Localization-associated immune phenotypes of clonally expanded tumor-infiltrating T cells and distribution of their target antigens in rectal cancer

The degree and type of T cell infiltration influence rectal cancer prognosis regardless of classical tumor staging. We asked whether clonal expansion and tumor infiltration are restricted to selected-phenotype T cells; which clones are accessible in peripheral blood; and what the spatial distribution of their target antigens is. From five rectal cancer patients, we isolated paired tumor-infiltrating T cells (TILs) and T cells from unaffected rectum mucosa (T(UM)) using 13-parameter FACS single cell index sorting. TCRαβ sequences, cytokine, and transcription factor expression were determined with single cell sequencing. TILs and T(UM) occupied distinct phenotype compartments and clonal expansion predominantly occurred within CD8(+) T cells. Expanded TIL clones identified by paired TCRαβ sequencing and exclusively detectable in the tumor showed characteristic PD-1 and TIM-3 expression. TCRβ repertoire sequencing identified 49 out of 149 expanded TIL clones circulating in peripheral blood and 41 (84%) of these were PD-1(-) TIM-3(-). To determine whether clonal expansion of predominantly tumor-infiltrating T cell clones was driven by antigens uniquely presented in tumor tissue, selected TCRs were reconstructed and incubated with cells isolated from corresponding tumor or unaffected mucosa. The majority of clones exclusively detected in the tumor recognized antigen at both sites. In summary, rectal cancer is infiltrated with expanded distinct-phenotype T cell clones that either i) predominantly infiltrate the tumor, ii) predominantly infiltrate the unaffected mucosa, or iii) overlap between tumor, unaffected mucosa, and peripheral blood. However, the target antigens of predominantly tumor-infiltrating TIL clones do not appear to be restricted to tumor tissue

Rapid single-cell identification of Epstein-Barr virus-specific T-cell receptors for cellular therapy

BACKGROUND AND AIMS: Epstein-Barr virus (EBV) is associated with solid and hematopoietic malignancies. After allogeneic stem cell transplantation, EBV infection or reactivation represents a potentially life-threatening condition with no specific treatment available in clinical routine. In vitro expansion of naturally occurring EBV-specific T cells for adoptive transfer is time-consuming and influenced by the donor's T-cell receptor (TCR) repertoire and requires a specific memory compartment that is non-existent in seronegative individuals. The authors present highly efficient identification of EBV-specific TCRs that can be expressed on human T cells and recognize EBV-infected cells. METHODS AND RESULTS: Mononuclear cells from six stem cell grafts were expanded in vitro with three HLA-B*35:01- or four HLA-A*02:01-presented peptides derived from six EBV proteins expressed during latent and lytic infection. Epitope-specific T cells expanded on average 42-fold and were single-cell-sorted and TCRαβ-sequenced. To confirm specificity, 11 HLA-B*35:01- and six HLA-A*02:01-restricted dominant TCRs were expressed on reporter cell lines, and 16 of 17 TCRs recognized their presumed target peptides. To confirm recognition of virus-infected cells and assess their value for adoptive therapy, three selected HLA-B*35:01- and four HLA-A*02:01-restricted TCRs were expressed on human peripheral blood lymphocytes. All TCR-transduced cells recognized EBV-infected lymphoblastoid cell lines. CONCLUSIONS: The authors' approach provides sets of EBV epitope-specific TCRs in two different HLA contexts. Resulting cellular products do not require EBV-seropositive donors, can be adjusted to cell subsets of choice with exactly defined proportions of target-specific T cells, can be tracked in vivo and will help to overcome unmet clinical needs in the treatment and prophylaxis of EBV reactivation and associated malignancies

Immune phenotypes and target antigens of clonally expanded bone marrow T cells in treatment-naïve multiple myeloma

