NK Cells Negatively Regulate CD8 T Cells to Promote Immune Exhaustion and Chronic Toxoplasma gondii Infection.

NK cells regulate CD4+ and CD8+ T cells in acute viral infection, vaccination, and the tumor microenvironment. NK cells also become exhausted in chronic activation settings. The mechanisms causing these ILC responses and their impact on adaptive immunity are unclear. CD8+ T cell exhaustion develops during chronic Toxoplasma gondii (T. gondii) infection resulting in parasite reactivation and death. How chronic T. gondii infection impacts the NK cell compartment is not known. We demonstrate that NK cells do not exhibit hallmarks of exhaustion. Their numbers are stable and they do not express high PD1 or LAG3. NK cell depletion with anti-NK1.1 is therapeutic and rescues chronic T. gondii infected mice from CD8+ T cell exhaustion dependent death, increases survival after lethal secondary challenge and alters cyst burdens in brain. Anti-NK1.1 treatment increased polyfunctional CD8+ T cell responses in spleen and brain and reduced CD8+ T cell apoptosis in spleen. Chronic T. gondii infection promotes the development of a modified NK cell compartment, which does not exhibit normal NK cell characteristics. NK cells are Ly49 and TRAIL negative and are enriched for expression of CD94/NKG2A and KLRG1. These NK cells are found in both spleen and brain. They do not produce IFNγ, are IL-10 negative, do not increase PDL1 expression, but do increase CD107a on their surface. Based on the NK cell receptor phenotype we observed NKp46 and CD94-NKG2A cognate ligands were measured. Activating NKp46 (NCR1-ligand) ligand increased and NKG2A ligand Qa-1b expression was reduced on CD8+ T cells. Blockade of NKp46 rescued the chronically infected mice from death and reduced the number of NKG2A+ cells. Immunization with a single dose non-persistent 100% protective T. gondii vaccination did not induce this cell population in the spleen, suggesting persistent infection is essential for their development. We hypothesize chronic T. gondii infection induces an NKp46 dependent modified NK cell population that reduces functional CD8+ T cells to promote persistent parasite infection in the brain. NK cell targeted therapies could enhance immunity in people with chronic infections, chronic inflammation and cancer.

Innate Lymphoid Cells in Protection, Pathology, and Adaptive Immunity During Apicomplexan Infection.

Apicomplexans are a diverse and complex group of protozoan pathogens including Toxoplasma gondii, Plasmodium spp., Cryptosporidium spp., Eimeria spp., and Babesia spp. They infect a wide variety of hosts and are a major health threat to humans and other animals. Innate immunity provides early control and also regulates the development of adaptive immune responses important for controlling these pathogens. Innate immune responses also contribute to immunopathology associated with these infections. Natural killer (NK) cells have been for a long time known to be potent first line effector cells in helping control protozoan infection. They provide control by producing IL-12 dependent IFNγ and killing infected cells and parasites via their cytotoxic response. Results from more recent studies indicate that NK cells could provide additional effector functions such as IL-10 and IL-17 and might have diverse roles in immunity to these pathogens. These early studies based their conclusions on the identification of NK cells to be CD3-, CD49b+, NK1.1+, and/or NKp46+ and the common accepted paradigm at that time that NK cells were one of the only lymphoid derived innate immune cells present. New discoveries have lead to major advances in understanding that NK cells are only one of several populations of innate immune cells of lymphoid origin. Common lymphoid progenitor derived innate immune cells are now known as innate lymphoid cells (ILC) and comprise three different groups, group 1, group 2, and group 3 ILC. They are a functionally heterogeneous and plastic cell population and are important effector cells in disease and tissue homeostasis. Very little is known about each of these different types of ILCs in parasitic infection. Therefore, we will review what is known about NK cells in innate immune responses during different protozoan infections. We will discuss what immune responses attributed to NK cells might be reconsidered as ILC1, 2, or 3 population responses. We will then discuss how different ILCs may impact immunopathology and adaptive immune responses to these parasites.

