APRIL Levels Research Articles

Non-invasive monitoring of adrenocortical activity in the Gould’s wattled bat (Chalinolobus gouldii)

Although bats are the second most species-rich mammalian order, very little is known about their endocrine physiology. Glucocorticoids (GCs) are commonly associated with the stress response, but also modulate vital physiological functions which help animals adapt to their environment. Understanding normal patterns of adrenocortical activity can provide valuable insights into a species’ fitness. Non-invasive hormone monitoring via faecal samples provides an integrated measure of adrenocortical activity while minimising stress on the animal but must be properly validated to ensure reliable results. The goal of this study was to validate an enzyme immunoassay for monitoring faecal glucocorticoid metabolites (FGMs) in a common Australian insectivorous bat species, the Gould’s wattled bat (Chalinolobus gouldii). We compared the performance of five assays for monitoring changes in FGMs following capture and transfer of C.gouldii from the wild to captivity. Four of the five assays detected a significant increase in FGMs following capture, but the magnitude of the increase and consistency across individuals differed considerably. We selected the UVM-69a assay as the best performing assay to then describe normative patterns of adrenocortical activity in the species. Males had higher FGM levels than females, and juveniles had higher FGM levels than adults. Individuals with poorer body condition had higher FGM levels. We also demonstrate seasonal patterns of FGMs with higher levels in March and April corresponding with reproductive up-regulation and lower levels in May and November. Our study is the first of its kind to examine adrenocortical activity in an Australian insectivorous bat and provides a valuable tool for studying this species. Understanding adrenal function in common species such as C.gouldii can shed light on the physiological mechanisms facilitating survival and success in changing environments.

Characterization of Inflammation/Immune-, Acute Phase-, Extracellular Matrix-, Adhesion-, and Serine Protease-Related Proteins in the Amniotic Fluid of Women With Early Preterm Prelabor Rupture of Membranes.

To determine whether altered concentrations of various inflammation/immune-, acute phase-, extracellular matrix-, adhesion-, and serine protease-related proteins in the amniotic fluid (AF) are independently associated with microbial invasion of the amniotic cavity and/or intra-amniotic inflammation (MIAC/IAI), imminent spontaneous preterm delivery (SPTD; ≤7 days), and major neonatal morbidity/mortality (NMM) in women with early preterm prelabor rupture of membranes (PPROM). This was a retrospective cohort study involving 111 singleton pregnant women with PPROM (24-31 weeks) undergoing amniocentesis to diagnose MIAC/IAI. The following proteins were measured in stored AF samples by enzyme-linked immunosorbent assay (ELISA): APRIL, DKK-3, Gal-3BP, IGFBP-2, IL-8, VDBP, lumican, MMP-2, MMP-8, SPARC, TGFBI, TGF-β1, E-selectin, ICAM-5, P-selectin, haptoglobin, hepcidin, SAA1, kallistatin, and uPA. Multivariate logistic regression analyses revealed that (i) elevated APRIL, IL-8, MMP-8, and TGFBI levels in the AF, reduced lumican and SPARC levels in the AF, and high percentages of samples above the lower limit of quantification for AF TGF-β1 and uPA were significantly associated with MIAC/IAI; (ii) elevated AF levels of IL-8 and MMP-8 were significantly associated with SPTD within 7 days; and (iii) elevated AF IL-6 levels were significantly associated with increased risk for major NMM, when adjusted for baseline covariates. ECM (lumican, SPRAC, TGFBI, and TGF-β1)- and serine protease (uPA)-associated proteins in the AF are involved in the regulation of the host response to infection/inflammation in the amniotic cavity, whereas AF inflammation (IL-8, MMP-8, and IL-6)-associated mediators are implicated in the development of preterm parturition and major NMM in early PPROM.

Characterization of the BCMA epitope bound by BCMA-CD3 T cell engager elranatamab.

