The immediate-early protein 1 of human herpesvirus 6B interacts with NBS1 and inhibits ATM signaling.

Viral infection often trigger an ATM serine/threonine kinase (ATM)-dependent DNA damage response in host cells that suppresses viral replication. Viruses evolved different strategies to counteract this antiviral surveillance system. Here, we report that human herpesvirus 6B (HHV-6B) infection causes genomic instability by suppressing ATM signaling in host cells. Expression of immediate-early protein 1 (IE1) phenocopies this phenotype and blocks homology-directed double-strand break repair. Mechanistically, IE1 interacts with NBS1, and inhibits ATM signaling through two distinct domains. HHV-6B seems to efficiently inhibit ATM signaling as further depletion of either NBS1 or ATM do not significantly boost viral replication in infected cells. Interestingly, viral integration of HHV-6B into the host's telomeres is not strictly dependent on NBS1, challenging current models where integration occurs through homology-directed repair. Given that spontaneous IE1 expression has been detected in cells of subjects with inherited chromosomally-integrated form of HHV-6B (iciHHV-6B), a condition associated with several health conditions, our results raise the possibility of a link between genomic instability and the development of iciHHV-6-associated diseases.

Inhibition of the CCR6-CCL20 axis prevents regulatory T cell recruitment and sensitizes head and neck squamous cell carcinoma to radiation therapy.

BACKGROUND:  Radioresistance of HNSCCs remains a major challenge for effective tumor control. Combined radiotherapy (RT) and immunotherapy (IT) treatment improved survival for a subset of patients with inflamed tumors or tumors susceptible to RT-induced inflammation. To overcome radioresistance and improve treatment outcomes, an understanding of factors that suppress anti-tumor immunity is necessary. In this regard, regulatory T cells (Tregs) are critical mediators of immune suppression in HNSCCs. In this study, we investigated how radiation modulates Treg infiltration in tumors through the chemokine CCL20. We hypothesized that radiation induces CCL20 secretion resulting in Treg infiltration and suppression of anti-tumor immunity. METHODS:  Human and mouse HNSCC cell lines with different immune phenotypes were irradiated at doses of 2 or 10 Gy. Conditioned media, RNA and protein were collected for assessment of CCL20. qPCR was used to determine CCL20 gene expression. In vivo, MOC2 cells were implanted into the buccal cavity of mice and the effect of neutralizing CCL20 antibody was determined alone and in combination with RT. Blood samples were collected before and after RT for analysis of CCL20. Tumor samples were analyzed by flow cytometry to determine immune infiltrates, including CD8 T cells and Tregs. Mass-spectrometry was performed to analyze proteomic changes in the tumor microenvironment after anti-CCL20 treatment. RESULTS:  Cal27 and MOC2 HNSCCs had a gene signature associated with Treg infiltration, whereas SCC9 and MOC1 tumors displayed a gene signature associated with an inflamed TME. In vitro, tumor irradiation at 10 Gy significantly induced CCL20 in Cal27 and MOC2 cells relative to control. The increase in CCL20 was associated with increased Treg migration. Neutralization of CCL20 reversed radiation-induced migration of Treg cells in vitro and decreased intratumoral Tregs in vivo. Furthermore, inhibition of CCL20 resulted in a significant decrease in tumor growth compared to control in MOC2 tumors. This effect was further enhanced after combination with RT compared to either treatment alone. CONCLUSION:  Our results suggest that radiation promotes CCL20 secretion by tumor cells which is responsible for the attraction of Tregs. Inhibition of the CCR6-CCL20 axis prevents infiltration of Tregs in tumors and suppresses tumor growth resulting in improved response to radiation.

Accurate calibration of a polymer gel dosimeter with a plastic scintillation detector.

PURPOSE: Three dimensional dose polymer gel dosimetry measurements provide unique information on sophisticated dose distributions. In this study, the authors propose a novel method to improve the accuracy of polymer gel dosimeters by inserting a plastic scintillation detector (PSD) to provide a dose reference. METHODS: PSD dosimeters were calibrated using chromatic deconvolution and then inserted into polyacrylanide gel (PAG) dosimeters. The gel and the PSDs were immersed into water and irradiated with 6 MV wedge filtered beams to obtain a wide range of dose variation. Calibration vials containing the same gel were also irradiated to generate a standard calibration curve. The distribution of magnetic nuclear transverse relaxation rate (R2) values of the gel was determined with a multislice multiecho MRI sequence at 1.5 T. Another calibration curve was obtained by assigning the R2 values in the gel surrounding the scintillators to the dose determined by the PSDs. A reference calibration point from a PSD located in a low dose gradient area served to correct the standard calibration method yielding three novel calibration methods. The results were compared with EBT2 GAFCHROMIC film measurements acquired in the same condition and with the Pinnacle3 treatment planning dose calculations, RESULTS: The mean absolute error of the standard calibration method ranged from 6.1 to 12.4%. The corresponding gamma index (3%/3 mm distance to agreement) criterion was satisfied for only 56% of the pixels in the middle slice of the gel compared to Pinnacle3 dose calculations and to EBT2 film measurements in the center part of the field. Calibration methods using a PSD reduced the mean absolute error to less than 4%; this value was under 2.6% for one of the three methods. In that case, 98% of the pixels satisfied the gamma index criterion. CONCLUSIONS: The accuracy of PAG dosimeters may be highly improved using one reference dose point measurement using a plastic scintillation detector. The best calibration procedure corrected the slope of the calibration curve derived from the calibration vials to match the R2 value around a PSD calibration, while keeping the R2 value at 0 Gy constant.

Octree indexing of DICOM images for voxel number reduction and improvement of Monte Carlo simulation computing efficiency.

The purpose of the present study is to introduce a compression algorithm for the CT (computed tomography) data used in Monte Carlo simulations. Performing simulations on the CT data implies large computational costs as well as large memory requirements since the number of voxels in such data reaches typically into hundreds of millions voxels. CT data, however, contain homogeneous regions which could be regrouped to form larger voxels without affecting the simulation's accuracy. Based on this property we propose a compression algorithm based on octrees: in homogeneous regions the algorithm replaces groups of voxels with a smaller number of larger voxels. This reduces the number of voxels while keeping the critical high-density gradient area. Results obtained using the present algorithm on both phantom and clinical data show that compression rates up to 75% are possible without losing the dosimetric accuracy of the simulation.