Mediated Diseases Research Articles

Is there an association between serum soluble interleukin-2 receptor levels and syndrome severity in persistent Complex Regional Pain Syndrome?

A potentially useful biomarker for Complex Regional Pain Syndrome (CRPS) is the serum soluble interleukin-2 receptor (sIL-2R) level, which is a marker for T-cell activation. Elevated serum sIL-2R levels have been described in CRPS patients compared to healthy controls. In T-cell mediated inflammatory diseases such as sarcoidosis and rheumatoid arthritis, the serum sIL-2R levels correlate with disease severity. In this study, we investigate whether an association exists between serum sIL-2R levels in CRPS patients and CRPS severity. A cross-sectional cohort study was conducted in a tertiary pain referral center in the Netherlands. Adult CRPS patients diagnosed by the IASP criteria were included between October 2018 until October 2022. The main study parameters were serum sIL-2R levels and the CRPS severity score. Fifty-three CRPS patients were included with a mean syndrome duration of 84 months (Q3 - Q1:180 - 48). The majority had persistent CRPS with a syndrome duration >1 year (n = 52, 98%). The median pain Numerical Rating Score (NRS) was 7 (Q3 - Q1: 8 - 5) and the mean CRPS severity score was 11 (SD ± 2.3). The median serum sIL-2R level was 330 U/mL (Q3 - Q1:451 - 256). No statistically significant correlation was observed between serum sIL-2R levels and the CRPS severity score (rs = 0.15, P = .28). Our findings suggest that serum sIL-2R levels cannot be used as a biomarker for syndrome severity in persistent CRPS (syndrome duration >1 year). Serial measurements of serum sIL-2R from early CRPS to persistent CRPS are needed to investigate whether serum sIL-2R levels can be used to monitor T-cell mediated inflammatory syndrome activity.

#3053 CORRELATION OF SYSTEMIC AND IN SITU BIOMARKERS OF COMPLEMENT ACTIVATION WITH HISTOPATHOLOGICAL FEATURES AND PROGNOSIS IN C3 GLOMERULOPATHY

Abstract Background and Aims C3 glomerulopathy is a rare complement mediated disease, resulting from complement alternative pathway (AP) activation. In addition to AP activation, there are emerging evidence for terminal pathway (TP) implication in kidney damage. A C3G histopathologic index has recently been proposed to evaluate both activity and chronicity parameters. Only a chronicity index > 4 has been independently validated as prognosis factor for end stage kidney disease. We aimed to determine the impact of local activation of the TP on disease phenotype. Method C3G histopathological index grading were evaluated via central review of 53 kidney biopsies from 43 patients carrying C3G. C5b-9 staining was performed on 53 FFPE kidney sections and quantified (scoring from grade 0 to 3 for C5b-9 deposits in glomeruli and from 0 to 2 in the tubulointerstitial compartment). Clinical data, C3 and sC5b-9 levels at the time of KB were retrospectively collected. Results KB were performed at diagnosis for n = 31. In 22 cases, KB was performed during follow up, under specific treatment (n = 10), without treatment (n = 7), or at disease relapse (n = 5). Twenty-five (47%) were children. Median[Q1-Q3] C3 level at KB was 623 mg/l [236-931]mg/l. Median[Q1-Q3] sC5b-9 level at KB, available in 16, was 374 ng/ml [203-1295]. Median [Q1-Q3] activity and chronicity index were respectively 9[6-12] and 1[0-6]. We confirmed the prognostic impact of chronicity index (p = 0.03) (Figure 1A) without impact of the activity index. Glomerular C5b-9 staining was positive in 47/53 (87%) (grade 1: n = 16 (30%); 2: n = 17 (32%); 3: n = 14 (26%)) and n = 40 (75%) had interstitial C5b-9 staining (grade 1: n = 28 (53%); 2: n = 12 (23%)). Soluble C5b-9 measurement correlated with intensity of C5b-9 glomerular deposit (coef corr = 0.69). As compared to patients with normal C3 level, patients with AP activation had higher proteinuria but kidney function at KB was similar. We found distinct histological features according to C3 plasmatic level (Table 1): patients with low C3 level had higher histological activity index (p = 0.03) and higher glomerular and tubulointerstitial C5b-9 staining (p = 0.001 and p = 0.002 respectively). C5b-9 glomeruli staining allows the identification of 3 groups of KB (1: no or low, 2: medium and 3: high C5b-9 staining) with distinct histological and clinical features. Activity index were higher in groups 2 & 3 (p = 0.05) and chronicity index in groups 1 & 3 (p = 0.003) (Table 2). Renal prognosis was poorer in patients with higher glomerular C5b-9 deposits (Figure 1B). Patients with chronicity index >4 and high C5b-9 staining had the poorest renal survival (Figure 1C). Conclusion Our results suggest that TP activation in glomeruli may have an impact on histological features, contributing therefore to renal prognosis. Deeper exploration of correlation between systemic and in situ complement activation and histological features could allow to better identify patients with worst prognosis or who could benefit from emerging complement inhibition therapeutics.

