FOSL1 Inhibits Type I Interferon Responses to Malaria and Viral Infections by Blocking TBK1 and TRAF3/TRIF Interactions

Innate immune response plays a critical role in controlling invading pathogens, but such an immune response must be tightly regulated. Insufficient or overactivated immune responses may lead to harmful or even fatal consequences. To dissect the complex host-parasite interactions and the molecular mechanisms underlying innate immune responses to infections, here we investigate the role of FOS-like antigen 1 (FOSL1) in regulating the host type I interferon (IFN-I) response to malaria parasite and viral infections. FOSL1 is known as a component of a transcription factor but was recently implicated in regulating the IFN-I response to malaria parasite infection. Here we show that FOSL1 can act as a negative regulator of IFN-I signaling. Upon stimulation with poly(I:C), malaria parasite-infected red blood cells (iRBCs), or vesicular stomatitis virus (VSV), FOSL1 “translocated” from the nucleus to the cytoplasm, where it inhibited the interactions between TNF receptor-associated factor 3 (TRAF3), TIR domain-containing adapter inducing IFN-β (TRIF), and Tank-binding kinase 1 (TBK1) via impairing K63-linked polyubiquitination of TRAF3 and TRIF. Importantly, FOSL1 knockout chimeric mice had lower levels of malaria parasitemia or VSV titers in peripheral blood and decreased mortality compared with wild-type (WT) mice. Thus, our findings have identified a new role for FOSL1 in negatively regulating the host IFN-I response to malaria and viral infections and have identified a potential drug target for controlling malaria and other diseases

Whole Genome PCR Scanning Reveals the Syntenic Genome Structure of Toxigenic Vibrio cholerae Strains in the O1/O139 Population

Vibrio cholerae is commonly found in estuarine water systems. Toxigenic O1 and O139 V. cholerae strains have caused cholera epidemics and pandemics, whereas the nontoxigenic strains within these serogroups only occasionally lead to disease. To understand the differences in the genome and clonality between the toxigenic and nontoxigenic strains of V. cholerae serogroups O1 and O139, we employed a whole genome PCR scanning (WGPScanning) method, an rrn operon-mediated fragment rearrangement analysis and comparative genomic hybridization (CGH) to analyze the genome structure of different strains. WGPScanning in conjunction with CGH revealed that the genomic contents of the toxigenic strains were conservative, except for a few indels located mainly in mobile elements. Minor nucleotide variation in orthologous genes appeared to be the major difference between the toxigenic strains. rrn operon-mediated rearrangements were infrequent in El Tor toxigenic strains tested using I-CeuI digested pulsed-field gel electrophoresis (PFGE) analysis and PCR analysis based on flanking sequence of rrn operons. Using these methods, we found that the genomic structures of toxigenic El Tor and O139 strains were syntenic. The nontoxigenic strains exhibited more extensive sequence variations, but toxin coregulated pilus positive (TCP+) strains had a similar structure. TCP+ nontoxigenic strains could be subdivided into multiple lineages according to the TCP type, suggesting the existence of complex intermediates in the evolution of toxigenic strains. The data indicate that toxigenic O1 El Tor and O139 strains were derived from a single lineage of intermediates from complex clones in the environment. The nontoxigenic strains with non-El Tor type TCP may yet evolve into new epidemic clones after attaining toxigenic attributes

Semiautomatic nonrigid registration for the prostate and pelvic MR volumes

Three-dimensional (3D) nonrigid image registration for potential applications in prostate cancer treatment and interventional magnetic resonance (iMRI) imaging-guided therapies were investigated. An almost fully automated 3D nonrigid registration algorithm using mutual information and a thin plate spline (TPS) transformation for MR images of the prostate and pelvis were created and evaluated. In the first step, an automatic rigid body registration with special features was used to capture the global transformation. In the second step, local feature points (FPs) were registered using mutual information. An operator entered only five FPs located at the prostate center, left and right hip joints, and left and right distal femurs. The program automatically determined and optimized other FPs at the external pelvic skin surface and along the femurs. More than 600 control points were used to establish a TPS transformation for deformation of the pelvic region and prostate. Ten volume pairs were acquired from three volunteers in the diagnostic (supine) and treatment positions (supine with legs raised). Various visualization techniques showed that warping rectified the significant pelvic misalignment by the rigid-body method. Gray-value measures of registration quality, including mutual information, correlation coefficient, and intensity difference, all improved with warping. The distance between prostate 3D centroids was 0.7 +/- 0.2 mm after warping compared with 4.9 +/- 3.4 mm with rigid-body registration. Semiautomatic nonrigid registration works better than rigid-body registration when patient position is changed greatly between acquisitions. It could be a useful tool for many applications in the management of prostate

High-field magnetic resonance imaging of the response of human prostate cancer to Pc 4-based photodynamic therapy in an animal model

