IgA levels when adoptively transferred. Thus, specific members of the intestinal microbiota and not the microbiota as such regulate gut homeostasis, by promoting the expression of immune-regulatory TGF-β and of mucosal IgA.

Background & Aims A substantial proportion patients with inflammatory bowel disease (IBD) have a primary non-response to infliximab; markers are needed to identify patients most likely to respond to treatment. We investigated whether production of tumor necrosis factor (TNF) by peripheral blood mononuclear cells (PBMCs) can be used as a marker to predict response. Methods We performed a prospective study of 41 adults with IBD (mean age, 38 years; 21 male; 21 with Crohn’s disease and 20 with ulcerative colitis) not treated with a biologic agent within the past 6 months; patients were given their first infusion of infliximab at a hospital or clinic in Berlin, Germany. We collected data on clinical scores, levels of C-reactive protein, and ultrasound results (Limberg scores) at baseline (before the first infusion) and after 6 weeks (3rd infliximab infusion). PMBCs were obtained from patients at baseline and 10 healthy individuals (controls) and incubated with lipopolysaccharide. We measured production of cytokines (TNF, interleukin 1 [IL1], IL6, IL8, IL10, IL12p70, and IL22) by ELISA and performed cytometric bead array and flow cytometry analyses. The primary endpoint was clinical response (decrease in Harvey Bradshaw Index scores of 2 or more or decrease in partial Mayo scores of 3 or more at week 6) in patients with PBMCs that produced high vs low levels of TNF. Results Responders had a shorter median disease duration (P = .018) and higher median Limberg score (P = .021), than nonresponders. Baseline PBMCs from responders produced significantly more TNF (P = .049) and IL6 (P = .028) than from nonresponders; a level of 500 pg/ml TNF identified responders with 82% sensitivity and 78% specificity. In patients with Crohn’s disease, this cutoff value (500 pg/ml TNF) identified responders with 100% sensitivity and 82% specificity; TNF levels above this level were independently associated with response to infliximab in multivariate analysis (odds ratio, 16.2; 95% CI, 1.8–148.7; P = .014). The percentage of TNF-positive cells was higher among CD14+ monocytes than lymphocytes after stimulation. Conclusions Production of a high level of TNF by PBMCs (specifically CD14+ cells) from patients with IBD can identify those most likely to have a clinical response to infliximab therapy. In patients with Crohn’s disease, a cutoff value of 500 pg/ml TNF identified responders with 100% sensitivity and 82% specificity.

Solitary intestinal lymphoid tissues such as cryptopatches (CPs) and isolated lymphoid follicles (ILFs) constitute steady-state activation hubs containing group 3 innate lymphoid cells (ILC3) that continuously produce interleukin (IL)-22. The outer surface of CPs and ILFs is demarcated by a poorly characterized population of CD11c+ cells. Using genome-wide single-cell transcriptional profiling of intestinal mononuclear phagocytes and multidimensional flow cytometry, we found that CP- and ILF-associated CD11c+ cells were a transcriptionally distinct subset of intestinal cDCs, which we term CIA-DCs. CIA-DCs required programming by CP- and ILF-resident CCR6+ ILC3 via lymphotoxin-β receptor signaling in cDCs. CIA-DCs differentially expressed genes associated with immunoregulation and were the major cellular source of IL-22 binding protein (IL-22BP) at steady state. Mice lacking CIA-DC-derived IL-22BP exhibited diminished expression of epithelial lipid transporters, reduced lipid resorption, and changes in body fat homeostasis. Our findings provide insight into the design principles of an immunoregulatory checkpoint controlling nutrient absorption.

Career situation of first and presenting author Post-doctoral fellow. Introduction Anti-TNF agents are widely used in rheumatoid arthritis (RA). Their effect on inflammation results from the neutralization of soluble TNF (sTNF), but also supposedly from the induction of reverse signaling through their binding to membrane TNF (tmTNF). Despite possible clinical relevance, reverse signaling has been described only in vitro but has not been proven in vivo. Objectives In this study we aim to demonstrate for the first time the existence of tmTNF reverse signaling in vivo and its importance in anti-TNF response during arthritis. Methods Triple transgenic mouse model (3TG), KO for TNFR1/TNFR2 and KI for tmTNF, thus secreting no sTNF was developed. To analyze reverse signaling, mice were injected either with etanercept (ETA, 10 mg/kg), an anti-mouse TNF antibody (MP6-XT22, rat IgG1, 10 mg/kg) or an anti-human IL17 antibody (secukinumab, SEC, 10 mg/kg) as a control. Daily clinical evaluation of K/BxN serum induced-arthritis was performed in 3TG as well as WT mice. Polarization of bone marrow-derived macrophages (BMDM) and cytokine production from non-arthritic WT and 3TG mice under the action of anti-TNF in vitro was evaluated by RT-qPCR, CBA and ELISA. Results In vivo, the administration of anti-TNF (ETA or MP6-XT22) decreased arthritic scores in WT mice (p=0.005) as well as in 3TG mice (p Conclusions Our work provides in vivo evidence for the involvement of reverse signaling in the anti-TNF-mediated modulation of arthritis. Reverse signaling is expected to result in the modulation of macrophage polarization from an inflammatory to an alternative functional phenotype in arthritic mice. Our data prompt us to consider new interpretation of the effects of anti-TNF in the treatment of RA. Disclosure of Interest None declared.