7546 Background: Elranatamab is a BCMA-CD3 bispecific antibody with an accelerated approval in the US and EU for relapsed/refractory multiple myeloma (RRMM). Elranatamab binds to CD3 on T cells and BCMA on MM tumor cells; this dual binding results in T cell activation, cytokine release, and tumor cell killing. Engagement of the BCMA surface receptor by its ligands, APRIL and BAFF, mediates plasma cell proliferation and survival. Elevated BCMA, as well as BAFF and APRIL are present in MM patients, yet in vitro functional assays have shown physiologically relevant soluble APRIL or BAFF concentrations did not significantly impact elranatamab activity. Moreover, emerging clinical data from relapsed patients has identified novel, yet relatively rare BCMA protein mutations that may impact BCMA ligand or targeted modality binding. Here we characterized the BCMA binding affinity, evaluated the impact on BCMA binding to BAFF and APRIL interactions, and identified the BCMA binding epitope. We also generated a model from crystal structure data to map reported BCMA mutations identified in patients. Methods: Elranatamab binding affinity to recombinant human BCMA and characterization of BAFF and APRIL interactions with elranatamab-bound BCMA were calculated via SPR at 37˚C with a classical sandwich format. The elranatamab BCMA binding epitope was identified by a co-crystal structure with a parental anti-BCMA Fab that has identical CDR sequences to elranatamab. Alpha-fold modeling was used to map identified BCMA mutations on the antibody/BCMA interface. Results: Elranatamab binds to BCMA with higher affinity (~38 pM) compared to reported binding affinities of BAFF (μM range) and APRIL (nM range) ligands. The identified elranatamab epitope largely overlaps with known ligand epitopes, supporting that elranatamab higher affinity BCMA binding blocks ligand binding. Alpha fold modeling of four recently reported rare BCMA mutations shows that they lie along the binding interface. Conclusions: Due to its high affinity for BCMA, elranatamab potency is unlikely to be impacted by elevated levels of soluble BAFF or APRIL in RRMM patients. Future studies are needed to characterize the impact of emerging MM patient BCMA mutations on the binding and function of anti-BCMA targeting modalities, particularly given the small extracellular domain of BCMA available for therapeutic targeting and the highly overlapping binding epitopes reported for approved BCMA agents.

Determining the Climatic Drivers for Wine Production in the Côa Region (Portugal) Using a Machine Learning Approach

The Côa region in inner-northern Portugal heavily relies on viticulture, which is a cornerstone of its economy and cultural identity. Understanding the intricate relationship between climatic variables and wine production (WP) is crucial for adapting management practices to changing climatic conditions. This study employs machine learning (ML), specifically random forest (RF) regression, to predict grapevine yields in the Côa region using high-resolution climate data for 2004–2020. SHAP (SHapley Additive exPlanations) values are used to potentially explain the non-linear relationships between climatic factors and WP. The results reveal a complex interplay between predictors and WP, with precipitation emerging as a key determinant. Higher precipitation levels in April positively impact WP by replenishing soil moisture ahead of flowering, while elevated precipitation and humidity levels in August have a negative effect, possibly due to late-season heavy rainfall damaging grapes or creating more favorable conditions for fungal pathogens. Moreover, warmer temperatures during the growing season and adequate solar radiation in winter months favor higher WP. However, excessive radiation during advanced growth stages can lead to negative effects, such as sunburn. This study underscores the importance of tailoring viticultural strategies to local climatic conditions and employing advanced analytical techniques such as SHAP values to interpret ML model predictions effectively. Furthermore, the research highlights the potential of ML models in climate change risk reduction associated with viticulture, specifically WP. By leveraging insights from ML and interpretability techniques, policymakers and stakeholders can develop adaptive strategies to safeguard viticultural livelihoods and stable WP in a changing climate, particularly in regions with a rich agrarian heritage, such as the Côa region.

Serum BAFF, APRIL, TWEAK, TNFSF18, TNFR2, and TNFRS12A levels in preschool children with autism spectrum disorder

Objectives A gradually increasing number of studies have reported that dysregulation in the immune system may be a contributing factor in the etiopathogenesis of autism spectrum disorder (ASD). However, only limited data has existed about the role of tumor necrosis factor (TNF) superfamily members in ASD. This study aimed to investigate whether serum levels of BAFF, APRIL, TWEAK, TNFSF18 TNFR2, and TNFRS12A are associated with ASD in preschool children. Methods A total of 65 (52 boys/13 girls) preschool children with ASD and 40 (28 boys/12 girls) healthy controls were included in the study. The severity of the ASD symptoms was assessed by the Autism Behavior Checklist and the Childhood Autism Rating Scale. Serum levels of immunologic parameters were measured by enzyme-linked immunosorbent assay. Results Analysis indicated that serum BAFF, TNFSF18, TNFR2, and TNFRS12A levels were significantly lower and serum APRIL and TWEAK levels were significantly higher in the ASD group than in the control group. However, with logistic regression only low BAFF levels and high APRIL levels had relationships with the presence of ASD. Conclusions These findings suggest that TNF superfamily members, especially the BAFF/APRIL system may play a role in the pathophysiology of ASD.