Discovery of Clinical Candidate 4-[2-(5-Amino-1H-pyrazol-4-yl)-4-chlorophenoxy]-5-chloro-2-fluoro-N-1,3-thiazol-4-ylbenzenesulfonamide (PF-05089771): Design and Optimization of Diaryl Ether Aryl Sulfonamides as Selective Inhibitors of NaV1.7.

A series of acidic diaryl ether heterocyclic sulfonamides that are potent and subtype selective NaV1.7 inhibitors is described. Optimization of early lead matter focused on removal of structural alerts, improving metabolic stability and reducing cytochrome P450 inhibition driven drug-drug interaction concerns to deliver the desired balance of preclinical in vitro properties. Concerns over nonmetabolic routes of clearance, variable clearance in preclinical species, and subsequent low confidence human pharmacokinetic predictions led to the decision to conduct a human microdose study to determine clinical pharmacokinetics. The design strategies and results from preclinical PK and clinical human microdose PK data are described leading to the discovery of the first subtype selective NaV1.7 inhibitor clinical candidate PF-05089771 (34) which binds to a site in the voltage sensing domain.

Discovery and Optimization of Selective Nav1.8 Modulator Series That Demonstrate Efficacy in Preclinical Models of Pain.

Voltage-gated sodium channels, in particular Nav1.8, can be targeted for the treatment of neuropathic and inflammatory pain. Herein, we described the optimization of Nav1.8 modulator series to deliver subtype selective, state, and use-dependent chemical matter that is efficacious in preclinical models of neuropathic and inflammatory pain.

Logical rules and the classification of integral-dimension stimuli.

A classic distinction in perceptual information processing is whether stimuli are composed of separable dimensions, which are highly analyzable, or integral dimensions, which are processed holistically. Previous tests of a set of logical-rule models of classification have shown that separable-dimension stimuli are processed serially if the dimensions are spatially separated and as a mixture of serial and parallel processes if the dimensions are spatially overlapping (Fific, Little, & Nosofsky, 2010; Little, Nosofsky, & Denton, 2011). In the current research, the logical-rule models are applied to predict response-time (RT) data from participants trained to classify integral-dimension color stimuli into rule-based categories. In dramatic contrast to the previous results for separable-dimension stimuli, analysis of the current data indicated that processing was best captured by a single-channel coactive model. The results converge with previous operations that suggest holistic processing of integral-dimension stimuli and demonstrate considerable generality for the application of the logical-rule models to predicting RT data from rule-based classification experiments.

Studies of implicit prototype extraction in patients with mild cognitive impairment and early Alzheimer's disease.

Studies of incidental category learning support the hypothesis of an implicit prototype-extraction system that is distinct from explicit memory (Smith, 2008). In those studies, patients with explicit-memory impairments due to damage to the medial-temporal lobe performed normally in implicit categorization tasks (Bozoki, Grossman, & Smith, 2006; Knowlton & Squire, 1993). However, alternative interpretations are that (a) even people with impairments to a single memory system have sufficient resources to succeed on the particular categorization tasks that have been tested (Nosofsky & Zaki, 1998; Zaki & Nosofsky, 2001) and (b) working memory can be used at time of test to learn the categories (Palmeri & Flanery, 1999). In the present experiments, patients with amnestic mild cognitive impairment or early Alzheimer's disease were tested in prototype-extraction tasks to examine these possibilities. In a categorization task involving discrete-feature stimuli, the majority of subjects relied on memories for exceedingly few features, even when the task structure strongly encouraged reliance on broad-based prototypes. In a dot-pattern categorization task, even the memory-impaired patients were able to use working memory at time of test to extract the category structure (at least for the stimulus set used in past work). We argue that the results weaken the past case made in favor of a separate system of implicit prototype extraction.

Primes and flankers: source confusions and discounting.