Monitoring immunomodulation strategies in type 1 diabetes.

Type 1 diabetes (T1D) is a T-cell mediated autoimmune disease. Short-term treatment with agents targeting T cells, B cells and inflammatory cytokines to modify the disease course resulted in a short-term pause in disease activity. Lessons learnt from these trials will be discussed in this review. It is expected that effective disease-modifying agents will become available for use in earlier stages of T1D. Progress has been made to analyze antigen-specific T cells with standardization of T cell assay and discovery of antigen epitopes but there are many challenges. High-dimensional profiling of gene, protein and TCR expression at single cell level with innovative computational tools should lead to novel biomarker discovery. With this, assays to detect, quantify and characterize the phenotype and function of antigen-specific T cells will continuously evolve. An improved understanding of T cell responses will help researchers and clinicians to better predict disease onset, and progression, and the therapeutic efficacy of interventions to prevent or arrest T1D.

Phase 1 dose-escalation, dose-expansion trial of intratumoral HER2- and HER3-primed dendritic cells injections for the treatment of early-stage TNBC and HR low positive breast cancer: DecipHER trial.

TPS629 Background: Patients with breast cancers (BCs) harboring low expression of hormone receptors (HRs) and human epidermal growth factor receptor-2 (HER2) have poor outcomes. Results from the KEYNOTE-522 trial showed that activation of the immune system using a PD1/PD-L1-targeted approach improves outcomes of patients with these high-risk tumors. Antigen-presenting cells (eg, dendritic cells [DCs]) are pivotal for robust cytotoxic responses due to broader activation of the adaptive immune system (ie, CD4+ and CD8+ Th1) against tumor-associated antigens (ie, HER2 or HER3) expressed by high-risk BCs. Methods: DecipHER is a dose-escalation, dose-expansion phase 1 trial designed to assess the safety and the preliminary efficacy of autologous, HER2- and HER3-primed DCs in combination with KEYNOTE 522 regimen in 30 patients. Patients with clinical stage cT1cN1/2 or cT2-4cN0-2, HR 20, HER2-negative BCs are eligible. Patients with inflammatory BC cancer and uncontrolled immune mediated diseases are excluded. After collection through aphaeresis, autologous DCs will be primed ex vivo against 6 HER2 and 8 HER3 immunogenic peptides. Participants will receive alternating ultrasound-guided intratumoral HER2 and HER3 DC injections administered twice a week for 8 doses in total starting 2 weeks prior to neoadjuvant therapy with KEYNOTE 522 regimen. The dose-escalation phase of the study has 3 planned cohorts (10-20, 30-50, 80-100 million DCs) and follows a 3+3 design (maximum n=18). The 12 additional patients enrolled will be treated at the maximum tolerated dose (MTD) in the dose-expansion cohort. The MTD will be defined as the highest dose level at which < 2 of 6 patients experience dose-limiting toxicities (grade ≥ 3 non-hematologic, ; grade ≥ 3 hematologic toxicity thought be at least possibly related to DCs ; any grade 4 nausea, vomiting, or diarrhea [or grade 3 if duration > 3 days]) during the 5 weeks following treatment initiation with DCs. Secondary endpoints include the absolute risk of adverse events, clinical and pathological responses, and recurrence free survival. Tumor tissue, blood and stool samples will be collected for correlative analyses. The study is open at H. Lee Moffitt Cancer Center. Clinical trial information: NCT05504707 .

