Liposome-Mediated Delivery of MERS Antigen Induces Potent Humoral and Cell-Mediated Immune Response in Mice

The advancements in the field of nanotechnology have provided a great platform for the development of effective antiviral vaccines. Liposome-mediated delivery of antigens has been shown to induce the antigen-specific stimulation of the humoral and cell-mediated immune responses. Here, we prepared dried, reconstituted vesicles (DRVs) from DPPC liposomes and used them as the vaccine carrier system for the Middle East respiratory syndrome coronavirus papain-like protease (DRVs-MERS-CoV PLpro). MERS-CoV PLpro emulsified in the Incomplete Freund’s Adjuvant (IFA-MERS-CoV PLpro) was used as a control. Immunization of mice with DRVs-MERS-CoV PLpro did not induce any notable toxicity, as revealed by the levels of the serum alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN) and lactate dehydrogenase (LDH) in the blood of immunized mice. Immunization with DRVs-MERS-CoV PLpro induced greater antigen-specific antibody titer and switching of IgG1 isotyping to IgG2a as compared to immunization with IFA-MERS-CoV PLpro. Moreover, splenocytes from mice immunized with DRVs-MERS-CoV PLpro exhibited greater proliferation in response to antigen stimulation. Moreover, splenocytes from DRVs-MERS-CoV PLpro-immunized mice secreted significantly higher IFN-γ as compared to splenocytes from IFA-MERS-CoV PLpro mice. In summary, DRVs-MERS-CoV PLpro may prove to be an effective prophylactic formulation to prevent MERS-CoV infection.

Glycan Activation of Clec4b Induces Reactive Oxygen Species Protecting against Neutrophilia and Arthritis

Animal models for complex diseases are needed to position and analyze the function of interacting genes. Previous positional cloning identified Ncf1 and Clec4b to be major regulators of arthritis models in rats. Here, we investigate epistasis between Ncf1 and Clec4b, two major regulators of arthritis in rats. We find that Clec4b and Ncf1 exert an additive effect on arthritis given by their joint ability to regulate neutrophils. Both genes are highly expressed in neutrophils, together regulating neutrophil availability and their capacity to generate reactive oxygen species. Using a glycan array, we identify key ligands of Clec4b and demonstrate that Clec4b-specific stimulation triggers neutrophils into oxidative burst. Our observations highlight Clec4b as an important regulator of neutrophils and demonstrate how epistatic interactions affect the susceptibility to, and severity of, autoimmune arthritis.

Anti-Inflammatory Effects of Immunostimulation in Patients with COVID-19 Pneumonia

Background: The effects of immunomodulators in patients with Coronavirus Disease 2019 (COVID-19) pneumonia are still unknown. We investigated the cellular inflammatory and molecular changes in response to standard-of-care + pidotimod (PDT) and explored the possible association with blood biomarkers of disease severity. Methods: Clinical characteristics and outcomes, neutrophil-to-lymphocyte ratio (NLR), plasma and cell supernatant chemokines, and gene expression patterns after SARS-CoV-2 and influenza (FLU) virus in vitro stimulation were assessed in 16 patients with mild-moderate COVID-19 pneumonia, treated with standard of care and PDT 800 mg twice daily (PDT group), and measured at admission, 7 (T1), and 12 (T2) days after therapy initiation. Clinical outcomes and NLR were compared with age-matched historical controls not exposed to PDT. Results: Hospital stay, in-hospital mortality, and intubation rate did not differ between groups. At T1, NLR was 2.9 (1.7–4.6) in the PDT group and 5.5 (3.4–7.1) in controls (p = 0.037). In the PDT group, eotaxin and IL-4 plasma concentrations progressively increased (p < 0.05). Upon SARS-CoV-2 and FLU-specific stimulation, IFN-γ was upregulated (p < 0.05), while at genetic transcription level, Pathogen Recognition Receptors (TRLs) were upregulated, especially in FLU-stimulated conditions. Conclusions: Immunomodulation exerted by PDT and systemic corticosteroids may foster a restoration in the innate response to the viral infection. These results should be confirmed in larger RCTs.

EVALUACIÓN DE LA CAPACIDAD ADYUVANTE DEL GINSENÓSIDO RG1 EN RATONES BALB/C INMUNIZADOS CON UN LISADO DE Staphylococcus aureus

Rg1 ginsenoside, the main active compound of Panax ginseng extract, could be a promising vaccine adjuvant against Staphylococcus aureusbovine mastitis. The aim was to evaluate the adjuvant effects of Rg1 combined with liposomes or alone against S. aureus lysate from bovine mastitis in mice. Female Balb/c mice were divided into 4 groups: S. aureus Lysate + IFA (Group I), S. aureus lysate + liposomes (Group II), S. aureus lysate + Rg1 (Group III) and S. aureus lysate + liposomes + Rg1 (Group IV). All mice were subcutaneously immunized with all formulations 3 times at 2-weeks intervals. Humoral response was evaluated by indirect IgG ELISA assay in plasma collected at the end of the assay. Cellular response was evaluated by MTT kit assay in spleen cells from immunized mice. Groups I and IV showed higher IgG values than groups II and III (p < 0.05). Groups I and II showed higher proliferation values than groups III and IV (p < 0.05). The Rg1-liposome combination increased S. aureus-specific IgG production. However, Rg1 was not able to stimulate lymphocyte proliferation from immunized mice, possibly due to the short S. aureus-specific stimulation time. New assays with different experimental conditions are needed to elucidate adjuvant effects of Rg1 in a murine model.

