Peripheral blood mononuclear cell transcriptome profile in a clinical trial with subcutaneous, grass pollen allergoid immunotherapy.

INTRODUCTION: Allergen-specific immunotherapy (AIT) is the only disease-modifying treatment in allergic airway diseases. Underlying immunological mechanisms and candidate biomarkers, which may be translated into predictive/surrogate measures of clinical efficacy, remain an active area of research. The aim of this study was to evaluate Pollinex Quattro (PQ) Grass AIT induced immunomodulatory mechanisms, based on transcriptome profiling of peripheral blood mononuclear cells. METHODS: 119 subjects with grass pollen induced seasonal allergic rhinitis (SAR) were randomized in a 2:2:1:1 ratio to receive a cumulative dose of PQ Grass as a conventional or extended pre-seasonal regimen, placebo, or placebo with MicroCrystalline Tyrosine. Gene expression analysis was an exploratory endpoint evaluated in a subgroup of 30 subjects randomly selected from the four treatment arms. Samples were collected at three time points: screening (baseline), before the start of the grass pollen season and at the end of the season. This study was funded by the manufacturer of PQ. RESULTS: Transcriptome analysis demonstrated that the most significant changes in gene expression, for both treatment regimens, were at the end of the grass pollen season, with the main Th1 candidate molecules (IL-12A, IFNγ) upregulated and Th2 signature cytokines downregulated (IL-4, IL-13, IL-9) (p < .05). Canonical pathways analysis demonstrated Th1, Th2, Th17 and IL-17 as the most significantly enriched pathways based on absolute value of activation z-score (IzI score ≥ 2, p < .05). Upstream regulator analysis showed pronounced inhibition of pro-inflammatory allergic molecules IgE, IL-17A, IL-17F, IL-25 (IL-17E) (IzI score ≥ 2, FDR < 0.05) and activation of pro-tolerogenic molecules IL-12A, IL-27, IL-35 (EBI3) at the end of the grass pollen season. CONCLUSION: Peripheral blood mononuclear cells transcriptome profile showed an inhibition of Th2, Th17 pro-inflammatory allergic responses and immune deviation towards Th1 responses. PQ Grass extended regimen exhibited a superior mechanistic efficacy profile in comparison with PQ conventional regimen.

Influence of antigen density and TLR ligands on preclinical efficacy of a VLP-based vaccine against peanut allergy.

BACKGROUND: Virus-like particle (VLP) Peanut is a novel immunotherapeutic vaccine candidate for the treatment of peanut allergy. The active pharmaceutical ingredient represents cucumber mosaic VLPs (CuMVTT -VLPs) that are genetically fused with one of the major peanut allergens, Ara h 2 (CuMVTT -Ara h 2). We previously demonstrated the immunogenicity and the protective capacity of VLP Peanut-based immunization in a murine model for peanut allergy. Moreover, a Phase I clinical trial has been initiated using VLP Peanut material manufactured following a GMP-compliant manufacturing process. Key product characterization studies were undertaken here to understand the role and contribution of critical quality attributes that translate as predictive markers of immunogenicity and protective efficacy for clinical vaccine development. METHOD: The role of prokaryotic RNA encapsulated within VLP Peanut on vaccine immunogenicity was assessed by producing a VLP Peanut batch with a reduced RNA content (VLP Peanut low RNA). Immunogenicity and peanut allergen challenge studies were conducted with VLP Peanut low RNA, as well as with VLP Peanut in WT and TLR 7 KO mice. Furthermore, mass spectrometry and SDS-PAGE based methods were used to determine Ara h 2 antigen density on the surface of VLP Peanut particles. This methodology was subsequently applied to investigate the relationship between Ara h 2 antigen density and immunogenicity of VLP Peanut. RESULTS: A TLR 7 dependent formation of Ara h 2 specific high-avidity IgG antibodies, as well as a TLR 7 dependent change in the dominant IgG subclass, was observed following VLP Peanut vaccination, while total allergen-specific IgG remained relatively unaffected. Consistently, a missing TLR 7 signal caused only a weak decrease in allergen tolerability after vaccination. In contrast, a reduced RNA content for VLP Peanut resulted in diminished total Ara h 2 specific IgG responses, followed by a significant impairment in peanut allergen tolerability. The discrepant effect on allergen tolerance caused by an absent TLR 7 signal versus a reduced RNA content is explained by the observation that VLP Peanut-derived RNA not only stimulates TLR 7 but also TLR 3. Additionally, a strong correlation was observed between the number of Ara h 2 antigens displayed on the surface of VLP Peanut particles and the vaccine's immunogenicity and protective capacity. CONCLUSIONS: Our findings demonstrate that prokaryotic RNA encapsulated within VLP Peanut, including antigen density of Ara h 2 on viral particles, are key contributors to the immunogenicity and protective capacity of the vaccine. Thus, antigenicity and RNA content are two critical quality attributes that need to be determined at the stage of manufacturing, providing robust information regarding the immunogenicity and protective capacity of VLP Peanut in the mouse which has translational relevance to the human setting.

