Functional Characterization of Neurotensin Receptors in Human Cutaneous T Cell Lymphoma Malignant Lymphocytes

Cutaneous T cell lymphomas are a clonal proliferation of CD4+ T lymphocytes primarily involving the skin. Mycosis fungoides is an epidermotropic CD4+ cutaneous T cell lymphoma, and a more aggressive form, Sezary syndrome, occurs when the malignant cells become nonepidermotropic. The role of neuropeptides in the growth and chemotaxis capacity of cutaneous T cell lymphoma cells remains unknown. In this report, we found that cutaneous T cell lymphoma cells, similarly to normal resting or activated peripheral lymphocytes, were able to bind neurotensin. We used an interleukin-2-dependent cutaneous T cell lymphoma malignant T cell line derived from cutaneous T cell lymphoma lesions in order to study the role of neurotensin in the proliferation and migration of these malignant cells. First, we determined that the malignant cells expressed neurotensin receptors on their cell membrane. Functional results indicated that neurotensin did not stimulate the growth of the cell line. In contrast, this neuropeptide inhibited the proliferation of the tumor cells in response to exogenous interleukin-2. Furthermore, we found that neurotensin enhanced both spontaneous and chemoattractant-induced migration of the malignant cells. This suggests that neurotensin in skin can play a role in the disease by locally limiting the growth of the cutaneous T cell lymphoma tumor cells in response to cytokines and by enhancing their chemotaxis capacity

Innate recognition of bacteria in human milk is mediated by a milk-derived highly expressed pattern recognition receptor, soluble CD14.

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A member of the tetra spans transmembrane protein superfamily is recognized by a monoclonal antibody raised against an HLA class I-deficient, lymphokine-activated killer-susceptible, B lymphocyte line. Cloning and preliminary functional studies.

International audienceThe IA4 mAb was identified among a series of antibodies raised in BALB/c mice after immunization against a HLA class I-deficient, lymphokine-activated killer (LAK)-susceptible EBV-B lymphocyte line. The IA4 antibody was selected because of its high expression, in the range of 10(5) to 25 x 10(5) sites/cell, on several B lymphocyte lines (EBV-transformed or Burkitt) and monocytic lines such as HL60 and U937, and because its expression was correlated with both target susceptibility to LAK lysis and reduced expression of HLA class I surface Ag on two pairs of EBV-B-transformed cell lines (721/721.134 and MM/10F2). Despite the strategy followed to raise the mAb and the correlation mentioned above, no direct role of the IA4 molecules in LAK susceptibility has been established, since the IA4 molecule is poorly expressed on the sensitive targets Daudi and K562; moreover, the IA4 antibody did not affect reproducibly the in vitro killing of positive target cells by LAK effectors. The IA4 antibody was poorly immunoprecipitating and the surface molecule recognized was identified by gene cloning following an expression strategy using a U937 cDNA library transfected in COS cells, and a screening strategy based on membrane expression of IA4 molecule. The IA4 cDNA is virtually identical to "R2," a mRNA species previously identified in activated human T cells by subtractive hybridization. The IA4 cDNA contains an open reading frame coding for a protein 267 amino acids long with four potential transmembrane domains and one large external hydrophilic domain of about 110 amino acids, possibly glycosylated. The encoded protein belongs to a family of surface molecules, the tetra spans transmembrane protein superfamily, all displaying the four transmembrane domains, expressed on various cell types including lymphocytes (CD9, CD37, CD53, TAPA-1), melanoma cells (ME491), and intestinal cells (CO-029). These molecules have been reported to be involved in cell activation and cell death. Surprisingly, the Schistosoma mansoni Ag Sm23 displays significant homologies with this family. The IA4 molecule is a widely distributed surface marker expressed on circulating lymphocytes and monocytes, newborn thymocytes, and the cell lines mentioned above. The IA4 molecule expression is up-regulated upon cell activation. Weakly expressed on resting peripheral T and B lymphocytes and large granular lymphocytes (NK), its expression roughly doubles after activation by PHA, staphylococcus aureus Cowan I, and IL-2, respectively.(ABSTRACT TRUNCATED AT 400 WORDS

In vitro functional evidence of different neurotensin-receptors modulating the motor response of human colonic muscle strips

1. The newly developed non-peptide neurotensin (NT)-receptor antagonists SR 48692 and SR 142948 were used to challenge NT responses of human colonic circular smooth muscle strips in vitro. The presence of NT(1) and NT(2) receptor transcripts in this tissue was tested by reverse transcriptase polymerase chain reaction (RT–PCR) analysis. 2. NT potently and dose-dependently contracted muscle strips, with significant regional differences in potency and efficacy between the transverse and distal colon: EC(50), 3.6 and 7.5 nM; the maximal effect was 70 and 55% of 0.1 mM carbachol. Colonic responses to NT in both segments were virtually the same in the presence of atropine (1 μM), levocabastine (10 μM) or tetrodotoxin (1 μM). 3. SR 142948 (10 nM–1 μM) competitively antagonized NT responses in the transverse and distal colon with similar affinities: pA(2) values 8.71 and 8.45, slopes 0.98 and 0.99. SR 48692 (10 nM–10 μM) antagonized the NT response competitively in the distal colon (pA(2) 6.55, slope 0.79) and non-competitively in the transverse colon (pA(2) 8.0, slope 0.51). Neither compound had any agonist effect. 4. The fact that the specific antagonists prevented NT-evoked atropine- and tetrodotoxin-insensitive mechanical responses of colonic muscle strips is highly consistent with the presence in these tissues of non-neuronal NT receptors, whose heterogeneity in the transverse segment is supported by the non-competitive antagonism of SR 48692. The finding of NT(1) receptor transcript in both transverse and distal colon suggests its identity with the lower affinity site disclosed functionally by SR 48692 in these segments

Rapporto tecnico sulle attività di campagna oceanografica “BANSIC 2013”

La campagna oceanografica BANSIC 2013 è stata condotta a bordo della N/O “Urania” dal 26 Giugno al 16 Luglio 2013 nell’ambito delle attività previste dal WP3 del progetto SSD-Pesca, finanziato dal MIUR su fondi MISE, a supporto della pesca italiana nelle Regioni Obiettivo 1, e dal progetto bandiera RITMARE (SP2_WP1_AZ1_UO01 e UO04) Obiettivi generali della campagna oceanografica sono stati lo studio delle relazioni tra le strutture oceanografiche a mesoscala (vortici verticali ed orizzontali, upwelling, ecc.) e le strutture spaziali dei fenomeni biologici relativi ai primi anelli della catena trofica (zooplancton, distribuzione e abbondanza di larve di piccoli pelagici e grandi pelagici), e lo sviluppo del dispositivo di Fishing Vessel Monitoring System denominato FOOS (Fishery Oceanography Observing System) da installare a bordo di imbarcazioni da pesca. In particolare, il campionamento di uova di acciuga è finalizzato all’applicazione del metodo DEPM (Daily Egg Production Method) per la stima dell’abbondanza dello stock riproduttore (SP2_WP1_AZ3_UO04). Il campionamento ittioplanctonico, che ha riguardato anche le acque Maltesi, è inserito anche nel piano di lavoro del progetto regionale MIPAF-FAO “MedSudMed” (“Assessment and Monitoring of the Fishery Resources and the Ecosystems in the Straits of Sicily”). La campagna ha visto anche l’avvio di una linea di ricerca in collaborazione con la Martin-Luther-Universität Halle-Wittenberg, avente come obiettivo i radiolari