NY-ESO-1 is a ''cancer-testis'' antigen expressed in epithelial ovarianHLA-DP4 ͉ peptide epitope ͉ tumor recognition ͉ vaccine T here is increasing evidence that the immune system has the ability to recognize tumor-associated antigens expressed in human malignancies and to induce antigen-specific humoral and cellular immune responses to these targets. In epithelial ovarian cancer (EOC), support for the role of immune surveillance of tumors comes from our recent observation indicating that the presence of intraepithelial CD8 ϩ -infiltrating T lymphocytes in tumors is associated with improved survival of patients with the disease (1). Although the majority of women with advanced-stage ovarian cancer respond to first-line chemotherapy, most of these responses are not durable, and Ͼ70% of patients die of recurrent disease within 5 years of diagnosis. Therefore, the development of strategies to enhance the potential of tumor antigen-specific CD8 ϩ T and CD4 ϩ T cells is urgently needed for extending remission rates in this disease. In this regard, cancer-testis antigens, a unique class of antigens that demonstrate high levels of expression in adult male germ cells but generally not in other normal adult tissues and aberrant expression in a variable proportion of a wide range of different cancer types, are promising candidates for immunotherapy. Among cancer-testis antigens, NY-ESO-1 (2) is one of the most spontaneously immunogenic tumor antigens described so far. Previously, we reported that NY-ESO-1 is a promising target for specific immunotherapy of EOC (3).Although the majority of cancer vaccine trials have focused on eliciting antigen-specific CD8 ϩ T cells, a growing body of evidence indicates that CD4 ϩ T cells play a pivotal role in orchestrating these responses. The multiple roles of antigen-specific CD4 ϩ T cells include the provision of help to CD8 ϩ T cells during the primary and secondary immune responses, direct cytolysis, and activation of B cells for production of tumor antigen-specific Abs. Therefore, we have focused on the NY-ESO-1 epitope, ESO 157-170 , a naturally processed helper epitope that is recognized by CD4 ϩ T cells in the context of HLA-DPB1*0401 and *0402 (4), prevalent MHC class II alleles present in Ϸ43-70% of Caucasians. Moreover, the NY-ESO-1 HLA-DP4 epitope has HLA-A2 (ESO 157-165 ) (5) and HLA-A24 (ESO 158-166 ) (6) motifs embedded in its natural sequence. In this study, we evaluated whether active immunization with ESO 157-170 would elicit NY-ESO-1-specific CD4 ϩ and CD8 ϩ T cell responses in ovarian cancer patients with minimal disease burden. In addition, we characterized NY-ESO-1-specific CD8 ϩ and CD4 ϩ T cell receptor (TCR) repertoires in conjunction with functional analysis of vaccine-elicited T cell clones.

The risks of cardiovascular disease and cancer were not elevated among postmenopausal women using vaginal estrogens, providing reassurance about the safety of treatment.

Autoantibodies, a hallmark of both autoimmunity and cancer, represent an easily accessible surrogate for measuring adaptive immune responses to cancer. Sera can now be assayed for reactivity against thousands of proteins using microarrays, but there is no agreed-upon standard to analyze results. We developed a set of tailored quality control and normalization procedures based on ELISA validation to allow patient comparisons and determination of individual cutoffs for specificity and sensitivity. Sera from 60 patients with pancreatic cancer, 51 patients with ovarian cancer, and 53 age-matched healthy donors were used to assess the binding of IgG antibodies against a panel of >8000 human antigens using protein microarrays and fluorescence detection. The resulting data interpretation led to the definition and ranking of proteins with preferred recognition by the sera from cancer patients in comparison with healthy donors, both by frequency and strength of signal. We found that 202 proteins were preferentially immunogenic in ovarian cancer sera compared to 29 in pancreatic cancer, with few overlaps. Correlates of autoantibody signatures with known tumor expression of corresponding antigens, functional pathways, clinical stage, and outcome were examined. Serological analysis of arrays displaying the complete human proteome (seromics) represents a new era in cancer immunology, opening the way to defining the repertoire of the humoral immune response to cancer. serum antibody | biomarkers | protein microarrays | serology | autoantigen

