A randomized, double-blinded Phase 3 study to demonstrate lot-to-lot consistency and to confirm immunogenicity and safety of the live-attenuated chikungunya virus vaccine candidate VLA1553 in healthy adults†.

BACKGROUND: The global spread of the chikungunya virus (CHIKV) increases the exposure risk for individuals travelling to or living in endemic areas. This Phase 3 study was designed to demonstrate manufacturing consistency between three lots of the single shot live-attenuated CHIKV vaccine VLA1553, and to confirm the promising immunogenicity and safety data obtained in previous trials. METHODS: This randomized, double-blinded, lot-to-lot consistency, Phase 3 study, assessed immunogenicity and safety of VLA1553 in 408 healthy adults (18-45 years) in 12 sites across the USA. The primary endpoint was a comparison of the geometric mean titre (GMT) ratios of CHIKV-specific neutralizing antibodies between three VLA1553 lots at 28 days post-vaccination. Secondary endpoints included immunogenicity and safety over 6 months post-vaccination. RESULTS: GMTs were comparable between the lots meeting the acceptance criteria for equivalence. The average GMT (measured by 50% CHIKV micro plaque neutralization test; µPRNT50) peaked with 2643 at 28 days post-vaccination and decreased to 709 at 6 months post-vaccination. An excellent seroresponse rate (defined as µPRNT50 titre ≥ 150 considered protective) was achieved in 97.8% of participants at 28 days post-vaccination and still persisted in 96% at 6 months after vaccination. Upon VLA1553 immunization, 72.5% of participants experienced adverse events (AEs), without significant differences between lots (related solicited systemic AE: 53.9% of participants; related solicited local AE: 19.4%). Overall, AEs were mostly mild or moderate and resolved without sequela, usually within 3 days. With 3.9% of participants experiencing severe AEs, 2.7% were classified as related, whereas none of the six reported serious adverse events was related to the administration of VLA1553. CONCLUSIONS: All three lots of VLA1553 recapitulated the safety and immunogenicity profiles of a preceding Phase 3 study, fulfilling pre-defined consistency requirements. These results highlight the manufacturability of VLA1553, a promising vaccine for the prevention of CHIKV disease for those living in or travelling to endemic areas.

Safety and immunogenicity of a single-shot live-attenuated chikungunya vaccine: a double-blind, multicentre, randomised, placebo-controlled, phase 3 trial.

BACKGROUND: VLA1553 is a live-attenuated vaccine candidate for active immunisation and prevention of disease caused by chikungunya virus. We report safety and immunogenicity data up to day 180 after vaccination with VLA1553. METHODS: This double-blind, multicentre, randomised, phase 3 trial was done in 43 professional vaccine trial sites in the USA. Eligible participants were healthy volunteers aged 18 years and older. Patients were excluded if they had history of chikungunya virus infection or immune-mediated or chronic arthritis or arthralgia, known or suspected defect of the immune system, any inactivated vaccine received within 2 weeks before vaccination with VLA1553, or any live vaccine received within 4 weeks before vaccination with VLA1553. Participants were randomised (3:1) to receive VLA1553 or placebo. The primary endpoint was the proportion of baseline negative participants with a seroprotective chikungunya virus antibody level defined as 50% plaque reduction in a micro plaque reduction neutralisation test (µPRNT) with a µPRNT50 titre of at least 150, 28 days after vaccination. The safety analysis included all individuals who received vaccination. Immunogenicity analyses were done in a subset of participants at 12 pre-selected study sites. These participants were required to have no major protocol deviations to be included in the per-protocol population for immunogenicity analyses. This trial is registered at ClinicalTrials.gov, NCT04546724. FINDINGS: Between Sept 17, 2020 and April 10, 2021, 6100 people were screened for eligibility. 1972 people were excluded and 4128 participants were enrolled and randomised (3093 to VLA1553 and 1035 to placebo). 358 participants in the VLA1553 group and 133 participants in the placebo group discontinued before trial end. The per-protocol population for immunogenicity analysis comprised 362 participants (266 in the VLA1553 group and 96 in the placebo group). After a single vaccination, VLA1553 induced seroprotective chikungunya virus neutralising antibody levels in 263 (98·9%) of 266 participants in the VLA1553 group (95% CI 96·7-99·8; p<0·0001) 28 days post-vaccination, independent of age. VLA1553 was generally safe with an adverse event profile similar to other licensed vaccines and equally well tolerated in younger and older adults. Serious adverse events were reported in 46 (1·5%) of 3082 participants exposed to VLA1553 and eight (0·8%) of 1033 participants in the placebo arm. Only two serious adverse events were considered related to VLA1553 treatment (one mild myalgia and one syndrome of inappropriate antidiuretic hormone secretion). Both participants recovered fully. INTERPRETATION: The strong immune response and the generation of seroprotective titres in almost all vaccinated participants suggests that VLA1553 is an excellent candidate for the prevention of disease caused by chikungunya virus. FUNDING: Valneva, Coalition for Epidemic Preparedness Innovation, and EU Horizon 2020.

