Skin infiltrating NK cells in cutaneous T-cell lymphoma are increased in number and display phenotypic alterations partially driven by the tumor

Cutaneous T-cell lymphomas (CTCL) are characterized by focal infiltration of malignant T cell clones in solitary skin lesions. Many CTCL patients experience an indolent disease, but some progress to advanced disease with high fatality. We hypothesized that natural killer (NK) cells participate in local control of tumor growth in CTCL skin. Immunohistochemistry and flow cytometry analysis of the density, localization, phenotype and function of NK cells in twenty-nine fresh or formalin-fixed skin biopsies from twenty-four CTCL patients and twenty-three biopsies from twenty healthy controls highlighted higher numbers of CD56+CD3- NK cells in CTCL skin. A reduced fraction of CTCL skin NK cells expressed the maturation marker CD57, the cytotoxic protein granzyme B and the activation marker CD69, indicating reduced tumor-killing abilities of the NK cells. Retained expression of immune checkpoint proteins or inhibitory proteins including PD1, TIM3, LAG3, CD73 and NKG2A and the activating receptors CD16 and NKp46 indicated maintained effector functions. Indeed, the capacity of NK cells to produce anti-tumor acting IFNγ upon PMA+ionomycin stimulation was similar in cells from CTCL and healthy skin. Co-cultures of primary human NK cells or the NK cell line NKL with CTCL cells resulted in reduced levels of granzyme B and CD69, indicating that close cellular interactions with CTCL cells induced the impaired functional NK cell phenotype. In conclusion, increased numbers of NK cells in CTCL skin exhibit a partially impaired phenotype in terms of activity. Enhancing NK cell activity with NK cell activating cytokines such as IL-15 or immune checkpoint blockade therefore represents a potential immunotherapeutic approach in CTCL.publishedVersio

CD49a Expression Defines Tissue-Resident CD8+ T Cells Poised for Cytotoxic Function in Human Skin

Tissue-resident memory T (Trm) cells form a heterogeneous population that provides localized protection against pathogens. Here, we identify CD49a as a marker that differentiates CD8(+) Trm cells on a compartmental and functional basis. In human skin epithelia, CD8(+)CD49a(+) Trm cells produced interferon-γ, whereas CD8(+)CD49a(−) Trm cells produced interleukin-17 (IL-17). In addition, CD8(+)CD49a(+) Trm cells from healthy skin rapidly induced the expression of the effector molecules perforin and granzyme B when stimulated with IL-15, thereby promoting a strong cytotoxic response. In skin from patients with vitiligo, where melanocytes are eradicated locally, CD8(+)CD49a(+) Trm cells that constitutively expressed perforin and granzyme B accumulated both in the epidermis and dermis. Conversely, CD8(+)CD49a(–) Trm cells from psoriasis lesions predominantly generated IL-17 responses that promote local inflammation in this skin disease. Overall, CD49a expression delineates CD8(+) Trm cell specialization in human epithelial barriers and correlates with the effector cell balance found in distinct inflammatory skin diseases

Skin infiltrating NK cells in cutaneous T-cell lymphoma are increased in number and display phenotypic alterations partially driven by the tumor

Cutaneous T-cell lymphomas (CTCL) are characterized by focal infiltration of malignant T cell clones in solitary skin lesions. Many CTCL patients experience an indolent disease, but some progress to advanced disease with high fatality. We hypothesized that natural killer (NK) cells participate in local control of tumor growth in CTCL skin. Immunohistochemistry and flow cytometry analysis of the density, localization, phenotype and function of NK cells in twenty-nine fresh or formalin-fixed skin biopsies from twenty-four CTCL patients and twenty-three biopsies from twenty healthy controls highlighted higher numbers of CD56+CD3- NK cells in CTCL skin. A reduced fraction of CTCL skin NK cells expressed the maturation marker CD57, the cytotoxic protein granzyme B and the activation marker CD69, indicating reduced tumor-killing abilities of the NK cells. Retained expression of immune checkpoint proteins or inhibitory proteins including PD1, TIM3, LAG3, CD73 and NKG2A and the activating receptors CD16 and NKp46 indicated maintained effector functions. Indeed, the capacity of NK cells to produce anti-tumor acting IFNγ upon PMA+ionomycin stimulation was similar in cells from CTCL and healthy skin. Co-cultures of primary human NK cells or the NK cell line NKL with CTCL cells resulted in reduced levels of granzyme B and CD69, indicating that close cellular interactions with CTCL cells induced the impaired functional NK cell phenotype. In conclusion, increased numbers of NK cells in CTCL skin exhibit a partially impaired phenotype in terms of activity. Enhancing NK cell activity with NK cell activating cytokines such as IL-15 or immune checkpoint blockade therefore represents a potential immunotherapeutic approach in CTCL

Studies on the effect of orexin on upper gastrointestinal function in rats and man