Effect of Kinesiophobia Mediated Disease Perception on Rehabilitation Exercise Adherence in Patients Undergoing Percutaneous Coronary Intervention

Effect of Kinesiophobia Mediated Disease Perception on Rehabilitation Exercise Adherence in Patients Undergoing Percutaneous Coronary Intervention

Role of Complement in Kidney Diseases - New Aspects

Complement is a central part of the immune system. In the human body, complement is responsible for recognition of infectious microbes, for coordinating the adaptive immune response, controlling homeotic reactions and for the non-inflammatory removal of modified self-cells and infectious microbes. Complement is also closely linked to another proteolytic cascade, the coagulation system. Defective activation and altered complement regulation drives pathology of several severe human kidney diseases.This manuscript summarizes the latest developments on the role of complement in kidney diseases, on new complement inhibitors and on recent complement targeting therapies. In particular focusing on diseases (1) atypical Hemolytic Uremic Syndrome, (2) C3 Glomerulopathy, (3) Anti Neutrophil Cytoplasmic Antibody Mediated Vasculitis, (4) IgA Nephropathy, (5) Membranous Glomerulopathy, (6) Systemic Lupus Erythematosus, (7) Transplant rejection and (8) COVID 19 Infection-Triggered Kidney Diseases. More excitement is generated in this field, as more and more complement mediated diseases can be treated. Several complement targeting compounds are approved by the EMA and FDA and an increasing number of new candidates are in late phase clinical trials. In addition, clinical guidelines are developed for Diagnosis and Therapy of complement mediated diseases, new biomarkers are evaluated in clinical studies, and diagnostic guidelines are in development. The recent Covid infections showed a clear link of complement in thrombo inflammation, which ultimately results in kidney damage. These aspects have increased further the focus of complement inhibitors in COVID infections.

Risk stratified monitoring for methotrexate toxicity in immune mediated inflammatory diseases: prognostic model development and validation using primary care data from the UK

ObjectiveTo develop and validate a prognostic model to inform risk stratified decisions on frequency of monitoring blood tests during long term methotrexate treatment.DesignRetrospective cohort study.SettingElectronic health records within the UK’s...

Immune Checkpoint Inhibitor-related Pancreatitis: A Case Series, Review of the Literature and an Expert Opinion.

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment of various malignancies, but are associated with serious adverse events like pancreatitis. Current guidelines are limited to the first step in treating acute ICI-related pancreatitis with steroids but lack treatment advices for steroid dependent pancreatitis. We describe a case series of 3 patients who developed ICI-related pancreatitis with chronic features such as exocrine insufficiency and pancreatic atrophy at imaging. Our first case developed after treatment with pembrolizumab. The pancreatitis responded well after discontinuation of immunotherapy but imaging showed pancreatic atrophy and exocrine pancreatic insufficiency persisted. Cases 2 and 3 developed after treatment with nivolumab. In both, pancreatitis responded well to steroids. However during steroid tapering, pancreatitis recurred and the latter developed exocrine pancreatic insufficiency and pancreatic atrophy at imaging. Our cases demonstrate resemblances with autoimmune pancreatitis based on clinical and imaging findings. In line, both diseases are T-cell mediated and for autoimmune pancreatitis azathioprine is considered as maintenance therapy. Guidelines of other T-cell mediated diseases like ICI-related hepatitis suggest tacrolimus. After adding tacrolimus in case 2 and azathioprine in case 3, steroids could be completely tapered and no new episodes of pancreatitis have occurred. These findings support the idea that the treatment modalities for other T-cell mediated diseases are worthwhile options for steroid dependent ICI-related pancreatitis.

