[Research consortium Neuroimmunology and pain in the research network musculoskeletal diseases]. / Forschungsverbund Neuroimmunologie und Schmerz (Neuroimpa) im Forschungsnetz Muskuloskelettale Erkrankungen.

BACKGROUND: The research consortium Neuroimmunology and Pain (Neuroimpa) explores the importance of the relationships between the immune system and the nervous system in musculoskeletal diseases for the generation of pain and for the course of fracture healing and arthritis. MATERIAL AND METHODS: The spectrum of methods includes analyses at the single cell level, in vivo models of arthritis and fracture healing, imaging studies on brain function in animals and humans and analysis of data from patients. RESULTS: Proinflammatory cytokines significantly contribute to the generation of joint pain through neuronal cytokine receptors. Immune cells release opioid peptides which activate opioid receptors at peripheral nociceptors and thereby evoke hypoalgesia. The formation of new bone after fractures is significantly supported by the nervous system. The sympathetic nervous system promotes the development of immune-mediated arthritis. The studies show a significant analgesic potential of the neutralization of proinflammatory cytokines and of opioids which selectively inhibit peripheral neurons. Furthermore, they show that the modulation of neuronal mechanisms can beneficially influence the course of musculoskeletal diseases. DISCUSSION: Interventions in the interactions between the immune system and the nervous system hold a great therapeutic potential for the treatment of musculoskeletal diseases and pain.

[Effector T cells]. / Effektor-T-Zellen.

BACKGROUND: Although the present understanding of the immunopathogenesis of rheumatoid inflammation is still incomplete, there is substantial evidence that effector CD4+ T helper (Th) cells play a central role. RESULTS: In recent years, in addition to the established Th cell subsets Th1 and Th2 cells, other subsets, such as Th9, Th17, Th22 and T follicular helper (Tfh) cells have been described. Defining the contribution of T cells in the initiation and maintenance of inflammation has been augmented by the identification of functionally distinct subsets of effector Th cells that can be classified based on their cytokine and transcription factor profiles. CONCLUSION: Increasing knowledge of the role of these various T cell populations in chronic inflammation provides a better understanding and insights into the pathogenic mechanisms and chronification of rheumatic diseases.

Bisphenols, but not phthalate esters, modulate gene expression in activated human MAIT cells in vitro.

One route of human exposure to environmental chemicals is oral uptake. This is primarily true for chemicals that may leach from food packaging materials, such as bisphenols and phthalate esters. Upon ingestion, these compounds are transported along the intestinal tract, from where they can be taken up into the blood stream or distributed to mucosal sites. At mucosal sites, mucosal immune cells and in the blood stream peripheral immune cells may be exposed to these chemicals potentially modulating immune cell functions. In the present study, we investigated the impact of three common bisphenols and two phthalate esters on mucosal-associated invariant T (MAIT) cells in vitro, a frequent immune cell type in the intestinal mucosae and peripheral blood of humans. All compounds were non-cytotoxic at the chosen concentrations. MAIT cell activation was only slightly affected as seen by flow cytometric analysis. Phthalate esters did not affect MAIT cell gene expression, while bisphenol-exposure induced significant changes. Transcriptional changes occurred in ∼ 25 % of genes for BPA, ∼ 22 % for BPF and ∼ 8 % for BPS. All bisphenols down-modulated expression of CCND2, CCL20, GZMB and IRF4, indicating an effect on MAIT cell effector function. Further, BPA and BPF showed a high overlap in modulated genes involved in cellular stress response, activation signaling and effector function suggesting that BPF may not be safe substitute for BPA.

[Bad memories: understanding and removal of pathogenic consequences of immunological memory in inflammatory rheumatic diseases. The IMPAM consortium]. / Schlechte Erinnerungen: pathogene Konsequenzen des immunologischen Gedächtnisses bei entzündlich rheumatischen Erkrankungen verstehen und beseitigen. Das IMPAM-Konsortium.

