BTLA+CD200+ B cells dictate the divergent immune landscape and immunotherapeutic resistance in metastatic vs. primary pancreatic cancer.

Response to cancer immunotherapy in primary versus metastatic disease has not been well-studied. We found primary pancreatic ductal adenocarcinoma (PDA) is responsive to diverse immunotherapies whereas liver metastases are resistant. We discovered divergent immune landscapes in each compartment. Compared to primary tumor, liver metastases in both mice and humans are infiltrated by highly anergic T cells and MHCIIloIL10+ macrophages that are unable to present tumor-antigen. Moreover, a distinctive population of CD24+CD44-CD40- B cells dominate liver metastases. These B cells are recruited to the metastatic milieu by Muc1hiIL18hi tumor cells, which are enriched >10-fold in liver metastases. Recruited B cells drive macrophage-mediated adaptive immune-tolerance via CD200 and BTLA. Depleting B cells or targeting CD200/BTLA enhanced macrophage and T-cell immunogenicity and enabled immunotherapeutic efficacy of liver metastases. Our data detail the mechanistic underpinnings for compartment-specific immunotherapy-responsiveness and suggest that primary PDA models are poor surrogates for evaluating immunity in advanced disease.

PD-L1 engagement on T cells promotes self-tolerance and suppression of neighboring macrophages and effector T cells in cancer.

Programmed cell death protein 1 (PD-1) ligation delimits immunogenic responses in T cells. However, the consequences of programmed cell death 1 ligand 1 (PD-L1) ligation in T cells are uncertain. We found that T cell expression of PD-L1 in cancer was regulated by tumor antigen and sterile inflammatory cues. PD-L1+ T cells exerted tumor-promoting tolerance via three distinct mechanisms: (1) binding of PD-L1 induced STAT3-dependent 'back-signaling' in CD4+ T cells, which prevented activation, reduced TH1-polarization and directed TH17-differentiation. PD-L1 signaling also induced an anergic T-bet-IFN-γ- phenotype in CD8+ T cells and was equally suppressive compared to PD-1 signaling; (2) PD-L1+ T cells restrained effector T cells via the canonical PD-L1-PD-1 axis and were sufficient to accelerate tumorigenesis, even in the absence of endogenous PD-L1; (3) PD-L1+ T cells engaged PD-1+ macrophages, inducing an alternative M2-like program, which had crippling effects on adaptive antitumor immunity. Collectively, we demonstrate that PD-L1+ T cells have diverse tolerogenic effects on tumor immunity.

Specialized dendritic cells induce tumor-promoting IL-10+IL-17+ FoxP3neg regulatory CD4+ T cells in pancreatic carcinoma.

The drivers and the specification of CD4+ T cell differentiation in the tumor microenvironment and their contributions to tumor immunity or tolerance are incompletely understood. Using models of pancreatic ductal adenocarcinoma (PDA), we show that a distinct subset of tumor-infiltrating dendritic cells (DC) promotes PDA growth by directing a unique TH-program. Specifically, CD11b+CD103- DC predominate in PDA, express high IL-23 and TGF-ß, and induce FoxP3neg tumor-promoting IL-10+IL-17+IFNγ+ regulatory CD4+ T cells. The balance between this distinctive TH program and canonical FoxP3+ TREGS is unaffected by pattern recognition receptor ligation and is modulated by DC expression of retinoic acid. This TH-signature is mimicked in human PDA where it is associated with immune-tolerance and diminished patient survival. Our data suggest that CD11b+CD103- DC promote CD4+ T cell tolerance in PDA which may underscore its resistance to immunotherapy.

Pharmacological Evaluation of Mentha spicata L. and Plantago major L., Medicinal Plants Used to Treat Anxiety and Insomnia in Colombian Caribbean Coast.

Anxiety disorders are highly prevalent, chronic, and disabling conditions that impose enormous health and economic costs both on individuals and on society. Medicinal plants are an invaluable source of bioactive metabolites that can be useful as new pharmacological treatment. Teas from Mentha spicata and Plantago major are employed by Colombian populations to treat stress and insomnia. This work was conducted to evaluate their anxiolytic and hypnotic properties. For this, we employed the Elevated Plus-Maze test and the sodium pentobarbital-induced hypnosis method using Wistar rats. Oral administration of M. spicata extract (1000 mg/Kg) significantly increased the exploration and time spent in the open arms, which indicates its anxiolytic activity. On the other hand, both M. spicata and P. major extracts (1000 mg/Kg) remarkably augmented the sleeping time induced by pentobarbital, suggesting a sedative and hypnotic effect of the plants extracts. In addition, the acute toxicological study demonstrated that the doses used did not induce mortality or toxicity effects at hepatic or renal level. The bioactivity seems to be related to several kinds of constituents, mainly phenolic compounds such as flavonoids and tannins. In conclusion, these results reinforce the potential use of these species in the therapy of anxiety.

Evaluation of beta-adrenergic receptor subtypes in the human prostate cancer cell line-LNCaP.

The present study was undertaken to determine the effects of catecholamines, agonists, and antagonists of beta-adrenergic receptors (AR) in the LNCaP cell line. Changes in cellular cyclic adenosine-3',5'-monophosphate (cAMP) levels were quantified by the use of a 6 cAMP response element (CRE)-luciferase reporter gene assay. LNCaP cells were transiently transfected with this gene construct, incubated in 96-well microtiter plates for 24 hr, and then treated with beta-AR agonists and/or antagonists for 4 hr. The rank order of potency for catecholamines and known beta-AR agonists was terbutaline(3.31 nM)>isoproterenol(8.31 nM)> or =fenoterol(15 nM)=epinephrine(16.2 nM)>norepinephrine(77.5 nM)>BRL-37344 [(R(*),R(*))-(+/-)4-[2-[(2-(3-chlorophenyl)-2-hydroxyethyl)amino]propyl]phenoxy acetic acid, sodium salt] (1000 nM)>dobutamine(1770 nM)>CGP12177 (4-[3-[(1,1-dimethylethyl)amino]-2-hydroxypropoxy]-1,3-dihydro-2H-benzimidazole-2-one hydrochloride) (inactive). The non-selective beta(1)-/-beta(2)-AR antagonists; propranolol and CGP 12177, at 10(-7)M, inhibited luciferase activity induced by these agonists by 80-96%. Propranolol blocked isoproterenol-induced luciferase responses in a competitive manner (K(B)=1.4 nM). In addition, isoproterenol-activated luciferase expression was blocked more potently by ICI 118,551 [(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethy) amino]-2-butanol], a beta(2)-AR antagonist than by ICI 89,406 [(+/-)-N-[2-[3-(2-cyanophenoxy-)]-2-hydroxypropylamino]ethyl-N-phenylurea], a beta(1)-AR antagonist, giving K(B) values of 1.07 and 161nM, respectively. These results suggest that the beta(2)-AR is the major subtype mediating catecholamine-induced cAMP changes in LNCaP cells.