A comparative study of different antiviral treatment protocols in HCV related cryoglobulinemic vasculitis

The treatment of HCV and its sequelae are used to be predominantly based on Interferon (IFN). However, this was associated with significant adverse events as a result of its immunostimulant capabilities. Since their introduction, the directly acting antiviral drugs (DAAs), have become the standard of care to treat of HCV and its complications including mixed cryoglobulinemic vasculitis (MCV). In spite of achieving sustained viral response (SVR), there appeared many reports describing unwelcome complications such as hepatocellular and hematological malignancies as well as relapses. Prolonged inflammation induced by a multitude of factors, can lead to DNA damage and affects BAFF and APRIL, which serve as markers of B-cell proliferation. We compared, head-to-head, three antiviral protocols for HCV–MCV treatment As regards the treatment response and relapse, levels of BAFF and APRIL among pegylated interferon α-based and free regimens (Sofosbuvir + Ribavirin; SOF–RIBA, Sofosbuvir + Daclatasvir; SOF–DACLA). Regarding clinical response HCV–MCV and SVR; no significant differences could be identified among the 3 different treatment protocols, and this was also independent form using IFN. We found no significant differences between IFN-based and free regimens DNA damage, markers of DNA repair, or levels of BAFF and APRIL. However, individualized drug-to-drug comparisons showed many differences. Those who were treated with IFN-based protocol showed decreased levels of DNA damage, while the other two IFN-free groups showed increased DNA damage, being the worst in SOF–DACLA group. There were increased levels of BAFF through follow-up periods in the 3 protocols being the best in SOF–DACLA group (decreased at 24 weeks). In SOF–RIBA, CGs relapsed significantly during the follow-up period. None of our patients who were treated with IFN-based protocol had significant clinico-laboratory relapse. Those who received IFN-free DAAs showed a statistically significant relapse of constitutional manifestations. Our findings suggest that IFN-based protocols are effective in treating HCV–MCV similar to IFN-free protocols. They showed lower levels of DNA damage and repair. We believe that our findings may offer an explanation for the process of lymphoproliferation, occurrence of malignancies, and relapses by shedding light on such possible mechanisms.

#1422 Exploring BAFF, APRIL, and clinical characteristics in IgA nephropathy: a decade-long study in the Bulgarian population

Abstract Background and Aims IgA nephropathy (IgAN) represents a significant global health concern, contributing substantially to chronic kidney disease and renal failure. Despite its widespread impact, the Bulgarian population lacks comprehensive epidemiological data and specific biomarkers for IgAN. This decade-long retrospective study, conducted at a tertiary Nephrology Clinic in Bulgaria, aims to explore the clinical characteristics and serum levels of BAFF and APRIL in biopsy-confirmed IgAN patients and healthy controls. Method The study includes 125 biopsy-confirmed IgAN patients and 30 healthy controls. Notably, 70.4% of IgAN patients were male, with an average age of 35.94 ± 11.91 years. Baseline characteristics reveal advanced disease stages at diagnosis, possibly linked to delayed biopsy (24.22 ± 32.9 months from initial symptoms) and the absence of a screening program. Analyses of serum biomarkers (C3, BAFF, and APRIL) are conducted to identify any significant variations between IgAN patients and healthy volunteers. Results The average baseline estimated glomerular filtration rate (eGFR) was 61.78 ± 27.72 ml/min/1.73 m², with a mean serum creatinine of 126 ± 67.83 μmol/l at the time of kidney biopsy. Among the patients, 51.2% had an eGFR below 60 ml/min, indicating CKD stage 3 or worse according to the KDIGO classification Binary logistic regression analysis identifies anemia as a significant contributor to disease progression, with a risk influence approximately 3.3 times higher for GFR decline over time. Anemia emerges as a crucial predictor for serum creatinine doubling, carrying a higher risk than hyperuricemia. Analyses of C3 (p = 0.658), BAFF (p = 0.432), and APRIL (p = 0.227), demonstrated no significant differences in their serum levels between IgAN patients and healthy volunteers. Conclusion This study provides comprehensive insights into IgAN in the Bulgarian population, addressing the dearth of epidemiological data and biomarkers. Anemia emerges as a significant predictor for disease progression, emphasizing its potential clinical relevance. However, C3, BAFF, and APRIL exhibit no notable variations, underscoring the need for additional biomarker exploration to enhance understanding of IgAN dynamics in this population.

