RÉSUMÉ
Adoptive T cell therapy with T cells expressing affinity-enhanced TCRs has shown promising results in phase 1/2 clinical trials for solid and hematological tumors. However, depth and durability of responses to adoptive T cell therapy can suffer from an inhibitory tumor microenvironment. A common immune-suppressive agent is TGF-ß, which is secreted by tumor cells and cells recruited to the tumor. We investigated whether human T cells could be engineered to be resistant to inhibition by TGF-ß. Truncating the intracellular signaling domain from TGF-ß receptor (TGFßR) II produces a dominant-negative receptor (dnTGFßRII) that dimerizes with endogenous TGFßRI to form a receptor that can bind TGF-ß but cannot signal. We previously generated specific peptide enhanced affinity receptor TCRs recognizing the HLA-A*02-restricted peptides New York esophageal squamous cell carcinoma 1 (NY-ESO-1)157-165/l-Ag family member-1A (TCR: GSK3377794, formerly NY-ESO-1c259) and melanoma Ag gene A10254-262 (TCR: ADP-A2M10, formerly melanoma Ag gene A10c796). In this article, we show that exogenous TGF-ß inhibited in vitro proliferation and effector functions of human T cells expressing these first-generation high-affinity TCRs, whereas inhibition was reduced or abolished in the case of second-generation TCRs coexpressed with dnTGFßRII (e.g., GSK3845097). TGF-ß isoforms and a panel of TGF-ß-associated genes are overexpressed in a range of cancer indications in which NY-ESO-1 is commonly expressed, particularly in synovial sarcoma. As an example, immunohistochemistry/RNAscope identified TGF-ß-positive cells close to T cells in tumor nests and stroma, which had low frequencies of cells expressing IFN-γ in a non-small cell lung cancer setting. Coexpression of dnTGFßRII may therefore improve the efficacy of TCR-transduced T cells.
Sujet(s)
Lymphocytes T CD8+/immunologie , Carcinome épidermoïde/thérapie , Tumeurs hématologiques/thérapie , Immunothérapie adoptive/méthodes , Mélanome/thérapie , Récepteur de type II du facteur de croissance transformant bêta/métabolisme , Récepteurs aux antigènes des cellules T/métabolisme , Récepteurs chimériques pour l'antigène/métabolisme , Sarcome synovial/thérapie , Facteur de croissance transformant bêta/métabolisme , Antigènes néoplasiques/immunologie , Carcinome épidermoïde/immunologie , Lignée cellulaire tumorale , Génie génétique , Antigène HLA-A2/métabolisme , Tumeurs hématologiques/immunologie , Humains , Tolérance immunitaire , Mélanome/immunologie , Protéines membranaires/immunologie , Protéines tumorales/immunologie , Fragments peptidiques/immunologie , Récepteur de type II du facteur de croissance transformant bêta/génétique , Récepteurs aux antigènes des cellules T/génétique , Récepteurs chimériques pour l'antigène/génétique , Sarcome synovial/immunologie , Spécificité antigénique des récepteurs des lymphocytes T , Microenvironnement tumoralRÉSUMÉ
Chimeric Antigen Receptor (CAR)-redirected T cells show great efficacy in the patient-specific therapy of hematologic malignancies. Here, we demonstrate that a DARPin with specificity for CD4 specifically redirects and triggers the activation of CAR engineered T cells resulting in the depletion of CD4+ target cells aiming for elimination of the human immunodeficiency virus (HIV) reservoir.
Sujet(s)
Répétition ankyrine , Lymphocytes T CD4+/virologie , Infections à VIH/immunologie , VIH (Virus de l'Immunodéficience Humaine)/isolement et purification , Immunothérapie adoptive , Déplétion lymphocytaire/méthodes , Peptides/pharmacologie , Lymphocytes T CD4+/effets des médicaments et des substances chimiques , Lymphocytes T CD4+/immunologie , Relation dose-réponse (immunologie) , Évaluation préclinique de médicament , Gammaretrovirus/génétique , Vecteurs génétiques/génétique , Cellules HEK293 , Infections à VIH/virologie , Humains , Activation des lymphocytes , Peptides/composition chimique , Anticorps à chaîne unique/immunologie , Transduction génétiqueRÉSUMÉ
The haemolymph lipoprotein of the scorpion, Pandinus imperator was isolated and characterised. Contrary to the lipoproteins of insects and the discoidal HDL-lipoproteins of a crayfish and polychaete, the Pandinus lipoprotein consists of three instead of two apoproteins (apoPiLp I = 230 kDa, apoPiLp II = 130 kDa and apoPiLp III = 120 kDa). The apolipoproteins are arranged in varying stoichiometries as judged by cross-linking experiments. In lipoprotein samples from individual animals, the two smaller subunits occurred in a 1:1 stoichiometry, while the relative amount of the 230 kDa peptide varied. The lipoprotein is a slightly heart-shaped HDL with a diameter of approximately 15 nm. It is present in two densities of 1100 and 1190 kg/m(3), of which the latter is by far more abundant. The native molecular mass was estimated to be approximately 500 kDa. The lipid content was determined as 33.5% and consists of approximately 70% neutral lipids and approximately 30% phospholipids. Strikingly, 42.5% of the phospholipids is phosphatidylserine while phosphatidylcholine and phosphatidylethanolamine account for 55.1% and approximately 2.3%, respectively. Carbohydrate analysis suggests the presence of only high-mannose-type N-glycans. N-glycan profiling shows glycans corresponding to a size of 8.0-11.5 hexose units.