Multiple myeloma is a hematologic malignancy of monoclonal plasma cells that accumulate in the bone marrow. Despite their clinical and pathophysiological relevance, the roles of bone marrow infiltrating T cells in treatment-naïve patients are incompletely understood. We investigated whether clonally expanded T cells i) were detectable in multiple myeloma bone marrow, ii) showed characteristic immune phenotypes, and iii) whether dominant clones recognized antigens selectively presented on multiple myeloma cells. Single-cell index sorting and T-cell receptor (TCR)αβ sequencing of bone marrow T cells from 13 treatment-naïve patients showed dominant clonal expansion within CD8+ cytolytic effector compartments, and only a minority of expanded T-cell clones expressed the classical immune checkpoint molecules PD 1, CTLA 4, or TIM 3. To identify their molecular targets, TCRs of 68 dominant bone marrow clones from five selected patients were re-expressed and incubated with multiple myeloma and non multiple myeloma cells from corresponding patients. Only one out of 68 TCRs recognized antigen presented on multiple myeloma cells. This TCR was HLA-C-restricted, self-peptide-specific, and could be activated by multiple myeloma cells of multiple patients. The remaining dominant T-cell clones did not recognize multiple myeloma cells and were, in part, specific for antigens associated with chronic viral infections. In conclusion, we showed that dominant bone marrow T-cell clones in treatment naïve patients rarely recognize antigens presented on multiple myeloma cells and exhibit low expression of classical immune checkpoint molecules. Our data provide experimental context for experiences from clinical immune checkpoint inhibition trials and will inform future T cell-dependent therapeutic strategies

Satellite Determination Of Api Gravity And Sara Components Of Offshore Petroleum Seeps

