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Background: In Anorexia Nervosa (AN), association of weight gain recovery above WHO’s lower limits, and classical nutritional markers normalization appears to be mandatory, but not always sufficient, to restore menses. This may worry both patient and practitioner and disrupt medical care. Therefore, this study was designed to evaluate the ability of anthropometric and hormonal factors to predict menses resumption in anorexia nervosa once normal weight was normalized. Method: AN patients recovering a normal BMI but not their menses (ANRec) were evaluated twice, at visit 1, and 6 months later if bodyweight was maintained over this period (visit 2). The 46 ANRec patients reaching visit 2 were separated into two groups: 20 with persistent amenorrhea (PA-ANRec) and 26 with recovered menses (RM-ANRec). Anthropometric and plasma levels of several hormones, Luteinizing Hormone (LH) pulse and LH response to gonadotrophin releasing hormone (GnRH) injection were then compared between groups at visit 1. Results: RM-ANRec presented with higher plasma level of follicular Stimulating hormone (FSH), Estradiol, inhibin B, LH/GnRH and ghrelin compared to PA-ANRec. ROC curves analyses showed that LH pulses’ number ≥2, LH/GnRH ≥ 33 UI/l and Inhibin B >63 pg/ml predicted menses resumption with high specificity (87, 100 and 100% respectively) and sensitivity (82, 80 and 79% respectively). Conclusions: LH pulses’ number≥2, LH/GnRH≥33UI/l or Inhibin B≥63pg/ml can predict accurately menses resumption in still amenorrheic weight-normalized AN patients if they do not lose weight nor practice intense exercise. Negative parameters could help clinician and patient to maintain efforts to achieve individual metabolic set-point.

Adoptive cell transfer (ACT) of tumor-specific T lymphocytes represents a relevant therapeutic strategy to treat metastatic melanoma patients. Ideal T-cells should combine tumor specificity and reactivity with survival in vivo, while avoiding autoimmune side effects. Here we report results from a Phase I/II clinical trial (NCT02424916, performed between 2015 and 2018) in which 6 metastatic HLA-A2 melanoma patients received autologous antigen-specific T-cells produced from PBMC, after peptide stimulation in vitro, followed by sorting with HLA-peptide multimers and amplification. Each patient received a combination of Melan-A and MELOE-1 polyclonal specific T-cells, whose specificity and anti-tumor reactivity were checked prior to injection, with subcutaneous IL-2. Transferred T-cells were also characterized in terms of functional avidity, diversity and phenotype and their blood persistence was evaluated. An increase in specific T-cells was detected in the blood of all patients at day 1 and progressively disappeared from day 7 onwards. No serious adverse events occurred after this ACT. Clinically, five patients progressed and one patient experienced a partial response following therapy. Melan-A and MELOE-1 specific T-cells infused to this patient were diverse, of high avidity, with a high proportion of T lymphocytes co-expressing PD-1 and TIGIT but few other exhaustion markers. In conclusion, we demonstrated the feasibility and safety of ACT with multimer-sorted Melan-A and MELOE-1 specific T cells to metastatic melanoma patients. The clinical efficacy of such therapeutic strategy could be further enhanced by the selection of highly reactive T-cells, based on PD-1 and TIGIT co-expression, and a combination with ICI, such as anti-PD-1. Supplementary Information The online version contains supplementary material available at 10.1007/s00262-021-02961-0.

Summary Background & aims Patients with anorexia nervosa (AN) have low serum IGF-I levels that may contribute to a lower bone mineral mass. We investigated the effects of a fermented, protein-fortified, dairy product on serum IGF-I levels in patients with AN during an in-hospital refeeding program. Methods In this multicenter, randomized, double-blind, placebo-controlled, clinical trial conducted at 3 university hospitals and 3 private clinics in France and Switzerland, 62 women recently admitted with confirmed AN and with a baseline low serum IGF-I level were randomized to 2 daily isocaloric fresh cheese pots containing either 15 g/150 g or 3 g/150 g (controls) of protein for 4 weeks. The primary outcome was the change in IGF-I levels. Results In the primary intention-to-treat analysis, mean serum IGF-I levels increased during the intervention phase from 22.9 ± 1.5 to 28.6 ± 1.3 nmol/L (means ± SEM) (+20.2%) in the intervention group and from 20.2 ± 1.2 to 25.7 ± 1.5 nmol/L (+16.8%) in controls. In a preplanned analysis of covariance with repeated measures, the between-group difference was close to statistical significance ( P = 0.071). In a post-hoc mixed-regression model analysis, the difference was statistically significant (4.9 nmol/l increase; P = 0.003), as was the change of the ratio IGF-I/IGF-BP3 ( P = 0.004 ). There was no between-group difference in biochemical markers of bone turnover (osteocalcin, P1NP, CTX) or in serum parathyroid hormone level. Serum calcium levels slightly increased during the intervention phase in the higher protein group ( P = 0.02). IGF-BP2 decreased significantly more in the intervention group during the follow up period at week 4 after supplements cessation (P = 0.019). Conclusions Intake of a fermented, protein-fortified, isocaloric dairy product during 4 weeks may slightly increase serum IGF-I levels in women with AN, without significant changes in bone turnover markers. Clinical Trial Registration Number: NCT01823822 (www.clinicaltrials.gov).

International audience; There is now a consensus that efficient peptide vaccination against cancer requires that peptides should (i) be exclusively presented by professional APC and (ii) stimulate both CD4 and CD8-specific T cell responses. To this aim, in recent trials, patients were vaccinated with pools of synthetic long peptides (SLP) (15-30 aa long) composed of a potential class I epitope(s) elongated at both ends with native antigen sequences to also provide a potential class II epitope(s). Using MELOE-1 as a model antigen, we present an alternative strategy consisting in linking selected class I and class II epitopes with an artificial cathepsin-sensitive linker to improve epitope processing and presentation by DC. We provide evidence that some linker sequences used in our artificial SLPs (aSLPs) could increase up to 100-fold the cross-presentation of class I epitopes to CD8-specific T cell clones when compared to cross-presentation of the corresponding native long peptide. Presentation of class II epitopes were only slightly increased. We confirmed this increased cross-presentation after in vitro stimulation of PBMC from healthy donors with aSLP and assessment of CD8-specific responses and also in vivo following aSLP vaccination of HLA*A0201/HLA-DRB0101 transgenic mice. Finally, we provide some evidence that vaccination with aSLP could inhibit the growth of transplanted tumors in mice. Our data thus support the use of such aSLPs in future cancer vaccination trials to improve anti-tumor CD8 T cell responses and therapeutic efficacy. ARTICLE HISTORY

