Exploring the Moderation Effect of Educational Stage on Visual Magnocellular Functioning Linked to Reading: A Study in French Primary School Children.

Many studies have investigated the visual magnocellular system functioning in dyslexia. However, very little is known on the relationship between the visual magnocellular system functioning and reading abilities in typical developing readers. In this study, we aimed at studying this relationship and more specifically the moderation effect of educational stage on this link. We thus tested 82 French typical developing readers (40 beginning readers-Grade 1 and 42 advanced readers-Grade 5) with reading tests and a coherent dot motion task measuring the visual magnocellular functioning. Results indicate positive correlations between visual magnocellular functioning and reading for beginning readers but not for advanced readers. Moreover, moderation analyses confirm that reading proficiency moderates the relationship between magnocellular system functioning and reading outcomes. We concluded that the relationship between visual magnocellular pathway functioning and reading abilities in typical developing readers could depend on reading proficiency.

Attentional Processing of Letter Strings by Children.

Reading a letter string requires attentional orienting toward the beginning of the string (left-dominant orientation), followed by orienting along the string. These attentional-orienting processes differ according to the lexicality of the letter string: Sequential processes apply when reading nonwords or pseudowords, while words can be processed more globally. The aim of this study was to evaluate the development of these attentional processes involved in reading. We conducted two experiments in 6- (first grade), 7- (second grade), and 9-year-old (fourth grade) children, using a procedure that required the detection of a letter (Experiment 1) or a nonletter (Experiment 2) target in a string of five characters. The target character could occur in the second (left) or fourth (right) position in the string. Results showed an advantage for left nonletter targets as early as age 6 and of left letter targets as early as age 7. In 6-year-olds, only good readers detected a left letter target faster than a right letter target; others detected a right letter target faster. Thus, dominant orienting toward the beginning of the letter string is not fully developed in children before the second year of reading. A possibility is that beginning readers have difficulties inhibiting an attention-orienting bias toward the right visual field in linguistic tasks. The results also showed that the lexicality effect on these attentional processes develops gradually until the fourth year of reading. We believe that the procedure used in this study will be very valuable for evaluating attentional difficulties during reading acquisition.

Uncertainty in fast task-irrelevant perceptual learning boosts learning of images in women but not men.

A key tenet of models of reinforcement learning is that learning is most desirable in the times of maximum uncertainty. Here we examine the role of uncertainty in the paradigm of fast task-irrelevant perceptual learning (fast-TIPL), where stimuli that are consistently presented at relevant points in times (e.g., with task targets or rewards) are better encoded than when presented at other times. We manipulated two forms of uncertainty, expected uncertainty and unexpected uncertainty (Yu & Dayan, 2005), and compared fast-TIPL under uncertainty with fast-TIPL under no uncertainty. Results indicate a larger fast-TIPL effect under uncertainty than under no uncertainty without a difference between expected and unexpected uncertainty. However, interestingly, this effect of uncertainty on fast-TIPL was found in women but not in men. In men, equivalent fast-TIPL was observed under no uncertainty and uncertainty, whereas in women, confirming previous results (Leclercq & Seitz, 2012b), no fast-TIPL was observed in the no-uncertainty condition, but fast-TIPL was observed in the uncertainty conditions. We discuss how these results imply differences in attention or neuromodulatory processes between men and women.

Encoding of episodic information through fast task-irrelevant perceptual learning.

The mechanisms guiding our learning and memory processes are of key interest to human cognition. While much research shows that attention and reinforcement processes help guide the encoding process, there is still much to know regarding how our brains choose what to remember. Recent research of task-irrelevant perceptual learning (TIPL) has found that information presented coincident with important events is better encoded even if participants are not aware of its presence (see Seitz & Watanabe, 2009). However a limitation of existing studies of TIPL is that they provide little information regarding the depth of encoding supported by pairing a stimulus with a behaviorally relevant event. The objective of this research was to understand the depth of encoding of information that is learned through TIPL. To do so, we adopted a variant of the "remember/know" paradigm, recently reported by Ingram, Mickes, and Wixted (2012), in which multiple confidence levels of both familiar (know) and remember reports are reported (Experiment 1), and in which episodic information is tested (Experiment 2). TIPL was found in both experiments, with higher recognition performance for target-paired than for distractor-paired images. Furthermore, TIPL benefitted both "familiar" and "remember" reports. The results of Experiment 2 indicate that the most confident "remember" response was associated with episodic information, where participants were able to access the location of image presentation for these items. Together, these results indicate that TIPL results in a deep enhancement in the encoding of target-paired information.

