Welcome to the Autoimmunity 2021 Congress Calendar

Background and Aims

Background and Aims: Understanding the molecular mechanisms underlying immune disorder is helpful for improving therapeutic strategies for treating RA. We report the molecular programmes of isolated Treg cells controlling treatment-naïve RA patients using single-cell RNA sequencing (scRNA-seq).

Methods

Methods: PBMCs were isolated from early treatment-naïve RA patients. 1×10⁶ cells were cultured with anti-CD3 and anti-CD28in the presence or absence of As₂O₃ (0.1 µM). Then, Treg cells isolated by FACS from PBMCs . Treg cells were performed by single-cells analysis to gain variance gene.

Results

Results: To define populations and identify genome-wide gene expression, we isolated Treg with or without As₂O₃ (0.1 µM) from treatment-naïve RA patients and performed scRNA-seq. We performed a volcano plot visualization of gene expression between the cells of early treatment-naive RA patients.

Conclusions

Conclusions: In conclusion, four novel mRNAs were used to generate a valuable target gene, which can serve as an independent indicator to predict the immune status of treatment-naïve RA patients due to their influences on the distinct signaling pathways.

Background and Aims

A recent study demonstrated that regulatory B cells (Bregs) play a crucial role in autoimmune diseases. However, the molecular mechanisms underlying the regulatory function of Bregs remain controversial.

Methods

In this study, we collected human peripheral blood and utilized mouse autoimmune disease models to identify the frequency and function of Bregs in autoimmune diseases such as IBD and Hashimoto thyroiditis.

Results

We found an increased population of Bregs with promoted suppressive function in autoimmune diseases. Moreover, via gene expression microarrays and flow cytometry, we found that HSP70 expression was significantly up-regulated in Bregs especially in diseases. In vitro and in vivo studies explored that inhibiting HSP70 expression changed the regulatory function of Bregs. Furthermore, adoptively transferred Hsp70 overexpressed B cells effectively rescue diseases. Mechanistically, HSP70 modulated Breg suppressive function via directly producing secreted HSP70 and targeting conventional T cells.

Conclusions

Our findings revealed that HSP70 is a novel factor that moderates Breg function, indicating a viable therapy aimed to enhance Breg-mediated production of HSP70.

Background and Aims

Autoimmune diseases as a collective are generally secondary to a dysregulation of the innate and/or adaptive immune systems leading to organ and/or systemic disease in a genetically and environmentally predisposed host. There is no cure for autoimmune diseases currently, but therapy is geared towards controlling the chronic systemic disease towards remission, supporting affected organs and preventing acute flares using corticosteroids, disease modifying antirheumatic drugs (DMARDs) and more recently biologic therapies.

Methods

Review of literature for evidence supporting the theory behind the immune system reboot therapy (ISRT) and its potential to cure autoimmune diseases.

Results

In theory, ISRT has the potential to cure immune mediated diseases which requires the elimination of all existing immune system memory without a bone marrow stem cell transplant, by allowing the pluripotent stem cells of the host to reboot, without reproducing the disease mimicking the original stem cell at birth since autoimmune disease are not congenital. One way in which such can be obtained is through a monitored water-only based prolonged fast of up to 30 days.

Conclusions

ISRT has a potential to completely cure immune mediated diseases. A prolonged water-only based monitored fast seems to be a potential mechanism to achieve this ideal, at least in theory.

Background and Aims

This study was designed to propose a simple “Fast Track algorithm” (Figure 1) for worldwide capillaroscopists of any level of experience to differentiate “scleroderma patterns” from “non-scleroderma patterns” on capillaroscopy (Figure 2) and to assess its inter-rater reliability.

Methods

Based on existing definitions to categorise capillaroscopic images as “scleroderma patterns” and taking into account the real life variability of capillaroscopic images described standardly according to the EULAR Study Group on Microcirculation in Rheumatic Diseases (EULAR SG MC/RD), a fast track decision tree, the “Fast Track algorithm” was created by the principal expert (VS) to facilitate swift categorisation of an image as “non-scleroderma pattern (category 1)” or “scleroderma pattern (category 2)”. Mean inter-rater reliability between all raters (experts/attendees) of the 8^th EULAR capillaroscopy course (Genoa, 2018) and, as external validation, of the 8^th EUSTAR course on systemic sclerosis (SSc) (Nijmegen, 2019) versus the principal expert, as well as reliability between the rater pairs themselves was assessed by mean Cohen’s and Light’s kappa coefficients.