Therapeutic strategies using anti–PD-1–blocking antibodies reported unparalleled effectiveness for melanoma immunotherapy, but deciphering immune responses modulated by anti–PD-1 treatment remains a crucial issue. Here, we analyzed the composition and functions of the large Melan-A–specific T-cell repertoire in the peripheral blood of 9 melanoma patients before and after 2 months of treatment with anti–PD-1. We observed amplification of Melan-A–specific Vß subfamilies undetectable before therapy (thereafter called emerging Vß subfamilies) in responding patients, with a predominant expansion in patients with a complete response. These emerging Vß subfamilies displayed a higher functional avidity for their cognate antigen than Vß subfamilies not amplified upon anti–PD-1 therapy and could be identified by a sustained coexpression of PD-1 and TIGIT receptors. Thus, in addition to the emergence of neoantigen-specific T cells previously documented upon anti–PD-1 therapy, our work describes the emergence of high-avidity Melan-A–specific clonotypes as a surrogate marker of treatment efficacy. Cancer Res; 77(24); 7083–93. ©2017 AACR.

Depressive, anxiety and obsessive symptoms frequently co-occur with anorexia nervosa (AN). The relationship between these clinical manifestations and the biological changes caused by starvation is not well understood. It has been hypothesised that reduced availability of tryptophan (TRP) could reduce serotonin activity and thus trigger these comorbid symptoms. The aim of this study, during re-feeding in individuals with AN, was to analyse covariations across measures of nutritional status, depressive and anxiety symptoms, and peripheral serotonin markers. Depressive and anxiety symptoms, nutritional status and serotonin markers--whole blood serotonin content, plasma TRP and the ratio between TRP and large neutral amino acids--were assessed for 42 AN participants at admission to inpatient treatment and after re-feeding. Biological measures were compared to those obtained in 42 non-eating disordered subjects. For those with AN, psychological, nutritional and biological parameters improved significantly during hospitalisation. Levels of serotonin markers were significantly lower in the AN group compared to the control group, at admission and at discharge. Increase in the TRP/LNAA ratio was correlated with a decrease in depressive symptoms. In addition, there was a positive correlation between serotonin levels and symptoms of both anxiety and depression at discharge. We speculate that enhanced TRP availability during re-feeding, as a result of the increase in the TRP/LNAA ratio, could restore serotonin neurotransmission and lead to a decrease in depressive symptoms. The association between serotonin and anxiety and depressive symptoms would be consistent with numerous observations indicating abnormal functioning of the serotoninergic system in AN.

Introduction Constitutional thinness (CT) is an underweight state characterized by normal menstruations and no change in feeding behaviour. Thinness is the only resemblance between Anorexia Nervosa (AN) and CT. Removal of amenorrhea from the new DSM 5 definition of AN might result in misdiagnosis between these two populations. The objective of this study was to compare CT, AN and Control subjects in terms of biological, anthropometric, and psychological markers in order to better distinguish AN from CT subjects. Materials and methods Body composition, nutritional markers, pituitary hormones, bone markers and psychological scores were evaluated in three groups of young women: fifty-six CT, forty restrictive-type AN and fifty-four Control subjects. For every marker, a receiver Operator Characteristics (ROC) curve was calculated to evaluate the accuracy of differentiation between AN and CT groups. Results For most studied parameters, CT subjects were similar to Controls but dramatically different from AN subjects. DEBQ Restrained Eating subscale score was identified by ROC data analysis as the only psychological parameter tested to successfully differentiate AN from CT. Free-T3 and Leptin were shown to be powerful markers to differentiate AN and CT populations as they were highly specific and sensitive ones. Conclusion The exclusive use of psychological testing criteria is not always sufficient to differentiate AN and CT patients. Minimally, additional testing of Free T3 levels, which is cheap and widely accessible for general practitioners, should be completed to avoid misdiagnosis which could result in the implementation of ineffective treatment plans and social stigmatization for CT women.

CD137/TNFR9/41BB was originally described as a surface molecule present on activated T and NK cells. However, its expression is broader among leukocytes, and it is also detected on hypoxic endothelial cells and inflamed blood vessels, as well as in atherosclerotic lesions. Here, we demonstrate that lymphatic endothelial cells (LECs) up-regulate CD137 expression from undetectable baseline levels on stimulation with TNF-α, LPS, and IL-1β. CD137 cross-linking with an agonistic mAb results in NF-κB nuclear translocation, followed by up-regulation of VCAM and a 3-fold increase in the production of the chemokine CCL21. Accordingly, there is a 50% increase in CCR7-dependent migration toward conditioned medium from activated LECs on CD137 cross-linking with the agonistic mAb or the natural ligand (CD137L). Such an enhancement of cell migration is also observed with monocyte-derived dendritic cells transmigrating across CD137-activated LEC monolayers. Using explanted human dermal tissue, we found that inflamed skin contains abundant CD137(+) lymphatic vessels and that ex vivo incubation of explanted human dermis with TNF-α induces CD137 expression in lymphatic capillaries. More interestingly, treatment with CD137 agonistic antibody induces CCL21 expression and DC accumulation close to lymphatic vessels. Collectively, our results demonstrate that the inflammatory function of lymphatic vessels can be regulated by CD137.

Cognitive deficits are commonly reported in schizophrenia and have a significant impact on the daily life of patients and on their social and work inclusion. Cognitive remediation therapies (CRT) may enhance the capabilities of schizophrenia patients. Although social and work integration is the ultimate goal of CRT, previous studies have failed to carry out a detailed assessment of the effects on everyday life.Fifty-nine schizophrenia patients were randomised into two groups (remediation or usual treatment) to test the effects of a new remediation programme, which included both rehearsal and strategy learning, on cognitive functions. An ecological test was used to evaluate its transfer to daily living skills.Cognitive improvements are revealed in CRT patients, mainly in memory and executive functions. Patients showing some deficiencies to perform the ecological test had better scores after the CRT. Moreover, they significantly improve their social activity scores.CRT would facilitate mental load monitoring by enhancing or reallocating cognitive resources, facilitating the patient's organisation and autonomy. The rehearsal learning approach improves the ability to carry out automatic operations that are less demanding in terms of cognitive resources, thereby increasing the resources available for acquisition and efficient use of strategies provided during the strategy learning approach.