A single bout of aerobic exercise does not alter inhibitory control preparatory set cerebral hemodynamics: Evidence from the antisaccade task.

A single bout of exercise improves executive function (EF) and is a benefit - in part -attributed to an exercise-mediated increase in cerebral blood flow enhancing neural efficiency. Limited work has used an event-related protocol to examine postexercise changes in preparatory phase cerebral hemodynamics for an EF task. This is salient given the neural efficiency hypothesis' assertion that improved EF is related to decreased brain activity. Here, event-related transcranial Doppler ultrasound was used to measure pro- (saccade to target) and antisaccades (saccade mirror-symmetrical target) preparatory phase middle cerebral artery velocity (MCAv) prior to and immediately after 15-min of aerobic exercise. Antisaccades produced longer reaction times (RT) and an increased preparatory phase MCAv than prosaccades - a result attributed to greater EF neural activity for antisaccades. Antisaccades selectively produced a postexercise RT reduction (ps < 0.01); however, antisaccade preparatory phase MCAv did not vary from pre- to postexercise (p=0.53) and did not correlate with the antisaccade RT benefit (p = 0.31). Accordingly, results provide no evidence that improved neural efficiency indexed via functional hyperemia is linked to a postexercise EF behavioural benefit. Instead, results support an evolving view that an EF benefit represents the additive interplay between interdependent exercise-mediated neurophysiological changes.

The next generation virus-like particle platform for the treatment of peanut allergy.

BACKGROUND: Allergy to peanut is one of the leading causes of anaphylactic reactions among food allergic patients. Immunization against peanut allergy with a safe and protective vaccine holds a promise to induce durable protection against anaphylaxis caused by exposure to peanut. A novel vaccine candidate (VLP Peanut), based on virus-like particles (VLPs), is described here for the treatment of peanut allergy. METHODS AND RESULTS: VLP Peanut consists of two proteins: a capsid subunit derived from Cucumber mosaic virus engineered with a universal T-cell epitope (CuMVTT ) and a CuMVTT subunit fused with peanut allergen Ara h 2 (CuMVTT -Ara h 2), forming mosaic VLPs. Immunizations with VLP Peanut in both naïve and peanut-sensitized mice resulted in a significant anti-Ara h 2 IgG response. Local and systemic protection induced by VLP Peanut were established in mouse models for peanut allergy following prophylactic, therapeutic, and passive immunizations. Inhibition of FcγRIIb function resulted in a loss of protection, confirming the crucial role of the receptor in conferring cross protection against peanut allergens other than Ara h 2. CONCLUSION: VLP Peanut can be delivered to peanut-sensitized mice without triggering allergic reactions, while remaining highly immunogenic and offering protection against all peanut allergens. In addition, vaccination ablates allergic symptoms upon allergen challenge. Moreover, the prophylactic immunization setting conferred the protection against subsequent peanut-induced anaphylaxis, showing the potential for preventive vaccination. This highlights the effectiveness of VLP Peanut as a prospective break-through immunotherapy vaccine candidate toward peanut allergy. VLP Peanut has now entered clinical development with the study PROTECT.