Development of endogenous orienting of attention in school-age children.

Orienting of attention was investigated in 6-, 8-, and 10-year-olds and in young adults, in a spatial cueing experiment comparing nonpredictive, predictive, and counterpredictive cues (in different blocks). A larger positive orienting effect (advantage of valid over invalid cues) in the predictive than in the nonpredictive condition occurred in all groups, showing efficient endogenous orienting of attention. However, this effect was larger in 6-year-olds, as if the ability to distribute attention between the different locations (and not only to orient to the most probable location) developed between 6 and 8 years. Moreover, only 10-year-olds and adults showed a significant negative orienting effect (advantage of invalid cues) in the counterpredictive condition, indicating inhibition of attentional capture by goal-irrelevant stimuli. Therefore, our results indicate a large change in the modulation of endogenous orienting between 6 and 10 years.

Enhancement from targets and suppression from cues in fast task-irrelevant perceptual learning.

Task-irrelevant perceptual learning (TIPL) refers to the phenomenon where the stimulus features of a subject's task are learned when they are consistently presented at times when behaviorally relevant events occur. In this article, we addressed two points concerning TIPL. First, we address the question, are all behaviorally relevant events equal in their impact on encoding processes? Second, we address the hypothesis that TIPL involves mechanisms of the alerting attentional system. Two experiments of fast-TIPL were conducted in which the attentional state of participants was manipulated by using an alerting cue (visual or auditory) that informed participants of the arrival of an upcoming target. Images were presented with task-related stimuli (cues, targets and distractors) and subjects were tested on their memory of those images. Results indicate that memory for target-paired images was enhanced and cue-paired images were suppressed relative to that of distractor-paired images. The alerting cue increased the ability to recall target-paired images presented after this cue, although this result depended on the proportion of cued trials in a session. These results demonstrate a complex interplay between task-elements and the encoding of stimuli paired with them where both enhancement and suppression of task-paired stimuli can be found depending whether those stimuli are paired with task-targets or cues.

Fast-TIPL occurs for salient images without a memorization requirement in men but not in women.

Recent research of task-irrelevant perceptual learning (TIPL) demonstrates that stimuli that are consistently presented at relevant point in times (e.g. with task-targets or rewards) are learned, even in the absence of attention to these stimuli. However, different research paradigms have observed different results for how salient stimuli are learned; with some studies showing no learning, some studies showing positive learning and others showing negative learning effects. In this paper we focused on how the level of processing of stimuli impacts fast-TIPL. We conducted three different experiments in which the level of processing of the information paired with a target was manipulated. Our results indicated that fast-TIPL occurs when participants have to memorize the information presented with the target, but also when they just have to process this information for a secondary task without an explicit memorization of those stimuli. However, fast-TIPL does not occur when participants have to ignore the target-paired information. This observation is consistent with recent models of TIPL that suggest that attentional signals can either enhance or suppress learning depending on whether those stimuli are distracting or not to the subjects' objectives. Our results also revealed a robust gender effect in fast-TIPL, where male subjects consistently show fast-TIPL, whereas the observation of fast-TIPL is inconsistent in female subjects.

The impact of orienting attention in fast task-irrelevant perceptual learning.