The angiogenic factor Midkine (MDK) is overexpressed in various human malignant tumors, although its expression is low or undetectable in normal adult tissues. Its expression in tumors and its detection in plasma have been associated with poor disease outcome, whereas its blockade was found to contribute to tumor regression. By weekly stimulation of T lymphocytes harvested in HLA-A2 healthy donors, we derived CD8 T cell lines specific for several MDK peptides. The T cell response was mostly dominated by two nonamer peptides localized in the signal peptide and in the C-terminal part of the protein, as assessed by IFN-γ ELISPOT and HLA-A2 tetramer labeling. Peptide-specific T cell lines recognized cells transfected with an MDK-encoded plasmid and tumor cell lines naturally expressing the MDK protein, but not untransfected cells. T cell presentation of the two MDK epitopes was found to be TAP dependent. Experiments performed in HLA-A2 transgenic mice demonstrated the capacity of the two identified CD8 T cell epitopes to elicit a cytotoxic response. Altogether, our data show that the secreted MDK protein is a candidate vaccine for multiple cancers.

Summary Constitutional thinness (CT) and anorexia nervosa (AN) are two categories of severely underweight subjects. Some appetite-regulating hormones display opposite levels in AN and CT. While levels of ghrelin, an orexigenic hormone, fit with the normal food intake in CT, the lack of efficacy of increased ghrelin levels in AN is not clear. Obestatin is a recently described peptide derived from the preproghrelin gene, reported to inhibit appetite in contrast to ghrelin. The aim of this study was to determine whether the circadian profile of obestatin, total and acylated ghrelin levels is different in CT subjects when compared with AN patients. Six-points circadian profiles of plasma obestatin, acylated ghrelin, total ghrelin and other hormonal and nutritional parameters were evaluated in four groups of young women: 10 CT, 15 restricting-type AN, 7 restored from AN and 9 control subjects. Obestatin circadian levels were significantly higher inAN (p < 0.0001) while no difference was found between CTand control subjects. Acylated and total ghrelin were found increased in AN. Acylated ghrelin/obestatin and total ghrelin/obestatin were found decreased in AN compared to CTor C subjects (p < 0.05). The percentage of acylated ghrelin was found decreased in CT group (p < 0.05). The decreased ghrelin/obestatin ratio found in AN might participate in the restraint in nutriment intake of these patients. In contrast, in CTa lower percentage of acylated over total ghrelin might be considered in the aetiology of this condition.

Resume Le statut du premier objet investi lors de la naissance de la vie psychique constitue une enigme complexe, tant scientifique que philosophique et humaine. Les auteurs en proposent une vision a la fois personnelle et inspiree du vaste champ metapsychologique jusqu’alors destine a apporter un support descriptif, voire explicatif, a cette question. Ils s’appuient pour cela sur la base d’un trepied conceptuel regroupant les processus de representation pictographique de type « contenant », le role reflecteur de l’objet externe et la bidimensionnalite. Leur modelisation decrit l’objet primaire sous la forme d’une experience psychique d’indifferenciation entre le sujet en devenir et son environnement, initiee au sein d’un cadre originaire pour se decliner ensuite graduellement et de maniere individuellement variable jusqu’a une ultime trace primaire.

// Maud Charpentier 1 , Mikael Croyal 2, 3 , Delphine Carbonnelle 1 , Agnes Fortun 1 , Laetitia Florenceau 1, 4 , Catherine Rabu 1 , Michel Krempf 2, 3, 4 , Nathalie Labarriere 1, 4 , Francois Lang 1 1 CRCNA, INSERM, CNRS, Universite d’Angers, Universite de Nantes, Nantes, France 2 UMR INRA 1280, CHU, Nantes, France 3 West Human Nutrition Research Center, CHU, Nantes, France 4 CHU, Nantes, France Correspondence to: Francois Lang, email: francois.lang@univ-nantes.fr Keywords: melanoma, IRES, long non coding RNA, tumor antigens, immunotherapy Received: June 07, 2016 Accepted: July 20, 2016 Published: July 29, 2016 ABSTRACT MELOE-1 and MELOE-2, two highly specific melanoma antigens involved in T cell immunosurveillance are produced by IRES-dependent translation of the long « non coding » and polycistronic RNA, meloe . In the present study, we document the expression of an additional ORF, MELOE-3, located in the 5′ region of meloe . Data from in vitro translation experiments and transfection of melanoma cells with bicistronic vectors documented that MELOE-3 is exclusively translated by the classical cap-dependent pathway. Using a sensitive tandem mass spectrometry technique, we detected the presence of MELOE-3 in total lysates of both melanoma cells and normal melanocytes. This contrasts with our previous observation of the melanoma-restricted expression of MELOE-1 and MELOE-2. Furthermore, in vitro stimulation of PBMC from 6 healthy donors with overlapping peptides from MELOE-1 or MELOE-3 revealed a very scarce MELOE-3 specific T cell repertoire as compared to the abundant repertoire observed against MELOE-1. The poor immunogenicity of MELOE-3 and its expression in melanocytes is consistent with an immune tolerance towards a physiologically expressed protein. In contrast, melanoma-restricted expression of IRES-dependent MELOE-1 may explain its high immunogenicity. In conclusion, within the MELOE family, IRES-dependent antigens represent the best T cell targets for immunotherapy of melanoma.