Career situation of first and presenting author Post-doctoral fellow. Introduction The human intestine is colonized with billions of microorganisms, which form the gut microbiota, consisting of up to 1000 different bacterial species. Recent studies have implicated the intestinal microbiota in the pathogenesis of chronic inflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, systemic sclerosis, and Sjogren’s syndrome. Yet, we still lack the knowledge which bacteria of the gut microbiota induce, promote or inhibit chronic inflammatory inflammation. Objectives The aim of our work is to identify members of the intestinal microbiota with pro- or anti-inflammatory properties for targeted and therapeutic manipulation of the microbiota in chronic inflammatory diseases. Methods We have developed high-resolution microbiota flow cytometry which allows us to analyze the microbiota on a single cell level. This provides a non-invasive, fast and efficient diagnostic tool to visualize dramatic changes of microbiota composition in inflammatory diseases, fast and efficiently, and isolate distinct bacteria for functional and molecular analyses. Results We have identified bacteria belonging to the genus Anaeroplasma, which enhances the levels of mucosal IgA. Adoptive transfer of Anaeroplasma increases the numbers of IgA+ germinal center B cells in the Peyer’s patches and of IgA-secreting plasma cells in the lamina propria of the small intestine leading to significantly enhanced mucosal IgA levels. Anaeroplasma controls IgA expression presumably its ability to induce expression of the regulatory cytokine TGF-β in T cells, as we show here. Conclusions The anti-inflammatory properties of Anaeroplasma to induce the anti-inflammatory cytokine TGF-β, thereby also strengthening the intestinal barrier by enhancing mucosal IgA, qualify Anaeroplasma as potent probiotic for the prevention and treatment of chronic inflammation. Disclosure of Interest None declared.

Background & Aims Unregulated activity of interleukin (IL) 22 promotes intestinal tumorigenesis in mice. IL22 binds the antagonist IL22 subunit alpha 2 (IL22RA2, also called IL22BP). We studied whether alterations in IL22BP contribute to colorectal carcinogenesis in humans and mice. Methods We obtained tumor and nontumor tissues from patients with colorectal cancer (CRC) and measured levels of cytokines by quantitative polymerase chain reaction, flow cytometry, and immunohistochemistry. We measured levels of Il22bp messenger RNA in colon tissues from wild-type, Tnf–/–, Lta–/–, and Ltb–/– mice. Mice were given azoxymethane and dextran sodium sulfate to induce colitis and associated cancer or intracecal injections of MC38 tumor cells. Some mice were given inhibitors of lymphotoxin beta receptor (LTBR). Intestine tissues were analyzed by single-cell sequencing to identify cell sources of lymphotoxin. We performed immunohistochemistry analysis of colon tissue microarrays from patients with CRC (1475 tissue cores, contained tumor and nontumor tissues) and correlated levels of IL22BP with patient survival times. Results Levels of IL22BP were decreased in human colorectal tumors, compared with nontumor tissues, and correlated with levels of lymphotoxin. LTBR signaling was required for expression of IL22BP in colon tissues of mice. Wild-type mice given LTBR inhibitors had an increased tumor burden in both models, but LTBR inhibitors did not increase tumor growth in Il22bp–/– mice. Lymphotoxin directly induced expression of IL22BP in cultured human monocyte–derived dendritic cells via activation of nuclear factor κB. Reduced levels of IL22BP in colorectal tumor tissues were associated with shorter survival times of patients with CRC. Conclusions Lymphotoxin signaling regulates expression of IL22BP in colon; levels of IL22BP are reduced in human colorectal tumors, associated with shorter survival times. LTBR signaling regulates expression of IL22BP in colon tumors in mice and cultured human dendritic cells. Patients with colorectal tumors that express low levels of IL22BP might benefit from treatment with an IL22 antagonist.

