Chimeric Rabies Virus-Like Particles Containing Membrane-Anchored GM-CSF Enhances the Immune Response against Rabies Virus

Rabies remains an important public health threat in most developing countries. To develop a more effective and safe vaccine against rabies, we have constructed a chimeric rabies virus-like particle (VLP), which containing glycoprotein (G) and matrix protein (M) of rabies virus (RABV) Evelyn-Rokitnicki-Abelseth (ERA) strain, and membrane-anchored granulocyte-macrophage colony-stimulating factor (GM-CSF), and it was named of EVLP-G. The immunogenicity and protective efficacy of EVLP-G against RABV were evaluated by intramuscular administration in a mouse model. The EVLP-G was successfully produced in insect cells by coinfection with three recombinant baculoviruses expressing G, M, and GM-CSF, respectively. The membrane-anchored GM-CSF possesses a strong adjuvant activity. More B cells and dendritic cells (DCs) were recruited and/or activated in inguinal lymph nodes in mice immunized with EVLP-G. EVLP-G was found to induce a significantly increased RABV-specific virus-neutralizing antibody and elicit a larger and broader antibody subclass responses compared with the standard rabies VLP (sRVLP, consisting of G and M). The EVLP-G also elicited significantly more IFN-γ- or IL-4-secreting CD4+ and CD8+ T cells than the sRVLP. Moreover, the immune responses induced by EVLP-G protect all vaccinated mice from lethal challenge with RABV. These results suggest that EVLP-G has the potential to be developed as a novel vaccine candidate for the prevention and control of animal rabies

Chimeric Rabies Virus-Like Particles Containing Membrane-Anchored GM-CSF Enhances the Immune Response against Rabies Virus

Rabies remains an important public health threat in most developing countries. To develop a more effective and safe vaccine against rabies, we have constructed a chimeric rabies virus-like particle (VLP), which containing glycoprotein (G) and matrix protein (M) of rabies virus (RABV) Evelyn-Rokitnicki-Abelseth (ERA) strain, and membrane-anchored granulocyte-macrophage colony-stimulating factor (GM-CSF), and it was named of EVLP-G. The immunogenicity and protective efficacy of EVLP-G against RABV were evaluated by intramuscular administration in a mouse model. The EVLP-G was successfully produced in insect cells by coinfection with three recombinant baculoviruses expressing G, M, and GM-CSF, respectively. The membrane-anchored GM-CSF possesses a strong adjuvant activity. More B cells and dendritic cells (DCs) were recruited and/or activated in inguinal lymph nodes in mice immunized with EVLP-G. EVLP-G was found to induce a significantly increased RABV-specific virus-neutralizing antibody and elicit a larger and broader antibody subclass responses compared with the standard rabies VLP (sRVLP, consisting of G and M). The EVLP-G also elicited significantly more IFN-γ- or IL-4-secreting CD4+ and CD8+ T cells than the sRVLP. Moreover, the immune responses induced by EVLP-G protect all vaccinated mice from lethal challenge with RABV. These results suggest that EVLP-G has the potential to be developed as a novel vaccine candidate for the prevention and control of animal rabies

Multi-Sequence Fusion Network via Single- Sequence CycleGANs for Improved Synthetic CT in Nasopharyngeal Carcinoma Treatment Planning

To investigate the effect of different MR sequences on the accuracy of Cycle-consistent Generative Adversarial Network (CycleGAN) based synthetic CT (sCT) generation in nasopharyngeal carcinoma (NPC). In this work, three sequences of MR, included T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), and T1 contrast-enhanced weighted imaging (T1WIC), and planning CT (PCT) images of 151 patients with NPC were collected. The number of training, verification, and test sets were 108, 16, and 27, respectively. Three unsupervised CycleGAN-based models were trained using different sequences (single channels) as inputs, and the synergistic fusion model were used multiple channels. To assess the precision of these models, we evaluated key metrics such as mean error (ME), mean absolute error (MAE), structural similarity index (SSIM), peak signal-to-noise ratio (PSNR), and dose distribution, comparing the PCT with the sCT generated by each model. Overall, The SCTT2 image generated by T2WI model achieved superior results than those of T1WIC and T1WI for a single-sequence model. Compared to the SCTT2, the quality of the SCTFusion image generated by the fusion model was further improved, with significant differences in the MAE, SSIM, and PNSR metrics (P<0.05), the average MAE, SSIM, and PNSR in the body were ( $112.97\pm 9.69$ vs. $90.99\pm 9.64$ , P<0.05) HU, ( $0.86\pm 0.02$ vs. $0.89\pm 0.02$ , P<0.05), and ( $21.66\pm 0.64$ vs. $23.28\pm 0.87$ , P<0.05), respectively. Meanwhile, the average gamma passing rates (3%, 3 mm) and the average absolute dose discrepancies were 99.45% $\pm ~1.05$ % and 0.73%±0.73% for the fusion model. In conclusion, our findings reveal that the CycleGAN model, particularly when employing fusion MR sequences as input, offers the highest accuracy in synthetic CT generation. Notably, T2WI images stand out as a viable option for sCT prediction in clinical settings where acquisition sequences or times are limited

Vitamin D Receptor Negatively Regulates Bacterial-Stimulated NF-κB Activity in Intestine

Vitamin D receptor (VDR) plays an essential role in gastrointestinal inflammation. Most investigations have focused on the immune response; however, how bacteria regulate VDR and how VDR modulates the nuclear factor (NF)-κB pathway in intestinal epithelial cells remain unexplored. This study investigated the effects of VDR ablation on NF-κB activation in intestinal epithelia and the role of enteric bacteria on VDR expression. We found that VDR−/− mice exhibited a pro-inflammatory bias. After Salmonella infection, VDR−/− mice had increased bacterial burden and mortality. Serum interleukin-6 in noninfected VDR+/+ mice was undetectable, but was easily detectable in VDR−/− mice. NF-κB p65 formed a complex with VDR in noninfected wild-type mouse intestine. In contrast, deletion of VDR abolished VDR/P65 binding. P65 nuclear translocation occurred in colonic epithelial cells of untreated VDR−/− mice. VDR deletion also elevated NF-κB activity in intestinal epithelia. VDR was localized to the surface epithelia of germ-free mice, but to crypt epithelial cells in conventionalized mice. VDR expression, distribution, transcriptional activity, and target genes were regulated by Salmonella stimulation, independent of 1,25-dihydroxyvitamin D3. Our study demonstrates that commensal and pathogenic bacteria directly regulate colonic epithelial VDR expression and location in vivo. VDR negatively regulates bacterial-induced intestinal NF-κB activation and attenuates response to infection. Therefore, VDR is an important contributor to intestinal homeostasis and host protection from bacterial invasion and infection