Constitutional thinness (CT) is characterized by a low and stable body mass index (BMI) without any hormonal abnormality. To understand the weight steadiness, energetic metabolism was evaluated. Seven CT, seven controls, and six anorexia nervosa (AN) young women were compared. CT and AN had a BMI 2. Four criteria were evaluated: 1) energy balance including diet record, resting metabolic rate (RMR) (indirect calorimetry), total energy expenditure (TEE) (doubly labeled water), physical activity; 2) body composition (dual-energy X-ray absorptiometry); 3) biological markers (leptin, IGF-I, free T3); 4) psychological profile of eating behavior. The normality of free T3 (3.7 ± 0.5 pmol/l), IGF-I (225 ± 93 ng/ml), and leptin (8.3 ± 3.4 ng/ml) confirmed the absence of undernutrition in CT. Their psychological profiles revealed a weight gain desire. TEE (kJ/day) in CT (8,382 ± 988) was not found significantly different from that of controls (8,793 ± 845) and AN (8,001 ± 2,152). CT food intake (7,565 ± 908 kJ/day) was found similar to that of controls (7,961 ± 1,452 kJ/day) and higher than in AN (4,894 ± 703 kJ/day), thus explaining the energy metabolism balance. Fat-free mass (FFM) (kg) was similar in CT and AN (32.5 ± 2.9 vs. 34.1 ± 1.9) and higher in controls (37.8 ± 1.6). While RMR absolute values (kJ/day) were lower in CT (4,839 ± 473) than in controls (5,576 ± 209), RMR values adjusted for FFM were the highest in CT. TEE-to-FFM ratio was also higher in CT than in controls. Energetic metabolism balance maintains a stable low weight in CT. An increased energy expenditure-to-FFM ratio differentiates CT from controls and could account for the resistance to weight gain observed in CT.

Resume L'avenement de la vie psychique chez le nourrisson met le chercheur au defi de rendre compte de l'objet originaire que les auteurs relient aux conditions originaires de cette emergence. Les reponses freudiennes, qui ont ete largement critiquees par la suite, renvoient a des theories « archeologiques » et a des modeles de transmission phylogenetique. Dans cet article, les auteurs proposent un modele base sur les connaissances actuelles, celui d'un « dispositif originaire ». Selon leurs hypotheses, mere et enfant sont structurellement et mutuellement impliques dans un « ecart originaire » qui exige d'etre regule. Du cote maternel, cette regulation est devolue a la preoccupation maternelle primaire et a l'interdit de l'inceste. Du cote de l'enfant, elle appartient a des schemes preformes physiologiques et instinctifs, a des competences sensorielles et interactives precoces ainsi qu'au narcissisme primaire. Sont de ce fait definis le contexte et les experiences conduisant a la construction de l'objet primaire qui, selon les auteurs, marque le veritable point d'entree dans la vie psychique.

Objective: To determine whether the presence of anxiety disorders is related to depressive comorbidity in subjects with eating disorders (ED), while taking into account certain variables that may be related to depression (subjects’ age, ED duration, prior incidents of anorexia nervosa in bulimic subjects, inpatient or outpatient status, nutritional state [as measured by body mass index]). Method: We evaluated the frequency of depressive disorders in 271 subjects presenting with a diagnosis of either anorexia nervosa or bulimia, using the Mini International Neuropsychiatric Interview, DSM-IV version. Results: A multivariate analysis reveals that anxiety disorders do not all have the same influence in terms of risk of onset of major depressive episode in anorexics and bulimics when adjusted on variables related to depression. Conclusion: Depression in subjects with ED can be explained in part by comorbidity with obsessive-compulsive disorder, generalized anxiety, social phobia, and panic disorder. D 2006 Published by Elsevier Inc.

The interaction between 4-1BB ligand (CD137L), a member of the tumor necrosis factor superfamily, and its receptor 4-1BB provides a co-stimulatory signal for T lymphocyte proliferation and survival. However, the structure of 4-1BBL has not been thoroughly investigated, and none of the human recombinant 4-1BBL molecules available have been described as capable of co-stimulating T cells. The present work provides a model of the three-dimensional structure of the tumor necrosis factor homology domain of 4-1BBL and describes the production of a recombinant human soluble 4-1BBL whose originality lies in that it contains the whole extracellular tail preceding the tumor necrosis factor homology domain and an AviTag peptide (AviTag-4-1BBL) allowing enzymatic biotinylation and multimerization via streptavidin. We provide evidence that this chimeric protein exists as a homotrimer, whereas commercial FLAG-tagged 4-1BBL does not. This resulted in a much higher affinity for 4-1BB (1.2 nM) as compared with FLAG-4-1BBL (55.2 nM). We demonstrate that the single extracellular cysteine residue in the tail (Cys-51) could form a disulfide bond, both in our recombinant protein and in physiologically expressed 4-1BBL. The mutation of this cysteine residue exerted no effect on trimerization but increased the dissociation rate of AviTag-4-1BBL from 4-1BB. In its soluble form, AviTag-4-1BBL did not stimulate purified T cells but dramatically inhibited proliferation of peripheral blood mononuclear cells stimulated with anti-CD3 mAb. In contrast, a very significant co-stimulatory effect was observed on purified T cells once AviTag-4-1BBL was immobilized onto streptavidin beads. In addition, we show that the cross-linking of two trimeric AviTag-4-1BBL molecules was the minimum step required to elicit significant costimulatory activity.

Resume L’objectif etait de repondre a la question suivante : existe-il des differences entre les groupes diagnostiques de troubles du comportement alimentaire (TCA) pour la prevalence des troubles depressifs et des troubles anxieux, en tenant compte des differences cliniques observees entre les groupes (âge actuel des sujets, duree d’evolution du TCA, patient hospitalier ou ambulatoire, Body Mass Index) ? Nous avons evalue la frequence des troubles anxieux et des troubles depressifs chez 271 sujets presentant un diagnostic actuel d’anorexie mentale ou de boulimie, en utilisant le Mini International Neuropsychiatric Interview (MINI), version DSM IV. Nous avons compare les prevalences entre les sous-types d’anorexiques (AN-R et AN-BN) et entre les sous-types de boulimiques (BN-P et BN-NP), et enfin entre les anorexiques mentales et les boulimiques en ajustant sur les variables definies ci-dessus. La comorbidite actuelle ou vie entiere des troubles anxieux et depressifs ne differe ni entre les AN-R et les AN-BN, ni entre les BN-P et les BN-NP. Seuls les diagnostics actuels d’agoraphobie et de trouble obsessionnel compulsif sont significativement plus frequents chez les AN que chez les BN. La plus grande frequence de la comorbidite entre trouble obsessionnel compulsif et AN en comparaison avec la BN, connue de longue date, n’est pas remise en question ; la plupart des autres troubles anxieux et des troubles depressifs sont aussi frequents dans tous les types diagnostiques de TCA.