IL-10-Engineered Dendritic Cells Modulate Allogeneic CD8+ T Cell Responses.

Tolerogenic dendritic cells (tolDC) play a central role in regulating immune homeostasis and in promoting peripheral tolerance. These features render tolDC a promising tool for cell-based approaches aimed at inducing tolerance in T-cell mediated diseases and in allogeneic transplantation. We developed a protocol to generate genetically engineered human tolDC overexpressing IL-10 (DCIL-10) by means of a bidirectional lentiviral vector (LV) encoding for IL-10. DCIL-10 promote allo-specific T regulatory type 1 (Tr1) cells, modulate allogeneic CD4+ T cell responses in vitro and in vivo, and are stable in a pro-inflammatory milieu. In the present study, we investigated the ability of DCIL-10 to modulate cytotoxic CD8+ T cell responses. We demonstrate that DCIL-10 reduces allogeneic CD8+ T cell proliferation and activation in primary mixed lymphocyte reactions (MLR). Moreover, long-term stimulation with DCIL-10 induces allo-specific anergic CD8+ T cells without signs of exhaustion. DCIL-10-primed CD8+ T cells display limited cytotoxic activity. These findings indicate that stable over-expression of IL-10 in human DC leads to a population of cells able to modulate cytotoxic allogeneic CD8+ T cell responses, overall indicating that DCIL-10 represent a promising cellular product for clinical applications aimed at inducing tolerance after transplantation.

Tolerogenic IL-10-engineered dendritic cell-based therapy to restore antigen-specific tolerance in T cell mediated diseases

Tolerogenic dendritic cells play a critical role in promoting antigen-specific tolerance via dampening of T cell responses, induction of pathogenic T cell exhaustion and antigen-specific regulatory T cells. Here we efficiently generate tolerogenic dendritic cells by genetic engineering of monocytes with lentiviral vectors co-encoding for immunodominant antigen-derived peptides and IL-10. These transduced dendritic cells (designated DCIL−10/Ag) secrete IL-10 and efficiently downregulate antigen-specific CD4+ and CD8+ T cell responses from healthy subjects and celiac disease patients in vitro. In addition, DCIL−10/Ag induce antigen-specific CD49b+LAG-3+ T cells, which display the T regulatory type 1 (Tr1) cell gene signature. Administration of DCIL−10/Ag resulted in the induction of antigen-specific Tr1 cells in chimeric transplanted mice and the prevention of type 1 diabetes in pre-clinical disease models. Subsequent transfer of these antigen-specific T cells completely prevented type 1 diabetes development. Collectively these data indicate that DCIL−10/Ag represent a platform to induce stable antigen-specific tolerance to control T-cell mediated diseases.

Insights into the expanding intestinal phenotypic spectrum of SOCS1 haploinsufficiency and therapeutic options

