Inflammatory cytokine signalling in vulvovaginal candidiasis: a hot mess driving immunopathology.

Protective immunity to opportunistic fungal infections consists of tightly regulated innate and adaptive immune responses that clear the infection. Immune responses to infections of the vaginal mucosa by Candida species are, however, an exception. In the case of vulvovaginal candidiasis (VVC), the inflammatory response is associated with symptomatic disease, rather than that it results in pathogen clearance. As such VVC can be considered an inflammatory disease, which is a significant public health problem due to its predominance as a female-specific fungal infection. Particularly, women with recurrent VVC (RVVC) suffer from a significant negative impact on their quality of life and mental health. Knowledge of the inflammatory pathogenesis of (R)VVC may guide more effective diagnostic and therapeutic options to improve the quality of life of women with (R)VVC. Here, we review the immunopathogenesis of (R)VVC describing several elements that induce an inflammatory arson, starting with the activation threshold established by vaginal epithelial cells that prevent unnecessary ignition of inflammatory responses, epithelial and inflammasome-dependent immune responses. These inflammatory responses will drive neutrophil recruitment and dysfunctional neutrophil-mediated inflammation. We also review the, sometimes controversial, findings on the involvement of adaptive and systemic responses. Finally, we provide future perspectives on the potential of some unexplored cytokine axes and discuss whether VVC needs to be subdivided into subgroups to improve diagnosis and treatment.

Elevated memory T-cell conversion in a preclinical mouse model of hemophilia A.

One of the major challenges in the choice of the best therapeutic approach for the treatment of patients affected by hemophilia A (HA) is the definition of criteria predicting the formation of factor VIII (FVIII) neutralizing antibodies, called inhibitors. Both genetic and environmental elements influencing the immune response toward FVIII have been identified but still not all the factors causing the pathological rejection of FVIII have been identified. Since there is a connection between coagulation and inflammation, here we assessed the role played by the FVIII deficiency in shaping the humoral and cellular response toward an antigen other than FVIII itself. To this aim, we challenged both HA and wild-type (WT) mice with either FVIII or ovalbumin (OVA) and followed antigen-specific antibody level, immune cell population frequency and phenotype up to 9 weeks after the last antigen booster. The activation threshold was evaluated in vitro by stimulating the murine T cells with a decreasing dose of α-CD3. The humoral response to FVIII was similar between the two groups while both the in vivo and in vitro experiments highlighted an antigen-independent sensitivity of HA compared with WT T cells causing an increase in memory T-cell conversion and proliferation capability.

Retinoid X receptor gamma dictates the activation threshold of group 2 innate lymphoid cells and limits type 2 inflammation in the small intestine.

Group 2 innate lymphoid cells (ILC2s) are crucial in promoting type 2 inflammation that contributes to both anti-parasite immunity and allergic diseases. However, the molecular checkpoints in ILC2s that determine whether to immediately launch a proinflammatory response are unknown. Here, we found that retinoid X receptor gamma (Rxrg) was highly expressed in small intestinal ILC2s and rapidly suppressed by alarmin cytokines. Genetic deletion of Rxrg did not impact ILC2 development but facilitated ILC2 responses and the tissue inflammation induced by alarmins. Mechanistically, RXRγ maintained the expression of its target genes that support intracellular cholesterol efflux, which in turn reduce ILC2 proliferation. Furthermore, RXRγ expression prevented ILC2 response to mild stimulations, including low doses of alarmin cytokine and mechanical skin injury. Together, we propose that RXRγ expression and its mediated lipid metabolic states function as a cell-intrinsic checkpoint that confers the threshold of ILC2 activation in the small intestine.

MicroRNA-29 specifies age-related differences in the CD8+ T cell immune response.

MicroRNAs (miRNAs) have emerged as critical regulators of cell fate in the CD8+ T cell response to infection. Although there are several examples of miRNAs acting on effector CD8+ T cells after infection, it is unclear whether differential expression of one or more miRNAs in the naive state is consequential in altering their long-term trajectory. To answer this question, we examine the role of miR-29 in neonatal and adult CD8+ T cells, which express different amounts of miR-29 only prior to infection and adopt profoundly different fates after immune challenge. We find that manipulation of miR-29 expression in the naive state is sufficient for age-adjusting the phenotype and function of CD8+ T cells, including their regulatory landscapes and long-term differentiation trajectories after infection. Thus, miR-29 acts as a developmental switch by controlling the balance between a rapid effector response in neonates and the generation of long-lived memory in adults.

