Neoantigen-reactive CD8+ T cells affect clinical outcome of adoptive transfer with tumor-infiltrating lymphocytes in melanoma

BACKGROUND: Neoantigen-driven recognition and T cell-mediated killing contribute to tumor clearance following adoptive cell therapy (ACT) with Tumor-Infiltrating Lymphocytes (TILs). Yet, how diversity, frequency, and persistence of expanded neoepitope-specific CD8+ T cells derived from TIL infusion products affect patient outcome is not fully determined. METHODS: Using barcoded pMHC multimers, we provide a comprehensive mapping of CD8+ T cells recognizing neoepitopes in TIL infusion products and blood samples from 26 metastatic mela-noma patients who received ACT. RESULTS: We identified 106 neoepitopes within TIL infusion products corresponding to 1.8% of all predicted neoepitopes. We observed neoepitope-specific recognition to be virtually devoid in TIL infusion products given to patients with progressive disease outcome. Moreover, we found that the frequency of neoepitope-specific CD8+ T cells in TIL infusion products correlated with in-creased survival, and that detection of engrafted CD8+ T cells in post-treatment (i.e. originating from the TIL infusion product) were unique to responders of TIL-ACT. Finally, we found that a transcriptional signature for lymphocyte activity within the tumor microenvironment was associated with a higher frequency of neoepitope-specific CD8+ T cells in the infusion product. CONCLUSIONS: These data support previous case studies of neoepitope-specific CD8+ T cells in melanoma, and indicate that successful TIL-ACT is associated with an expansion of neoepitope-specific CD8+ T cells. FUNDING: NEYE Foundation; European Research Council; Lundbeck Foundation Fellowship; Carlsberg Foundation

Identifying Canadian Freshwater Fishes through DNA Barcodes

BACKGROUND: DNA barcoding aims to provide an efficient method for species-level identifications using an array of species specific molecular tags derived from the 5' region of the mitochondrial cytochrome c oxidase I (COI) gene. The efficiency of the method hinges on the degree of sequence divergence among species and species-level identifications are relatively straightforward when the average genetic distance among individuals within a species does not exceed the average genetic distance between sister species. Fishes constitute a highly diverse group of vertebrates that exhibit deep phenotypic changes during development. In this context, the identification of fish species is challenging and DNA barcoding provide new perspectives in ecology and systematics of fishes. Here we examined the degree to which DNA barcoding discriminate freshwater fish species from the well-known Canadian fauna, which currently encompasses nearly 200 species, some which are of high economic value like salmons and sturgeons. METHODOLOGY/PRINCIPAL FINDINGS: We bi-directionally sequenced the standard 652 bp "barcode" region of COI for 1360 individuals belonging to 190 of the 203 Canadian freshwater fish species (95%). Most species were represented by multiple individuals (7.6 on average), the majority of which were retained as voucher specimens. The average genetic distance was 27 fold higher between species than within species, as K2P distance estimates averaged 8.3% among congeners and only 0.3% among concpecifics. However, shared polymorphism between sister-species was detected in 15 species (8% of the cases). The distribution of K2P distance between individuals and species overlapped and identifications were only possible to species group using DNA barcodes in these cases. Conversely, deep hidden genetic divergence was revealed within two species, suggesting the presence of cryptic species. CONCLUSIONS/SIGNIFICANCE: The present study evidenced that freshwater fish species can be efficiently identified through the use of DNA barcoding, especially the species complex of small-sized species, and that the present COI library can be used for subsequent applications in ecology and systematics

Efficacy and Safety of Olmesartan Medoxomil 40 mg/Hydrochlorothiazide 12.5 mg Combination Therapy versus Olmesartan Medoxomil 40 mg Monotherapy in Patients with Moderate to Severe Hypertension

Current hypertension guidelines recommend using two antihypertensive agents when blood pressure (BP) control is not achieved with one single agent. Objective: This study was designed to assess the antihypertensive benefit of the olmesartan medoxomil 40mg/hydrochlorothiazide (HCTZ) 12.5mg combination versus olmesartan medoxomil 40 mg monotherapy in patients with moderate to severe hypertension. Methods: This was a randomized, double-blind, parallel-group, up-titration, multicentre, multinational, phase III study. Following a 2-week single-blind placebo run-in phase, 846 hypertensive patients with mean seated systolic BP (SeSBP) of 160200mmHg and mean seated diastolic BP (SeDBP) of 100120mmHg were randomized (1 : 2 ratio) to receive double-blind treatment with olmesartan medoxomil 40 mg or olmesartan medoxomil 40mg/HCTZ 12.5 mg for 8 weeks (phase A). At week 8, patients not reaching BP goal (<140/90 mmHg; <130/80mmHg in patients with diabetes mellitus) were up-titrated from olmesartan medoxomil 40mg to olmesartan medoxomil 40mg/HCTZ 12.5mg or from olmesartan medoxomil 40 mg/HCTZ 12.5mg to olmesartan medoxomil 40 mg/HCTZ 25mg for an additional 8 weeks (phase B). Patients on goal continued their initial treatment. The primary efficacy parameter was the change in mean SeDBP during phase A. Results: Olmesartan medoxomil 40 mg/HCTZ 12.5mg reduced mean SeDBP significantly more (-18.9mmHg) than olmesartan medoxomil 40mg (-15.8mmHg) after 8 weeks of double-blind treatment (difference: -3.1mmHg, p < 0.0001). Olmesartan medoxomil 40mg/HCTZ 12.5mg also reduced mean SeSBP significantly more than olmesartan medoxomil 40mg (-5.4 mmHg, p < 0.0001). As a result, BP goal rates at week 8 were significantly higher with olmesartan medoxomil 40mg/HCTZ 12.5 mg than with olmesartan medoxomil 40mg (58.5% vs 44.3%; odds ratio 1.88; 95% CI 1.32, 2.54). During phase B, mean BP reductions were greater in patients up-titrated from olmesartan medoxomil 40 mg to olmesartan medoxomil 40 mg/HCTZ 12.5mg than in those continuing on olmesartan medoxomil 40 mg (SeDBP: 9.3mmHg vs 0.5 mmHg; SeSBP: 12.4mmHg vs 0.5 mmHg). Similarly, mean BP reductions were greater in patients up-titrated from olmesartan medoxomil 40 mg/HCTZ 12.5mg to olmesartan medoxomil 40 mg/HCTZ 25mg than in those continuing on olmesartan medoxomil 40 mg/HCTZ 12.5mg (SeDBP: 8.0mmHg vs 0.3 mmHg; SeSBP: 12.1mmHg vs 0.4 mmHg). In patients not on goal at week 8, addition of HCTZ 12.5mg to olmesartan medoxomil 40 mg or up-titration from olmesartan medoxomil 40mg/HCTZ 12.5mg to olmesartan medoxomil 40 mg/HCTZ 25 mg brought additional patients to goal at week 16 (38.8% vs 36.9%). All treatments were well tolerated. Conclusion: The olmesartan medoxomil 40mg/HCTZ 12.5mg combination is superior to olmesartan medoxomil 40mg monotherapy in reducing SeDBP and SeSBP and increasing BP goal rates after 8 weeks. Patients not on goal at week 8 with olmesartan medoxomil 40mg or olmesartan medoxomil 40mg/HCTZ 12.5mg benefited from adding HCTZ 12.5mg or up-titrating to olmesartan medoxomil 40 mg/HCTZ 25 mg, respectively, confirming that uptitration is a clinically meaningful way to improve BP control