PurposeHyper activation of the JAK-STAT signaling underlies the pathophysiology of many human immune–mediated diseases. Herein, the study of 2 adult patients with SOCS1 haploinsufficiency illustrates the severe and pleomorphic consequences of its impaired regulation in the intestinal tract.MethodsTwo unrelated adult patients presented with gastrointestinal manifestations, one with Crohn’s disease-like ileo-colic inflammation refractory to anti-TNF and the other with lymphocytic leiomyositis causing severe chronic intestinal pseudo-occlusion. Next-generation sequencing was used to identify the underlying monogenic defect. One patient received anti-IL-12/IL-23 treatment while the other received the JAK1 inhibitor, ruxolitinib. Peripheral blood, intestinal tissues, and serum samples were analyzed before-and-after JAK1 inhibitor therapy using mass cytometry, histology, transcriptomic, and Olink assay.ResultsNovel germline loss-of-function variants in SOCS1 were identified in both patients. The patient with Crohn-like disease achieved clinical remission with anti-IL-12/IL-23 treatment. In the second patient with lymphocytic leiomyositis, ruxolitinib induced rapid resolution of the obstructive symptoms, significant decrease of the CD8+ T lymphocyte muscular infiltrate, and normalization of serum and intestinal cytokines. Decreased frequencies of circulating Treg cells, MAIT cells, and NK cells, with altered CD56bright:CD16lo:CD16hi NK subtype ratios were not modified by ruxolitinib.ConclusionSOCS1 haploinsufficiency can result in a broad spectrum of intestinal manifestations and need to be considered as differential diagnosis in cases of severe treatment-refractory enteropathies, including the rare condition of lymphocytic leiomyositis. This provides the rationale for genetic screening and considering JAK inhibitors in such cases.

Combinatorial inhibition of Tec kinases BTK and ITK is beneficial in ameliorating murine sclerodermatous chronic graft versus host disease.

Graft-versus-host disease (GVHD) is the major factor limiting the widespread use of potentially curative allogeneic hematopoietic stem cell transplant (allo-HSCT). Chronic GVHD is characterized by the activation of alloreactive donor immune cells, especially B- and T-cells, leading to tissue damage and pathogenic fibrosis. In this study, we used highly specific next-generation inhibitors of ITK (PCYC-274), BTK (PCYC-804), and ibrutinib-like BTK/ITK inhibitors (PCYC-914 and PCYC-401) in the B10.D2 → BALB/C model of murine sclerodermatous cGVHD. From the third week onward, allogeneic recipients in each group of respective Tec kinase inhibitors were treated three times weekly with inhibitors at doses of 10 and 30 mg/kg or with saline control via oral gavage. Overall, we found that selective BTK inhibition was less effective than combined ITK/BTK or ITK inhibition in lengthening survival and reducing symptoms of cGVHD. ITK inhibition was most efficacious, with PCYC-274 and PCYC-401 demonstrating a nearly 50 percent reduction in GVHD scoring even at the 10 mg/kg dose, while 30 mg/kg of these compounds almost completely ameliorated GVHD symptomology. BTK/ITK and ITK-treated mice showed significant reductions in overall pathology. Significant reductions in dermal thickness and fibrosis were shown for all treatment groups. There was evidence of mixed Th1 and Th2 cytokine profiles in the skin of mice with dermal cGVHD, as both IFN-gamma and IL-4 were upregulated in the allogeneic control group, while kinase inhibition significantly reduced levels of these cytokines. Using an in vitro model of T-cell polarization, Th1 cell production of TNF-alpha and IFN-gamma were partially blocked by ITK. Th2 cell production of IL-4 was almost completely blocked synergistically by ITK and BTK inhibition. BTK-specific inhibition was unable to block either Th1 or Th2 cytokine production. Taken together, these results confirm previous reports that ITK-focused inhibition inhibits Th1 and Th2 cells. Additionally, the compound's effects on T-cell proliferation were tested by CFSE assay. Pure ITK inhibition was most effective at blocking T-cell proliferation, with no proliferation in PCYC-274-treated cells even at 0.1uM. PCYC-401 and PCYC-914 showed some inhibition at lower doses, with complete inhibition evident at 10uM. PCYC-804 was only partially able to block proliferation even at 10uM. In conclusion, we observed substantial benefit for differential inhibition of Tec kinases in GVHD, with ITK being most efficacious and Th1 cells being more resistant to inhibition, matching the previously reported findings of a Th2 to Th1 selective pressure in cells treated with ibrutinib. Our data warrants the further development of ITK and ITK/BTK inhibitors with specific inhibitory ratios to improve the treatment of GVHD and other T-cell mediated diseases.