The immunosuppressive properties of a benzamide derivative, JM34, previously characterized as an anti-inflammatory compound are described. The immunosuppressive potential of JM34 was evidenced by inhibition of PBMC proliferation in vitro with an IC50 of 20 microM. In contrast with classical immunosuppressive drugs, JM34 affected neither cytokine production nor IL-2R expression from activated T cell clones, and displayed only moderate inhibition of IL-2-induced or anti-CD3/anti-CD28-induced proliferation. We investigated its effects on dendritic cells (DC) in vitro. Addition of JM34 during DC maturation inhibited the expression of some maturation markers: specifically, MHC molecule up-regulation was totally inhibited and CD83 expression was significantly reduced, while up-regulation of CD86, CD80 or CD40 was less affected. Moreover, JM34-treated DC showed impaired IL-12 but not IL-10 secretion, and a markedly reduced ability to present antigens to naive T lymphocytes in vitro. We provide evidence that these JM34-induced alterations of DC were associated with a marked inhibition of NF-kappaB nuclear translocation. Finally, JM34 inhibited delayed type hypersensitivity dose dependently in mice. In conclusion, our data suggest that JM34 inhibited T lymphocyte activation mainly by targeting DC, and thus may represent a new class of therapeutic agents in the fields of transplantation and autoimmune diseases.

We postulated that dynamic alterations in QT interval adaptation could characterize patients with anorexia nervosa (AN) and could be restored after weight gain. To assess ventricular repolarization features, we evaluated the QT dynamicity along RR intervals from 24-hour electrocardiographic data of patients with AN before and after refeeding. Ten young women with AN (19 +/- 3 years) were included in the study. The QT/RR slope was found significantly enhanced compared with normals (-1.82 +/- 0.62 vs -1.40 +/- 0.30; p0.05). This slope returned to normal range values after refeeding. The QT/RR slope was significantly correlated with the body mass index (r = 0.59; p0.007) in the patient group.

Alexithymia, depressive feelings, and dependency are interrelated dimensions that are considered potential "risk factors" for addictive disorders. The aim of this study was to investigate the relationships between these dimensions and to define a comprehensive model of addiction in a large sample of addicted subjects, whether affected by an eating disorder or presenting an alcohol- or a drug use-related disorder. The participants in this study were gathered from a multicenter collaborative study on addictive behaviors conducted in several psychiatric departments in France, Switzerland, and Belgium between January 1995 and March 1999. The clinical sample was composed of 564 patients (149 anorexics, 84 bulimics, 208 alcoholics, 123 drug addicts) of both genders with a mean age of 27.3 +/- 8 years. A path analysis was conducted on the 564 dependent patients and 518 matched controls using the scores of the Toronto Alexithymia Scale, the Depressive Experiences Questionnaire, and the Interpersonal Dependency Inventory. Statistical analyses showed good adjustment (Goodness of Fit Index = 0.977) between the observable data and the assumed model, thus supporting the hypothesis that a depressive dimension, whether anaclitic or self-critical, can facilitate the development of dependency in vulnerable alexithymic subjects. This result has interesting clinical implications because identifying specific patterns of relationships leading from alexithymia to dependency can provide clues to the development of targeted strategies for at-risk subjects.

Women who were referred with an eating disorder (ED) were compared with a matched normal control group to answer the following questions: What are the frequencies of anxiety disorders in cases of anorexia and bulimia nervosa diagnosed according to DSM-IV criteria? Are anxiety disorders significantly more frequent among women with an eating disorder than among women from the community? We assessed the frequencies of six specific anxiety disorders among 271 women with a current diagnosis of anorexia or bulimia nervosa and 271 controls, using the Mini-International Neuropsychiatric Interview, French DSM-IV version. A lifetime comorbidity with at least one anxiety disorder was found in 71% of both the anorexic and the bulimic subjects, significantly higher than the percentage of controls with an anxiety disorder. The prevalence was significantly higher in the eating disorder groups than in controls for most types of anxiety disorder, and between 41.8 and 53.3% of comorbid cases had an anxiety disorder preceding the onset of the eating disorder. Anxiety disorders are significantly more frequent in subjects with eating disorders than in volunteers from the community, a finding that has important etiological and therapeutic implications.

Despite its negative regulatory role on tumor-specific T cells, Programmed cell death 1 (PD-1) is also a marker of activated tumor-infiltrating T cells. In cancer, PD-1 blockade partially reverses T cell dysfunction allowing the amplification of tumor reactive T cells. Here, we investigated the role of PD-1 signaling on effector/memory human T cells specific for shared melanoma antigens, derived from blood. We documented for the first time the existence of melanoma-specific T cell clones unable to express PD-1. This stable feature was due to the persistent methylation of the PDCD1 promoter. These PD-1neg clones were of lower avidity than their PD-1pos counterparts, suggesting that high-affinity-specific T cell clones unable to express PD-1 are not or rarely present in peripheral blood, as they are probably eliminated by negative selection, due to their high reactivity. We also documented the existence of such PD-1neg T cell clones in melanoma tumor-infiltrating lymphocytes (TIL), which also exhibited a lower functional avidity than PD-1pos TIL clones. This clearly shows that PD-1 expression identifies antigen-specific T cell clonotypes of high functional avidity. Finally, we demonstrated that PD-1 blockade during the in vitro selection process of Melan-A-specific T cells favored the amplification of higher avidity T cell clonotypes. This preferential amplification of high-avidity memory T cells upon PD-1 blockade resonates with the expansion of reactive T cells, including neo-antigen-specific T cells observed in anti-PD-1-treated patients. This feature should also be a useful biomarker of clinical efficiency, while providing new insights for adoptive transfer treatments.

To modulate alloreactivity after hematopoietic stem cell transplantation, suicide gene–expressing donor T cells can be administered with an allogeneic T-cell–depleted bone marrow graft. Immune competence of such cells is a critical issue. The impact of the ex vivo gene transfer protocol (12-day culture period including CD3/interleukin-2 [IL-2] activation, retroviral-mediated gene transfer, and G418-based selection) on the anti–Epstein-Barr virus (EBV) potential of gene-modified cells has been examined. Cytotoxic (pCTL) and helper (pTh) cell precursor limiting dilution assays, interferon-γ enzyme-linked immunospot, or fluorescence-activated cell sorter analysis after tetrameric HLA-A2/EBV peptide complexes revealed that the frequency of anti-EBV T cells was lower in gene-modified cells (GMCs) than in similarly cultured but untransduced T cells and was even lower than in fresh peripheral blood mononuclear cells, demonstrating both an effect of the culture and of the transduction or selection. The culture-dependent loss of EBV-reactive cells resulted from the preferential induction of activation-induced cell death in tetramer+ cells. Replacing the initial CD3/IL-2 activation by CD3/CD28/IL-2 partially restored the anti-EBV response of GMCs by reducing the initial activation-induced cell death and enhancing the proliferation of EBV-tetramer+cells. Moreover, the G418 selection, and not the transduction, was directly toxic to transduced tetramer+ cells. Replacing the G418 selection by an immunomagnetic selection significantly prevented the selection-dependent loss of EBV-specific cells. Overall, ex vivo gene modification of primary T cells can result in a significant reduction in EBV-reactive T cells through both culture-dependent and selection-dependent mechanisms. Improving immune functions of GMCs through modifications of the cell culture conditions and transduction/selection processes is critical for further clinical studies.