Task-irrelevant perceptual learning (TIPL) refers to the phenomenon where the stimulus features are learned when they are consistently presented at behaviorally relevant times (e.g., with task targets or rewards). Studies on the role of attention in TIPL have found that attention negatively impacts this type of learning; however, these studies involved stimuli that were completely irrelevant to the subjects and that, when noticed, were distracting to the subjects' task. Here, we asked whether attention would have a beneficial impact on learning in the case where the target-paired stimuli were relevant to a secondary task that subjects were required to perform. We conducted three experiments in adult subjects, using the fast-TIPL paradigm (which allows one to study TIPL with as little as a single trial of exposure). The results from Experiments 1 and 2 showed that fast-TIPL occurred for the target-paired stimuli but that the manipulation of attention increased performance for stimuli presented after the target. Experiment 3 was conducted to address whether the direction of attention positively or negatively impacted fast-TIPL and to better control for the effects of attention. The results of this experiment demonstrate that in the case of fast-TIPL, exogenously directed attention aids in the memorization of target-paired stimuli. Overall, our results demonstrate that attention operates in a beneficial manner in fast-TIPL, where the target-paired stimuli are relevant to a secondary task that subjects perform.

Fast task-irrelevant perceptual learning is disrupted by sudden onset of central task elements.

The basic phenomenon of task-irrelevant perceptual learning (TIPL) is that the stimulus features of a subject's task will be learned when they are consistently presented at times of reward or behavioral success. Recent progress in studies of TIPL has been made by the discovery of a fast form of TIPL (fast-TIPL), which can be observed with as little as a single trial of exposure. In the present study, we investigated the task-conditions required to observe fast-TIPL. We had participants perform a target detection task at fixation while scenes to memorize were presented peripherally. In some experiments the target was presented in a sequence of distractors (Experiments 2 and 4) and in others alone (Experiments 1 and 3). In each experiment we assessed whether learning for target-paired scenes was greater than that of nontarget-paired scenes. The results indicated an enhanced memorization for scenes paired with the targets in the experiments where the target was presented with distractors, but not in the experiments where distractors were not presented. We hypothesized that without the presentation of distractors the onset of the target was sudden and this may have exogenously drawn attention to the center of the display disrupting TIPL. This sudden onset hypothesis was experimentally confirmed in Experiment 5. We conclude that fast-TIPL, with its rapid time-course, and its production of learning for supraliminally presented stimuli, shows great promise as an efficient paradigm through which to understand mechanisms of learning.

Clinical-grade preparation of human natural regulatory T-cells encoding the thymidine kinase suicide gene as a safety gene.

BACKGROUND: Human CD4+CD25+FOXP3+ natural regulatory T-cells (nTreg) have a great therapeutic potential for the induction of tolerance in allo-transplanted patients or for the control of severe auto-immune diseases. However, clinical-grade production of nTreg remains difficult to achieve because of the absence of a truly specific surface marker and of their low frequency that implies a need for their ex vivo expansion. Furthermore, safety issues should be taken into consideration due to the risk of either uncontrolled nTreg-induced immunosuppression or uncontrolled proliferation of autoreactive contaminating T-cells particularly in an auto-immune context. METHODS: We compared different clinical-grade conditions for immuno-magnetic selection and ex vivo expansion of nTreg. For safety, expanded cells were genetically modified with retroviral vectors co-expressing human CD90 and HSV1 thymidine kinase. The CD90 surface marker and thymidine kinase allow for selection and elimination of transduced cells by ganciclovir, respectively. RESULTS: We showed that (i) nTreg could be enriched in a one step using CD25 microbeads, were functionally suppressive and mainly FOXP3+; (ii) using anti-CD28- and anti-CD3-coated beads, interleukin-2 and rapamycin, nTreg were expanded 150-200-fold after 3 weeks. Under these clinical-grade conditions, they remained suppressive, and no major alteration of the TCR repertoire was observed; (iii) after efficient retroviral transduction and CD90 selection, nTreg maintained their suppressive activity; (iv) transduced nTreg could be eliminated by ganciclovir upon activation. CONCLUSIONS: The efficient procedure reported here for the preparation of nTreg, whose safety has been ensured, is now applicable for further clinical trials.

Initial depletion of regulatory T cells: the missing solution to preserve the immune functions of T lymphocytes designed for cell therapy.