Transcriptional regulatory signatures of systemic diseases in periodontitis with dyslipidemia

BackgroundPeriodontitis is a chronic bacterial infection of tooth that increases the risk of systemic diseases like diabetes, cancer, and cardiovascular diseases. Although it has been have found altered lipids in periodontitis patients, its gene regulation is largely unknown. This study aimed to examine the lipid meditated regulatory network in periodontitis that may helpful in early detection of periodontal mediated systemic diseases. MethodsWe employed a high-throughput gene expression data of 1) patients with periodontitis (n = 6); 2) the periodontitis patients with dyslipidemia (n = 6); and 3) healthy control group (n = 6). The over represented (DEGs) genes in SET-A (control vs. periodontitis) and SET-B (control vs. periodontitis with dyslipidemia) was identified. The protein interaction network was generated for the over represented genes in both the conditions. The constructed network was dissected into multiple regulatory clusters, containing over expressed transcription factors with its interacting proteins. Further the behavior of the clusters was determined through gene ontology and molecular pathways. ResultsOn expression analysis, 751 in SET-A and 561 in SET-B were over expressed compared to healthy control. Using over expressed genes, protein interaction networks were constructed for SET-A and SET-B, respectively. Topological analysis revel the difference in the complexity of both the network. Four regulatory clusters (ESR1, FOS, RUNX2, and SP1) from SET-A and six (ESR1, ESR2, FOS, JDP2, PBX1, and TAL1) from the SET-B network was extracted. Each cluster displayed a variety of molecular mechanism associated with immune system, cell cycle, and signal processing. Clusters from SET-B showed diverse regulatory pattern in associated with cancer, neurological, psychiatric and metabolic diseases. ConclusionOur findings demonstrate difference in regulatory patterns between periodontitis and periodontitis with dyslipidemia. These finding may provide evidence for dyslipidemia mediated periodontitis contribute to progressive systemic diseases. Further experiments are required to validate these regulators as biomarkers and drug targets.

Tuning peptide specificity for T cell tolerance in Type 1 diabetes

Abstract Type 1 Diabetes (T1D) is a classical T-cell mediated autoimmune disease, and fundamental data have implicated insulin as the dominant autoantigen in T1D disease. In the NOD mouse model of T1D, notable studies have found that mice lacking native insulin expression but with an altered insulin sequence to maintain blood glucose levels are protected from insulitis and diabetes. Growing evidence also indicates how the insulin peptide is bound within MHC Class II (peptide register) is essential in determining the strength of interactions and recognition of autoreactive T-cells. We have uncovered a unique peptide binding characterized by the dominant insulin epitope InsB:9–23. The majority of InsB:9–23-specific CD4 T-cells in the periphery recognize insulin bound in the unusual register 3, and knocking in this single amino acid variation into just one copy of the insulin gene in NOD mice, confers protection against T1D. In addition, these mice were protected from T1D, even when treated with checkpoint inhibitor blockade, unlike their wild-type littermates. Introducing this superagonist epitope into the NOD mouse has allowed us to address several key questions surrounding pathways of peptide generation, presentation by MHC molecules, and recognition by auto-reactive pathogenic T cells. In addition, we have been able to model how central and peripheral tolerance mechanisms can be altered when major epitopes in the Insulin gene are mutated, allowing us to explore means of dominant tolerance to understand the potential for translation into treatments for T1D. Supported by grants from NIH (R01 DK133443-01A)

The investigation of selective sphingosine-1 phosphate receptor 1&4 modulator to treat alopecia areata mouse model