The transcription factor HMGB2 indirectly regulates APRIL expression and Gd-IgA1 production in patients with IgA nephropathy

Background IgA nephropathy (IgAN) is the most common primary glomerulonephritis worldwide. Proliferation-inducing ligand (APRIL) was identified as an important cause of glycosylation deficiency of IgA1 (Gd-IgA1), which can ‘trigger’ IgAN. Our previous study indicated that high migration group protein B2 (HMGB2) in peripheral blood mononuclear cells from patients with IgAN was associated with disease severity, but the underlying mechanism remains unclear. Materials and methods The location of HMGB2 was identified by immunofluorescence. qRT-PCR and Western blotting were used to measure HMGB2, HMGA1, and APRIL expression. Gd-IgA1 levels were detected by enzyme-linked immunosorbent assay (ELISA). In addition, we used DNA pull-down, protein profiling, and transcription factor prediction software to identify proteins bound to the promoter region of the APRIL gene. RNA interference and coimmunoprecipitation (Co-IP) were used to verify the relationships among HMGB2, high mobility group AT-hook protein 1 (HMGA1), and APRIL. Results HMGB2 expression was greater in IgAN patients than in HCs and was positively associated with APRIL expression in B cells. DNA pull-down and protein profiling revealed that HMGB2 and HMGA1 bound to the promoter region of the APRIL gene. The expression levels of HMGA1, APRIL, and Gd-IgA1 were downregulated after HMGB2 knockdown. Co-IP indicated that HMGB2 binds to HMGA1. The Gd-IgA1 concentration in the supernatant was reduced after HMGA1 knockdown. HMGA1 binding sites were predicted in the promoter region of the APRIL gene. Conclusion HMGB2 expression is greater in IgAN patients than in healthy controls; it promotes APRIL expression by interacting with HMGA1, thereby inducing Gd-IgA1 overexpression and leading to IgAN.

Indoor Radon Surveying and Mitigation in the Case-Study of Celleno Town (Central Italy) Located in a Medium Geogenic Radon Potential Area

Indoor radon surveying and remediation were implemented in a single-family home affected by high levels of indoor radon in the Celleno municipality (central Italy) with the aim of identifying the contribution of radon sources, evaluating the factors affecting radon entry into the building, and reducing radon risk. Average radon levels were relatively low at the ground floor (286 ± 202 Bq m−3) and first floor (167 ± 84 Bq m−3) in autumn when the temperature was still warm and the windows were open, but increased up to 2776 ± 1768 Bq m−3 and 970 ± 202 Bq m−3 in the first half of December, when the heating system was on and the windows were closed. The inner walls of the pilot room at the ground floor, semi buried on one side, were then treated with a waterproof product (a silane terminated polymer) and the average radon was halved (1475 ± 1092 Bq m−3) in the following month, which was still characterised by winter conditions. Radon entry in the room was identified and sealed with the same product, and a radon accumulation space behind a NE-SW oriented wall was naturally ventilated, reducing radon below the reference level in April with northerly winds conditions.

The paradox of spring: Thyroid and glucocorticoid responses to cold temperatures and food availability in free living Carneddau ponies

In seasonal environments, maintaining a constant body temperature poses challenges for endotherms. Cold winters at high latitudes, with limited food availability, create opposing demands on metabolism: upregulation preserves body temperature but depletes energy reserves. Examining endocrine profiles, such as thyroid hormone triiodothyronine (T3) and glucocorticoids (GCs), proxies for changes in metabolic rate and acute stressors, offer insights into physiological trade-offs. We evaluated how environmental conditions and gestation impact on faecal hormone metabolites (fT3Ms and fGCMs) from late winter to spring in a free-living population of Carneddau ponies. Faecal T3Ms were highest in late February and March, when temperatures were lowest. Then, fT3Ms concentrations decreased throughout April and were at the lowest in May before increasing towards the end of the study. The decline in fT3M levels in April and May was associated with warmer weather but poor food availability, diet diversity and diet composition. On the other hand, fGCM levels did not display a clear temporal pattern but were associated with reproductive status, where pregnant and lactating females had higher fGCM levels as compared to adult males and non-reproductive females. The temporal profile of fT3Ms levels highlights metabolic trade-offs in a changing environment. In contrast, the ephemeral but synchronous increase in fGCM concentrations across the population suggest a shared experience of acute stressors (i.e., weather, disturbance or social). This multi-biomarker approach can evaluate the role of acute stressors versus energy budgets in the context of interventions, reproduction, seasonality and environmental change, or across multiple scales from individuals to populations.

Clinical and Epidemiological Study of IgA Nephropathy in the Bulgarian Population: Insights into Disease Presentation and Potential Biomarkers.

