Immunological and Clinical Effects of Vaccines Targeting p53-Overexpressing Malignancies

Approximately 50% of human malignancies carry p53 mutations, which makes it a potential antigenic target for cancer immunotherapy. Adoptive transfer with p53-specific cytotoxic T-lymphocytes (CTL) and CD4+ T-helper cells eradicates p53-overexpressing tumors in mice. Furthermore, p53 antibodies and p53-specific CTLs can be detected in cancer patients, indicating that p53 is immunogenic. Based on these results, clinical trials were initiated. In this paper, we review immunological and clinical responses observed in cancer patients vaccinated with p53 targeting vaccines. In most trials, p53-specific vaccine-induced immunological responses were observed. Unfortunately, no clinical responses with significant reduction of tumor-burden have occurred. We will elaborate on possible explanations for this lack of clinical effectiveness. In the second part of this paper, we summarize several immunopotentiating combination strategies suitable for clinical use. In our opinion, future p53-vaccine studies should focus on addition of these immunopotentiating regimens to achieve clinically effective therapeutic vaccination strategies for cancer patients

ImmunoGlobulin galaxy (IGGalaxy) for simple determination and quantitation of immunoglobulin heavy chain rearrangements from NGS

Background: Sequence analysis of immunoglobulin heavy chain (IGH) gene rearrangements and frequency analysis is a powerful tool for studying the immune repertoire, immune responses and immune dysregulation in health and disease. The challenge is to provide user friendly, secure and reproducible analytical services that are available for both small and large laboratories which are determining VDJ repertoire using NGS technology. Results: In this study we describe ImmunoGlobulin Galaxy (IGGalaxy)- a convenient web based application for analyzing next-generation sequencing results and reporting IGH gene rearrangements for both repertoire and clonality studies. IGGalaxy has two analysis options one using the built in igBLAST algorithm and the second using output from IMGT; in either case repertoire summaries for the B-cell populations tested are available. IGGalaxy supports multi-sample and multi-replicate input analysis for both igBLAST and IMGT/HIGHV-QUEST. We demonstrate the technical validity of this platform using a standard dataset, S22, used for benchmarking the performance of antibody alignment utilities with a 99.9 % concordance with previous results. Re-analysis of NGS data from our samples of RAG-deficient patients demonstrated the validity and user friendliness of this tool. Conclusions: IGGalaxy provides clinical researchers with detailed insight into the repertoire of the B-cell population per individual sequenced and between control and pathogenic genomes. IGGalaxy was developed for 454 NGS results but is capable of analyzing alternative NGS data (e.g. Illumina, Ion Torrent). We demonstrate the use of a Galaxy virtual machine to determine the VDJ repertoire for reference data and from B-cells taken from immune deficient patients. IGGalaxy is available as a VM for download and use on a desktop PC or on a server

Asymptotic Conformal Yano--Killing Tensors for Schwarzschild Metric

The asymptotic conformal Yano--Killing tensor proposed in J. Jezierski, On the relation between metric and spin-2 formulation of linearized Einstein theory [GRG, in print (1994)] is analyzed for Schwarzschild metric and tensor equations defining this object are given. The result shows that the Schwarzschild metric (and other metrics which are asymptotically ``Schwarzschildean'' up to O(1/r^2) at spatial infinity) is among the metrics fullfilling stronger asymptotic conditions and supertranslations ambiguities disappear. It is also clear from the result that 14 asymptotic gravitational charges are well defined on the ``Schwarzschildean'' background.Comment: 8 pages, latex, no figure

Harmonization of the intracellular cytokine staining assay

Active immunotherapy for cancer is an accepted treatment modality aiming to reinforce the T-cell response to cancer. T-cell reactivity is measured by various assays and used to guide the clinical development of immunotherapeutics. However, data obtained across different institutions may vary substantially making comparative conclusions difficult. The Cancer Immunotherapy Immunoguiding Program organizes proficiency panels to identify key parameters influencing the outcome of commonly used T-cell assays followed by harmonization. Our successes with IFNγ-ELISPOT and peptide HLA multimer analysis have led to the current study on intracellular cytokine staining (ICS). We report the results of three successive panels evaluating this assay. At the beginning, 3 out of 9 participants (33 %) were able to detect >6 out of 8 known virus-specific T-cell responses in peripheral blood of healthy individuals. This increased to 50 % of the laboratories in the second phase. The reported percentages of cytokine-producing T cells by the different laboratories were highly variable with coefficients of variation well over 60 %. Variability could partially be explained by protocol-related differences in background cytokine production leading to sub-optimal signal-to-noise ratios. The large number of protocol variables prohibited identification of prime guidelines to harmonize the assays. In addition, the gating strategy used to identify reactive T cells had a major impact on assay outcome. Subsequent harmonization of the gating strategy considerably reduced the variability within the group of participants. In conclusion, we propose that first basic guidelines should be applied for gating in ICS experiments before harmonizing assay protocol variables