Aims: To define the morphological localisation of orexin A (OXA) in the gut, and study the effect of endogenous and peripherally administered OXA on fasting small bowel motility, gastric emptying and acid secretion, hormones involved in glucose homeostasis and pharmacokinetic profile of OXA in rats and humans. Methods: The distribution of OXA, orexin receptors 1 and 2 (OX1 R, OX2R), neuronal nitric oxide synthase (nNOS) and various peptides were studied with immunocytochemistry in rat and human gut. In rats, the migrating myoelectric complex (MMC) was studied during intravenous (IV) infusion of OXA or the selective OX1 R-antagonist SB-334867-A. In separate sets of experiments, the rats were pretreated with vagotomy or the NOS-antagonist Nomeganitro-L-arginine (L-NNA). Gastric acid secretion and emptying of a 51Cr-labelled liquid nutrient or non-nutrient were studied in rats equipped with a gastric fistula and subjected to IV infusion of OXA or SB-334867-A. In humans, gastric emptying was studied of a 99mTclabelled omelette during IV infusion of OXA. Simultaneously, appetite ratings were measured using visual analogue scale (VAS) ratings. Plasma concentrations of OXA, leptin, insulin, glucagon, gastrin and glucose were analysed with radioimmunoassay (RIA). Results: OXA- and OXRs-like immunoreactivity were found in nerve fibres and neurones in myenteric and submucosal ganglia, the circular muscle, the mucosa and endocrine cells in the stomach, duodenum and pancreas in both rats and humans. OXA was co-expressed with nNOS in myenteric neurons and circular muscle in rats and humans. IV OXA inhibited fasting small bowel motility in rats through OX1 R and nitric oxide (NO), independent of the vagus nerve. SB-334867-A inhibited gastric emptying and acid secretion independent of gastrin in rats. Gastric retention in humans increased during IV infusion of OXA without affecting appetite ratings. Plasma levels of leptin decreased and insulin increased in humans and OXA has a half-life in plasma of 27.1 min in rats. Conclusions: Peripheral IV OXA affects upper gastrointestinal function in rats and humans. The activity of peripheral OXA may involve several pathways, including the central nervous system (CNS), intrinsic circuits of the enteric nervous system (ENS), vago-vagal reflexes or interactions with other gut hormones and NO. In humans, IV OXA affects hormones implicated in the regulation of glucose and energy homeostasis. The long half-life of OXA in plasma needs to be taken into account when interpreting results on metabolic and gut function after peripheral administration of OXA. Antagonists against OXRs may become a target in the treatment of obesity, diabetes and motility disorders of the gut

Short Trail Running Race: Beyond the Classic Model for Endurance Running Performance

International audiencePURPOSE:This study aimed to examine the extent to which the classical physiological variables of endurance running performance (maximal oxygen uptake (V˙O2max), %V˙O2max at ventilatory threshold (VT), and running economy (RE)) but also muscle strength factors contribute to short trail running (TR) performance.METHODS:A homogeneous group of nine highly trained trail runners performed an official TR race (27 km) and laboratory-based sessions to determine V˙O2max, %V˙O2max at VT, level RE (RE0%) and RE on a +10% slope, maximal voluntary concentric and eccentric knee extension torques, local endurance assessed by a fatigue index (FI), and a time to exhaustion at 87.5% of the velocity associated with V˙O2max. A simple regression method and commonality analysis identifying unique and common coefficients of each independent variable were used to determine the best predictors for the TR race time (dependent variable).RESULTS:Pearson correlations showed that FI and V˙O2max had the highest correlations (r = 0.91 and r = -0.76, respectively) with TR performance. The other selected variables were not significantly correlated with TR performance. The analysis of unique and common coefficients of relative V˙O2max, %V˙O2max at VT, and RE0% provides a low prediction of TR performance (R = 0.48). However, adding FI and RE on a +10% slope (instead of RE0%) markedly improved the predictive power of the model (R = 0.98). FI and V˙O2max showed the highest unique (49.8% and 20.4% of total effect, respectively) and common (26.9% of total effect) contributions to the regression equation.CONCLUSIONS:The classic endurance running model does not allow for meaningful prediction of short TR performance. Incorporating more specific factors into TR such as local endurance and gradient-specific RE testing procedures should be considered to better characterize short TR performance

DataSheet_1_Skin infiltrating NK cells in cutaneous T-cell lymphoma are increased in number and display phenotypic alterations partially driven by the tumor.docx

Cutaneous T-cell lymphomas (CTCL) are characterized by focal infiltration of malignant T cell clones in solitary skin lesions. Many CTCL patients experience an indolent disease, but some progress to advanced disease with high fatality. We hypothesized that natural killer (NK) cells participate in local control of tumor growth in CTCL skin. Immunohistochemistry and flow cytometry analysis of the density, localization, phenotype and function of NK cells in twenty-nine fresh or formalin-fixed skin biopsies from twenty-four CTCL patients and twenty-three biopsies from twenty healthy controls highlighted higher numbers of CD56+CD3- NK cells in CTCL skin. A reduced fraction of CTCL skin NK cells expressed the maturation marker CD57, the cytotoxic protein granzyme B and the activation marker CD69, indicating reduced tumor-killing abilities of the NK cells. Retained expression of immune checkpoint proteins or inhibitory proteins including PD1, TIM3, LAG3, CD73 and NKG2A and the activating receptors CD16 and NKp46 indicated maintained effector functions. Indeed, the capacity of NK cells to produce anti-tumor acting IFNγ upon PMA+ionomycin stimulation was similar in cells from CTCL and healthy skin. Co-cultures of primary human NK cells or the NK cell line NKL with CTCL cells resulted in reduced levels of granzyme B and CD69, indicating that close cellular interactions with CTCL cells induced the impaired functional NK cell phenotype. In conclusion, increased numbers of NK cells in CTCL skin exhibit a partially impaired phenotype in terms of activity. Enhancing NK cell activity with NK cell activating cytokines such as IL-15 or immune checkpoint blockade therefore represents a potential immunotherapeutic approach in CTCL.</p