Results

Mean Cohen’s kappa was 1/0.96 for the 6 experts/135 attendees of the 8^th EULAR capillaroscopy course and 1/0.94 for the 3 experts/85 attendees of the 8^th EUSTAR SSc course. Light’s kappa was 1/0.92 at the 8^th EULAR capillaroscopy course, and 1/0.87 at the 8^th EUSTAR SSc course.

Conclusions

For the first time, a clinical expert based fast track decision algorithm has been developed to differentiate a “non-scleroderma” from a “scleroderma pattern” on capillaroscopic images, demonstrating excellent reliability when applied by capillaroscopists with varying levels of expertise versus the principal expert.

Background and Aims

Distinguishing autoimmune myositis from other muscle disorders with prominent intramuscular inflammation such as muscular dystrophies, is often not easy yet highly relevant for guiding therapeutic decisions and for offering founded prognoses. We investigated the biomarker potential of growth differentiation factor-15 (GDF-15), a cytokine associated with inflammation, to differentiate myositis of autoimmune origin.

Methods

Sera from 35 myositis patients were collected, who had been diagnosed with immune-mediated necrotizing myopathy (n=21), sporadic inclusion body myositis (n=10), polymyositis (n=3), and dermatomyositis (n=1). GDF-15 protein levels were determined using a commercial enzyme-linked immunosorbent assay, and compared with the levels in sera from healthy controls (n=10), and patients with limb girdle muscular dystrophy or mitochondrial myopathies (n=8). Muscle tissue GDF-15 expression patterns were studied using double immunofluorescent staining.

Results

Serum levels of GDF-15 were significantly higher in myositis patients (625±358 pg/ml) than in healthy controls (326±204 pg/ml) (Mann-Whitney U test p=0.01), with no difference between patient subgroups (p=0.1). No correlation was found between GDF-15 levels and blood CK values (p=0.2). Patients with dystrophies displayed substantially lower levels (127±41 pg/ml), while patients with mitochondrial disease displayed the highest mean levels (1312±1393 pg/ml). GDF-15 could be localized to skeletal muscle fibers, with strong sarcoplasmic staining observed in subsets of small CD56 positive regenerating or denervated fibers, and in the inclusions present in sporadic inclusion body myositis fibers.

Conclusions

GDF-15 serum levels could represent a potential biomarker for myositis, with expression by skeletal muscle fibers being a probable source of the cytokine.

Background and Aims

Systemic sclerosis (SSc) is an autoimmune disease characterized by fibrosis and vascular abnormalities. No drug significantly influences the course of SSc, many patients resistant to standard treatment, it is suspected the addition of Physalis angulata Linn (Ciplukan herb) extract can improve the SSc skin fibrosis. The aim of this study was to evaluate the effect of Ciplukan extract on skin fibrosis based on MRSS, CRP, P1NP in SSc with standard therapy.

Methods

Double-blind randomized clinical trial was performed in SSc with stable disease at RSCM and RSHS (November 2015−March 2017) who met the inclusion criteria, continued the standard therapy. The subjects were randomly divided into two groups: the study group (Ciplukan 3 x 250 mg/day for 12 weeks) and the placebo group. MRSS, CRP, P1NP were performed every 4 weeks until the end of the study.

Results

Fifty-nine subjects, divided into two groups: 29 subjects (in the study group); 30 subjects (in the placebo group), the average age of 41 ± 9 years, the proportion of women: male = 9: 1. There was a significant improvement of skin fibrosis in the study group with a relative decrease in MRSS (35.9%) compared with placebo (6.3%; p <0.001), a relative decrease in P1NP (17.8%) versus placebo (0.7%; p = 0.002), and relative decrease in CRP (16,12%) versus placebo (3,29%; p = 0.181). The adverse effect was mild gastrointestinal symptoms.

Conclusions

Ciplukan extract with dose 3 x 250 mg for 12 weeks as adjuvant therapy on SSc with standard therapy, clinically and statistically improvement of skin fibrosis.

Background and Aims

The role of helminth treatment in autoimmune diseases is constantly growing. Systemic lupus erythematosus (SLE) is a multi-system autoimmune disease with challenging treatment options. Tuftsin phosphorylcholine (TPC) is a novel helminth-based compound that modulates the host immune network.

This study was conducted to evaluate the potential value of TPC in ameliorating lupus nephritis in a murine model and specifically to compare the efficacy of TPC to the existing first-line therapy for SLE: corticosteroids (methylprednisolone).

Methods

Lupus-prone NZBxW/F1mice were treated with TPC (5 µg/mouse), methylprednisolone (MP; 5mg/body weight) or PBS (control) three times per week once glomerulonephritis, defined as proteinuria of grade > 100 mg/dL, was established. Levels of anti-dsDNA autoantibodies were evaluated by ELISA, splenic cytokines were measured in-vitro, and the kidney microscopy was analyzed following staining.