IgA nephropathy (IgAN) is the most common glomerulonephritis worldwide and a leading cause of chronic kidney disease and renal failure. However, the Bulgarian population has limited epidemiological data and biomarkers for IgAN. In this retrospective monocentric analysis, we investigated all the patients with biopsy-proven IgAN over 10 years in a tertiary Bulgarian institution. From the analysis of 762 kidney biopsies, the diagnosis of primary IgAN was established in 125, with an average age of 35.94 ± 11.91 years. Our study aimed to assess the clinical characteristics, histological features, and potential biomarkers of IgAN in the Bulgarian population. We evaluated parameters such as proteinuria, hematuria, serum creatinine, and glomerular filtration rate (GFR). In fifty IgAN patients and 30 healthy controls, serum levels of Gd-IgA1, IgA, C3, BAFF, and APRIL using ELISA were examined. The results revealed significant differences in serum concentrations of Gd-IgA1 (p < 0.001), Gd-IgA1/IgA (p = 0.022), IgA (p = 0.014), and IgA/C3 (p = 0.047) between patients and controls. However, no correlation was found between Gd-IgA1, IgA, Gd-IgA1/IgA, and IgA/C3 and chronic kidney disease progression. Our study reports evidence of the diagnostic value of Gd-IgA1 and contributes to the understanding of IgAN in the Bulgarian population and suggests potential biomarkers for disease diagnosis and prognosis.

Disease-specific plasma protein profiles in patients with fever after traveling to tropical areas.

Fever is common among individuals seeking healthcare after traveling to tropical regions. Despite the association with potentially severe disease, the etiology is often not determined. Plasma protein patterns can be informative to understand the host response to infection and can potentially indicate the pathogen causing the disease. In this study, we measured 49 proteins in the plasma of 124 patients with fever after travel to tropical or subtropical regions. The patients had confirmed diagnoses of either malaria, dengue fever, influenza, bacterial respiratory tract infection, or bacterial gastroenteritis, representing the most common etiologies. We used multivariate and machine learning methods to identify combinations of proteins that contributed to distinguishing infected patients from healthy controls, and each other. Malaria displayed the most unique protein signature, indicating a strong immunoregulatory response with high levels of IL10, sTNFRI and II, and sCD25 but low levels of sCD40L. In contrast, bacterial gastroenteritis had high levels of sCD40L, APRIL, and IFN-γ, while dengue was the only infection with elevated IFN-α2. These results suggest that characterization of the inflammatory profile of individuals with fever can help to identify disease-specific host responses, which in turn can be used to guide future research on diagnostic strategies and therapeutic interventions.

Impact of different genetic mutations on granulocyte development and G-CSF responsiveness in congenital neutropenia

AbstractCongenital neutropenia (CN) is a genetic disorder characterized by persistent or intermittent low peripheral neutrophil counts, thus increasing susceptibility to bacterial and fungal infections. Various forms of CN, caused by distinct genetic mutations, exhibit differential responses to granulocyte colony–stimulating factor (G-CSF) therapy, with the underlying mechanisms not fully understood. This study presents an in-depth comparative analysis of clinical and immunological features in 5 CN patient groups (severe congenital neutropenia [SCN]1, SCN3, cyclic neutropenia [CyN], warts, hypogammaglobulinaemia, infections and myelokathexis [WHIM], and Shwachman-Bodian-Diamond Syndrome [SBDS]) associated with mutations in ELANE, HAX1, CXCR4, and SBDS genes. Our analysis led to the identification of 11 novel mutations in ELANE and 1 each in HAX1, CXCR4, and G6PC3 genes. Investigating bone marrow (BM) granulopoiesis and blood absolute neutrophil count after G-CSF treatment, we found that SCN1 and SCN3 presented with severe early-stage disruption between the promyelocyte and myelocyte, leading to a poor response to G-CSF. In contrast, CyN, affected at the late polymorphonuclear stage of neutrophil development, showed a strong G-CSF response. WHIM, displaying normal neutrophil development, responded robustly to G-CSF, whereas SBDS, with moderate disruption from the early myeloblast stage, exhibited a moderate response. Notably, SCN1 uniquely impeded neutrophil development, whereas SCN3, CyN, WHIM, and SBDS also affected eosinophils and basophils. In addition, SCN1, SCN3, and CyN presented with elevated serum immunoglobulins, increased BM plasma cells, and higher A Proliferation-Inducing Ligand levels. Our study reveals a strong correlation between the stage and severity of granulocyte development disruption and the efficacy of G-CSF therapy.