Effects of Multisensory Context on Tofu and Soy Sauce Evaluation and Consumption

We examined the effects of an informative pitch and multisensory contexts as potential factors influencing individuals’ experience of tofu with soy sauce and the amount consumed outside the lab. Two hundred and sixteen participants watched one of two pitches (promoting either vegetarian diets or exercise) and were guided into one of three multisensory contexts (‘sustainable’, ‘meat’, or ‘neutral’ theme). Participants rated the aroma and appearance of soy sauce and the taste of tofu dipped in it using the intuitive ‘one touch’ EmojiGrid valence and arousal measuring tool. Our results showed that the ‘meat’ context increased arousal ratings for soy sauce and the tendency to consume more tofu relative to the other contexts. Pitch did not influence affective ratings or amounts consumed. We conclude that the multisensory context has the potential to positively affect peoples’ choices and perceptions of plant-based and sustainable food and promote its consumption

Antibodies against p53 are associated with poor prognosis of colorectal cancer.

Mutation of the p53 gene is a common event in colorectal cancer. This alteration can result in cellular accumulation of p53 and may also induce p53 antibodies. Accumulation of p53 in tumour cells has been associated with poor prognosis of colorectal cancer. We tested preoperative sera from 255 patients with colorectal cancer by enzyme-linked immunosorbent assay (ELISA). A total of 70.2% had reactivity that was higher than the 'low' control serum. Employing a cut-off level of 10% of the 'high' control sample, 25.5% of the patients were positive for p53 antibodies. The presence of p53 antibodies correlated with the following prognostic factors: histological differentiation grade, shape of the tumour, and tumour invasion into blood vessels. Patients with p53 antibodies were shown to have decreased survival and decreased disease-free survival. Specifically for patients with cancer stage A and B1 the presence of p53 antibodies selected a subgroup with poor prognosis

Prospects for high-z cluster detections with Planck, based on a follow-up of 28 candidates using MegaCam@CFHT

The Planck catalogue of SZ sources limits itself to a significance threshold of 4.5 to ensure a low contamination rate by false cluster candidates. This means that only the most massive clusters at redshift z>0.5, and in particular z>0.7, are expected to enter into the catalogue, with a large number of systems in that redshift regime being expected around and just below that threshold. In this paper, we follow-up a sample of SZ sources from the Planck SZ catalogues from 2013 and 2015. In the latter maps, we consider detections around and at lower significance than the threshold adopted by the Planck Collaboration. To keep the contamination rate low, our 28 candidates are chosen to have significant WISE detections, in combination with non-detections in SDSS/DSS, which effectively selects galaxy cluster candidates at redshifts $z\gtrsim0.5$. By taking r- and z-band imaging with MegaCam@CFHT, we bridge the 4000A rest-frame break over a significant redshift range, thus allowing accurate redshift estimates of red-sequence cluster galaxies up to z~0.8. After discussing the possibility that an overdensity of galaxies coincides -by chance- with a Planck SZ detection, we confirm that 16 of the candidates have likely optical counterparts to their SZ signals, 13 (6) of which have an estimated redshift z>0.5 (z>0.7). The richnesses of these systems are generally lower than expected given the halo masses estimated from the Planck maps. However, when we follow a simplistic model to correct for Eddington bias in the SZ halo mass proxy, the richnesses are consistent with a reference mass-richness relation established for clusters detected at higher significance. This illustrates the benefit of an optical follow-up, not only to obtain redshift estimates, but also to provide an independent mass proxy that is not based on the same data the clusters are detected with, and thus not subject to Eddington bias.Comment: 13 pages, 7 figures. Accepted for publication in A&

Extended weekly dose-dense paclitaxel/carboplatin is feasible and active in heavily pre-treated platinum-resistant recurrent ovarian cancer

There is increasing evidence of the efficacy of dose-dense therapy in the management of platinum-resistant/refractory ovarian cancer. We report our experience of extended weekly carboplatin and paclitaxel in this population group. Twenty patients with platinum-resistant/refractory ovarian cancer received carboplatin AUC 3 and paclitaxel 70 mg m−2 on day 1, 8, 15 q 4 weekly for six planned cycles. Toxicity was assessed using Common Toxicity Criteria. Response was evaluated using radiological and CA125 criteria. Median age was 61 years (range 40–74 years). Median number of prior therapies is three (range 1–8). Response rate was 60% by radiological criteria (RECIST) and 76% by CA125 assessment. Grade 3 toxicities consisted of neutropenia (29% of patients) and anaemia (5%). One patient experienced grade 4 neutropenia. No grade 3/4 thombocytopaenia was reported. Fatigue, nausea and peripheral neuropathy were the most frequent non-hematological side effects. Median progression-free survival was 7.9 months and overall survival was 13.3 months. The dynamics of response to dose-dense therapy were as rapid as with front-line therapy within the same patient. This dose-dense regimen can be extended to at least 18 weekly cycles over 6 months and is well tolerated with high response rates in heavily pre-treated, platinum-resistant ovarian cancer. It forms a highly active and tolerable cytotoxic scaffold to which molecular-targeted therapies can be added in platinum-resistant ovarian cancer