Molecular mechanism underlying modulation of TRPV1 heat activation by polyols.

Sensing noxiously high temperatures is crucial for living organisms to avoid heat-induced injury. The TRPV1 channel has long been known as a sensor for noxious heat. However, the mechanism of how this channel is activated by heat remains elusive. Here we found that a series of polyols including sucrose, sorbitol, and hyaluronan significantly elevate the heat activation threshold temperature of TRPV1. The modulatory effects of these polyols were only observed when they were perfused extracellularly. Interestingly, mutation of residues E601 and E649 in the outer pore region of TRPV1 largely abolished the effects of these polyols. We further observed that intraplantar injection of polyols into the hind paws of rats reduced their heat-induced pain response. Our observations not only suggest that the extracellular regions of TRPV1 are critical for the modulation of heat activation by polyols, but also indicate a potential role of polyols in reducing heat-induced pain sensation.

M-wave and H-reflex recruitment curves in boys and men.

The aim of the present study was to check whether the M-wave and H-reflex recruitment curves differ between prepubertal boys and men. Eleven boys (9-11 yr) and eleven men (18-35 yr) were magnetically stimulated at the tibial nerve in a prone position. M-wave and H-reflex maximal amplitudes (Hmax; Mmax ; Hmax /Mmax ), thresholds, regression slopes (Hslp ; Mslp ; Hslp /Mslp ) were extracted from M-wave and H-reflex recruitment curves and compared between the two age groups. Overall, no significant difference in M-wave and H-reflex recruitment curve parameters was found between the two populations. Nevertheless, the size of the M-wave associated with maximal H-reflex amplitude was lower in boys as compared to men when expressed relative to maximal M-wave amplitude (MHmax /Mmax : 0.18 ± 0.06 vs. 0.31 ± 0.13; p < .05). This result suggests that the development of peripheral nerve was completed in 9 to 11-year-old boys and did not affect the M-wave and H-reflex recruitment curves parameters. In neuromuscular function studies, it implies that Hmax /Mmax and Hslp /Mslp could be used indifferently to compare spinal motoneuron excitability between 9-11-year-old boys and men. Conversely, evoking H-reflexes at a given percentage of Mmax may bias the comparison between boys and men.

Plexin-Bs enhance their GAP activity with a novel activation switch loop generating a cooperative enzyme.

Plexins receive guidance cues from semaphorin ligands and transmit their signal through the plasma membrane. This family of proteins is unique amongst single-pass transmembrane receptors as their intracellular regions interact directly with several small GTPases, which regulate cytoskeletal dynamics and cell adhesion. Here, we characterize the GTPase Activating Protein (GAP) function of Plexin-B1 and find that a cooperative GAP activity towards the substrate GTPase, Rap1b, is associated with the N-terminal Juxtamembrane region of Plexin-B1. Importantly, we unveil an activation mechanism of Plexin-B1 by identifying a novel functional loop which partially blocks Rap1b entry into the plexin GAP domain. Consistent with the concept of allokairy developed for other systems, Plexin-B activity is increased by an apparent substrate-mediated cooperative effect. Simulations and mutagenesis suggest the repositioned JM conformation is stabilized by the new activation switch loop when the active site is occupied, giving rise to faster enzymatic turnover and cooperative behavior. The biological implications, essentially those of a threshold behavior for cell migration, are discussed.

Comparison of Subthreshold 577 and 810 nm Micropulse Laser Effects on Heat-Shock Protein Activation Kinetics: Implications for Treatment Efficacy and Safety.