Aspheric surface measurement by absolute wavelength scanning interferometry with model-based retrace error correction.

This paper presents a non-nulling absolute interferometric method for fast and full-area measurement of aspheric surfaces without the necessity of any mechanical movement. Several single frequency laser diodes with some degree of laser tunability are used to achieve an absolute interferometric measurement. The virtual interconnection of three different wavelengths makes it possible to accurately measure the geometrical path difference between the measured aspheric surface and the reference Fizeau surface independently for each pixel of the camera sensor. It is thus possible to measure even in undersampled areas of the high fringe density interferogram. After measuring the geometrical path difference, the retrace error associated with the non-nulling mode of the interferometer is compensated for using a calibrated numerical model (numerical twin) of the interferometer. A height map representing the normal deviation of the aspheric surface from its nominal shape is obtained. The principle of absolute interferometric measurement and numerical error compensation are described in this paper. The method was experimentally verified by measuring an aspheric surface with a measurement uncertainty of λ/20, and the results were in good agreement with the results of a single-point scanning interferometer.

Optically modified second harmonic generation in silicon oxynitride thin films via local layer heating.

Strong second harmonic generation (SHG) in silicon nitride has been extensively studied-among others, in terms of laser-induced SHG enhancement in Si3N4 waveguides. This enhancement has been ascribed to the all-optical poling induced by the coherent photogalvanic effect. Yet, an analogous process for Si3N4 thin films has not been reported. Our article reports on the observation of laser-induced threefold SHG enhancement in Si3N4 thin films. The observed enhancement has many features similar to all-optical poling, such as highly nonlinear power dependence, cumulative effect, or connection to the Si3N4-Si interface. However, identical experiments for low-oxygen silicon oxynitride thin films lead to complex behavior, including laser-induced SHG reduction. Following a thorough experimental study, including the effects of repetition rate or pulse length, the observed results were ascribed to heat-induced SHG variation. In addition to revealing a new mechanism of laser-induced SHG variation, our results also provide a means to identify this mechanism.

Squared Focal Intensity Distributions for Applications in Laser Material Processing.

Tailored intensity profiles within the focal spot of the laser beam offer great potential for a well-defined control of the interaction process between laser radiation and material, and thus for improving the processing results. The present paper discusses a novel refractive beam-shaping element that provides different squared intensity distributions converted from the Gaussian output beam of the utilized femtosecond (fs) laser. Using the examples of surface structuring of stainless-steel on the micro- and nano-scale, the suitability of the beam-shaping element for fs-laser material processing with a conventional f-Theta lens is demonstrated. In this context, it was shown that the experimental structuring results are in good agreement with beam profile measurements and numerical simulations of the beam-shaping unit. In addition, the experimental results reveal the improvement of laser processing in terms of a significantly reduced processing time during surface nano-structuring and the possibility to control the ablation geometry during the fabrication of micro-channels.

Receptor-independent modulation of cAMP-dependent protein kinase and protein phosphatase signaling in cardiac myocytes by oxidizing agents.