The Research Consortium IMPAM (IMprinting of the PAthogenic Memory for rheumatic inflammation) has recently been funded by the Federal Ministry of Education and Research in Germany. Within this consortium ten different research groups, coordinated by the German Rheumatism Research Center (DRFZ) and the University Hospital Jena, will examine the molecular dialogue between immune system memory cells and mesenchymal cells in chronic rheumatic diseases, such as rheumatoid arthritis or ankylosing spondylitis. The consortium's aim is to understand and modulate these interactions therapeutically, such that the pathogenic imprinting of proinflammatory memory cells can be extinguished and the anti-inflammatory capacity of the patients' regulatory cells can be restored.

MAIT cell activation is reduced by direct and microbiota-mediated exposure to bisphenols.

Oral uptake is the primary route of human bisphenol exposure, resulting in an exposure of the intestinal microbiota and intestine-associated immune cells. Therefore, we compared the impact of bisphenol A (BPA), bisphenol F (BPF) and bisphenol S (BPS) on (i) intestinal microbiota, (ii) microbiota-mediated immunomodulatory effects and (iii) direct effects on mucosal-associated invariant T (MAIT) cells in vitro. We acutely exposed human fecal microbiota, Bacteroides thetaiotaomicron and Escherichia coli to BPA and its analogues BPF and BPS referring to the European tolerable daily intake (TDI), i.e. 2.3 µg/mL, 28.3 µg/mL and 354.0 µg/mL. Growth and viability of E. coli was most susceptible to BPF, whereas B.thetaiotaomicron and fecal microbiota were affected by BPA > BPF > BPS. At 354.0 µg/mL bisphenols altered microbial diversity in compound-specific manner and modulated microbial metabolism, with BPA already acting on metabolism at 28.3 µg/mL. Microbiota-mediated effects on MAIT cells were observed for the individual bacteria at 354.0 µg/mL only. However, BPA and BPF directly modulated MAIT cell responses at low concentrations, whereby bisphenols at concentrations equivalent for the current TDI had no modulatory effects for microbiota or for MAIT cells. Our findings indicate that acute bisphenol exposure may alter microbial metabolism and impact directly on immune cells.

[Intestinal microbiota as regulator of the immune system]. / Darmflora als Regulator des Immunsystems.

The intestinal microbiota contributes to the regulation of the intestinal immune system and protection against intestinal infections. Recent studies revealed that the locally restricted intestinal microbiota affects systemic immunity and influences the induction of autoimmunity.

[Th1, Th17 and Th1+17 cells]. / Th1-, Th17- und Th1+17-Zellen.

T helper cells contribute to the induction and maintenance of rheumatic inflammation through the secretion of cytokines. The analysis of Th1 cells expressing interferon-γ, Th17 cells expressing interleukin-17 and the newly described Th1+17 cells could give insight into the pathophysiological mechanisms of rheumatic diseases. This could lead to the development of novel, targeted therapeutic strategies.

T-bet expression by Th cells promotes type 1 inflammation but is dispensable for colitis.

The transcription factor T-bet is highly expressed by Th cells isolated from the inflamed intestine of Crohn's disease patients, and has been regarded a critical driver of murine T cell-induced colitis. However, we show here that T-bet expression by Th cells is not required for the manifestation of T-cell-induced colitis in the presence of segmented filamentous bacteria and Helicobacter hepaticus. T-bet expression by Th cells controls their survival and localization, their repertoire of chemokine and chemokine receptor expression, the accumulation of monocytes and macrophages in the inflamed colon, and their differentiation to the M1 type, i.e., type 1 inflammation. Nevertheless, T-bet-deficient Th cells efficiently induce colitis, as reflected by weight loss, diarrhea, and colon histopathology. T-bet-deficient Th cells differentiate into Th1/17 cells, able to express IFN-γ and IL-17A upon restimulation. While neutralization of IL-17A exacerbated colitis induced by wild-type or T-bet-deficient Th cells, neutralization of IFN-γ completely abolished colitis.