A single bout of passive exercise mitigates a mental fatigue-induced inhibitory control deficit.

Sustained cognitive effort associated with the psychomotor vigilance task (PVT) increases objective and subjective measures of mental fatigue and elicits a post-PVT inhibitory control deficit. In contrast, passive exercise wherein an individual's limbs are moved via an external force (i.e., mechanically driven cycle ergometer flywheel) provides a postexercise inhibitory control benefit linked to an exercise-based increase in cerebral blood flow. Here, we examined whether passive exercise performed concurrently with the PVT 'blunts' an inhibitory control deficit. On separate days, participants (N = 27) completed a 20 min PVT protocol (control condition) and same duration PVT protocol paired with passive cycle ergometry (passive exercise condition). Prior to (i.e., baseline), immediately after and 30 min after each condition inhibitory control was assessed via the antisaccade task. Antisaccades require a goal-directed eye movement (i.e., saccade) mirror-symmetrical to a target and provide an ideal tool for evaluating task-based changes in inhibitory control. PVT results showed that vigilance (as assessed via reaction time: RT) during control and passive exercise conditions decreased from the first to last 5 min of the protocol and increased subjective ratings of mental fatigue. As well, in the control condition, immediate (but not 30-min) post-intervention antisaccade RTs were longer than their baseline counterparts-a result evincing a transient mental fatigue-based inhibitory control deficit. For the passive exercise condition, immediate and 30-min post-intervention antisaccade RTs were shorter than their baseline counterparts and this result was linked to decreased subjective ratings of mental fatigue. Thus, passive exercise ameliorated the selective inhibitory control deficit associated with PVT-induced mental fatigue and thus provides a potential framework to reduce executive dysfunction in vigilance-demanding occupations.

Cerebral blood flow and immediate and sustained executive function benefits following single bouts of passive and active exercise.

Passive exercise occurs when an individual's limbs are moved via an external force and is a modality that increases cerebral blood flow (CBF) and provides an immediate postexercise executive function (EF) benefit. To our knowledge, no work has examined for how long passive exercise benefits EF. Here, healthy young adults (N = 22; 7 female) used a cycle ergometer to complete three 20-min conditions: passive exercise (via mechanically driven flywheel), a traditional light intensity (37 W) "active" exercise condition (i.e., via volitional pedalling) and a non-exercise control condition. An estimate of CBF was obtained via transcranial Doppler ultrasound measurement of middle cerebral artery blood velocity (MCAv) and antisaccades (i.e., saccade mirror-symmetrical to a target) were completed prior to and immediately, 30- and 60-min following each condition to assess EF. Passive and active exercise increased MCAv; however, the increase was larger in the latter condition. In terms of antisaccades, passive and active exercise provided an immediate postexercise reaction time benefit. At the 30-min assessment, the benefit was observed for active but not passive exercise and neither produced a benefit at the 60-min assessment. Thus, passive exercise provided an evanescent EF "boost" and is a finding that may reflect a smaller cortical hemodynamic response.

Distinct cortical haemodynamics during squat-stand and continuous aerobic exercise do not influence the magnitude of a postexercise executive function benefit.

A single bout of aerobic exercise benefits executive function (EF). A potential mechanism for this benefit is an exercise-mediated increase in cerebral blood flow (CBF) that elicits vascular endothelial shear-stress improving EF efficiency. Moderate intensity continuous aerobic exercise (MCE) asymptotically increases CBF, whereas continuous body weight squat-stand exercise (SSE) provides a large amplitude oscillatory response. Some work has proposed that an increase in CBF oscillation amplitude provides the optimal shear-stress for improving EF and brain health. We examined whether a large amplitude oscillatory CBF response associated with a single bout of SSE imparts a larger postexercise EF benefit than an MCE cycle ergometer protocol. Exercise changes in middle cerebral artery velocity (MCAv) were measured via transcranial Doppler ultrasound to estimate CBF, and pre- and postexercise EF was assessed via the antisaccade task. MCE produced a steady state increase in MCAv, whereas SSE produced a large amplitude MCAv oscillation. Both conditions produced a postexercise EF benefit that null hypothesis and equivalence tests showed to be comparable in magnitude. Accordingly, we provide a first demonstration that a single bout of SSE benefits EF; however, the condition's oscillatory CBF response does not impart a larger benefit than a time- and intensity-matched MCE protocol.