Abstract Alopecia areata (AA) is a T cell mediated autoimmune disease characterized phenotypically by hair loss. NKG2D +CD8 +T cells is involved in the pathogenesis of AA. Sphingosine-1 phosphate (S1P) is a biologically active metabolite of cell membrane sphingolipids, which is essential for immune cell transport, and is known to be mainly involved in immune and inflammatory reactions. Due to involvement of S1P receptor (S1PR) in the immune system, S1PR modulator is being investigated for application in multiple sclerosis and other conditions. Among the five S1PRs, S1PRs 1&4 modulator (NXC736, Nextgen Bioscience, South Korea) was used to confirm its efficacy in the treatment of AA. AA lesion size measurement, disease-free ratio, immunohistochemistry and flow cytometry were used for analysis method. After oral administration of NXC736 for 84 days to AA mice, it was confirmed that the AA lesion area and disease-free ratio were significantly decreased compared to control group, and the infiltration of CD8 +T cells in the lesion skin was also significantly decreased. In addition, NKG2D level was decreased in effector CD8 +T cells. According to this result, we speculate that S1PR 1&4 modulator can be used as a promising treatment for AA. Supported by Nextgen Bioscience

A computational pipeline for drug discovery in immunometabolism against autoimmune diseases

Abstract We developed a computational pipeline to study immunometabolism for drug discovery in immune mediated diseases. Our pipeline integrates RNA-seq and proteomics data to build genome scale metabolic models and integrate these models with external drug databases and patient data to perform metabolic flux analyses for identifying drug targets and repurposing. We applied our pipeline to explore the metabolism of B cells in the context of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). We constructed metabolic models of B cells, performed enzymatic inhibition of targets of existing drugs and compounds, and integrated the model's results with differentially expressed genes in RA and SLE patients. Using a computational scoring method, we ranked the inhibited targets based on their ability to stimulate the reactions of differentially expressed genes in diseases to have metabolic flux in opposite directions of their up and down-regulation. As a result, we identified 25 B cell drug targets against RA and 23 B cell drug targets against SLE. We validated these targets by comparing them against the current treatment options for these diseases. For RA, five of the identified targets are genes that are already targeted by existing drugs used to treat RA. Our top-ranked targets for RA are adenine phosphoribosyltransferase, galactosidase beta 1, and aldehyde dehydrogenase 5 family member A1. For SLE, the top-ranked new targets are geranylgeranyl diphosphate synthase 1, glutathione-disulfide reductase, and galactosidase beta 1. Our pipeline provides a streamlined approach for identifying drug targets and predicting repurposable drugs and can facilitate the investigation of novel indications for diseases to improve global health. Supported by Layman Seed grant from University of Nebraska Foundation, and NIH grant R35GM119770.

Immune system perturbations in patients with severe long COVID

Abstract SARS-CoV-2 infections have different outcomes ranging from mild to severe acute infection as well as post-infectious multi-system inflammatory syndromes (MIS-C/A). Post-acute sequelae of COVID-19 (PASC) or long COVID is another outcome, which is poorly understood and range from mild to very severe persistent organ dysfunction and damages. To maximize our chances of identifying underlying mechanisms of long COVID we focus on the most severe cases with objective measures of disease such as lung damages, microvascular and autonomic nerve dysfunction. We are testing the following hypotheses that I) long COVID is a super-antigen mediated disease, II) an autoimmune condition driven by pathogenic autoantibodies, and/or III) a disease caused by viral persistence. A cohort of over 100 long COVID patients, 24 recovered long COVID patients as well as 9 COVID convalescent controls have been recruited thus far and blood samples analyzed for plasma proteome (Olink assays), immune cell composition and phenotypes (mass cytometry), T-cell repertoire (scTCR-seq), innate cell activation (circulating nucleosomes), and auto-antibody analyses (Luminex arrays) in order to understand immune perturbations in severe long COVID patients. Unlike children with hyperinflammatory MIS-C, we have not found any evidence of superantigen-mediated T-cell activation in long COVID, but instead substantial clonal expansion of memory T cell clones with effector memory states and shared clonality across patients. Additionally, cytokine perturbations such as elevated levels of IL8 and elevated levels of circulating nucleosomes indicate persistent activation of neutrophils in severe long COVID.