ObjectivesNeutralisation of tumour necrosis factor (TNF) is widely used as a therapy for rheumatoid arthritis (RA). However, this therapy is only effective in less than a half of patients and is associated with several side effects. We hypothesised that TNF may possess non-redundant protective and immunomodulatory functions in vivo that cannot be blocked without a cost. The present work aimed to identify cellular sources of protective and pathogenic TNF, and its molecular forms during autoimmune arthritis.MethodsMice lacking TNF expression by distinct cell types, such as myeloid cells and T or B lymphocytes, were subjected to collagen-induced arthritis (CIA) and collagen antibody-induced arthritis. Mice lacking soluble TNF production were also employed. The severity and incidence of the disease, as well as humoral and cellular responses were assessed.ResultsMyeloid cell-derived TNF contributes to both induction and pathogenesis of autoimmune arthritis. Conversely, T cell-derived TNF is protective during the induction phase of arthritis via limiting of interleukin-12 production by dendritic cells and by subsequent control of autoreactive memory T cell development, but is dispensable during the effector phase of arthritis. B cell-derived TNF mediates severity of CIA via control of pathogenic autoantibody production.ConclusionsDistinct TNF-producing cell types may modulate disease development through different mechanisms, suggesting that in arthritis TNF ablation from restricted cellular sources, such as myeloid cells, while preserving protective TNF functions from other cell types may be superior to pan-anti-TNF therapy.

It is well accepted that Th17 cells are a highly plastic cell subset that can be easily directed toward the Th1 phenotype in vitro and also in vivo during inflammation. However, there is an ongoing debate regarding the reverse plasticity (conversion from Th1 to Th17). We show here that ectopic ROR-γt expression can restore or initiate IL-17 expression by non-classic or classic Th1 cells, respectively, while common pro-Th17 cytokine cocktails are ineffective. This stability of the Th1 phenotype is at least partially due to the presence of a molecular machinery governed by the transcription factor Eomes, which promotes IFN-γ secretion while inhibiting the expression of ROR-γt and IL-17. By using a mouse model of T cell-dependent colitis we demonstrate that Eomes controls non-classic Th1 cell development also in vivo and promotes their pathogenic potential. Eomes expression associates to a highly inflammatory phenotype also in patients with juvenile idiopathic arthritis. Indeed, it favors the acquisition of a cytotoxic signature, and promotes the development of IFN-γ+ GM-CSF+ cells that have been described to be pathogenic in chronic inflammatory disorders.

Hintergrund Gegenwartig basiert die Behandlung von Autoimmunerkrankungen auf der Beeinflussung allgemeiner Kontrollmechanismen, einschlieslich derer, die durch immunregulatorische Zytokine vermittelt werden. Dieser Ansatz ist nicht kurativ und kann unerwunschte Nebenwirkungen haben, da die einzelnen Zytokine jeweils zahlreiche nutzbringende Funktionen haben, die nicht redundant sind, d. h., nicht von anderen Zytokinen ubernommen werden konnen.

Foxp3

Broad-spectrum antibiotics are widely used with patients in intensive care units (ICUs), many of whom develop hospital-acquired infections with Pseudomonas aeruginosa. Although preceding antimicrobial therapy is known as a major risk factor for P. aeruginosa-induced pneumonia, the underlying mechanisms remain incompletely understood. Here we demonstrate that depletion of the resident microbiota by broad-spectrum antibiotic treatment inhibited TLR-dependent production of a proliferation-inducing ligand (APRIL), resulting in a secondary IgA deficiency in the lung in mice and human ICU patients. Microbiota-dependent local IgA contributed to early antibacterial defense against P. aeruginosa. Consequently, P. aeruginosa-binding IgA purified from lamina propria culture or IgA hybridomas enhanced resistance of antibiotic-treated mice to P. aeruginosa infection after transnasal substitute. Our study provides a mechanistic explanation for the well-documented risk of P. aeruginosa infection following antimicrobial therapy, and we propose local administration of IgA as a novel prophylactic strategy.

Although in normal lamina propria (LP) large numbers of eosinophils are present, little is known about their role in mucosal immunity at steady state. Here we show that eosinophils are needed to maintain immune homeostasis in gut-associated tissues. By using eosinophil-deficient ΔdblGATA-1 and PHIL mice or an eosinophil-specific depletion model, we found a reduction in immunoglobulin A(+) (IgA(+)) plasma cell numbers and in secreted IgA. Eosinophil-deficient mice also showed defects in the intestinal mucous shield and alterations in microbiota composition in the gut lumen. In addition, TGF-β-dependent events including class switching to IgA in Peyer's patches (PP), the formation of CD103(+) T cells including Foxp3(+) regulatory (Treg), and also CD103(+) dendritic cells were disturbed. In vitro cultures showed that eosinophils produce factors that promote T-independent IgA class switching. Our findings show that eosinophils are important players for immune homeostasis in gut-associated tissues and add to data suggesting that eosinophils can promote tissue integrity.