Immunological effects of adjuvanted low-dose allergoid allergen-specific immunotherapy in experimental murine house dust mite allergy.

BACKGROUND: Native allergen extracts or chemically modified allergoids are routinely used to induce allergen tolerance in allergen-specific immunotherapy (AIT), although mechanistic side-by-side studies are rare. It is paramount to balance optimal dose and allergenicity to achieve efficacy warranting safety. AIT safety and efficacy could be addressed by allergen dose reduction and/or use of allergoids and immunostimulatory adjuvants, respectively. In this study, immunological effects of experimental house dust mite (HDM) AIT were investigated applying high-dose HDM extract and low-dose HDM allergoids with and without the adjuvants microcrystalline tyrosine (MCT) and monophosphoryl lipid A (MPL) in a murine model of HDM allergy. METHODS: Cellular, humoral, and clinical effects of the different AIT strategies were assessed applying a new experimental AIT model of murine allergic asthma based on physiological, adjuvant-free intranasal sensitization followed by subcutaneous AIT. RESULTS: While low-dose allergoid and high-dose extract AIT demonstrated comparable potency to suppress allergic airway inflammation and Th2-type cytokine secretion of lung-resident lymphocytes and draining lymph node cells, low-dose allergoid AIT was less effective in inducing a potentially protective IgG1 response. Combining low-dose allergoid AIT with MCT or MCT and dose-adjusted MPL promoted Th1-inducing mechanisms and robust B-cell activation counterbalancing the allergic Th2 immune response. CONCLUSION: Low allergen doses induce cellular and humoral mechanisms counteracting Th2-driven inflammation by using allergoids and dose-adjusted adjuvants. In light of safety and efficacy improvement, future therapeutic approaches may use low-dose allergoid strategies to drive cellular tolerance and adjuvants to modulate humoral responses.

The unidirectional prosaccade switch-cost: no evidence for the passive dissipation of an oculomotor task-set inertia.

Cognitive flexibility is a core component of executive function and supports the ability to 'switch' between different tasks. Our group has examined the cost associated with switching between a prosaccade (i.e., a standard task requiring a saccade to veridical target location) and an antisaccade (i.e., a non-standard task requiring a saccade mirror-symmetrical to veridical target) in predictable (i.e., AABB) and unpredictable (e.g., AABAB…) switching paradigms. Results have shown that reaction times (RTs) for a prosaccade preceded by an antisaccade (i.e., task-switch trial) are longer than when preceded by its same task-type (i.e., task-repeat trial), whereas RTs for antisaccade task-switch and task-repeat trials do not differ. The asymmetrical switch-cost has been attributed to an antisaccade task-set inertia that proactively delays a subsequent prosaccade (i.e., the unidirectional prosaccade switch-cost). A salient question arising from previous work is whether the antisaccade task-set inertia passively dissipates or persistently influences prosaccade RTs. Accordingly, participants completed separate AABB (i.e., A = prosaccade, B = antisaccade) task-switching conditions wherein the preparation interval for each trial was 'short' (1000-2000 ms; i.e., the timeframe used in previous work), 'medium' (3000-4000 ms) and 'long' (5000-6000 ms). Results demonstrated a reliable prosaccade switch-cost for each condition (ps < 0.02) and two one-sided test statistics indicated that switch cost magnitudes were within an equivalence boundary (ps < 0.05). Hence, null and equivalence tests demonstrate that an antisaccade task-set inertia does not passively dissipate and represents a temporally persistent feature of oculomotor control.

Evaluating the efficacy of an iPad® app in determining a single bout of exercise benefit to executive function.