Impact of the COVID‐19 pandemic on enrollment at United States Alzheimer’s Disease Research Centers

AbstractBackgroundImpacts of the COVID‐19 pandemic have been disproportionate by race/ethnicity and sex. We aimed to characterize the impact of the pandemic on enrollment in US Alzheimer’s Disease Research Centers (ADRCs) overall and by race/ethnicity and sex using National Alzheimer’s Coordinating Center (NACC) data.MethodsWe included data on N = 8875 participants (mean age, 69.4 years; 59% female; 9% Hispanic/Latino; 14% Black/African American) from 29 ADRCs with ≥5 years of data and study visits after 2020 (e.g., baseline visits from January 2017‐December 2021). We used interrupted time series (ITS) analyses to assess the relationship of the pandemic on enrollment and calculated projected date of recovery of pre‐pandemic enrollment overall and by race, ethnicity, and sex.ResultsEnrollment dropped to 0% of pre‐pandemic levels in April 2020; through December 2021, enrollment recovered to only 46% of pre‐pandemic levels. Participants enrolled during the pandemic (vs pre‐pandemic) were less likely to be: women, cognitively normal, and able to live independently. They were slightly younger, slightly more educated, and more likely to: be married, be referred by a doctor, nurse, or other professional, and come in for a clinical or mixed clinical and research study evaluation (p’s&lt;0.05). The two groups did not vary by race, ethnicity, living situation (alone or with others), or co‐participant characteristics. ITS demonstrated that the pandemic caused an immediate 79% drop in enrollment rate, with an estimated post‐pandemic increase in enrollment of 0.92%/month, leading to a projected recovery date of January 2026. Due to parallel pre‐ and post‐pandemic enrollment slopes among Black/African American participants, we could not predict a recovery date for this group. Projected recovery varied by ethnicity with Hispanic participant enrollment recovering earlier than non‐Hispanic participant enrollment (January 2022 vs. April 2027), largely because Hispanic participant enrollment was declining nearly 1.5%/month pre‐pandemic. Recovery date projections also varied by sex, with female participant enrollment recovering sooner than male participant enrollment (June 2025 vs. May 2027).ConclusionsStrategies to speed enrollment recovery should be developed and implemented, with special attention given to Black/African American and Hispanic enrollment. Funders and researchers should be aware of and account for ongoing COVID‐19 impact on ADRD research enrollment.

Follicular DNA Damage and Pesticide Exposure Among Latinx Children in Rural and Urban Communities

The intersectional risks of children in United States immigrant communities include environmental exposures. Pesticide exposures and their biological outcomes are not well characterized in this population group. We assessed pesticide exposure and related these exposures to DNA double-strand breaks (DSBs) in Latinx children from rural, farmworker families (FW; N = 30) and from urban, non-farmworker families (NFW; N = 15) living in North Carolina. DSBs were quantified in hair follicular cells by immunostaining of 53BP1, and exposure to 72 pesticides and pesticide degradation products were determined using silicone wristbands. Cholinesterase activity was measured in blood samples. DSB frequencies were higher in FW compared to NFW children. Seasonal effects were detected in the FW group, with highest DNA damage levels in April–June and lowest levels in October–November. Acetylcholinesterase depression had the same seasonality and correlated with follicular DNA damage. Organophosphate pesticides were more frequently detected in FW than in NFW children. Participants with organophosphate detections had increased follicular DNA damage compared to participants without organophosphate detection. Follicular DNA damage did not correlate with organochlorine or pyrethroid detections and was not associated with the total number of pesticides detected in the wristbands. These results point to rural disparities in pesticide exposures and their outcomes in children from vulnerable immigrant communities. They suggest that among the different classes of pesticides, organophosphates have the strongest genotoxic effects. Assessing pesticide exposures and their consequences at the individual level is key to environmental surveillance programs. To this end, the minimally invasive combined approach used here is particularly well suited for children.

Initial Report of Povetacicept, an Enhanced Dual BAFF/APRIL Antagonist, in Autoimmune Cytopenias: The RUBY-4 Study

