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Lapuente, D.

Publications and source records attributed to Lapuente, D..

4 recordsLinked to original sources

A confirmatory, dual-centric non-human primate study on the efficacy of novel oropharyngeal spray immunization with an adenoviral vector vaccine against RSV -- Important lessons learned

In a confirmatory study, we evaluated the immunogenicity and protective efficacy of a heterologous prime-boost vaccination strategy against respiratory syncytial virus (RSV) in non-human primates. Building on prior evidence of protective mucosal immunity induced by intramuscular DNA priming followed by an oropharyngeal adenoviral boost, we conducted a randomized, blinded, dual-centre study across two European primate research facilities. Rhesus macaques received a codon-optimized RSV-F DNA vaccine via electroporation, followed by two mucosal administrations of a recombinant adenovirus serotype 5 vector encoding the same antigen. Control groups included animals vaccinated with irrelevant influenza antigens and a comparator group mimicking natural immunity induced by primary RSV infection. Systemic and mucosal immune responses, including RSV-F-specific antibodies and tissue-resident memory T cells, were monitored longitudinally. Here, we detected robust immune responses, but with some variability between the two centres. However, following experimental RSV challenge performed 22 weeks after the final immunization, RSV-vaccinated animals demonstrated markedly reduced viral replication in both upper and lower respiratory tracts. However, unexpected RSV-specific immunity in the control group at one single study site prevented confirmation of the predefined primary endpoint. Overall, these results support the potential of mucosal adenoviral boosting following DNA priming to induce protective immunity against RSV, while highlighting challenges associated with multi-centre preclinical vaccine studies.

immunology↗

A Novel Secretome Rewrites the Immune Response in Viral Acute Respiratory Distress Syndrome

BackgroundHyperinflammatory syndromes such as viral acute respiratory distress syndrome (ARDS) demand immunotherapies that are safe and effective at suppressing cytokine storm. As a novel treatment for ARDS, PRS CK STORM is proposed as a next-generation cell-free secretome derived from co-cultures of M2 macrophages and mesenchymal stromal cells. MethodsWe performed a double-blind, randomized controlled trial to test the effects of PRS CK STORM in K18-hACE2 transgenic mice infected intranasally with 105 PFU of SARS-CoV-2 BE.1.1. This was complemented by a mechanistic analysis of this secretome using transcriptomic and COX2 enzymatic activity studies, as well as a compositional analysis of its proteomic and miRNA profile. ResultsIn a lethal SARS-CoV-2 ARDS mouse model, PRS CK STORM significantly improved lung histopathology to a degree on par with corticosteroids, while stimulating angiogenesis in damaged lung tissue. Early and late cytokine profiling showed marked reductions in IFN-{gamma}. In response to PRS CK STORM treatment, transcriptomic analyses in inflamed macrophages revealed robust downregulation of key proinflammatory drivers (MyD88, TRAF6, IKK2, NF-{kappa}B, COX2). In vitro enzymatic assays confirmed potent, dose-dependent inhibition of COX2, with high inter-batch reproducibility. A compositional analysis revealed this secretome to be rich in anti-inflammatory miRNAs, immune-modulating proteins, and regenerative factors. ConclusionsPRS CK STORM operates as a multi-target immune recalibrator, enabling broad downregulation of pathological inflammation while promoting tissue repair. Its off-the-shelf, GMP-manufactured format ensures reproducibility and scalability, offering a novel resolution pharmacology approach for cytokine storm syndromes originated from ARDS and beyond.

immunology↗

Alcohol-sourced acetate reduces T cell filamentous actin branching and migration

Alcohol is among the most widely consumed dietary substances. Excessive alcohol consumption damages the liver, heart and brain. Alcohol also has strong immunoregulatory properties. Here we report how alcohol impairs T cell function via acetylation of cortactin, a protein that binds filamentous actin and facilitates branching. Upon alcohol consumption, acetate, the metabolite of alcohol, accumulates in lymphoid organs. T cells exposed to acetate, exhibit increased acetylation of cortactin. Acetylation of cortactin inhibits filamentous actin binding and hence reduces T cell migration, immune synapse formation and activation. While mutated, acetylation-resistant cortactin rescued the acetate-induced inhibition of T cell migration, primary mouse cortactin knock-out T cells exhibited impaired migration. Furthermore, acetate-induced cytoskeletal changes effectively inhibited activation, proliferation, and immune synapse formation in T cells in vitro and in vivo in an influenza infection model in mice. Together these findings reveal cortactin as a possible target for mitigation of T cell driven autoimmune diseases.

immunology↗

Protective mucosal immunity against SARS-CoV-2 after heterologous systemic RNA-mucosal adenoviral vector immunization

Several effective SARS-CoV-2 vaccines are currently in use, but in the light of waning immunity and the emergence of novel variants, effective boost modalities are needed in order to maintain or even increase immunity. Here we report that intranasal vaccinations with adenovirus 5 and 19a vectored vaccines following a systemic DNA or mRNA priming result in strong systemic and mucosal immunity in mice. In contrast to two intramuscular injections with an mRNA vaccine, the mucosal boost with adenoviral vectors induced high levels of IgA and tissue-resident memory T cells in the respiratory tract. Mucosal neutralization of virus variants of concern was also enhanced by the intranasal boosts. Importantly, priming with mRNA provoked a more comprehensive T cell response consisting of circulating and tissue-resident memory T cells after the boost, while a DNA priming induced mostly mucosal T cells. Concomitantly, the intranasal boost strategies provided protection against symptomatic disease. Therefore, a mucosal booster immunization after mRNA priming is a promising approach to establish mucosal immunity in addition to systemic responses.

immunology↗