Purpose: To compare the safety and efficacy of 810 versus 577 nm laser wavelengths for micropulse subthreshold (sublethal) laser treatment by mathematical analysis. Methods: Two different representative laser parameter sets for micropulsed subthreshold diode laser treatment, one employing 810 nm and the other 577 nm, are compared with regard to efficacy by analysis of the kinetics of laser-induced heat-shock protein (HSP) activation; and for safety, by scaling law analysis. Results: Kinetics analysis of laser-induced HSP activation shows that the primary therapeutic effect of laser is thermal incitement of a long-term wavelength-independent increase in the rate of HSP-mediated protein repair specific to sick and dysfunctional cells, rather than from short-term increases in free intracellular HSP concentrations. Scaling law analysis of the same 810 and 577 nm laser parameters, however, finds treatment safety highly wavelength-sensitive, favoring 810 over 577 nm. Conclusions: Mathematical analyses of the effects retinal laser-induced HSP activation provide important insights into the mechanism of action and the importance of wavelength selection in modern retinal laser therapy. Our analyses find 810 and 577 nm to be equally effective, but 810 nm having a significantly wider therapeutic range/safety margin, and thus less likely to cause inadvertent, and thus unpredictable, laser-induced retinal damage, than 577 nm. Translational Relevance: Mathematical analysis of enzyme reaction kinetics provides important insights into the mechanism of action and clinical implications of wavelength selection in modern retinal laser therapy.

Determining Whether Agonist Density or Agonist Number Is More Important for Immune Activation via Micoparticle Based Assay.

It is unknown if surface bound toll-like-receptor (TLR) agonists activate cells via density or total molecular number. To answer this question, we developed a TLR agonist surface conjugated polystyrene microparticle (MP) system. Using a library of MPs with varying TLR agonist density and number, we simultaneously observed innate immune cell MP uptake and TNFα expression using ImageStream flow cytometry on a cell by cell basis. The data shows that total TLR number and not density drives cellular activation with a threshold of approximately 105-106 TLR agonists. We believe that this information will be crucial for the design of particulate vaccine formulations.

Secrecy Performance Analysis of Wireless Powered Sensor Networks Under Saturation Nonlinear Energy Harvesting and Activation Threshold.

In this paper, we investigate the impact of saturation nonlinear energy harvesting (EH) and activation threshold on the multiuser wireless powered sensor networks (WPSNs) from the physical layer security (PLS) perspective. In particular, for improving the secrecy performance, the generalized multiuser scheduling (GMS) scheme is exploited, in which the Kth strongest sensor is chosen based on the legitimate link. For evaluating the impact of various key parameters on the security of system, we obtain the exact closed-form expressions for secrecy outage probability (SOP) under linear EH (LEH), saturation nonlinear EH (SNEH) and saturation nonlinear EH with activation threshold (SNAT), respectively, and solve the maximization problem of secure energy efficiency (SEE). Simulation results demonstrate that: (1) the number of source sensors, the EH efficiency and the transmit power of power beacon (PB) all have positive impact on SOP, and the smaller generalized selection coefficient is advantageous for secrecy performance; (2) LEH is an ideal situation for SNEH when the saturation threshold is large enough and SNEH is a special situation for SNAT when the activation threshold is low enough; (3) the time-switching factor and the activation threshold both have an important impact on the secrecy performance, which are worth considering carefully.

Management of Fertigation in Horticultural Crops through Automation with Electrotensiometers: Effect on the Productivity of Water and Nutrients.

Water and nutrient requirements of horticultural crops are influenced by different factors such as: Type of crop, stage of development and production system. Although greenhouse horticultural crops are more efficient in the use of water and fertilizers compared to other production systems, it is necessary increase efficiency for which individualized fertigation strategies must be designed for each greenhouse. The automation of fertigation based on the level of soil moisture allows optimization of management. The objective of this work was to determine the influence of the activation command of fertigation with electrotensiometers and the characteristics of the greenhouse on the productivity of the crop and the efficiency of use of water and nutrients in a sweet pepper crop. The trial was developed in two greenhouses. Four treatments were studied, combination of who two-factor: Soil matric potential (SMP) (SMP-10: Automatic activation of irrigation to -10 kPa and SMP-20: Automatic activation of irrigation to -20 kPa) and greenhouse characteristics (G1 and G2). The nutritive solution applied was the same in all treatments. The yield and volume of water and nutrients applied were determined, calculating the productivity of the water (WP), as well as productivity the nutrients. The fertigation activation threshold of -10 kPa presented the best results, increasing the yield and conserving WP and nutrient productivity with respect to -20 kPa in both greenhouses. The automation of irrigation with electrotensiometers allowed the application of different volume of fertigation demanded by the crop in each greenhouse, equalizing the WP and nutrient productivity without producing drainage. The pepper crop in the greenhouse G1 presented greater vegetative development, higher yield and demanded a greater volume of fertigation than G2 regardless of the activation threshold. This was due to the fact that the soil matric potential after irrigation in greenhouse G1 was closer to zero, being able to conclude that not only the soil matric potential threshold of irrigation activation has an influence on crop, but also the potential registered after irrigation. Soil matric potentials closer to zero are more productive.