The contraction and relaxation of the heart is controlled by stimulation of the ß1-adrenoreceptor (AR) signaling cascade, which leads to activation of cAMP-dependent protein kinase (PKA) and subsequent cardiac protein phosphorylation. Phosphorylation is counteracted by the main cardiac protein phosphatases, PP2A and PP1. Both kinase and phosphatases are sensitive to intramolecular disulfide formation in their catalytic subunits that inhibits their activity. Additionally, intermolecular disulfide formation between PKA type I regulatory subunits (PKA-RI) has been described to enhance PKA's affinity for protein kinase A anchoring proteins, which alters its subcellular distribution. Nitroxyl donors have been shown to affect contractility and relaxation, but the mechanistic basis for this effect is unclear. The present study investigates the impact of several nitroxyl donors and the thiol-oxidizing agent diamide on cardiac myocyte protein phosphorylation and oxidation. Although all tested compounds equally induced intermolecular disulfide formation in PKA-RI, only 1-nitrosocyclohexalycetate (NCA) and diamide induced reproducible protein phosphorylation. Phosphorylation occurred independently of ß1-AR activation, but was abolished after pharmacological PKA inhibition and thus potentially attributable to increased PKA activity. NCA treatment of cardiac myocytes induced translocation of PKA and phosphatases to the myofilament compartment as shown by fractionation, immunofluorescence, and proximity ligation assays. Assessment of kinase and phosphatase activity within the myofilament fraction of cardiac myocytes after exposure to NCA revealed activation of PKA and inhibition of phosphatase activity thus explaining the increase in phosphorylation. The data suggest that the NCA-mediated effect on cardiac myocyte protein phosphorylation orchestrates alterations in the kinase/phosphatase balance.

Tuning the resolution and depth of field of a lens using an adjustable ring beam illumination.

A pair of axicons with an adjustable separation between them is used to generate a variable diameter ring beam with high efficiency. This beam illuminates a lens to produce quasi-diffraction-free beams with a tunable spot size and depth of field. We studied the generated beam characteristics while changing either the ring diameter or its thickness. Such a scheme has applications in adjustable imaging, including nondiffracting beam microscopy, material processing with an irradiance above a certain threshold value, and particle trapping/manipulation.

Flat-field illumination for quantitative fluorescence imaging.

The uneven illumination of a Gaussian profile makes quantitative analysis highly challenging in laser-based wide-field fluorescence microscopy. Here we present flat-field illumination (FFI) where the Gaussian beam is reshaped into a uniform flat-top profile using a high-precision refractive optical component. The long working distance and high spatial coherence of FFI allows us to accomplish uniform epi and TIRF illumination for multi-color single-molecule imaging. In addition, high-throughput borderless imaging is demonstrated with minimal image overlap.

Measuring the spatiotemporal electric field of tightly focused ultrashort pulses with sub-micron spatial resolution.

We demonstrate a powerful and practical spectral interferometer with near-field scanning microscopy (NSOM) probes for measuring the spatiotemporal electric field of tightly focused ultrashort pulses with high spatial and spectral resolution. Our measurements involved numerical apertures as high as 0.44 and yielded the spatiotemporal field at and around the foci produced by two microscope objectives and several different lenses. For the first time, we measure the spatiotemporal field of the Bessel-like X-shaped pulse caused by spherical aberrations and a "fore-runner pulse" due to chromatic aberrations. We observed spatial features smaller than 1 microm and verified these results with non-paraxial simulations.

Erbium fiber laser based on intracore femtosecond-written fiber Bragg grating.

We report the inscription of fiber Bragg gratings (FBGs) into a nonphotosensitive Er-doped fiber by using a phase-mask scanning technique with near IR femtosecond laser pulses. A grating of 40 mm length with a period of 1.075 microm was realized. We measured transmission losses of -18.9 dB at lambda=1554.5 nm with a FWHM bandwidth of 0.15 nm. By pumping the fiber containing the fabricated FBG, it was possible to realize a fiber laser with an output power of 38 mW, a slope efficiency of 21.1%, and low noise (SNR=60 dB).

Hybrid optics for focusing ultrashort laser pulses.

We present a specially designed hybrid optics comprising refraction and diffraction effects for tight spatial and temporal focusing of ultrashort laser pulses. Both aims can be put into practice by having a high numerical aperture (NA=0.45) and low internal dispersion at the same time. We are presenting what we believe is the first experimental realization of such a hybrid short plus focusing optics. The focusing properties are compared with a commonly used microscope objective (20 X, NA = 0.45) in theory and experimentally.

Ultra-short pulse propagation in complex optical systems.

In application of ultra-short laser pulses the pulse parameters have to be controlled accurately. Hence the manipulation of the propagation behavior of ultra-short pulses requires for specially designed optics. We have developed a tool for the simulation of ultra-short laser pulse propagation through complex real optical systems based on a combination of ray-tracing and wave optical propagation methods. For the practical implementation of the approach two commercially available software packages have been linked together, which are ZEMAX and Virtual Optics Lab. The focussing properties of different lenses will be analyzed and the results are demonstrated.