Crude oils can be classified based on their API gravity or their SARA components (i.e. saturated, aromatics, resins, and asphaltenes). This work evaluates the possibility to infer such oil's characteristics by means of reflectance spectroscopy and multispectral and hyperspectral imagery. Reflectance spectra (0.35-2.5μm) of oils and films of oil on water were measured under laboratory conditions using a high-resolution spectroradiometer. Multivariate statistics (i.e. principal component and partial least square analysis) were used to evaluate these spectra, taking into consideration both the spectral resolution at laboratory measurements (2150 bands) and the spectral resolution offered by orbital sensors (i.e. Hyperion (220 bands) and ASTER (9 bands)). The statistical approach yielded predictive models based on the correlation of oil composition and its spectral response, allowing remote assessment of the oil quality of a particular seepage from the Campos Basin (Brazil). This oil has a known API gravity ranging between 19-22, 40-49% mass/mass of saturated, 33-25 of aromatics, 20-28 of resins and 3-1.5 of asphaltenes. The remotely retrieved values based on the spectral response of the seep are within the actual range and are remarkably similar: API gravity of 19.6 and 45.38% mass/mass of saturated, 26.91 of aromatics, 24.61 of resins and 2.14 of asphaltenes. This result indicates the potential of this methodology, first proposed in this work, for the indirect inference of API gravity and SARA composition based on remote sensing data and techniques. © 2012 Sociedade Brasileira de Geofísica.304419430Abrams, M., Hook, S., Ramachandran, B., (2002) ASTER User Handbook., p. 135. , Version 2. Jet Propulsion LaboratoryAlboudwarej, J.H., Felix, J., Taylor, S., Highlighting Heavy Oil (2006) Oilfield Review, pp. 34-53. , http://www.slb.com.br, Available on Accessed on June 01, 2012Aske, N., Kallevik, H., Sjöblom, J., Determination of Saturate, Aromatic, Resin, and Asphaltenic (SARA) Components in Crude Oils by Means of Infrared and Near-Infrared Spectroscopy (2001) Energy & Fuel, (15), pp. 1304-1312Beebe, K.R., Kowalski, B.R., An introduction to Multivariate Calibration and Analysis (1987) Anal Chem., 59 (17), pp. 1007A-1017ABentz, C.B., Lorenzzetti, J.A., Kampel, M., Politano, A.T., Lucca, E.V.L., Contribuição de dados ASTER, CBERS, R99/SIPAM e OrbiSAR-1 para monitoramento oceânico-Resultados do Projeto FITOSAT (2007) Anais XIII Simpósio Brasileiro de Sensoriamento Remoto, pp. 3755-3762. , In: Florianópolis, SC, Brasil, INPEBorin, J.E., (2003) Estudo Geoquímico de óleo do Campo de Marlim, Bacia de Campos, Síntese de Biomarcadores Esteranos Aromáticos., p. 139. , M. Sc. dissertation. Instituto de Química, Universidade Estadual de Campinas, SP(2003) Relatório Técnico-Atendimento 178/03., , http://www.cetesb.sp.gov.br/emergencia/acidentes/vazamento/publicacoes/13.pdf, CETESB (COMPANHIA AMBIENTAL DO ESTADO DE SÃO PAULO). Operação TEBAR VII-Píer Sul. Available on Accessed on June 01, 2012Cloutis, E.A., Spectra Reflectance Properties of Hydrocarbons: Remote Sensing Implications (1989) Science, 245, pp. 165-168. , Washington: American Association for the Advancement of ScienceGama, R., (2008) Caracterização Geoquímica de Óleos Biodegradados dos Campos de Marlim Sul e Roncador, na Bacia de Campos., p. 188. , M. Sc. dissertation, Universidade Federal do Rio de Janeiro, RJGeladi, P., Kowalski, B.R., Partial Least-Square Regression: A Tutorial (1986) Anal. Chim. Acta, 185, pp. 1-17Hannisdal, A., Hemmingsen, P.V., Sjoblom, J., Group Type Analysis of Heavy Crude Oils Using Vibrational Spectroscopy in Combination with Multivariate Analysis (2005) Ind. Eng. Chem. Res., 44, pp. 1349-1357Hidajat, K., Chong, S.M., Quality Characterization of crude oils by partial least square calibration of NIR spectral profiles (2000) J. Near Infrared Spectrosc, 8, pp. 53-59Hunt, G.R., (1979) Petroleum Geochemistry and Geology., p. 617. , W.H. Freeman and Company, San Francisco(2001) ACORN User's Guide. Analytical Imaging and Geophysics., p. 64. , IMSPEC. Boulder, CO, USAIwasaki, A., Fujisada, H., Akao, H., Shindou, O., Akagi, S., Enhancement of spectral separation performance for ASTER/SWIR (2001) Proceedings of SPIE, The International Society for Optical Engineering, 4486. , 4-2-50Iwasaki, A., Tonooka, H., Validation of crosstalk correction algorithm for ASTER/SAWIR (2005) IEEE T Geosci Remote Sensing, 43 (12), pp. 2747-2751Kallevik, H., Kvalheimom, S.J., Öblom, J., Quantitative Determination of Asphaltenes and Resins in Solution by Means of Near-Infrared Spectroscopy: Correlations to Emulsion Stability (2000) J. Colloid Interf. Sci., 225, pp. 494-504(2012), http://laqqa.iqm.unicamp.br/, LAQQA (LABORAT ÓRIO DE QUIMIOMETRIA EM QUÍMICA ANALÍTICA). Available on Accessed on June 01, 2012Lima, A.A., Farah, M.A., Rajagopal, K., A Viscosidade De Óleo Cru (2007) 4o PDPETRO (ABPG), , In: Campinas, SP 3.1.0174-1-1-3.1.0174-1-8Lammoglia, T., Souza Filho, C.R., Spectroscopic characterization of oils yielded from Brazilian offshore basins: Potential applications of remote sensing Rem (2011) Sen. of Environment, 10 (17), pp. 2525-2535. , Volume 115Lammoglia, T., Souza Filho, C.R., Mapping and characterization of the API gravity of offshore hydrocarbon seepages using multispectral ASTER data (2012) Rem. Sen. of Environment, 123, pp. 381-389Lorenzzetti, J.A., Kampel, M., Bentz, C.M., Torres Jr., A.R., A Meso-scale Brazil current frontal eddy: Observations by ASAR, RADARSAT-1 complemented with visible and infrared sensors, in situ data and numerical modeling (2006) SEASAR, , In: Frascati, Italy, SP-613Mello, M.R., Koutsoukos, E.A.M., Mohriak, W.U., Bacoccoli, G., Selected petroleum systems in Brazil (1994) The petroleum system-From source to trap, pp. 499-512. , In: MAGOON LB & DOW WG (Eds.). AAPG Memoir 60Milani, E.J., Araújo, L.M., Recursos Minerais Energéticos: Petróleo (2003) Geologia, Tectônica e Recursos Minerais do Brasil, pp. 541-576. , In: BIZZI LA, SCHOBBENHAUS C, VIDOTTI RM & GONÇALVES JH (Eds.). CPRM, Brasília, BRPasquini, C., Bueno, A.F., Characterization of petroleum using near-infrared spectroscopy: Quantitative modeling for the true boiling point curve and specific gravity (2007) Fuel, 86, pp. 1927-1934Pearlman, J., Segal, C., Liao, L., Carman, S., Folkman, M., Browne, B., Ungar, S., (2000) Development and Operations of the EO-1 Hyperion Imaging Spectrometer., p. 11. , http://eo1.gsfc.nasa.gov/miscPages/TechForumOther/Yperion%20SPIE%20Publication.pdf, Available on Accessed on Dec. 01, 2007Silverstein, R.M., Webster, F.X., (1998) Infrared Spectroscopy In Spectrometric Identification of Organic Compounds., pp. 70-105. , New York, Ed. Wiley(2012) THE UNSCRAMBLER., , http://www.camo.com/, Available on Accessed on June 01, 2012Thomas, E.J., O Petróleo (2001) Fundamentos de Engenharia de Petróleo., pp. 1-13. , In: Editora Interciência 2edTissot, B.P., Welte, D.H., (1984) Petroleum formation and occurrence, p. 699. , Heidelbergvan der Meer, F., Analysis of spectral absorption features in hyperspectral imagery (2004) Int. J. Earth Observ., 5 (1), pp. 55-68Wettle, M., Daniel, P.J., Logan, G.A., Thankappan, M., Assessing the effect of hydrocarbon oil type and thickness on a remote sensing signal: A sensitivity study based on the optical properties of two different oil types and the HYMAP and Quickbird sensors (2009) Remote Sens. Environ., 113, pp. 2000-2010Winkelmann, K.H., (2005) On the applicability of imaging spectrometry for the detection and investigation of contaminated sites with particular consideration given to the detection of fuel hydrocarbon contaminants in soil., p. 236. , Doctore thesis. Universidade de Cottbus-AlemanhaZílio, E.L., Pinto, U.B., Identificação e Distribuição dos Principais Grupos de Compostos Presentes nos Petróleos Brasileiros (2002) Boletim Técnico. PETROBRAS, Rio de Janeiro, 45 (1), pp. 21-2