Objectifs: Syn thèse des dif fi cultés de tra duc tion des tests, de scrip tion des méth odes de tra duc tion et des procédu res de vali da tion des tests. Ap pli ca tion à un ques tion naire de personnalité. Méthodologie: Le ques tion naire de per son nal ité de Freiburg (FPI-R) a été traduit et les tech niques de vali da tion d'un test lui ont été ap pli quées: rétro tra duc tion, pré test et révi sion par un comité d'ex perts soig neuse ment choisi. Résultats: Au moyen de la re vue de la docu men ta tion, les dif fi cultés de la tra duc tion du FPI-R sont explici tées, en par ticu lier les diffé rents types d'équiva lence langue source/langue ci ble (séman tique, tech nique, idio ma tique, liée à l'expé ri ence et con cep t uelle). Les procédu res sta tis tiques de vali da tion ne sont abordées que dans leurs prin ci pes. Conclusion: La méth ode cou rante al liant tra duc tion et rétro tra duc tion est in suffi sante et doit s'ac com pag ner au mini mum d'un pré test et d'une révi sion à chaque étape par un comité d'ex perts. La pré sence d'ex perts mono lingues sem ble in dis pen sa ble pour met tre à jour les moin dres détails de la langue ci ble, là où les bi lingues peu vent éch ouer.

The aim of the study was to determine the comorbidity of dependent personality disorder (DPD) and separation anxiety disorder (SAD) in large nonclinical and clinical samples. Using a cross-sectional method, the relationship between these disorders was studied in 784 healthy subjects and in 708 subjects presenting with various addictive behaviors. DPD was significantly associated with SAD in healthy subjects, alcoholics, drug abusers and anorectic and bulimic patients. The comorbidities of SAD and DPD suggest that specific therapeutic strategies could be discussed. Moreover, longitudinal studies are required to determine if SAD constitutes a risk factor for DPD.

This paper describes a qualitative immunoreactivity cholecystokinin (CCK) characterization in 24 hours urine specimens. Urine was subjected to ethanol precipitation, followed by ion exchange chromatography, and immunoblotting with 2 antibodies. HPLC was developed to authenticate qualitatively the presence of CCK revealed by immunoblotting. The results obtained by HPLC showed this method was more specific. HPLC allowed us to distinguish, through anti-CCK reactive fractions, a single peak which reacted only with CCK antibodies and not with gastrin antibodies.

Efficient detection and immunomagnetic sorting of specific T cells using multimers of MHC class I and peptide with reduced CD8 binding

OBJECTIVE The present study was conducted in order to describe the variations and circadian rhythm of biochemical markers of bone remodelling at baseline and after weight gain in patients with aneroxia nervosa (AN). SUBJECTS We studied 9 women (mean age 21 years, range: 16–30) with established AN who remained amenorrhoeic during the study and with a low body mass index (BMI) after refeeding and 6 female controls (mean age 20 years, range, 18–24 and BMI: 20.6 ± 1.1 kg/m2). Refeeding was not associated with any other intervention or treatment, especially oestrogen replacement or hormonal contraception. Serum levels of oestradiol remained below 70 pmol/l before and after refeeding. MEASUREMENTS During the study, PTH and 25-hydroxyvitamin D measurements were performed. Markers of bone formation: serum intact osteocalcin (iBGP) and serum intact BGP + fragments (iBGP+F) and markers of bone resorption: urine C-teloptide of type I collagen (uCTX) and serum C-telopeptide ofvtype 1 collagen (s-CTX) were measured. RESULTS At baseline, PTH and 25 OH-vitamin D concentrations were within the normal range in AN patients and no significant variation was observed after refeeding. Bone formation markers were found to be significantly different at baseline between AN patients and controls. After refeeding, iBGP and iBGP+F levels increased by 172% and 154%, respectively, to values no different from controls. Intact BGP and iBGP+F exhibited a significant circadian variation in controls (P

A series of N-pyridinyl(methyl)fluorobenzamides issued from 2,4-dimethyl-6-aminopyridine and 3-aminomethylpyridine were synthesized and evaluated as inhibitors of TNF-α production. Although less active than the corresponding phthalimides, several pentafluorobenzamides exhibited significant activity at 10 μM. N-(4,6-dimethylpyridin-2-yl)pentafluorobenzamide was selected for a preliminary in-vivo assay. Although its inhibitory activity against carrageenan-induced rat paw oedema was moderate, it induced a significant reduction in ear thickness in the PMA-induced mouse ear-swelling test (49± 6% inhibition after a dose of 0.1 mM kg−1, p.o.).

Accumulating evidence that serum levels of soluble class I HLA molecules (sHLA-I) can, under various pathological conditions, correlate with disease stage and/or patient survival, has stimulated interest in defining whether sHLA-I can exert immunological functions. However, despite a mounting number of publications suggesting the ability of sHLA-I to affect immune effectors in vitro, the precise underlying mechanism still remains controversial. In this article, we address potential functions of both classical and nonclassical sHLA-I, using soluble recombinant HLA-I/peptide monomers, and clearly demonstrate their ability to trigger Ag-specific activation of CD8 T cells in vitro. Furthermore, we provide strong evidence that this behavior results from the passive transfer of peptides from monomers to T cell–bound HLA-I molecules, allowing for fratricide representation and activation. Hence, we proposed a unifying model of T cell activation by HLA-I/peptide monomers, reappraising the potential involvement of sHLA-I molecules in the immune response.