Results

TPC and MP treatments significantly improved lupus nephritis and prolonged survival in NZBxW/F1 mice. TPC-treated mice showed a significantly decreased level of proteinuria (p<0.001) and anti-dsDNA antibodies (p<0.001) as compared to PBS-treated mice. Moreover, TPC and MP inhibited the production of the pro-inflammatory cytokines IFN-γ, IL-1β, and IL-6 (p<0.001) and enhanced expression of the anti-inflammatory cytokine IL-10 (p<0.001). Finally, microscopy analysis of the kidneys demonstrated that TPC-treated mice maintained normal structure equally to MP-treated mice.

Conclusions

This data indicates that the small molecule named TPC hinders lupus development in genetically lupus-prone mice equally as methylprednisolone in most of the cases. Hence, TCP may be employed as a therapeutic potential for lupus nephritis.

Background and Aims

Unlimited cytokine production regulates chronic inflammation and joint destruction in patients with inflammatory joint diseases. Chronic inflammation in such patients is induced by the imbalance between the pro- and antiinflammatory cytokines.Potential triggers for this process are the pro-inflammatory cytokines TNF-α and IL-6. The aim of the study is to assess the correlation between their levels and the activity indices in patients with inflammatory joint diseases .

Methods

213 patients with RA, AS and PsA and a control group of 31 healthy subjects were studied.All patients were tested for TNF-α and IL-6 prior to treatment initiation with blockers of TNF-α.Disease indices were used to evaluate disease activity.

Results

There is strong correlation between serum levels of TNF-α and IL-6 prior treatment in patients with RA(r_s=-0.018;p=0.001). Similar relationship exists between the tested cytokines and the index of activity and HAQ-DI (r_s=0.033;p=0.003). (R_s=-0.025;p=0.001)Between the levels of TNF-α and IL-6 prior treatment initiation in patients with AS has a strong correlation(r_s=-0.010;p=0.004). Relationship exists between the cytokines and activity index BASFI, ASDAS and BASDAI, respectively for TNF-α (r_s=-0.176;p=0.041),(r_s=0.342;p=0.001),(r_s=367;p=0.001) and for IL-6(r_s=0.406,p=0.016),(r_s=-0.083;p=0.002),(r_s=263;p=0.001). Between levels of TNF-α and IL-6 prior treatment in patients with PsA no significant correlation was observed(r_s=-0.031;p =0.827).There was also no significant correlation between DAPSA index and IL-6 (r_s=-0.187;p=0.237) and between IL-6 and HAQ-DI (r_s=-0.035;p =0.827) .

Conclusions

Between the serum levels of TNF -α and IL -6, and the indices for activity in patients with inflammatory joint disease there is a strong correlation relationship, which reflects the importance of these cytokines for clinical evaluation of patients.

Background and Aims

Immune checkpoint inhibitors have introduced a new and heterogeneous class of immune-related adverse effects, with the endocrine system being a predominant target for autoimmunity. Autoimmune hypothalamic-pituitary-adrenal (HPA) axis diseases induced by checkpoint inhibitors are being increasingly recognized. We aimed to characterize the spectrum of HPA axis endocrinopathies.

Methods

A retrospective cohort study all patients who received immunotherapy (Ipilimumab, Nivolumab, Pembrolizumab, Atezolizumab, Avelumab and/or Durvalumab) between January 2015 and October 2018 in a large tertiary cancer center. Patients were characterized for HPA axis abnormalities based on clinical and pituitary axes evaluation. The risk for developing HPA endocrinopathies was compared by log- rank test, by the time since checkpoint inhibitors initiation. Additionally, the risk for developing HPA endocrinopathies after adjusting for covariates was assessed using multivariable logistic regression analysis.

Results

Among 1615 patients, fourteen (0.87%) patients developed isolated adrecocorticotrophic hormone deficiency (IAD), six (0.37%) - hypophysitis and no case of adrenalitis was identified. IAD presented with mild and non-specific symptoms, mainly asthenia. In multivariable analysis, exposure to both PD-1/PD-L1 and Ipilimumab and female gender were associated with an increased odds ratio (OR) for developing IAD (6.98 [95% CI 2.38-20.47, p<0.001] and 3.67 [95% CI 1.13-11.84, p=0.03]), respectively.

Conclusions

Isolated ACTH Deficiencty, a rare disease before the immunotherapy era, has become a predominant checkpoint related HPA axis autoimmune injury. Despite its life threatening potential, IAD may be missed due to its subtle presentation. Patients exposed to Ipilimumab and PD-1/PD-L1 in combination or sequentially and women have an increased risk for developing IAD.