Mapping and characterization of the API gravity of offshore hydrocarbon seepages using multispectral ASTER data

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The objective of this work is the qualitative remote characterization (API gravity degree) of oil seepages on the ocean surface. In order to achieve this goal, multispectral data acquired by the Advanced Spaceborne Thermal Emission and Reflection (ASTER) sensor are employed. ASTER registered tracts of oil along oceanic sectors of the Campos Basin (Brazil) and the Bay of Campeche (Gulf of Mexico) in several occasions. Numerous evidences indicate that these oil patches bear a straight link to oceanic seepages. The delimitation and segmentation of these seepages in the ASTER imagery are accomplished making use of an unsupervised, neural network fuzzy-clustering algorithm. Spectra representative of the seepages are extracted from atmospherically-corrected ASTER data pixels (9 bands spanning from visible to shortwave infrared wavelengths) contained in the classified segments. The ASTER spectra are checked against a predictive (o)API partial least square regression model. This model is established on the basis of oil spectra of known (o)API varying from 13 to 47 yielded by laboratory measurements. Considering this model, API gravity degrees of 19.6+/-1.37 and 15.9+/-2.9 are remotely estimated for the seepage in the Campos Basin and the Bay of Campeche, respectively. Oils produced from Campos and Campeche fields typically show (o)API varying from 17-24 and 12-16.5. correspondingly. These results indicate the potential of the methodology proposed and of ASTER data and alike to remotely infer physical-chemical properties of hydrocarbons, since a close match was verified between predicted and true API gravity degrees for both study areas. The data, methods and experience gained in this research can be operationally tested in offshore oil exploration and, likewise, be adapted to environmental monitoring of oil spills in coastal regions. (C) 2012 Elsevier Inc. All rights reserved.123381389Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Effects of Dietary Fat on Hormone and Metabolite Concentrations and Follicular Development in Unilateral Ovariectomized Estrous Cycling Brahman Cows

Last updated: 5/11/201