CD300c costimulates IgE-mediated basophil activation, and its expression is increased in patients with cow's milk allergy.

BACKGROUND: Basophils express high-affinity IgE receptors (FcεRI), which play an essential role in allergic diseases. It is important to characterize new cell-surface receptors that modulate IgE-mediated basophil activation threshold to design promising immunomodulatory therapies. OBJECTIVES: We sought to analyze the expression of CD300 receptors on human basophils and their implication in IgE-mediated basophil activation processes. METHODS: Blood samples from healthy subjects and patients with cow's milk allergy were collected through the Basque Biobank under an institutional review board-approved protocol. PBMCs were obtained by means of density centrifugation, basophils were purified with a specific isolation kit, and phenotypic and functional studies were performed by using flow cytometry. RESULTS: We demonstrate that basophils express the activating receptor CD300c, which is specifically upregulated in response to IL-3. CD300c works as a costimulatory molecule during IgE-mediated basophil activation, as shown by a significant increase in degranulation and cytokine production when basophils are activated in the presence of CD300c cross-linking compared with activation through the IgE/FcεRI axis alone. Coligation of FcεRI and CD300c increased intracellular calcium mobilization and phosphorylation of signaling intermediates evoked only by FcεRI ligation. We show that the natural ligands of CD300c, phosphatidylserine and phosphatidylethanolamine, modulate IgE-mediated basophil activation. Furthermore, we have observed that CD300c expression in children with cow's milk allergy is increased compared with that in healthy control subjects and that the intensity of expression correlates with the severity of the hypersensitivity symptoms. CONCLUSION: CD300c could be considered a biomarker and therapeutic target in patients with IgE-mediated allergic diseases because it seems to be involved in the modulation of IgE-mediated basophil activation.

Controlling Depth of Cellular Quiescence by an Rb-E2F Network Switch.

Quiescence is a non-proliferative cellular state that is critical to tissue repair and regeneration. Although often described as the G0 phase, quiescence is not a single homogeneous state. As cells remain quiescent for longer durations, they move progressively deeper and display a reduced sensitivity to growth signals. Deep quiescent cells, unlike senescent cells, can still re-enter the cell cycle under physiological conditions. Mechanisms controlling quiescence depth are poorly understood, representing a currently underappreciated layer of complexity in growth control. Here, we show that the activation threshold of a Retinoblastoma (Rb)-E2F network switch controls quiescence depth. Particularly, deeper quiescent cells feature a higher E2F-switching threshold and exhibit a delayed traverse through the restriction point (R-point). We further show that different components of the Rb-E2F network can be experimentally perturbed, following computer model predictions, to coarse- or fine-tune the E2F-switching threshold and drive cells into varying quiescence depths.

High mTOR activity is a hallmark of reactive natural killer cells and amplifies early signaling through activating receptors.

NK cell education is the process through which chronic engagement of inhibitory NK cell receptors by self MHC-I molecules preserves cellular responsiveness. The molecular mechanisms responsible for NK cell education remain unclear. Here, we show that mouse NK cell education is associated with a higher basal activity of the mTOR/Akt pathway, commensurate to the number of educating receptors. This higher activity was dependent on the SHP-1 phosphatase and essential for the improved responsiveness of reactive NK cells. Upon stimulation, the mTOR/Akt pathway amplified signaling through activating NK cell receptors by enhancing calcium flux and LFA-1 integrin activation. Pharmacological inhibition of mTOR resulted in a proportional decrease in NK cell reactivity. Reciprocally, acute cytokine stimulation restored reactivity of hyporesponsive NK cells through mTOR activation. These results demonstrate that mTOR acts as a molecular rheostat of NK cell reactivity controlled by educating receptors and uncover how cytokine stimulation overcomes NK cell education.