Objectives Despite the increasing prevalence of Type 2 diabetes mellitus (T2DM) among children and adolescents, little is known about their risk of developing diabetic retinopathy (DR). We sought to identify risk factors for DR in youth with DM, to compare DR rates for youth with Type 1 diabetes mellitus (T1DM) and T2DM, and to assess whether adherence to DR screening guidelines promoted by the American Academy of Ophthalmology, American Academy of Pediatrics, and American Diabetes Association adequately capture youth with DR. Design Retrospective observational longitudinal cohort study. Participants Youth aged ≤ 21 years with newly diagnosed T1DM or T2DM enrolled in a large U.S. managed care network. Main Outcome Measure Hazard ratios (HR) with 95% confidence intervals (CIs) for developing DR. Methods In this study of youth aged ≤ 21 years with newly diagnosed T1DM or T2DM enrolled in a large U.S. managed care network who were under ophthalmic surveillance, we identified the incidence and timing of DR onset for youth with T1DM and T2DM. Kaplan-Meier survival curves assessed the timing of initial diagnosis of DR for youth with each type of diabetes. Multivariable Cox proportional hazard regression modeling identified factors associated with the hazard of developing DR. Model predictors were age and calendar year at initial diabetes mellitus diagnosis, sex, race/ethnicity, net worth, and glycosylated hemoglobin (HbA1c). Results Among the 2240 youth with T1DM and 1768 youth with T2DM, 20.1% and 7.2% developed DR, over a median follow-up of 3.2 and 3.1 years, respectively. Survival curves demonstrated that youth with T1DM developed DR faster than youth with T2DM (P<0.0001). For every one-point increase in HbA1c, the hazard for DR increased by 20% (HR=1.20, CI 1.06–1.35) and 30% (HR=1.30, CI 1.08–1.56) among youth with T1DM and T2DM, respectively. Current guidelines suggest ophthalmic screening begin 3–5 years after initial DM diagnosis, at which point in our study, over 18% of youth with T1DM had already had received ≥1 DR diagnosis. Conclusions Youth with T1DM or T2DM exhibit a considerable risk for DR and should undergo regular screenings by eye-care professionals to ensure timely DR diagnosis and limit progression to vision-threatening disease.

Background Vitamin D has anti-inflammatory and anti-microbial properties that, together with its influence on bone health, may confer periodontal benefit. Methods We investigated cross-sectional associations (1997–2000) between plasma 25-hydroxyvitamin D concentrations [25(OH)D] and periodontal measure among 920 postmenopausal women. Chronic measures of disease were defined based on: 1) alveolar crestal height (ACH) measures from intraoral radiographs and tooth loss, and the 2) Center for Disease Control and Prevention (CDC)/American Academy of Periodontology (AAP) criteria using measures of clinical attachment level (CAL) and probing pocket depth (PD). Acute oral inflammation was assessed by the % of gingival sites that bled upon assessment with a probe. Logistic regression was used to estimate the odds ratios (OR) and 95% confidence intervals (CIs) for periodontal disease among participants with adequate ([25(OH)D]≥50 nmol/L) compared to deficient/inadequate ([25(OH)D]<50 nmol/L) vitamin D status adjusted for age, dental visit frequency, and body mass index. Results No association was observed between vitamin D status and periodontal disease defined by ACH and tooth loss (adjusted OR=0.96, 95% CI: 0.68–1.35). In contrast, women with adequate compared to deficient/inadequate vitamin D status had a 33% lower odds (95% CI: 5%–53%) of periodontal disease defined using the CDC/AAP definition and a 42% lower odds (95% CI: 21%-58%) of having ≥50% of gingival sites that bled. Conclusion Vitamin D status was inversely associated with gingival bleeding, an acute measure of oral health and inflammation and inversely associated with clinical categories of chronic periodontal disease that incorporated PD, an indicator of oral inflammation. However, vitamin D was not associated with chronic periodontal disease based on measures of ACH in combination with tooth loss.