We examined the efficacy and feasibility of an iPad® app used at-home in identifying a postexercise benefit to executive function. The iPad® app required simple reaching movements mirror-symmetrical to an exogenously presented target (i.e., antipointing) and is a task that lab-based behavioral and neuroimaging work has shown to provide a valid measure of the response inhibition component of executive function. Fifty English-speaking individuals (18 female, age range 18-26 years of age) completed the iPad® app before and immediately after a 20-min session of heavy-intensity aerobic exercise, and on a separate day completed the app prior to and following a 20-min non-exercise control condition. Results showed antipointing reaction times (RTs) in the exercise condition decreased by an average of 18 ms postexercise (p < 0.001) with an observed large effect size (dz = 0.90), whereas control condition pre- and post-assessment RTs did not reliably differ (p = 0.12, dz = 0.22) and were within an equivalence boundary (p < 0.005). Further, pre-assessment exercise and control condition antipointing RTs were within an equivalence boundary (p < 0.05). Accordingly, a simple iPad® app provides the requisite resolution to detect subtle executive function benefits derived from a single bout of exercise.

Visually guided saccades and acoustic distractors: no evidence for the remote distractor effect or global effect.

A remote visual distractor increases saccade reaction time (RT) to a visual target and may reflect the time required to resolve conflict between target- and distractor-related information within a common retinotopic representation in the superior colliculus (SC) (i.e., the remote distractor effect: RDE). Notably, because the SC serves as a sensorimotor interface it is possible that the RDE may be associated with the pairing of an acoustic distractor with a visual target; that is, the conflict related to saccade generation signals may be sensory-independent. To address that issue, we employed a traditional RDE experiment involving a visual target and visual proximal and remote distractors (Experiment 1) and an experiment wherein a visual target was presented with acoustic proximal and remote distractors (Experiment 2). As well, Experiments 1 and 2 employed no-distractor trials. Experiment 1 RTs elicited a reliable RDE, whereas Experiment 2 RTs for proximal and remote distractors were shorter than their no distractor counterparts. Accordingly, findings demonstrate that the RDE is sensory specific and arises from conflicting visual signals within a common retinotopic map. As well, Experiment 2 findings indicate that an acoustic distractor supports an intersensory facilitation that optimizes oculomotor planning.

'Delaying' a saccade: Preparatory phase cortical hemodynamics evince the neural cost of response inhibition.

Minimally delayed (MD) saccades require inhibition of a prepotent response until a target is extinguished, and unlike the more extensively studied antisaccade task, do not require the additional cognitive component of vector inversion (i.e., 180° target spatial transposition). Here, participants completed separate blocks of MD and prepotent stimulus-driven saccades (i.e., respond at target onset) while cortical hemodynamics were measured via functional transcranial Doppler ultrasound. MD saccades produced longer and more variable reaction times (RT). In turn, MD and stimulus-driven saccade preparatory phase cortical hemodynamics increased and decreased, respectively, relative to baseline and the two conditions differed from one another throughout the preparatory phase. The longer RTs and increased cortical hemodynamics of MD saccades is taken to evince response complexity and the increased neural activity to accommodate response inhibition. To our knowledge, such findings provide the first work to examine the neural foundations of MD saccades.

Vaccine against peanut allergy based on engineered virus-like particles displaying single major peanut allergens.