A Minimum Epitope Overlap between Infections Strongly Narrows the Emerging T Cell Repertoire.

Many infections are caused by pathogens that are similar, but not identical, to previously encountered viruses, bacteria, or vaccines. In such re-infections, pathogens introduce known antigens, which are recognized by memory T cells and new antigens that activate naive T cells. How preexisting memory T cells impact the repertoire of T cells responding to new antigens is still largely unknown. We demonstrate that even a minimum epitope overlap between infections strongly increases the activation threshold and narrows the diversity of T cells recruited in response to new antigens. Thus, minimal cross-reactivity between infections can significantly impact the outcome of a subsequent immune response. Interestingly, we found that non-transferrable memory T cells are most effective in raising the activation threshold. Our findings have implications for designing vaccines and suggest that vaccines meant to target low-affinity T cells are less effective when they contain a strong CD8 T cell epitope that has previously been encountered.

IL-2 sensitivity and exogenous IL-2 concentration gradient tune the productive contact duration of CD8(+) T cell-APC: a multiscale modeling study.

BACKGROUND: The CD8(+) T cell immune response fights acute infections by intracellular pathogens and, by generating an immune memory, enables immune responses against secondary infections. Activation of the CD8(+) T cell immune response involves a succession of molecular events leading to modifications of CD8(+) T cell population. To understand the endogenous and exogenous mechanisms controlling the activation of CD8(+) T cells and to investigate the influence of early molecular events on the long-term cell population behavior, we developed a multiscale computational model. It integrates three levels of description: a Cellular Potts model describing the individual behavior of CD8(+) T cells, a system of ordinary differential equations describing a decision-making molecular regulatory network at the intracellular level, and a partial differential equation describing the diffusion of IL-2 in the extracellular environment. RESULTS: We first calibrated the model parameters based on in vivo data and showed the model's ability to reproduce early dynamics of CD8(+) T cells in murine lymph nodes after influenza infection, both at the cell population and intracellular levels. We then showed the model's ability to reproduce the proliferative responses of CD5(hi) and CD5(lo) CD8(+) T cells to exogenous IL-2 under a weak TCR stimulation. This stressed the role of short-lasting molecular events and the relevance of explicitly describing both intracellular and cellular scale dynamics. Our results suggest that the productive contact duration of CD8(+) T cell-APC is influenced by the sensitivity of individual CD8(+) T cells to the activation signal and by the IL-2 concentration in the extracellular environment. CONCLUSIONS: The multiscale nature of our model allows the reproduction and explanation of some acquired characteristics and functions of CD8(+) T cells, and of their responses to multiple stimulation conditions, that would not be accessible in a classical description of cell population dynamics that would not consider intracellular dynamics.

Dynamic tuning of lymphocytes: physiological basis, mechanisms, and function.

Dynamic tuning of cellular responsiveness as a result of repeated stimuli improves the ability of cells to distinguish physiologically meaningful signals from each other and from noise. In particular, lymphocyte activation thresholds are subject to tuning, which contributes to maintaining tolerance to self-antigens and persisting foreign antigens, averting autoimmunity and immune pathogenesis, but allowing responses to strong, structured perturbations that are typically associated with acute infection. Such tuning is also implicated in conferring flexibility to positive selection in the thymus, in controlling the magnitude of the immune response, and in generating memory cells. Additional functional properties are dynamically and differentially tuned in parallel via subthreshold contact interactions between developing or mature lymphocytes and self-antigen-presenting cells. These interactions facilitate and regulate lymphocyte viability, maintain their functional integrity, and influence their responses to foreign antigens and accessory signals, qualitatively and quantitatively. Bidirectional tuning of T cells and antigen-presenting cells leads to the definition of homeostatic set points, thus maximizing clonal diversity.