Command and Control Innate lymphoid cells are vital for the development of gut-associated lymphoid tissues, maintenance of the epithelial barrier, and protection against intestinal microbes; their dysfunction can promote immune pathology. Immunoglobulin A (IgA) production is important for maintenance of the gut epithelial barrier and the composition of the gut microbiota. Through the generation of knockout mouse models, Kruglov et al. (p. 1243 ) were able to distinguish how soluble and membrane-bound lymphotoxins expressed by innate lymphoid cells in the gut specifically regulate IgA production and thereby control gut microbiota composition.

Currently, treatment of autoimmune diseases is based on manipulation of general control mechanisms, including those mediated by immunoregulatory cytokines. This approach is non-curative and may cause unwanted side effects due to numerous beneficial and non-redundant functions of a particular cytokine.Techniques of reverse genetics, such as conditional gene targeting, were employed to uncover the contributions of two proinflammatory and immunomodulatory cytokines, tumour necrosis factor (TNF) and interleukin 6 (IL-6), in various disease states.Several non-redundant functions of TNF from distinct cellular sources were identified. TNF from myeloid cells is pathogenic in several autoimmune diseases, whereas TNF produced by T cells showed non-redundant protective functions in experimental arthritis and in a Mycobacterium tuberculosis infection model. To test the idea of selective pharmacological inhibition of "bad" TNF produced by myeloid cells while sparing "good" TNF produced by T lymphocytes, a myeloid-specific TNF inhibitor (MYSTI) was designed-a recombinant mini-antibody with dual specificity that can bind to the surface molecule F4/80 on myeloid cells and to TNF. In vitro experiments confirmed retention of TNF on the surface of TNF-producing cells and in vivo experiments indicated that MYSTI can protect mice from lethal TNF-mediated hepatotoxicity. MYSTI is also effective in experimental arthritis.The proposed therapeutic strategy may be more effective than systemic anti-cytokine therapy in several human autoimmune diseases, as it would preserve the potentially beneficial effects of the same cytokine produced by other cell types. Such bispecific biological agents may become interesting tools for experimental studies and, eventually, drug development.

Correlation between the chemical structure of lipid A from various Gram-negative bacteria and biological activity of their lipopolysaccharide (LPS) as an agonist of the innate immune receptor Toll-like receptor 4 was investigated. Purified LPS species were quantitatively evaluated by their ability to activate the production of tumor necrosis factor (TNF) by murine bone marrow-derived macrophages in vitro. Wild-type LPS from plague-causing bacteria Yersinia pestis was compared to LPS from mutant strains with defects in acyltransferase genes (lpxM, lpxP) responsible for the attachment of secondary fatty acid residues (12:0 and 16:1) to lipid A. Lipid A of Y. pestis double ΔlpxM/ΔlpxP mutant was found to have the chemical structure that was predicted based on the known functions of the respective acyltransferases. The structures of lipid A from two members of the ancient psychrotrophic bacteria of the genus Psychrobacter were established for the first time, and biological activity of LPS from these bacteria containing lipid A fatty acids with shorter acyl chains (C10–C12) than those in lipid A from LPS of Y. pestis or E. coli (C12–C16) was determined. The data revealed a correlation between the ability of LPS to activate TNF production by bone marrow-derived macrophages with the number and the length of acyl chains within lipid A.

Tumor necrosis factor (TNF) is an important cytokine for host defense against pathogens but is also associated with the development of human immunopathologies. TNF blockade effectively ameliorates many chronic inflammatory conditions but compromises host immunity to tuberculosis. The search for novel, more specific human TNF blockers requires the development of a reliable animal model. We used a novel mouse model with complete replacement of the mouse TNF gene by its human ortholog (human TNF [huTNF] knock-in [KI] mice) to determine resistance to Mycobacterium bovis BCG and M. tuberculosis infections and to investigate whether TNF inhibitors in clinical use reduce host immunity. Our results show that macrophages from huTNF KI mice responded to BCG and lipopolysaccharide similarly to wild-type macrophages by NF-κB activation and cytokine production. While TNF-deficient mice rapidly succumbed to mycobacterial infection, huTNF KI mice survived, controlling the bacterial burden and activating bactericidal mechanisms. Administration of TNF-neutralizing biologics disrupted the control of mycobacterial infection in huTNF KI mice, leading to an increased bacterial burden and hyperinflammation. Thus, our findings demonstrate that human TNF can functionally replace murine TNF in vivo , providing mycobacterial resistance that could be compromised by TNF neutralization. This new animal model will be helpful for the testing of specific biologics neutralizing human TNF.