Biology subjects

Roestenberg, M.

Publications and source records attributed to Roestenberg, M..

5 recordsLinked to original sources

bioRxiv2026

O-GlcNAcylation and low glycolysis underpin Th2 polarization by dendritic cells

Activation of dendritic cells (DCs) is dependent on rewiring of their cellular metabolism. However, the metabolic requirements for DCs to prime T helper 2 (Th2) responses are still poorly understood. Using unbiased transcriptomics and non-targeted metabolomics we find that helminth antigen-conditioned human DCs suppress glycolysis while increasing hexosamine biosynthesis to fuel protein O-GlcNAcylation. Functionally, glycolytic inhibition of DCs selectively enhanced, while blocking O-GlcNAcylation impaired, Th2-priming capacity. In helminth infection and allergic challenge, Th2 responses were also attenuated in vivo in mice with specific deletion of O-GlcNAc Transferase (OGT) in CD11c-expressing cells. Mechanistically, through proteomic analysis and functional validation, we identified O-GlcNAcylation as a critical negative regulator of immune synapse formation by controlling cytoskeletal organization via Fascin-1 and Zyxin, thereby dampening TCR signalling to promote Th2 polarization. Altogether we reveal a novel metabolic program in DCs that governs Th2 polarization, that could potentially be harnessed to treat type 2 mediated inflammatory diseases.

immunology↗

bioRxiv2025

Manufacture of Clostridioides difficile spores for experimental infection of human volunteers

BackgroundInfections with C. difficile remain a significant global healthcare problem for which novel (preventive) treatment strategies are urgently needed. Controlled human infection models (CHIMs) can contribute to a better understanding of the disease and accelerate (novel) medicinal product development. MethodsA robust production process for C. difficile spores for experimental human administration, was implemented. Direct-release capsules containing viable C. difficile spores were formulated. The manufacturing process aligns with good manufacturing practices (GMP) guidelines, including quality controls and release by a qualified person (QP). ResultsSelected non-toxigenic and toxigenic C. difficile strains were used to create a master cell bank and working cell bank from which multiple batches of purified active substance were produced. Substance was then used to manufacture the final active product. The active product and its intermediate products passed all quality controls for identity, potency, purity and safety, and were individually released by the QP. Stability of active substance and product is confirmed up to 12 months. ConclusionAlthough there are no clear standardized international guidelines for challenge material production, we demonstrate that it is feasible to produce C. difficile human challenge material for small scale application in CHIM trials. This application of GMP principles to an unconventional production process of a bacterial spore-forming anaerobic challenge agent is an example for the production of challenge material as described in the auxiliary medicinal product guidelines of EMA and FDA.

microbiology↗

bioRxiv2025

Pre-clinical studies of Schistosoma mansoni vaccines: a scoping review

BackgroundSchistosomiasis is caused by infection with worms of the genus Schistosoma including S. mansoni. Over 200 million people are infected, sterile immunity does not naturally develop, and no vaccine is available. A vaccine could be a critical tool to achieve control and elimination. Numerous candidates have been tested in pre-clinical models, but there is not yet an approved vaccine. Methodology/Principal FindingsWe conducted a scoping review using a keyword search on Web of Science and a MeSH term search on PubMed. Articles were screened and included if they tested a defined vaccine candidate in a pre-clinical protection assay against S. mansoni between 1994-2024. Vaccine formulation, study design, and efficacy parameters from all articles were extracted. This data was summarised graphically, with the influence of different parameters appraised. A total of 141 candidate antigens were tested in 108 articles over the last 30 years, with most antigens tested only once and three (Sm-CatB, Sm-p80, and Sm-14) tested over 20 times. The median protective efficacy against worms was 35%. 10 antigens achieved over 60% efficacy, and only two (Sm-p80 and Sm-CatB) over 90%. Large variations in efficacy were observed with all repeatedly tested antigens, likely attributable to differing formulations and study designs. The effect of these varying parameters on the resultant efficacy was evaluated. ConclusionsA few vaccine candidates have achieved promising efficacy in pre-clinical studies. Most vaccines tested however have efficacy that falls short of that required for an impactful schistosomiasis vaccine. The diversity in study designs makes comparing vaccine targets a challenge. Use of consistent and optimized vaccine formulation (including adjuvant and platform) and study design parameters is critical to expedite the development of a schistosome vaccine. Author SummarySchistosomiasis, a major neglected tropical disease, is caused by infection with parasitic worms of the Schistosoma species, including Schistosoma mansoni. Individuals can be repeatedly re-infected, and there is no available vaccine. An initial stage of vaccine development is testing in a pre-clinical animal model. Here we have summarised tests of Schistosoma mansoni vaccines in the last three decades. 100+ vaccine candidates have been tested, with only 10 of these achieving efficacy of over 60%, and only 2 at over 90%. When the same vaccine candidate (antigen) is tested in a different formulation, or using different study design the efficacy varies greatly. We have summarised the formulations and study designs used, and highlighted how certain parameters affect efficacy. Finally, we have assembled a series of recommendations to researchers on how to perform vaccine tests in the future.

immunology↗

bioRxiv2024

Correlative humoral and cellular immunity to genetically attenuated malaria parasites in humans

Malaria caused by Plasmodium falciparum remains one of the major infectious diseases with a high burden in Sub-Saharan Africa. In spite of the advancements made in vaccine development and implementation in endemic countries, sterile and durable protection has not been achieved. Recently, we have shown the superior protective capacity of whole sporozoites attenuated to arrest late but not early during the liver stage development in a controlled human malaria infection study. Here we report the breadth of antigens targeted by hitherto understudied parasite liver stage immunity and convey the coherence between humoral and cellular immunity observed in our clinical study. Our findings uncover the underlying immunogenic differences between early- and late-liver stage arresting parasites and identify key liver-stage antigens for future vaccine development focused on inducing sterile immunity to malaria.

immunology↗

bioRxiv2020

Clustering and erratic movement patterns of syringe-injected versus mosquito-inoculated malaria sporozoites underlie decreased infectivity

Live attenuated malaria sporozoites are promising vaccine candidates, however, their efficacy critically depends on their capability to reach and infect the host liver. Administration via mosquito inoculation is by far the most potent method for inducing immunity, but highly unpractical. Here, we observed that intradermal syringe-injected Plasmodium berghei sporozoites (syrSPZ) were three-fold less efficient in migrating to and infecting mouse liver compared to mosquito-inoculated sporozoites (msqSPZ). This was related to a clustered dermal distribution (2-fold decreased median distance between syrSPZ vs msqSPZ) and, more importantly, a 1.4-fold significantly slower and more erratic movement pattern. These erratic movement patterns were likely caused by alteration of dermal tissue morphology (>15 m intercellular gaps) due to injection pressure and may critically decrease sporozoite infectivity. These results suggest that novel microvolume-based administration technologies hold promise for replicating the success of mosquito-inoculated live attenuated sporozoite vaccines.

microbiology↗