Introduction: BAFF and APRIL are cytokines with key roles in B cell development, survival, differentiation into antibody-secreting cells (ASCs; plasmablasts and plasma cells), and production of antibodies, making them promising targets for the treatment of autoantibody-associated diseases such as immune thrombocytopenia (ITP), warm autoimmune hemolytic anemia (wAIHA), and cold agglutinin disease (CAD). Patients with ITP or AIHA have elevated serum BAFF and APRIL levels and belimumab, a BAFF inhibitor, has exhibited encouraging efficacy in the treatment of lupus-associated ITP as well as alone or in combination with rituximab for the treatment of primary ITP. Due to their overlapping but non-redundant roles, inhibition of both BAFF and APRIL may be required for optimal efficacy in autoimmune cytopenias. In preclinical studies, dual BAFF/APRIL inhibition achieved superior suppression of ASCs and immunoglobulin levels vs inhibitors of BAFF or APRIL alone or an anti-CD20 antibody, suggesting the potential of this novel mechanism to modify the underlying pathogenesis of autoimmune cytopenias. Povetacicept (ALPN-303) is a TACI-Fc fusion protein engineered for potent dual BAFF/APRIL inhibition. In a mouse model of AIHA, povetacicept suppressed anti-erythrocyte autoantibodies and increased hematocrit (ASH 2022 abstract #3763). In a first-in-human study in healthy volunteers, povetacicept was well tolerated and reduced levels of circulating ASCs and immunoglobulins. This is an initial report of an open-label study of povetacicept in autoimmune cytopenias (RUBY-4; NCT05757570). Methods: RUBY-4 is a phase 1b, group-sequential, parallel-cohort study evaluating povetacicept in participants aged ≥18 years with a documented diagnosis of primary ITP, wAIHA, or CAD of ≥12 weeks in duration (Figure). Eligible participants must have sustained thrombocytopenia (ITP cohort) or sustained symptomatic anemia (wAIHA/CAD cohorts)-i.e., platelet count &amp;lt;30 × 10 9/L or hemoglobin ≤10 g/dL on 2 occasions ≥7 days apart during screening-and a history of failure or relapse on ≥2 prior treatments (including a thrombopoietin receptor agonist for the ITP cohort). Background therapy, including corticosteroids, is allowed but must remain at stable doses for the durations specified in the protocol. Participants receive povetacicept 240 mg subcutaneously once every 4 weeks for 24 weeks, with an optional 24-week treatment extension, followed by an 8-week off-treatment follow-up period. The primary objective is safety and tolerability; secondary objectives include pharmacokinetics, pharmacodynamics, immunogenicity, and efficacy. Results: As of 31 July 2023, 1 participant with ITP has enrolled and received 2 doses of povetacicept 240 mg without incident. Initial, preliminary observations include expected reductions in B cells and immunoglobulin levels. The RUBY-4 study continues to enroll in the US, Canada, Australia, and Turkey. At the time of presentation, available data including safety, pharmacokinetics, pharmacodynamics, and preliminary efficacy (e.g., platelet or hemoglobin response rate, duration of response, and time to response) findings will be reported. Conclusions: To our knowledge, povetacicept is the first dual BAFF/APRIL inhibitor to be evaluated for the treatment of autoimmune cytopenias. Previous preclinical and healthy volunteer studies suggest the potential of povetacicept for suppression of the pathogenic autoimmunity that mediates platelet/erythrocyte destruction in ITP, wAIHA, and CAD, which may lead to more durable responses and improved outcomes for patients.

A Phase 1, First-in-Human, Dose Escalation and Dose-Expansion Study of a BCMAxCD38xCD3 Targeting Trispecific Antibody ISB 2001 in Subjects with Relapsed/Refractory Multiple Myeloma

Background Despite advances, multiple myeloma (MM) remains an incurable disease and resistance mechanisms are emerging. ISB 2001 has been designed to overcome those resistance mechanisms inherent to current MM therapies such as monoclonal antibodies (e.g. daratumumab), BCMA-targeted therapies, proteasome inhibitors(PIs), or immunomodulatory drugs(IMiDs). ISB 2001 is a first in class Trispecific T cell engager (TCE) that redirects cytotoxic T cells to BCMA and/or CD38 expressing myeloma cells. Simultaneous targeting of the two tumor associated antigens (TAA) may increase binding to tumor cells through avidity even with heterogeneous or low expression of the targeted TAA (BCMA or CD38). Therefore, it may overcome tumor escape mechanisms associated with low tumor antigenic expression inherent to current MM targeted therapies. Preclinically, ISB 2001 has demonstrated improved activity when compared to other BCMA or CD38 targeted molecules either alone or in combination across different MM models (M. Pihlgren et al., Blood. 140, 858-859 (2022), M. Pihlgren et al., Cancer Res. 83, 2970-2970 (2023)). Study design and methods This first-in-human, multicenter, open label Phase 1 study is assessing the safety, tolerability, efficacy, pharmacokinetics (PK), and pharmacodynamics (PD) of ISB 2001 in relapsed/refractory multiple myeloma (RRMM) patients. This study will enroll subjects treated with prior IMiDs, PIs, and anti-CD38 therapies, or intolerant of, established therapies known to provide benefit (prior BCMA therapies, or prior transplantation and cellular therapies allowed).be conducted in two parts (Part 1: Dose escalation and Part 2: Dose expansion) (Figure 1). Criteria for enrollment in this study include patients with RRMM with measurable disease who have been treated with an anti-CD38 antibody, IMiDs, PIs either in combination or as a single agent, and /or intolerant of established therapies known to provide clinical benefit. ISB 2001 is being administered weekly by subcutaneous (SC) injection in 28-day cycles, with two step-up doses on cycle 1 day 1 (C1D1) and C1D4 before administering the full dose on C1D8. The first-in-human dose was derived by integrating both in vitro and in vivo preclinical data benchmarked to teclistamab utilizing a quantitative systems pharmacology (QSP) model guided approach. The fractionated step-up dosing before administering the full dose on C1D8 was included in every cohort preemptively to minimize the potential cytokine release syndrome (CRS) risk. The study will follow a rapid titration single patient dose escalation design until the completion of cohort 3 or until one of the safety conversion criteria are met, whichever comes first, after which the design will be converted to a conventional 3+3 dose escalation. The primary outcome measure is the number of DLTs during the first 28 days after the first study treatment (ie, Cycle 1) in each cohort. Once a potential safe and efficacious dose range is identified, the Part 2 expansion will start enrollment in two arms at two putative recommended Part 2 doses with 1:1 randomization with at least 20 patients per arm (FDA Project Optimus). The primary objective of Part 2 is to confirm safety and to select the recommended Phase 2 dose (RP2D). Secondary endpoints include PK parameters, immunogenicity incidence, ORR, CRR and DOR based on IMWG criteria. Exploratory endpoints include levels of cytokines, chemokines, sCD38, sBCMA, APRIL and BAFF in peripheral blood, immunophenotyping in peripheral blood and bone marrow, and MRD negative rate. Approximately 40 patients will be enrolled in Part 1 at sites in Australia, France and the United States. A total of approximately 40 evaluable patients will be enrolled in Part 2. The study is currently open for enrollment (Clinicaltrials.gov identifier: NCT05862012).

