ABSTRACT
Today, there is greater awareness on the association between oral diseases and respiration diseases after the outbreak of COVID-19. However, confusion regarding the oral health management and medical risk prevention for patients with chronic airway diseases has been remained among dental clinicians. Therefore, the dental experts of the Fifth General Dentistry Special Committee, Chinese Stomatological Association, combined with the experts of respiratory and critical care medicine, undertook the formation of consensus on the oral health management of patients with chronic airway diseases in order to help dental clinicians to evaluate medical risks and make better treatment decision in clinical practice. In the present consensus report, the relationship of oral diseases and chronic airway diseases, the oral health management and the treatment recommendations of patients with chronic airway diseases are provided.
Subject(s)
Humans , COVID-19 , Consensus , Oral Health , Oral MedicineABSTRACT
<p><b>OBJECTIVE</b>To observe the functional changes of dendritic cells (DCs) infected in vitro by 3 recombinant adenoviruses encoding Her2/neu extracellular first-receptor domain (Her2-ECDs), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM) proteins (rAdHer2-ECDs, rAdHer2-ECD and rAdHer2-TM, respectively).</p><p><b>METHODS</b>The expressions of the target proteins were detected with Western blotting. The level of both interleukin (IL)-12 in the supernatant of in vitro cultured DCs infected with recombined adenoviruses and interferon gamma (IFN-gamma) in the supernatant of the lymphocyte populations co-cultured with DCs were determined by enzyme-linked immunosorbent assay (ELISA). The capacity of the DCs to stimulate allogeneic T lymphocyte proliferation was assessed by mixed lymphocyte reaction, and the activity of cellular toxic T lymphocytes (CTL) were investigated by MTT assay.</p><p><b>RESULTS</b>Her2-ECDs, ECD and TM proteins were detected in the transfected DCs. Compared with the untransfected DCs, more abundant IL-12 production was detected in the supernatant of the DCs 5 days after transfection, but the IL-12 level showed no significant difference between the DCs infected with the 3 recombinant adenoviruses. IFN-gamma production increased gradually with passage of the time following DC-stimulated lymphocyte proliferation irrespective of infection of the DCs, and only the DCs infected with rAdHer2-TM seemed to result in significant difference in DC-mediated allogeneic T lymphocyte proliferation. The killing of breast cancer cell line with Her2 overexpression was more efficient with infected DCs priming autologous T lymphocyte to generate CTL than with uninfected DCs and those modified by SK-OV-3 cell fragment. CTL activity induced by rAdHer2-TM-infected DCs was the strongest, and breast cancer cell-killing activity was more efficient against cell line with Her2/neu-overexpression.</p><p><b>CONCLUSION</b>The DCs infected with the recombinant adenovirus encoding Her2/neu extracellular and transmembrane domains show enhanced anti-tumor effect and induce Her2/neu-specific CTL activity.</p>
Subject(s)
Female , Humans , Adenoviridae , Genetics , Blotting, Western , Breast Neoplasms , Genetics , Allergy and Immunology , Pathology , Cell Line, Tumor , Cell Proliferation , Cell Survival , Allergy and Immunology , Coculture Techniques , Cytotoxicity, Immunologic , Allergy and Immunology , Dendritic Cells , Cell Biology , Allergy and Immunology , Metabolism , Enzyme-Linked Immunosorbent Assay , Extracellular Matrix Proteins , Genetics , Metabolism , Interferon-gamma , Metabolism , Interleukin-12 , Metabolism , Lymphocyte Culture Test, Mixed , Lymphocytes , Cell Biology , Allergy and Immunology , Membrane Proteins , Genetics , Metabolism , Ovarian Neoplasms , Genetics , Allergy and Immunology , Pathology , Receptor, ErbB-2 , Genetics , Metabolism , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Metabolism , T-Lymphocytes, Cytotoxic , Cell Biology , Allergy and Immunology , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To construct eukaryotic expression vectors using recombinant adenovirus containing the gene fragments encoding Her2/neu extracellular first ligand-binding domain (Her2-ECD), full-length extracellular domain (Her2-ECD), and extracellular and transmembrane domain (Her2-TM).</p><p><b>METHODS</b>The cDNAs were amplified by RT-PCR and inserted into shuttle pAdTrack-CMV plasmids. Viral plasmids were obtained from homologous recombination in E. coli BJ5183, and transfected into 293 cells via liposome. Formation of viral plaque and expression of green fluorescent protein were observed by fluorescence microscopy, and the target proteins were detected by Western blotting.</p><p><b>RESULTS</b>The target cDNA fragments were amplified by PCR with expected lengths and the DNA sequences were confirmed against Genbank. Formation of viral plaque, expression of green fluorescent protein and the target proteins were detected in 293 cells transfected by the viral plasmids, which showed elevated expression of Her2/neu protein with the increase of multiplicity of infection (MOI).</p><p><b>CONCLUSION</b>The eukaryotic expression vectors using recombinant adenovirus have been successfully constructed for expression of Her2/neu extracellular and transmembrane domains.</p>