BACKGROUND: Peanut allergy is a severe and increasingly frequent disease with high medical, psychosocial, and economic burden for affected patients and wider society. A causal, safe, and effective therapy is not yet available. OBJECTIVE: We sought to develop an immunogenic, protective, and nonreactogenic vaccine candidate against peanut allergy based on virus-like particles (VLPs) coupled to single peanut allergens. METHODS: To generate vaccine candidates, extracts of roasted peanut (Ara R) or the single allergens Ara h 1 or Ara h 2 were coupled to immunologically optimized Cucumber Mosaic Virus-derived VLPs (CuMVtt). BALB/c mice were sensitized intraperitoneally with peanut extract absorbed to alum. Immunotherapy consisted of a single subcutaneous injection of CuMVtt coupled to Ara R, Ara h 1, or Ara h 2. RESULTS: The vaccines CuMVtt-Ara R, CuMVtt-Ara h 1, and CuMVtt-Ara h 2 protected peanut-sensitized mice against anaphylaxis after intravenous challenge with the whole peanut extract. Vaccines did not cause allergic reactions in sensitized mice. CuMVtt-Ara h 1 was able to induce specific IgG antibodies, diminished local reactions after skin prick tests, and reduced the infiltration of the gastrointestinal tract by eosinophils and mast cells after oral challenge with peanut. The ability of CuMVtt-Ara h 1 to protect against challenge with the whole extract was mediated by IgG, as shown via passive IgG transfer. FcγRIIb was required for protection, indicating that immune complexes with single allergens were able to block the allergic response against the whole extract, consisting of a complex allergen mixture. CONCLUSIONS: Our data suggest that vaccination using single peanut allergens displayed on CuMVtt may represent a novel therapy against peanut allergy with a favorable safety profile.

Increased cerebral blood flow supports a single-bout postexercise benefit to executive function: evidence from hypercapnia.

A single bout of aerobic exercise improves executive function; however, the mechanism for the improvement remains unclear. One proposal asserts that an exercise-mediated increase in cerebral blood flow (CBF) enhances the efficiency of executive-related cortical structures. To examine this, participants completed separate 10-min sessions of moderate- to heavy-intensity aerobic exercise, a hypercapnic environment (i.e., 5% CO2), and a nonexercise and nonhypercapnic control condition. The hypercapnic condition was included because it produces an increase in CBF independent of metabolic demands. An estimate of CBF was achieved via transcranial Doppler ultrasound and near-infrared spectroscopy that provided measures of middle cerebral artery blood velocity (BV) and deoxygenated hemoglobin (HHb), respectively. Exercise intensity was adjusted to match participant-specific changes in BV and HHb associated with the hypercapnic condition. Executive function was assessed before and after each session via antisaccades (i.e., saccade mirror-symmetrical to a target) because the task is mediated via the same executive networks that demonstrate task-dependent modulation following single and chronic bouts of aerobic exercise. Results showed that hypercapnic and exercise conditions were associated with comparable BV and HHb changes, whereas the control condition did not produce a change in either metric. In terms of antisaccade performance, the exercise and hypercapnic, but not control, conditions demonstrated improved postcondition reaction times (RT), and the magnitude of the hypercapnic and exercise-based increase in estimated CBF was reliably related to the postcondition improvement in RT. Accordingly, results evince that an increase in CBF represents a candidate mechanism for a postexercise improvement in executive function.NEW & NOTEWORTHY Single-bout aerobic exercise "boosts" executive function, and increased cerebral blood flow (CBF) has been proposed as a mechanism for the benefit. In this study, participants completed 10 min of aerobic exercise and 10 min of inhaling a hypercapnic gas, a manipulation known to increase CBF independently of metabolic demands. Both exercise and hypercapnic conditions improved executive function for at least 20 min. Accordingly, an increase in CBF is a candidate mechanism for the postexercise improvement in executive function.

A single bout of moderate intensity exercise improves cognitive flexibility: evidence from task-switching.

Executive function entails the core components of response  inhibition, working memory and cognitive flexibility. An accumulating literature has shown that a single bout of exercise improves the response inhibition  and working memory components of executive function; however, limited work has examined a putative exercise-related improvement to cognitive flexibility. To address this limitation, Experiment 1 entailed a 20-min session of moderate intensity aerobic exercise (via cycle ergometer), and pre- and post-exercise cognitive flexibility was examined via a task-switching paradigm involving alternating pro- and antisaccades (AABB: A = prosaccade, B = antisaccade). In Experiment 2, participants sat on the cycle ergometer without exercising (i.e., rest break) and the same AABB paradigm was examined pre- and post-break. We used an AABB pro- and antisaccade paradigm because previous work has shown that a prosaccade preceded by an antisaccade exhibits a reliable-and large magnitude-increase in reaction time, whereas the converse switch does not (i.e., the unidirectional prosaccade switch-cost). Experiment 1 showed a unidirectional prosaccade switch-cost pre-exercise (p = .012)-but not post-exercise (p = .30), and a two one-sided t test indicated that the latter comparison was within an equivalence boundary (p < .01). In contrast, Experiment 2 revealed a unidirectional prosaccade switch-cost at pre- and post-break assessments (ps < .01). Accordingly, our results indicate that a single bout of exercise improves cognitive flexibility and provides convergent evidence that exercise improves global components of executive function.