B cell phenotype and serum levels of interferons, BAFF, and APRIL in multisystem inflammatory syndrome in children associated with COVID-19 (MIS-C)

BackgroundMultisystem inflammatory syndrome in children associated with COVID-19 (MIS-C) is a late complication of pediatric COVID-19, which follows weeks after the original SARS-CoV-2 infection, regardless of its severity. It is characterized by hyperinflammation, neutrophilia, lymphopenia, and activation of T cells with elevated IFN-γ. Observing the production of autoantibodies and parallels with systemic autoimmune disorders, such as systemic lupus erythematodes (SLE), we explored B cell phenotype and serum levels of type I, II, and III interferons, as well as the cytokines BAFF and APRIL in a cohort of MIS-C patients and healthy children after COVID-19.ResultsWe documented a significant elevation of IFN-γ, but not IFN-α and IFN-λ in MIS-C patients. BAFF was elevated in MIS-C patient sera and accompanied by decreased BAFFR expression on all B cell subtypes. The proportion of plasmablasts was significantly lower in patients compared to healthy post-COVID children. We noted the pre-IVIG presence of ENA Ro60 autoantibodies in 4/35 tested MIS-C patients.ConclusionsOur work shows the involvement of humoral immunity in MIS-C and hints at parallels with the pathophysiology of SLE, with autoreactive B cells driven towards autoantibody production by elevated BAFF.Graphical

Alpha-Fetoprotein as a Factor of Differentiation and Functional Activity of Myeloid-Derived Suppressor Cells.

We studied the role of alpha-fetoprotein (AFP) in regulation of differentiation and functional activity of human myeloid-derived suppressor cells (MDSC) in vitro. To obtain MDSC, CD11b+ cells were isolated from the peripheral blood of healthy donors followed by cytokine induction (IL-1β+GM-CSF) into the MDSC phenotype. The cell functions were assessed by the expression of indoleamine 2,3-dioxygenase (IDO) and arginase-1 (Arg1) and cytokine profile of the cell cultures. Native AFP did not affect the total number of MDSC and the percentage of polymorphonuclear MDSC (PMN-MDSC), but increased the number of monocytic MDSC (M-MDSC). AFP did not change the expression of Arg1, but in low concentrations (10 and 50 U/ml) increased the number of IDO-containing cells. AFP modulated the cytokine profile of CD11b+ cells: it reliably decreased the level of IL-19 (50 and100 U/ml) and showed a tendency to decrease the levels of IL-34, MMP-2, sCD163, CHI3L1, OPN and to increase the levels of IL-29, IL-32, APRIL, PTX3, and sTNF-R1. Thus, we have demonstrated a regulatory effect of native AFP at the level of MDSC generated from CD11b+ cells under conditions of cytokine induction in vitro.

Children with extended oligoarticular and polyarticular juvenile idiopathic arthritis have alterations in B and T follicular cell subsets in peripheral blood and a cytokine profile sustaining B cell activation

ObjectivesThe main goal of this study was to characterise the frequency and phenotype of B, T follicular helper (Tfh) and T follicular regulatory (Tfr) cells in peripheral blood and the...