Expression hierarchy of T cell epitopes from melanoma differentiation antigens: unexpected high level presentation of tyrosinase-HLA-A2 Complexes revealed by peptide-specific, MHC-restricted, TCR-like antibodies.

Peptide Ags presented by class I MHC molecules on human melanomas and that are recognized by CD8(+) T cells are the subjects of many studies of antitumor immunity and represent attractive candidates for therapeutic approaches. However, no direct quantitative measurements exist to reveal their expression hierarchy on the cell surface. Using novel recombinant Abs which bind these Ags with a peptide-specific, MHC-restricted manner, we demonstrate a defined pattern of expression hierarchy of peptide-HLA-A2 complexes derived from three major differentiation Ags: gp100, Melan-A/Mart-1, and tyrosinase. Studying melanoma cell lines derived from multiple patients, we reveal a surprisingly high level of presentation of tyrosinase-derived complexes and moderate to very low expression of complexes derived from other Ags. No correlation between Ag presentation and mRNA expression was found; however, protein stability may play a major role. These results provide new insights into the characteristics of Ag presentation and are particularly important when such targets are being considered for immunotherapy. These results may shed new light on relationships between Ag presentation and immune response to cancer Ags

Lymph node-targeted immunotherapy mediates potent immunity resulting in regression of isolated or metastatic human papillomavirus-transformed tumors

PURPOSE: The goal of this study was to investigate the therapeutic potential of a novel immunotherapy strategy resulting in immunity to localized or metastatic human papillomavirus 16-transformed murine tumors. EXPERIMENTAL DESIGN: Animals bearing E7-expressing tumors were coimmunized by lymph node injection with E7 49-57 antigen and TLR3-ligand (synthetic dsRNA). Immune responses were measured by flow cytometry and antitumor efficacy was evaluated by tumor size and survival. In situ cytotoxicity assays and identification of tumor-infiltrating lymphocytes and T regulatory cells were used to assess the mechanisms of treatment resistance in bulky disease. Chemotherapy with cyclophosphamide was explored to augment immunotherapy in late-stage disease. RESULTS: In therapeutic and prophylactic settings, immunization resulted in a considerable expansion of E7 49-57 antigen-specific T lymphocytes in the range of 1/10 CD8(+) T cells. The resulting immunity was effective in suppressing disease progression and mortality in a pulmonary metastatic disease model. Therapeutic immunization resulted in control of isolated tumors up to a certain volume, and correlated with antitumor immune responses measured in blood. In situ analysis showed that within bulky tumors, T-cell function was affected by negative regulatory mechanisms linked to an increase in T regulatory cells and could be overcome by cyclophosphamide treatment in conjunction with immunization. CONCLUSIONS: This study highlights a novel cancer immunotherapy platform with potential for translatability to the clinic and suggests its potential usefulness for controlling metastatic disease, solid tumors of limited size, or larger tumors when combined with cytotoxic agents that reduce the number of tumor-infiltrating T regulatory cells

Reconstruction of human exposure to heavy metals using synchrotron radiation microbeams in prehistoric and modern humans

Objective Teeth can serve as records of environmental exposure to heavy metals during their formation. We applied a new technology — synchrotron radiation microbeams (SRXRF) — for analysis of heavy metals in human permanent teeth in modern and historical samples. Methods Each tooth was cut in half. A longitudinal section 200 μm in thickness was subjected to the determination of the heavy metal content by SRXRF or conventional analytical methods (ICP-MS analysis or reduction–aeration atomic absorption spectrometry). The relative concentrations of Pb, Hg, Cu and Zn measured by SRXRF were translated in concentrations (in g of heavy metal/g of enamel) using calibration curves by the two analytical methods. Results Concentrations in teeth in the modern females (n = 5) were 1.2 ± 0.5 μg/g (n = 5) for Pb; 1.7 ± 0.2 ng/g for Hg; 0.9 ± 1.1 μg/g for Cu; 150 ± 24.6 μg/g for Zn. The levels of Pb were highest in the teeth samples obtained from the humans of the Edo era (1603–1868 ad) (0.5–4.0 μg/g, n = 4). No trend was observed in this study in the Hg content in teeth during 3, 000 years. The concentrations of Cu were highest in teeth of two medieval craftsmen (57.0 and 220 μg/g). The levels of Zn were higher in modern subjects (P < 0.05) than those in the Jomon (~1000 bc) to Edo periods [113.2 ± 27.4 (μg/g, n = 11)]. Reconstruction of developmental exposure history to lead in a famous court painter of the Edo period (18th century) revealed high levels of Pb (7.1–22.0 μg/g) in his childhood. Conclusions SRXRF is useful a method for reconstructing human exposures in very long trends

Eye gaze and pupillary response in Angelman syndrome

BackgroundAngelman syndrome (AS) is a rare neurological disorder characterized by severe developmental disability, communication impairment, elevated seizure risk, and motor system abnormalities.AimsThe aims of this study were to determine the feasibility of social scene eye tracking and pupillometry measures in individuals with AS and to compare the performance of AS participants to individuals with idiopathic Autism Spectrum Disorder (ASD) and typically developing controls (TDC).Methods and proceduresIndividuals with AS and age- and gender- matched controls completed a social eye tracking paradigm. Neurobehavioral characterization of AS participants was completed via a battery of psychological testing and caregiver behavioral evaluations.Outcomes and resultsEight of seventeen recruited AS participants completed the eye tracking paradigm. Compared to TDC, AS subjects demonstrated significantly less preference for social scenes than geometric shapes. Additionally, AS subjects showed less pupil dilation, compared to TDC, when viewing social scenes versus geometric shapes. There was no statistically significant difference found between AS and ASD subjects in either social eye tracking or pupillometry.Conclusions and implicationsThe use of eye tracking and pupillometry may represent an innovative measure for quantifying AS-associated impairments in social salience