INTRODUCTION: High-field magnetic resonance imaging (MRI) is an emerging technique that provides a powerful, non-invasive tool for in vivo studies of cancer therapy in animal models. Photodynamic therapy (PDT) is a relatively new treatment modality for prostate cancer, the second leading cause of cancer mortality in American males. The goal of this study was to evaluate the response of human prostate tumor cells growing as xenografts in athymic nude mice to Pc 4-sensitized PDT. MATERIALS AND METHODS: PC-3, a cell line derived from a human prostate malignant tumor, was injected intradermally on the back flanks of athymic nude mice. Two tumors were initiated on each mouse. One was treated and the other served as the control. A second-generation photosensitizing drug Pc 4 (0.6 mg/kg body weight) was delivered to each animal by tail vein injection 48 hours before laser illumination (672 nm, 100 mW/cm(2), 150 J/cm(2)). A dedicated high-field (9.4 T) small-animal MR scanner was used for image acquisitions. A multi-slice multi-echo (MSME) technique, permitting noninvasive in vivo assessment of potential therapeutic effects, was used to measure the T2 values and tumor volumes. Animals were scanned immediately before and after PDT and 24 hours after PDT. T2 values were computed and analyzed for the tumor regions. RESULTS: For the treated tumors, the T2 values significantly increased (P < 0.002) 24 hours after PDT (68.2 ± 8.5 milliseconds), compared to the pre-PDT values (55.8 ± 6.6 milliseconds). For the control tumors, there was no significant difference (P = 0.53) between the pre-PDT (52.5 ± 6.1 milliseconds) and 24-hour post-PDT (54.3 ± 6.4 milliseconds) values. Histologic analysis showed that PDT-treated tumors demonstrated necrosis and inflammation that was not seen in the control. DISCUSSION: Changes in tumor T2 values measured by multi-slice multi-echo MR imaging provide an assay that could be useful for clinical monitoring of photodynamic therapy of prostate tumors

Registration of micro-PET and high-resolution MR images of mice for monitoring photodynamic therapy

We are investigating imaging techniques to study the rapid biochemical and physiological response of tumors to photodynamic therapy (PDT). Positron emission tomography (PET) can provide physiological and functional images of cancers. While MRI can provide high resolution anatomical images and generate serial, noninvasive, in vivo observations of morphological changes. In this study, we investigate image registration methods to combine MRI and micro-PET ( PET) images for improved tumor monitoring. We acquired high resolution MR and PET F-fluorodeoxyglucose (FDG) images from mice with RIF-1 tumors. We used rigid body registration with three translations and three angular variables. We used normalized mutual information as the similarity measure. To assess the quality of registration, we performed slice by slice review of both image volumes, manually segmented feature organs such as the left and right kidneys and the bladder in each slice, and computed the distance between corresponding centroids of the organs. We also used visual inspection techniques such as color overlay displays. Over 40 volume registration experiments were performed with MR and PET images acquired from three C3H mice. The color overlays showed that the MR images and the PET images matched well. The distance between corresponding centroids of organs was 1.5 ± 0.4 mm which is about 2 pixels of PET. In conclusion, registration of high resolution MR and PET images of mice may be useful to combine anatomical and functional information that could be used for the potential application in photodynamic therapy

Deformable and rigid registration of MRI and microPET images for photodynamic therapy of cancer in mice

We are investigating imaging techniques to study the tumor response to photodynamic therapy (PDT). Positron emission tomography (PET) can provide physiological and functional information. High-resolution magnetic resonance imaging (MRI) can provide anatomical and morphological changes. Image registration can combine MRI and PET images for improved tumor monitoring. In this study, we acquired high-resolution MRI and microPET F 18 -fluorodeoxyglucose (FDG) images from C3H mice with RIF-1 tumors that were treated with Pc 4-based PDT. We developed two registration methods for this application. For registration of the whole mouse body, we used an automatic three-dimensional, normalized mutual information algorithm. For tumor registration, we developed a finite element model (FEM)-based deformable registration scheme. To assess the quality of whole body registration, we performed slice-by-slice review of both image volumes; manually segmented feature organs, such as the left and right kidneys and the bladder, in each slice; and computed the distance between corresponding centroids. Over 40 volume registration experiments were performed with MRI and microPET images. The distance between corresponding centroids of organs was 1.5 ± 0.4 mm which is about 2 pixels of microPET images. The mean volume overlap ratios for tumors were 94.7% and 86.3% for the deformable and rigid registration methods, respectively. Registration of high-resolution MRI and microPET images combines anatomical and functional information of the tumors and provides a useful tool for evaluating photodynamic therapy

Finite element model-based tumor registration of microPET and high-resolution MR images for photodynamic therapy in mice

We are investigating imaging techniques to study the tumor response to photodynamic therapy (PDT). PET can provide physiological and functional information. High-resolution MRI can provide anatomical and morphological changes. Image registration can combine MRI and PET images for improved tumor monitoring. In this study, we acquired high-resolution MRI and microPET [ F]fluorodeoxyglucose (FDG) images from C3H mice with RIF-1 tumors that were treated with Pc 4-based PDT. For tumor registration, we developed a finite element model (FEM)-based deformable registration scheme. To assess the registration quality, we performed slice by slice review of both image volumes, computed the volume overlap ratios, and visualized both volumes in color overlay. The mean volume overlap ratios for tumors were 94.7% after registration. Registration of high-resolution MRI and microPET images combines anatomical and functional information of the tumors and provides a useful tool for evaluating photodynamic therapy