Visual identification of briefly presented target words is affected by the presence of nondiagnostic prime words that immediately precede the target, flanker words simultaneously presented adjacent to the target, and visual masks that immediately follow the target in the same location. Priming is duration dependent: In a forced choice target identification task, brief primes produce a strong preference to choose the primed alternative, whereas long primes have the opposite effect. The ROUSE model (Huber, Shiffrin, Lyle, & Ruys, Psychological Review 108:149-182, 2001) predicts this interaction by positing that prime features are confused with target features and that evidence regarding the prime features is discounted less for short primes and more for long primes, when both are compared with the optimal level. In the present study, we augmented the typical short-term priming experiment by adding flankers that appeared simultaneously with the target and remained for a short or long duration. In the experiment, we replicated previous priming effects and produced novel effects of flanker duration. ROUSE accounted for both the priming and flanker findings with the previously posited processes, but with different quantitative parameters for flankers: Relative to optimal levels of discounting, all flanker features were underdiscounted, but longer flankers were discounted more than short flankers.

The discovery of UK-369003, a novel PDE5 inhibitor with the potential for oral bioavailability and dose-proportional pharmacokinetics.

This paper describes our recent efforts to design and synthesise potent and selective PDE5 inhibitors and the use of in vitro predictors of clearance, absorption and permeability to maximise the potential for dose-proportional pharmacokinetics and good oral bioavailability in man. Optimisation of the preclinical profile resulted in the identification of UK-369003 (19a) and its nomination as a clinical candidate. The clinical pharmacokinetic and safety profile has enabled us to progress the compound to test its efficacy in patients with lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH) and a paper describing its efficacy has recently been published.

Response-time tests of logical-rule models of categorization.

A recent resurgence in logical-rule theories of categorization has motivated the development of a class of models that predict not only choice probabilities but also categorization response times (RTs; Fific, Little, & Nosofsky, 2010). The new models combine mental-architecture and random-walk approaches within an integrated framework and predict detailed RT-distribution data at the level of individual participants and individual stimuli. To date, however, tests of the models have been limited to validation tests in which participants were provided with explicit instructions to adopt particular processing strategies for implementing the rules. In the present research, we test conditions in which categories are learned via induction over training exemplars and in which participants are free to adopt whatever classification strategy they choose. In addition, we explore how variations in stimulus formats, involving either spatially separated or overlapping dimensions, influence processing modes in rule-based classification tasks. In conditions involving spatially separated dimensions, strong evidence is obtained for application of logical-rule strategies operating in a serial-self-terminating processing mode. In conditions involving spatially overlapping dimensions, preliminary evidence is obtained that a mixture of serial and parallel processing underlies the application of rule-based classification strategies. The logical-rule models fare considerably better than major extant alternative models in accounting for the categorization RTs.

Rule-based extrapolation: a continuing challenge for exemplar models.

Erickson and Kruschke (1998, 2002) demonstrated that in rule-plus-exception categorization, people generalize category knowledge by extrapolating in a rule-like fashion, even when they are presented with a novel stimulus that is most similar to a known exception. Although exemplar models have been found to be deficient in explaining rule-based extrapolation, Rodrigues and Murre (2007) offered a variation of an exemplar model that was better able to account for such performance. Here, we present the results of a new rule-plus-exception experiment that yields rule-like extrapolation similar to that of previous experiments, and yet the data are not accounted for by Rodrigues and Murre's augmented exemplar model. Further, a hybrid rule-and-exemplar model is shown to better describe the data. Thus, we maintain that rule-plus-exception categorization continues to be a challenge for exemplar-only models.

Attention and salience in associative blocking.

The associative learning effect called blocking has previously been found in many cue-competition paradigms where all cues are of equal salience. Previous research by Hall, Mackintosh, Goodall,and dal Martello (1977) found that, in animals, salient cues were less likely to be blocked. Crucially, they also found that when the to-be-blocked cue was highly salient, the blocking cue would lose some control over responding. The present article extends these findings to humans and suggests that shifts in attention can explain the apparent loss of control by the previously learned cue. A connectionist model that implements attentional learning is shown to fit the main trends in the data. Model comparisons suggest that mere forgetting, implemented as weight decay, cannot explain the results.