The Capacities of the Probiotic Strains L. helveticus MIMLh5 and L. acidophilus NCFM to Induce Th1-Stimulating Cytokines in Dendritic Cells Are Inversely Correlated with the Thickness of Their S-Layers

The surface layer (S-layer) of probiotic bacteria plays an important role in their interaction with the host immune system. In this article, Valentina Taverniti , Paolo D’Incecco , Stefano Farris , Peter Riber Jonsen , Helene Skovsted Eld , Juliane Sørensen, Laura Brunelli, Giacomo Mantegazza, Stefania Arioli and Hanne Frøkiær, investigated how the thickness of the S-layer influences the ability of Lactobacillus helveticus MIMLh5 and Lactobacillus acidophilus NCFM to stimulate Th1-related cytokine production in dendritic cells.

The results revealed an inverse correlation between S-layer thickness and the induction of interleukin-12, indicating that thinner S-layers are associated with a stronger immune-stimulating response. These findings provide new insights into the structure–function relationship of bacterial surface layers and their role in probiotic–host interactions.

Atomic force microscopy (AFM) was used for nanomechanical and morphological characterization of bacterial cells. Measurements were performed using a commercially available AFM instrument operated in contact resonance amplitude imaging (CRAI) mode. An Nanoworld Arrow-FMR force modulation AFM probe was used. This silicon AFM probe features a rectangular beam with a triangular free end and a tetrahedral tip (tip radius ~10 nm, tip height 10–15 μm), a spring constant of 2.8 N/m and a resonance frequency of 75 kHz. Images of 10 × 10 μm² and force–distance curves were recorded at multiple locations on the bacterial surface. Nanomechanical properties, including the elastic (Young’s) modulus, were determined by fitting approach curves to the Hertzian model with an indentation depth set to 2 nm.

figure S1: Schematic representation of the 4-step procedure for the AFM analysis of the bacteria surface

Figure S1: Schematic representation of the 4-step procedure for the AFM analysis of the bacteria surface: scanning of the surface in contact resonance amplitude (CRAI) mode (a); creation of the 10-point map of the nanomechanical test (b); generation of the force-distance curves (c); and fitting procedure for the extrapolation of the elastic modulus (d).     

 

Taverniti, V.; D’Incecco, P.; Farris, S.; Jonsen, P. R.; Eld, H. S.; Sørensen, J.; Brunelli, L.; Mantegazza, G.; Arioli, S.; Mora, D.; Guglielmetti, S.; Frøkiær, H.
The Capacities of the Probiotic Strains L. helveticus MIMLh5 and L. acidophilus NCFM to Induce Th1-Stimulating Cytokines in Dendritic Cells Are Inversely Correlated with the Thickness of Their S-Layers.
Biomolecules 2025, 15(7), 1012.
https://doi.org/10.3390/biom15071012

The article: The Capacities of the Probiotic Strains L. helveticus MIMLh5 and L. acidophilus NCFM to Induce Th1-Stimulating Cytokines in Dendritic Cells Are Inversely Correlated with the Thickness of Their S-Layers, is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Meet us at 63rd Annual Meeting of the Biophysical Society of Japan next week

NanoWorld AG CEO Manfred Detterbeck will be at the NanoAndMore Japan booth at the 63rd Annual Meeting of the Biophysical Society of Japan held from September 24– 26, 2025 at Nara Prefectural Convention Center.   Will we meet you there too?

cartoon of the NanoWorld AFM probes professor and robot dreaming of Nara Japan, dream bubble is showing the Nara skyline
Meet you at the NanoAndMore Japan booth at Nara Prefectural Convention Center next week

 

 

Skyline of Nara
JosepPerianes/Shutterstock.com

DNA Origami Nanovaccine Demonstrates Full Protection Against SARS-CoV-2 in Mice

A study recently published in Communications Biology (Nature Portfolio) by Esra Oktay, Farhang Alem, Keziah Hernandez, Michael Girgis, Christopher Green, Divita Mathur, Igor L. Medintz, Aarthi Narayanan, and Rémí Veneziano introduces an innovative DNA origami–based nanovaccine platform targeting the receptor-binding domain (RBD) of SARS-CoV-2. Their findings highlight the potential of rationally engineered DNA nanoparticles to elicit strong and durable immune protection.

The researchers designed wireframe DNA origami nanoparticles (DNA-NPs) functionalized with ten trimerized RBD antigens and CpG oligonucleotides as adjuvants. Trimer assembly was achieved by peptide nucleic acid (PNA)–mediated conjugation of protein G to RBD-Fc, enabling precise orientation on DNA-NP overhangs. Biophysical characterization employed dynamic light scattering, Förster resonance energy transfer (FRET)–based stability assays, and quantitative fluorescence labeling. Structural confirmation was obtained via atomic force microscopy (AFM). For high-resolution AFM imaging, the team employed NanoWorld® USC-F0.3-k0.3 ultra-short cantilevers, chosen for their 300 kHz resonance frequency, ~0.3 N/m force constant, and <10 nm tip radius. These probes provided enhanced stability and resolution, enabling accurate nanoscale visualization of the vaccine constructs. In murine models, the fully assembled RBD-DNA-NP with CpG provided complete protection following viral challenge. Vaccinated mice exhibited no weight loss or mortality, in stark contrast to control groups that received unconjugated components, bare origami scaffolds, or RBD alone—several of which experienced significant morbidity or mortality. Immunological assays revealed sustained neutralizing antibody activity two months post-immunization, with robust titers of RBD-specific IgG, IgM, and IgA, underscoring the durability of the immune response. This work demonstrates that DNA origami nanostructures can serve as an adaptable and effective vaccine delivery platform. By integrating precise antigen display with adjuvant loading, the approach provides a promising blueprint for next-generation nanovaccines. The use of advanced AFM probes, such as the NanoWorld® USC-F0.3-k0.3, further underscores the importance of high-precision nanometrology tools in validating nanoscale biomedical designs. Hydrodynamic diameter measurement of DNA-NPs with DLS
a Hydrodynamic diameter measurement of DNA-NPs with DLS. b Atomic force microscopy imaging of DNA-NPs formation and conjugation with 3-mer RBDs (scale bar: 20 nm). c Fluorescence-intensity based determination of PG and RBD stoichiometry on DNA origami NP. i) Tryptophan fluorescence emission was used to determine the total number of PG loaded on the surface of the DNA-NPs. ii) Second, the stoichiometry of RBD on NP was quantified via measuring the emission of Cy5 dyes conjugated to the RBD antigens. The bar graph represents the total coverage percentage for the PG (green bar) and the 3-mer RBD (orange bar) on the DNA-NP surface normalized to the number of conjugation sites available. Data are shown as the mean ± SD (n = 3 independent experiments).

This article contains images reused from Jia et al., Nature Communications 14, 1394 (2023), licensed under CC BY 4.0.