International audience; Our previous studies on melanoma antigens identified two new polypeptides, named MELOE-1 and MELOE-2, that are involved in immunosurveillance. Intriguingly, these antigens are coded by distinct open reading frames (ORF) of the meloe mRNA which is significantly expressed only in the melanocytic lineage. In addition, MELOE-1 and -2 specific T cell clones recognized melanoma cells but very poorly normal melanocytes suggesting differential translation of meloe in normal vs tumor cells. This prompted us to elucidate the mechanisms of translation of these antigens in melanoma cells. We first demonstrated that no splicing event or cryptic promoter could generate shorter meloe transcripts containing only one of the two ORFs. Triggering meloe RNA degradation with a siRNA close to the ORF coding for MELOE-2 abrogated expression of both MELOE-1 and MELOE-2, thus confirming that the two ORFs are always associated. Next we showed, in a bicistronic reporter system, that IRES activities could be detected upstream of MELOE-1 and MELOE-2 and finally confirmed their translation from full length meloe cDNA in melanoma cells with eGFP constructs. In conclusion, meloe is a polycistronic mRNA that generates both MELOE-1 and MELOE-2 antigens through IRES-dependent translation in melanoma cells and that may explain their tumor specificity.

A number of trials of adoptive transfer of tumor-specific T lymphocytes have been performed in the last 20 years in metastatic melanoma, with increasingly encouraging results as the relevant melanoma antigens were identified and the purity/specificity of injected T cells improved. We have previously described a sorting method of epitope-specific T lymphocytes that uses magnetic beads coated with HLA/peptide complexes and we suggested that this method could be applied to a clinical setting. In the present work, we provide a detailed description of the whole GMP process of sorting and amplification of clinical grade T cells specific for the melanoma antigens Melan-A and MELOE-1. All the reagents used in this process including the sorting reagent were produced in GMP conditions and we document the optimization of the different steps of the process such as peptide stimulation, sorting, and amplification. The optimized procedure, validated in 3 blank runs in a clinical setting, allowed the production of at least 108pure (>90%) Melan-A- and MELOE-1-specific T cells within 28 days starting with 100 mL of blood from metastatic melanoma patients. This GMP process is thus ready to be used in an upcoming phase I/II clinical trial on metastatic melanoma patients.

Summary Pharmacomodulation of the non-carboxylic NSAID N -(4,6-dimethylpyridin-2-yl)benzamide 1 led to the synthesis of structurally related furan, thiophene and pyrrole carboxamides 3–14 . The derivatives benzenethiocarboxamides 15–18 and heteroaryl-thiocarboxamides 19–22 were also prepared by oxygen/sulfur exchange; this reaction was more efficiently carried out by P 4 S 10 than by Lawesson's reagent. The 20 synthesized compounds were evaluated against peripheral edema by a foot-pad carrageenin-induced edema test. Amides 3–5, 8, 9, 11, 12 and 14 were most active, exhibiting > 90% inhibition after oral administration of 0.8 mmol·kg −1 . Two amides, 3 and 5 , were selected for evaluation of their inhibitory activity in PLA 2 -induced brain edema and were found to be more potent than dexamethasone after IP administration.

International audience; MELOE-1 is an overexpressed melanoma antigen containing a HLA-A2 restricted epitope, involved in melanoma immunosurveillance of patients adoptively transferred with tumour infiltrating lymphocytes (TIL). The use of the full-length antigen (46 aa) for anti-melanoma vaccination could be considered, subject to the presence of Th epitopes all along MELOE-1 sequence. Thus, in this study we evaluated in vitro the immunoprevalence of the different regions of MELOE-1 (i.e. their ability to induce CD4 T cell responses in vitro from PBMC). Stimulation of PBMC from healthy subjects with MELOE-1 induced the amplification of CD4 T cells specific for various regions of the protein in multiple HLA contexts, for each tested donor. We confirmed these results in a panel of melanoma patients, and documented that MELOE-1 specific CD4 T cells, were mainly Th1 cells, presumably favourable to the amplification of CD8 specific T cells. Using autologous DC, we further showed that these class II epitopes could be naturally processed from MELOE-1 whole protein and identified minimal epitopes derived from each region of MELOE-1, and presented in four distinct HLA contexts. In conclusion, vaccination with MELOE-1 whole polypeptide should induce specific Th1 CD4 responses in a majority of melanoma patients, stimulating the amplification of CD8 effector cells, reactive against melanoma cells.

Conventional CD8αβ participates in the activation of T cells by binding to the same peptide-MHC complex as does the TCR. A recent paper in Science shows, however, that the CD8αα form on iIELs binds TL and could alter signals from the TCR.

Introduction The impact of undernutrition on endocrine and exocrine gonadatrope function is poorly known in male anorexia nervosa (AN) patients. Aim The aim of this study was to compare the pituitary–gonadal function of male AN subjects with that of healthy controls, Kallmann syndrome (KS) patients, and female AN subjects. Methods Observational monocentric cross‐sectional study performed in 31 male and 25 female subjects with restrictive‐type AN, 22 male and 20 female controls, and nine male KS patients. Main Outcome Measures Hormonal parameters are as follows: follicule stimulating hormone (FSH), luteinizing hormone (LH), sex hormone binding globulin, estradiol, testosterone, inhibin B, thyroid hormones, growth hormone (GH), insulin‐like growth factor 1 (IGF‐1), cortisol, adrenocorticotropic hormone (ACTH), dehydroepiandrosterone sulfate, and leptin. Results Similar abnormalities of free T3, GH, IGF‐I, cortisol, and leptin were found in men as in AN women with equivalent undernutrition status when compared with corresponding controls. Low levels of LH, FSH were found in both male and female AN patients. In male AN, total testosterone was found lower than in controls but higher than in KS, while a lack of estradiol was noticed in AN women. Sex hormones variations were directly related to weight gain only in AN men. No relationship was found between sex hormones and leptin variation for both sexes. In AN men, inhibin B levels were similar to that of controls and did not correlate with testosterone levels. Conclusions Significant differences of undernutrition impact on gonadal status were noticed between male and female AN subjects, including partial preservation of testosterone release and probable preservation of exocrine function, according to the normal inhibin B levels. Galusca B, Leca V, Germain N, Frere D, Khalfallah Y, Lang F, and Estour B. Normal inhibin B levels suggest partial preservation of gonadal function in adult male patients with anorexia nervosa. J Sex Med 2012;9:1442–1447.