Microcrystalline Tyrosine and Aluminum as Adjuvants in Allergen-Specific Immunotherapy Protect from IgE-Mediated Reactivity in Mouse Models and Act Independently of Inflammasome and TLR Signaling.

Allergen immunotherapy (AIT) is the only modality that can modify immune responses to allergen exposure, but therapeutic coverage is low. One strategy to improve AIT safety and efficacy is the use of new or improved adjuvants. This study investigates immune responses produced by microcrystalline tyrosine (MCT)-based vaccines as compared with conventional aluminum hydroxide (alum). Wild-type, immune-signaling-deficient, and TCR-transgenic mice were treated with different Ags (e.g., OVA and cat dander Fel d 1), plus MCT or alum as depot adjuvants. Specific Ab responses in serum were measured by ELISA, whereas cytokine secretion was measured both in culture supernatants by ELISA or by flow cytometry of spleen cells. Upon initiation of AIT in allergic mice, body temperature and further clinical signs were used as indicators for anaphylaxis. Overall, MCT and alum induced comparable B and T cell responses, which were independent of TLR signaling. Alum induced stronger IgE and IL-4 secretion than MCT. MCT and alum induced caspase-dependent IL-1ß secretion in human monocytes in vitro, but inflammasome activation had no functional effect on inflammatory and Ab responses measured in vivo. In sensitized mice, AIT with MCT-adjuvanted allergens caused fewer anaphylactic reactions compared with alum-adjuvanted allergens. As depot adjuvants, MCT and alum are comparably effective in strength and mechanism of Ag-specific IgG induction and induction of T cell responses. The biocompatible and biodegradable MCT seems therefore a suitable alternative adjuvant to alum-based vaccines and AIT.

Pro- and antisaccade task-switching: response suppression-and not vector inversion-contributes to a task-set inertia.

Alternating between different tasks represents an executive function essential to activities of daily living. In the oculomotor literature, reaction times (RT) for a 'standard' and stimulus-driven (SD) prosaccade (i.e., saccade to target at target onset) are increased when preceded by a 'non-standard' antisaccade (i.e., saccade mirror-symmetrical to target at target onset), whereas the converse switch does not elicit an RT cost. The prosaccade switch-cost has been attributed to lingering neural activity-or task-set inertia-related to the antisaccade executive demands of response suppression and vector inversion. It is, however, unclear whether response suppression and/or vector inversion contribute to the prosaccade switch-cost. Experiment 1 of the present work had participants alternate (i.e., AABB paradigm) between minimally delayed (MD) pro- and antisaccades. MD saccades require a response after target extinction and necessitate response suppression for both pro- and antisaccades-a paradigm providing a framework to determine whether vector inversion contributes to a task-set inertia. In Experiment 2, participants alternated between SD pro- and MD antisaccades-a paradigm designed to determine if a switch-cost is selectively imparted when a SD and standard response is preceded by a non-standard response. Experiment 1 showed that RTs for MD pro- and antisaccades were refractory to the preceding trial-type; that is, vector inversion did not engender a switch-cost. Experiment 2 indicated that RTs for SD prosaccades were increased when preceded by an MD antisaccade. Accordingly, response suppression engenders a task-set inertia but only for a subsequent stimulus-driven and standard response (i.e., SD prosaccade). Such a result is in line with the view that response suppression is a hallmark feature of executive function.

Response suppression produces a switch-cost for spatially compatible saccades.