High-throughput Western Blot for in vitro and in vivo assessment of human Ig specific cleaving enzymes

Abstract Antibodies play a vital role in the body’s immune defense mechanism. However, pathogenic Ig antibodies are thought to contribute to several autoimmune conditions and transplant rejection. Proteases derived from human pathogens can specifically cleave IgG into F(ab’)2 and Fc fragments, thus allowing for rapid clearance of IgG. This unique trait suggests a novel opportunity to use these molecules to treat autoantibody mediated diseases. Using our IMPACT platform leveraging machine learning, we have engineered Ig proteases that specifically cleave IgG, and treatment with these proteases causes cleavage below the hinge region, which can result in the generation of the following degradation products: 1) Fab’2 and Fc fragments lacking effector function; or 2) single chain cleaved fragments with reduced effector functions. To detect these species, we developed a method using JESS, an automated high throughput western blot instrument. Using this system, we can detect intact, single cleaved and fully cleaved IgG in a time and dose dependent manner in both in vitro and in vivo assays. In conclusion, we successfully optimized a high throughput assay format that is highly specific and sensitive in evaluating the cleavage activity of different Ig proteases and could be a powerful tool to assess efficacy of IgG cleaving enzymes in the clinic.

Toward smartphone-based blood pressure monitoring

A single-centre, prospective longitudinal study to assess changes in maternal arterial stiffness and cardiac output parameters among low-risk healthy pregnant women.Thirty low-risk, healthy, pregnant women attending their routine antenatal dating ultrasound scan were recruited. Non-invasive assessment of arterial stiffness and cardiac output was undertaken at five gestational windows from 11 to 40 weeks of pregnancy. Data were analysed using a linear mixed model incorporating time and other relevant predictors as fixed effects, and patient as a random effect.Gestational age had a significant effect on all arterial stiffness parameters, including brachial augmentation index (AIx) (p = .001), aortic AIx (p = .002) and aortic pulse wave velocity (p = .002). The aortic AIx (%) reduced during pregnancy: the lowest mean (standard error, SE) was 4.07 (1.01) at 28 weeks before it increased to 7.04 (SE 1.64) at 40 weeks.Similarly, non-invasive assessments of cardiac output (p < .001), stroke volume (p = .014), heart rate (p < .001) and total peripheral resistance (p < .001) demonstrated significant changes with gestational age. Mean cardiac output (l/m) increased during pregnancy reaching a peak at 28 weeks gestation 6.66 (SE 0.28), but dropped thereafter to reach 5.71 (SE 0.25) around term.The current study provides pregnancy normograms for gestational changes in arterial stiffness and cardiac output parameters among low-risk, healthy pregnant women. Further work will be required to assess the risk of placental mediated diseases and pregnancy outcome among pregnant women with parameters outside the normal range.

Association between psoriasis, sleep, and dermatological quality of life: results of a cross-sectional study.

Psoriasis is a chronic relapsing inflammatory T-cell mediated disease who affects patients' daily activities and life quality. The association between sleep quality, dermatological quality of life (QoL) and psoriasis severity has been poorly investigated to date. The aim of this study is to investigate how sleep quality impacts on the severity of psoriasis, and to assess whether the different therapies used for psoriasis affect the dermatological QoL. We conducted a cross-sectional study on 152 adult patients based on specific questionnaires about the sleep quality (PSQI) and the dermatological quality of life (DLQI). Patients were divides into three groups according to severity (mild, moderate and severe) and therapy (group 1: no current therapy or exclusive use of topical drugs, group 2: use of conventional systemic drugs and group 3: biologics). The outcomes were expressed in the form of an Odd Ratio (OR) and for each variable it was commented whether the OR obtained was statistically significant or not. Inferential statistics comparing patients' DLQI showed that patients in group 3 and group 1 had comparable results. The OR obtained allowed us to state that those not taking biological drugs have a 4-fold higher risk of developing severe psoriasis than those taking them as therapy. No statistical difference was highlighted about sleep quality. This emphasizes that adequate therapy with biologic drugs allows patients with severe psoriasis to have a comparable QoL to those who are not impaired enough to require systemic or biologic therapy.