International audience; We have designed a cytometry-based competition assay to evaluate peptide binding to empty recombinant HLA class II molecules. The efficiency of this assay was evaluated using recombinant HLA-DP0401 molecules (HLA-DP) produced in insect cells and 13 peptides from human telomerase reverse transcriptase (hTERT). We demonstrate that our method allowed accurate measurements of peptide Ki values and can thus discriminate strong, moderate and poor HLA-DP binders. In parallel, we showed that among hTERT peptides, the most immunodominant in healthy individuals were those with moderate affinity for HLA-DP while no T cell response could be evidenced against peptides with very strong or very low affinities for HLA-DP. This strongly suggests that the precise determination of peptide affinity with our method can improve HLA class II epitope prediction.

International audience; Adoptive transfer of T cells remains a promising approach in melanoma. Initial clinical trials performed with polyclonal tumor-infiltrating lymphocyte gave limited clinical results. Nonetheless, encouraging results have been reported in adjuvant setting (stage III melanoma), and when tumor-infiltrating lymphocytes were associated with lymphodepleting regimens. Specificity of adoptive cell therapy has been achieved with the infusion of antigen specific cytotoxic T-lymphocyte clones, associated with some clinical responses. Antigen specificity can also be obtained by the allogeneic transfer of high-avidity T-cell receptors into autologous T cells. We propose an alternative strategy based on the selection of antigen-specific T cells with magnetic beads coated with HLA-peptide multimers. Future improvements of adoptive melanoma immunotherapy may be achieved by its association with other therapeutic strategies such as targeted therapy against signaling pathways.

International audience; The activity of brain regions of the so-called default mode network (DMN) attenuates during the performance of goal-directed tasks. These activity decreases (named task-induced deactivations; TID) are though to reflect the reallocation of cognitive resources from the DMN to areas implicated in the execution of the task. Recently, DMN activity suppression has been studied in schizophrenia patients. Although these works showed that TID are altered in schizophrenia, they also revealed inconsistent findings. We hypothesized that reallocation of resources is altered in schizophrenia patients and is context or task specific.We investigated TID using functional MRI in 26 schizophrenic patients and 13 control subjects while performing two different goal-directed tasks (the Hayling Sentence Completion Test and the N-Back task). Both whole brain and region of interest conjunction analyses were conducted to investigate brain areas commonly deactivated in the two tasks (task unspecific deactivations).Task-unspecific deactivations were not observed in the schizophrenia group, although these were strongly significant in the control group. Differences between patient and control participants were observed in different regions of the DMN depending whether the subjects performed the Hayling or the N-back task.These results suggest that reallocation of cognitive resources is altered in our patient sample. Moreover, TID were task-unspecific indicating that resources reallocation is context dependant in schizophrenia. DMN activity attenuates differently in schizophrenia patients depending on the cognitive processes involved in the task.

We previously reported a significant increase in percentages of peripheral blood gamma delta+ T cells in islet cell antibody (ICA) positive relatives of patients with insulin-dependent diabetes (IDD). In the present study, we further characterized this T cell abnormality in a larger group of ICA+ subjects and report that (1) Percentages of gamma delta+ T lymphocytes were significantly increased only in subjects with high ICA titers (> or = 20 JDF units) (P = 0.005) and resulted from an increase in absolute numbers of gamma delta+ T lymphocytes. (2) In these subjects, the increase in gamma delta+ T lymphocytes was associated with an increase in the V gamma 9 V delta 2 subpopulation (r = 0.99). (3) In these same subjects, high percentages of gamma delta+ T lymphocytes were associated with normal beta cell function while low percentages were associated with diminished insulin response. Using 65 microU/ml as the threshold of abnormal intravenous glucose tolerance test (IVGTT) response, percentages of gamma delta+ T lymphocytes could significantly predict IVGTT status in these subjects (P < 0.01). A longitudinal follow-up further suggested that the development of an abnormal IVGTT response and progression to diabetes was associated with a decrease in percentages of gamma delta+ T lymphocytes while patients whose gamma delta+ T cell percentages remained high retained normal beta cell function. Our data therefore suggest that gamma delta+ T lymphocytes and more specifically V gamma 9 V delta 2 T cells are implicated in the autoimmune process leading to diabetes and may have a regulatory role. The monitoring of their percentages in the blood of patients at risk for diabetes may be useful as an additional predictor of diabetes development.

N-Aryl-3-(indol-3-yl)propanamides were synthesized and their immunosuppressive activities were evaluated. This study highlighted the promising potency of 3-[1-(4-chlorobenzyl)-1H-indol-3-yl]-N-(4-nitrophenyl)propanamide 15 which exhibited a significant inhibitory activity on murine splenocytes proliferation assay in vitro and on mice delayed-type hypersensitivity (DTH) assay in vivo.

Context: The relevance of hormonal assessment in anorexia nervosa (AN) management is still unclear. The short-term physical risk during undernutrition period of the disease is partially predicted by anthropometric and electrolytic parameters.Objective: The objective of the study was to evaluate hormonal profiles in a large cohort of AN and their relationship with critical states.Design and Setting: This was an observational monocentric cross-sectional study performed in the endocrinological unit.Patients and Other Participants: Participants included 210 young female subjects with restrictive-type AN and 42 female controls of comparable age.Main Outcome Measures: The following hormonal parameters were measured: thyroid hormones, GH, IGF-I, cortisol, oestradiol, FSH, LH, SHBG, dehydroepiandrosterone sulfate, plasma metanephrines, and bone markers. Their relation with registered short-term evolution of AN subjects after hormonal assessment was evaluated.Results: Except for metanephrines and dehydroepiandrosterone sulfate, most of the hormonal abnormalities previously reported in AN were confirmed. The manifestation of these hormonal abnormalities started below different body mass index (BMI) levels, ranging between 17 and 15 kg/m2, even though an important percentage of normal values for every parameter was still noticed for very low BMIs. All patients who developed critical states during the 3 months after the hormonal assessment presented with BMI less than 15 kg/m2 and a very increased level of cortisol, GH, and increased values of metanephrines.Conclusions: The hormonal response to undernutrition is heterogeneous in a large population with restrictive AN. In clinical practice, metanephrines, GH, and/or cortisol data could be used as important predictors for severe short-term outcome.