Executive function supports the rapid alternation between tasks for online reconfiguration of attentional and motor goals. The oculomotor literature has found that a prosaccade (i.e., saccade to veridical target location) preceded by an antisaccade (i.e., saccade mirror symmetrical to a target) elicits an increase in reaction time (RT), whereas the converse switch does not. This switch-cost has been attributed to the antisaccade task's requirement of inhibiting a prosaccade (i.e., response suppression) and transforming a target's coordinate (i.e., vector inversion)-executive processes thought to contribute to a task-set inertia that proactively interferes with the planning of a subsequent prosaccade. It is, however, unclear whether response suppression and vector inversion contribute to a task-set inertia or whether the phenomenon relates to a unitary component (e.g., response suppression). Here, the same stimulus-driven (SD) prosaccades (i.e., respond at target onset) as used in previous work were used with minimally delayed (MD) prosaccades (i.e., respond at target offset) and arranged in an AABB paradigm (i.e., A = SD prosaccade, B = MD prosaccade). MD prosaccades provide the same response suppression as antisaccades without the need for vector inversion. RTs for SD task-switch trials were longer and more variable than their task-repeat counterparts, whereas values for MD task-switch and task-repeat trials did not reliably differ. Moreover, SD task-repeat and task-switch movement times and amplitudes did not vary and thus demonstrate that a switch-cost is unrelated to a speed accuracy trade-off. Accordingly, results suggest the executive demands of response suppression is sufficient to engender the persistent activation of a non-standard task-set that selectively delays the planning of a subsequent SD prosaccade.

Safety Evaluation of PQ Birch Allergy Immunotherapy to Support Product Development.

PQ Birch represents an allergen-specific immunotherapy for the treatment of birch pollinosis. It consists of native birch pollen extract chemically modified with glutaldehyde adsorbed to L-tyrosine in its microcrystalline form with addition of the adjuvant Monophosphoryl Lipid A (MPL®). A nonclinical safety testing strategy was designed based upon interpretation of current legislation and regulatory intelligence and comprised genotoxicity studies (bacterial reverse mutation and Chinese hamster ovary micronucleus assays), a rat repeat dose toxicology study and a rabbit local tolerance study. No safety findings of concern were found. Thus, no evidence of genotoxicity was found. Relatively minor, immunostimulatory effects were seen following repeated subcutaneous dosing (once every 2 weeks for 13 weeks) as reversible increased white cell count (notably neutrophils), increased globulin level (resulting in decreased albumin/globulin [A/G] ratio) and increased fibrinogen, as well as minor dose site reaction in the form of inflammatory cell infiltrate. These findings are likely due to the immunostimulatory nature of MPL® and/or the presence of L-tyrosine within the adjuvanted vaccine. Similar dose site inflammatory changes to the injected formulation were also noted in the rabbit local tolerance study.

New toxicity testing of PQ grass allergy immunotherapy to support product development.

PQ Grass represents an allergen-specific immunotherapy for pre-seasonal treatment of patients with seasonal allergic rhinitis (or rhinoconjunctivitis) with or without mild-to-moderate bronchial asthma. It consists of a native pollen extract for 13 grass species, chemically modified with glutaraldehyde, and adsorbed to l-tyrosine in a microcrystalline form with addition of the adjuvant Monophosphoryl Lipid A (MPL® ). Previous non-clinical safety testing, including rat repeat dose toxicity in adult and juvenile animals, rat reproductive toxicity and rabbit local tolerance studies showed no safety findings of concern. A new Good Laboratory Practice compliant rat subcutaneous repeat dose toxicity study to evaluate a higher clinical dose and modified posology (once every 2 weeks for 13 weeks) showed no signs of toxicity. As seen in previous studies, relatively minor, immunostimulatory effects were seen such as reversible increased white cell count (notably neutrophils), increased globulin level (resulting in decreased A/G ratio) and increased fibrinogen as well as minor dose site reaction in the form of inflammatory cell infiltrate. These findings are likely due to the immunostimulatory nature of MPL and/or the presence of l-tyrosine within the adjuvanted vaccine. This new toxicity study with PQ Grass therefore supports longer posology with higher dose levels.