Background and Aims

To characterize the repertoires of targeted autoantigens in chronic inflammatory demyelinating polyneuropathy (CIDP) for systemic peculiarities, we detected the autoantigens of patients and controls and analyzed them in a holistic way.

Methods

We screened 43 human sera, comprising 22 CIDP patients, 12 patients with other neuropathies (ONP), and 9 apparently healthy controls (HC) via HuProt protein microarrays testing about 16,000 distinct human bait proteins in parallel. Autoantigen repertoires were analyzed via bioinformatical approaches of autoantigenomics: principle component and hierarchical cluster analysis, analysis of the repertoire sizes in disease groups and clinical subgroups, overrepresentation analyses using databases like those of Gene Ontology, Reactome Pathway, and pathway representation.

Results

1) The cohort of CIDP patients was heterogeneous regarding their autoantigen repertoires; 2) the number of the targeted antigens per CIDP patient depended on the clinical situation: intravenous immunoglobulin therapy responders targeted three times more autoantigens than non-responders; 3) a significant part of the autoantibody set specifically targeted proteins involved in anchoring junction components, motile cilium proteins, glycoprotein metabolic processes, and certain signaling pathway proteins; 4) ≥9 out of 23 central signaling proteins of the growth factor receptor pathway are targeted by CIDP-specific antibodies.

Conclusions

The repertoire of targeted autoantigens of CIDP patients differs systematically from those of controls. Considering systemic autoantigenomic approaches in addition to single antibody approaches may help to understand the disease and to discover novel options for improved diagnosis and prognosis.

Background and Aims

Sarcoidosis is one of granulomatous diseases. Granuloma formation is a result of lymphocytes activation. In different studies the changes in B cells and T cells subsets in blood and broncho-alveolar fluid were found. Alteration of B cells and T cells subsets might be the result of T follicular helpers (Tfh) activity. Nowadays there is no data about the role of T follicular helpers in sarcoidosis.

The aim of this study was to determine the peripheral blood Tfh in patients with sarcoidosis.

Methods

In a prospective comparative study conducted in 2016-2018 we analyzed peripheral blood circulating Tfh cell subsets in 34 patients with first diagnosed sarcoidosis and 30 healthy donors using multicolor flow cytometry. The statistical comparisons of data were performed using the Mann–Whitney U test and ROC analysis with GraphPad Prism Version 6.0 (USA).

Results

According to Mann-Whitney U test in sarcoidosis patients in comparison with healthy subjects the level of Tfh1 in effector and central memory was found to be decreased and the level of Tf2, Tfh17 increased (p<0.01). According to the ROC analysis the difference significant for diagnostic purpose wasn`t found (AUC ≤0.7).

Conclusions

In sarcoidosis group were found an increase of Tfh2 and Tfh17 cells, that induce antibody secretion in B cells, and a decrease in Tfh1 cells, that regulate the apoptosis of B cells. This results might be the additional confirmation of autoimmune origin of sarcoidosis.

Background and Aims

Sarcoidosis and tuberculosis have several similar clinical and pathogenetic characteristics, what makes some researchers consider a common pathogenesis for these diseases. HLA-genotypes are known as a genetic predisposition factor for many diseases and are studied for sarcoidosis and tuberculosis patients. However there is no comparative study of genetic predisposition for sarcoidosis or tuberculosis development.

The aim of this study was to analyze the relationship between HLA genotypes and the development of sarcoidosis and tuberculosis.

Methods

Original (n=19) and review articles (n=14) published in various online databases from 1960 to 2019 were studied. The papers were selected according to key words: sarcoidosis, tuberculosis, HLA-genotype, genetic predisposition.

Results

The study results showed opposite effects of the HLA genotypes associated with a predisposition to the development of sarcoidosis or tuberculosis. It was revealed, the genotypes predisposing to the development of sarcoidosis (HLA-DRB1*03/07/15) have protective properties against the development of tuberculosis. Moreover, genotypes causing its development (HLA-DRB1*04) have a protective effect on the development of sarcoidosis. Some of these results might be explained by the weak affinity of MHC molecules for mycobacterial antigens in patients with HLA-DRB1*04 genotype and the strong affinity of MHC proteins for bacterial antigens in individuals with HLA-DRB1*03: 01 genotype

Conclusions

The HLA system may determine the development of the immune response in contact with mycobacteria. It was shown that HLA-DRB1*04 genotype predispose to tuberculosis development, while the HLA-DRB1*03/07/15 genotypes to sarcoidosis. Determining the HLA genotypes may result in assessing a risk degree for developing tuberculosis or sarcoidosis in the foci of mycobacterial infection.