We investigated the causes of the altered functionality of T cells cultured under conditions designed for cell and gene therapy and the strategies to prevent their defects. We first showed that human T cells cultured for 6 days with anti-CD3 +/- anti-CD28 antibodies and interleukin-2 presented a 50% decrease of their proliferative responses to allogeneic or recall antigens. Similarly, day-6 cultured murine T cells completely lost their capacity to reject allogeneic skin grafts and to provoke graft-versus-host disease (GVHD) when infused into irradiated semi-allogeneic mice. Interestingly, injection of higher amounts of cultured T cells restored GVHD induction. Moreover, depletion of CD25+ cells prior to T-cell cultures can prevent these deficiencies both in mice and humans. Therefore, we demonstrated that culture conditions used for T-cell therapy preferentially activated and expanded regulatory T cells (Treg's). Thus, we showed that dividing cells sorted from T-cell cultures strongly suppressed the proliferation of autologous T cells in response to allogeneic stimulation. An increased detection of Foxp3 at mRNA and protein levels in the cultures confirmed the Treg expansion. Overall, we demonstrate that T-cell cultures promote Treg expansion over effector T cells, leading to deleterious immune functions, and that this imbalance can be prevented by an initial depletion of CD25+ cells.

Efficient transduction and selection of human T-lymphocytes with bicistronic Thy1/HSV1-TK retroviral vector produced by a human packaging cell line.

BACKGROUND: T-cells expressing the HSV1-TK suicide gene can be used for the control of graft-versus-host disease following allogeneic stem cell transplantation. To develop clinical trials based on such a strategy, we have generated under good manufacturing procedures a novel 'split genome' human packaging cell line (1704 cells). METHODS: To minimize the risk of generating replication-competent retroviruses, pol was truncated to remove sequences overlapping with env. To improve retroviral infection and selection of transduced T-cells, high titers of GALV-pseudotyped retroviral particles harboring a bicistronic Thy1-IRES-TK vector coding for the CD90 GPI-anchored membrane molecule were produced by 1704 cells. RESULTS: Using 1704 cell supernatant and an optimized transduction protocol, approximately 50% of primary T-cells were transduced and could then be purified (approximately 95%) using clinical-grade immunomagnetic beads directed against CD90. Over 96% of these OKT3/IL-2-activated CD90(+)-selected T-cells were killed by ganciclovir. Cell proliferation and cytokine production of transduced T-cells and HLA-restricted cytotoxicity of transduced T-cell clones were identical to those of their non-transduced counterparts cultured under the same conditions. CONCLUSIONS: GALV-pseudotyped retroviral particles harboring a bicistronic Thy1-IRES-TK vector allow efficient transduction and rapid selection of human T-cells under conditions applicable for clinical trials using the new human 1704 packaging cell line.

Highly active antiretroviral therapy corrects hematopoiesis in HIV-1 infected patients: interest for peripheral blood stem cell-based gene therapy.

OBJECTIVES: To study, in asymptomatic HIV-1-infected (HIV+) patients, whether peripheral blood hematopoietic progenitor/stem cells (PBPC) mobilized by granulocyte colony stimulating factor (G-CSF), can be used as a source of cells for retroviral gene therapy. DESIGN: PBPC from two groups of HIV+ patients (treated or untreated by highly active antiretroviral therapy) and from seronegative donors were mobilized with G-CSF. METHODS: PBPC collected by leukapheresis were enriched for CD34 cells, immunophenotypically and functionally characterized, cultured and infected with retroviral vectors. HIV proviral integration was studied on fresh and cultured cells. RESULTS: G-CSF moderately and transiently increased the viral load in untreated patients only, and induced in both groups of HIV+ patients mobilization of percentages and numbers of CD34 cells comparable to those of seronegative volunteers. The most immature CD34 cell subset, the clonogenic progenitor and long-term culture initiating cells were significantly decreased in leukapheresis products and CD34-enriched fractions from untreated HIV+ patients but not in those from treated HIV+ patients. Cell cycle activation and growth factor responses of CD34 cells from both groups of HIV+ patients were not different from those of the control group. Culture and retroviral infection of CD34 cells from HIV+ patients did not enhance HIV replication, and yielded transduction levels similar to those obtained using CD34 cells from seronegative donors. CONCLUSIONS: G-CSF-mobilized PBPC can be safely used for HIV retroviral gene therapy in asymptomatic treated patients while highly active antiretroviral therapy would control the G-CSF-induced increase in viral load and correct the defective hematopoiesis observed in untreated patients, without inhibiting the retroviral transduction of PBPC.