It has been reported that levo-1-methyl tryptophan (L-1MT) can block indoleamine-2,3-dioxygenase (IDO) expressed by human dendritic cells (DC), whereas dextro-1-methyl tryptophan (D-1MT) is inefficient. However, whether L-1MT or D-1MT can efficiently reverse IDO-induced arrest of human T-cell proliferation has not been clarified. Here, we show a marked immunosuppressive effect of IDO derived from INDOtransfected 293 cell, IDO + ovarian cancer cells, and monocytederived DCs on CD4 + Th1 cells, CD8 + T cells, and natural killer cells derived from peripheral blood, ascites, and tumors of ovarian cancer patients. We found that, whereas L-1MT and D/L-1MT can restore proliferation of tumor-derived and peripheral blood T-cell subsets, D-1MT does not effectively restore IDO-induced arrest of T-cell proliferation. Although D-1MT inhibited kynurenine production at high concentrations, L-1MT was more effective in abrogating kynurenine generation and tryptophan depletion, whereas tryptophan was completely depleted by IDO even in the presence of high amounts of D-1MT. Together, the results indicate that, whereas the generation of tryptophan metabolites (kynurenines) by IDO is important in mediating suppression of T-cell proliferation, the degree to which tryptophan depletion is restored by 1MT is also critical in overcoming IDO-induced arrest of T-cell proliferation. [Cancer Res 2009;69(13):5498-504]

Multiplexing arrays increase the throughput and decrease sample requirements for studies employing multiple biomarkers. The goal of this project was to examine the performance of Multiplex arrays for measuring multiple protein biomarkers in saliva and serum. Specimens from the OsteoPerio ancillary study of the Women’s Health Initiative Observational Study were used. Participants required the presence of at least 6 teeth and were excluded based on active cancer and certain bone issues but were not selected on any specific condition. Quality control (QC) samples were created from pooled serum and saliva. Twenty protein markers were measured on five multiplexing array panels. Sample pretreatment conditions were optimized for each panel. Recovery, lower limit of quantification (LLOQ) and imprecision were determined for each analyte. Statistical adjustment at the plate level was used to reduce imprecision estimates and increase the number of usable observations. Sample pre-treatment improved recovery estimates for many analytes. The LLOQ for each analyte agreed with manufacturer specifications except for MMP-1 and MMP-2 which were significantly higher than reported. Following batch adjustment, 17 of 20 biomarkers in serum and 9 of 20 biomarkers in saliva demonstrated acceptable precision, defined as <20% coefficient of variation (<25% at LLOQ). The percentage of cohort samples having levels within the reportable range for each analyte varied from 10% to 100%. The ratio of levels in saliva to serum varied from 1∶100 to 28∶1. Correlations between saliva and serum were of moderate positive magnitude and significant for CRP, MMP-2, insulin, adiponectin, GM-CSF and IL-5. Multiplex arrays exhibit high levels of analytical imprecision, particularly at the batch level. Careful sample pre-treatment can enhance recovery and reduce imprecision. Following statistical adjustments to reduce batch effects, we identified biomarkers that are of acceptable quality in serum and to a lesser degree in saliva using Multiplex arrays.

Purpose Few prospective studies have reported on relationships between objective periodontal disease (PD) measures and cancer risk. This association was examined in 1,337 postmenopausal women participating in the Buffalo OsteoPerio Study. Methods Oral alveolar crestal bone height (ACH) was measured using oral radiographs. Incident cancers were adjudicated with medical records. Hazard ratios (HR) and 95% confidence intervals (CI) for associations between ACH and incident cancer outcomes were estimated using Cox proportional hazards models. Results There were 203 confirmed total incident cancer cases during follow-up (12.2±4.2 years). After adjusting for age and smoking, there were no statistically significant associations between ACH-defined PD categories and total cancer risk (mild/moderate vs. none: HR=1.33, 95%CI: 0.91–1.94; severe vs. none: HR=1.20, 95%CI: 0.77–1.86). ACH-defined PD categories were not associated with common site-specific cancers. Whole mouth mean and worst site ACH (per 1mm loss) were significantly associated with increased risk of lung (adjusted HR=1.81, 95% CI: 1.30–2.54; adjusted HR=1.34, 95% CI: 1.08–1.66, respectively), but not total or other site-specific cancer. Smoking status modified the associations between continuous ACH variables and total cancer risk; measures of PD were associated with total cancer among smokers but not never-smokers (interaction p=0.02 and p<0.01 for whole mouth mean and worst site ACH, respectively). Conclusions ACH-defined PD was associated with total cancer risk in ever but not never-smoking postmenopausal women. Whole mouth mean and worst site ACH were associated with increased lung cancer risk. However, these results need to be interpreted cautiously given the small number of lung cancer cases (n=18). Further research utilizing a larger sample is warranted to confirm the relationships among oral bone loss, site-specific cancers, and total cancer.