Human protective response induced by meningococcus B vaccine is mediated by the synergy of multiple bactericidal epitopes.

4CMenB is the first broad coverage vaccine for the prevention of invasive meningococcal disease caused by serogroup B strains. To gain a comprehensive picture of the antibody response induced upon 4CMenB vaccination and to obtain relevant translational information directly from human studies, we have isolated a panel of human monoclonal antibodies from adult vaccinees. Based on the Ig-gene sequence of the variable region, 37 antigen-specific monoclonal antibodies were identified and produced as recombinant Fab fragments, and a subset also produced as full length recombinant IgG1 and functionally characterized. We found that the monoclonal antibodies were cross-reactive against different antigen variants and recognized multiple epitopes on each of the antigens. Interestingly, synergy between antibodies targeting different epitopes enhanced the potency of the bactericidal response. This work represents the first extensive characterization of monoclonal antibodies generated in humans upon 4CMenB immunization and contributes to further unraveling the immunological and functional properties of the vaccine antigens. Moreover, understanding the mechanistic nature of protection induced by vaccination paves the way to more rational vaccine design and implementation.

Dynamic behaviour of multilamellar vesicles under Poiseuille flow.

Surfactant solutions exhibit multilamellar surfactant vesicles (MLVs) under flow conditions and in concentration ranges which are found in a large number of industrial applications. MLVs are typically formed from a lamellar phase and play an important role in determining the rheological properties of surfactant solutions. Despite the wide literature on the collective dynamics of flowing MLVs, investigations into the flow behavior of single MLVs are scarce. In this work, we investigate a concentrated aqueous solution of linear alkylbenzene sulfonic acid (HLAS), characterized by MLVs dispersed in an isotropic micellar phase. Rheological tests show that the HLAS solution is a shear-thinning fluid with a power law index dependent on the shear rate. Pressure-driven shear flow of the HLAS solution in glass capillaries is investigated using high-speed video microscopy and image analysis. The so obtained velocity profiles provide evidence for a power-law fluid behaviour of the HLAS solution and images show a flow-focusing effect of the lamellar phase in the central core of the capillary. The flow behavior of individual MLVs shows analogies with that of unilamellar vesicles and emulsion droplets. Deformed MLVs exhibit typical shapes of unilamellar vesicles, such as parachute and bullet-like. Furthermore, MLV velocity follows the classical Hetsroni theory for droplets provided that the power law shear dependent viscosity of the HLAS solution is taken into account. The results of this work are relevant for the processing of surfactant-based systems in which the final properties depend on the flow-induced morphology, such as cosmetic formulations and food products.

High numerical aperture holographic microscopy reconstruction with extended z range.

A holographic microscopy reconstruction method compatible with a high numerical aperture microscope objective (MO) up to NA=1.4 is proposed. After off-axis and reference field curvature corrections, and after selection of the +1 grating order holographic image, a phase mask that transforms the optical elements of the holographic setup into an afocal device is applied in the camera plane. The reconstruction is then made by the angular spectrum method. The field is first propagated in the image half-space from the camera to the afocal image of the MO optimal plane (the plane for which the MO has been designed) by using a quadratic kernel. The field is then propagated from the MO optimal plane to the object with the exact kernel. Calibration of the reconstruction is made by imaging a calibrated object such as a USAF resolution target for different positions along z. Once the calibration is done, the reconstruction can be made with an object located in any plane z. The reconstruction method has been validated experimentally with a USAF target imaged with a NA=1.4 microscope objective. Near-optimal resolution is obtained over an extended range (±50 µm) of z locations.