Effect of central and peripheral cone- and rod-specific stimulation on the pupillary light reflex

Purpose To assess the effect of central and peripheral stimulation on the pupillary light reflex. The aim was to detect possible differences between cone- and rod-driven reactions. Methods Relative maximal pupil constriction amplitude (relMCA) and latency to constriction onset (latency) to cone- and rod-specific stimuli of 30 healthy participants (24 ± 5 years (standard deviation)) were measured using chromatic pupil campimetry. Cone- and rod-specific stimuli had different intensities and wavelengths according to the Standards in Pupillography. Five filled circles with radii of 3°, 5°, 10°, 20° and 40° and four rings with a constant outer radius of 40° and inner radii of 3°, 5°, 10° and 20° were used as stimuli. Results For cone-and rod-specific stimuli, relMCA increased with the stimulus area for both, circles and rings. However, increasing the area of a cone-specific ring by minimizing its inner radius with constant outer radius increased relMCA significantly stronger than the same did for a rod-specific ring. For cones and rods, a circle stimulus with a radius of 40° created a lower relMCA than the summation of the relMCAs to the corresponding ring and circle stimuli which combined create a 40° circle-stimulus. Latency was longer for rods than for cones. It decreased with increasing stimulus area for circle stimuli while it stayed nearly constant with increasing ring stimulus area for cone- and rod-specific stimuli. Conclusion The effect of central stimulation on relMCA is more dominant for cone-specific stimuli than for rod-specific stimuli while latency dynamics are similar for both conditions.

A Rapid and Simple Multiparameter Assay to Quantify Spike-Specific CD4 and CD8 T Cells after SARS-CoV-2 Vaccination: A Preliminary Report

mRNA and Adenovirus vaccines for COVID-19 are used to induce humoral and cell-mediated immunity, with the aim to generate both SARS-CoV-2 B and T memory cells. In present study, we described a simple assay to detect and quantify Spike-specific CD4+ and CD8+ T cell responses induced by vaccination in healthy donors and in subjects with B cell compart impairment, in which antibody response is absent due to primary immunodeficiencies or CD20 depleting therapy. We detect and quantified memory T cell immune responses against SARS-CoV-2 evocated by vaccination in both groups, irrespective to the humoral response. Furthermore, we identified TNF-α as the main cytokine produced by T memory cells, after antigen-specific stimulation in vitro, that could be considered, other than IFN-γ, an additional biomarker of induction of T memory cells upon vaccination. Further studies on the vaccine-induced T cell responses could be crucial, not only in healthy people but also in immunocompromised subjects, where antigen specific T cells responses play a protective role against SARS-CoV-2.

tACS competes with ongoing oscillations for control of spike-timing in the primate brain.

Transcranial alternating current stimulation (tACS) is a promising but controversial method for modulating neural activity noninvasively. Much of the controversy revolves around the question of whether tACS can generate electric fields that are strong enough to entrain neuronal spiking activity. Here we show that what matters is not the electric field strength per se, but rather the strength of the stimulation relative to ongoing oscillatory entrainment. We recorded from single neurons in the cortex and subcortex of behaving non-human primates, while applying tACS at different frequencies and amplitudes. When neuronal activity was weakly locked to ongoing oscillations, tACS readily entrained single-neuron activity to specific stimulation phases. In contrast, neurons that were strongly locked to ongoing oscillations usually exhibited decreased entrainment during low-amplitude tACS. As this reduced entrainment is a property of many oscillating systems, attempts to impose an external rhythm on spiking activity may often yield precisely the opposite effect.

Selective electrical stimulation with currentfield modulation

Selective electrical stimulation using a multielectrode array is a promising technique that can potentially bring electrical stimulation treatment modalities a step forward. A microcontroller-controlled electrical stimulator system delivering a single pulse was designed, suitable for current-field modulation. The goal is to make electrical stimulation with surface electrodes more specific. A graphical user interface (GUI) was developed to control stimulation parameters and current-field within a multi-electrode array wirelessly. The stimulator generates arbitrary biphasic waveforms with a 5-bit resolution and high temporal precision (<10 μs) and was demonstrated to stimulate posterior lumbar root fibers in transcutaneous spinal cord stimulation (tSCS) treatment selectively. Current-field modulation throughout a sixteen-electrode array was achieved. The system has the goal to improve control of stimulation conditions in electrophysiological studies and time-dependent and site-specific stimulation patterns for neuromodulation applications. A novel feature is the current-field modulation ability of the stimulator for surface electrode arrays.