Abnormal 201Tl myocardial single photon emission computed tomography in energetic male patients with myocardial bridge.

Myocardial bridge is a relatively benign condition where a major coronary artery is bridged by a band of muscle and narrows during systole, particularly during rapid heart rates. Its clinical presentation and electrocardiogram (ECG) changes overlap with that of coronary artery disease. 201Tl myocardial perfusion imaging is thus frequently prescribed for further evaluation. This retrospective study was carried out to determine the 201Tl image patterns in patients with myocardial bridge. A total of 17 male patients (aged from 30 to 63 years) who had a positive exercise ECG and angiographic evidence of myocardial bridge in the mid-third of the left anterior descending coronary artery were recruited. Most of them were robust and received routine physical check-ups. They had no known heart disease or medication that affected cardiac function. The patients' clinical presentations, echocardiograph and exercise ECG findings were analysed. 201Tl single photon emission computed tomography (SPECT) was performed by intravenous injection of 201Tl (111 MBq) immediately following stress (treadmill or dipyridamole induced) and 4 h after stress, using a fixed, right angle camera equipped with a low energy, general purpose collimator. The images were interpreted independently by two experienced nuclear medicine physicians. Nine of the 17 patients had anterior chest pain during exercise. All patients had an abnormal ECG during exercise, including ST-T wave depression in leads II, III and aVF, and v4-6. Except for eight patients revealing reversible perfusion defect (R), 16 of the 17 patients also exhibited a partial reversible perfusion defect (PR) or a significant reverse redistribution (RR) scan pattern in the anterior or inferior walls of the left ventricle. Myocardial bridge should be taken into consideration in energetic male patients who had abnormal exercise ECGs and the corresponding patterns of Tl SPECT abnormalities including R, PR and RR.

Circumvention of MHC class II restriction by genetic immunization.

The fate of T cell responses to peptide-based vaccination is subject to constraints by the major histocompatibility complex (MHC), MHC restriction. Using as a model system of T and B cell epitopes from the circumsporozoite protein of Plasmodium falciparum malaria parasite, we show that vaccination by somatic transgene immunization readily primes Balb/c mice (H-2(d)) a strain previously reported to be non-responder to immunization with a synthetic peptide vaccine encompassing these epitopes. Following genetic vaccination Balb/c mice developed a primary T cell response comparable to that of the responder strain C57Bl/6 (H-2(b)). Following booster immunization on day 45 Balb/c mice responded with a typical T cell memory response. Priming induced the formation of specific antibodies, which rose sharply after booster immunization. These findings suggests that genetic immunization can circumvent MHC class II restriction.

Spontaneous transgenesis of human B lymphocytes.

DNA can cross the cell membrane by natural means, but the functional relevance of this phenomenon has not been fully elucidated. Here, we analyzed spontaneous transgenesis of human B cells using plasmid DNA coding for a functional immunoglobulin (Ig) heavy chain gene under the control of a B-cell-specific promoter. Using polymerase chain reaction (PCR), reverse transcriptase-PCR, and flow cytometry in combination, spontaneous transgenesis was documented in Burkitt's lymphoma cell lines, Epstein-Barr virus-transformed cell lines, and peripheral blood B lymphocytes of the mature naïve phenotype (IgM(+)/IgD(+)/CD27(-)). By immunoelectron microscopy, the internalized DNA was seen in the lysosomes/late endosomes and in the cytosol proximal to the nucleus. Importantly, spontaneously transgenic B cells processed and presented to major histocompatibility complex (MHC)-restricted T lymphocytes a peptide expressed in the transgenic product. This is the first demonstration that primary B lymphocytes possess a program for the spontaneous internalization of DNA, which in turn imparts the cell with new immunological functions. As spontaneous transgenesis is obtained using a nonviral vector, does not require prior cell activation, and is not associated with chromosomal integration, the findings reported here open new possibilities for genetic manipulations of mature naïve B lymphocytes for therapy and vaccination.