Inactivation of HIV-1 in Polarized Infant Tonsil Epithelial Cells by Human Beta-Defensins 2 and 3 Tagged with the Protein Transduction Domain of HIV-1 Tat

Mother-to-child transmission (MTCT) of HIV-1 may occur during pregnancy, labor, and breastfeeding; however, the molecular mechanism of MTCT of virus remains poorly understood. Infant tonsil mucosal epithelium may sequester HIV-1, serving as a transient reservoir, and may play a critical role in MTCT. Innate immune proteins human beta-defensins 2 (hBD-2) and -3 may inactivate intravesicular virions. To establish delivery of hBD-2 and -3 into vesicles containing HIV-1, we tagged hBDs with the protein transduction domain (PTD) of HIV-1 Tat, which facilitates an efficient translocation of proteins across cell membranes. Our new findings showed that hBD-2 and -3 proteins tagged with PTD efficiently penetrated polarized tonsil epithelial cells by endocytosis and direct penetration. PTD-initiated internalization of hBD-2 and -3 proteins into epithelial cells led to their subsequent penetration of multivesicular bodies (MVB) and vacuoles containing HIV-1. Furthermore, PTD played a role in the fusion of vesicles containing HIV-1 with lysosomes, where virus was inactivated. PTD-initiated internalization of hBD-2 and -3 proteins into ex vivo tonsil tissue explants reduced the spread of virus from epithelial cells to CD4+ T lymphocytes, CD68+ macrophages, and CD1c+ dendritic cells, suggesting that this approach may serve as an antiviral strategy for inactivating intraepithelial HIV-1 and reducing viral MTCT.

Immunodiagnostics, immunotherapy and immunoprophylaxis in otorhinolaryngological practice. Report 3. Diagnostic approaches to the palatine tonsils functional state assessment for selecting a treatment strategy for patients with chronic tonsillitis (analytical summary)

Experimental work carried out over the past 20 years to study the activity of various functional groups in tonsillar cells in patients with chronic tonsillitis (CT) under local exposure to various immune response modifiers showed that: - tonsillar cells from CT patients in 69.5% of cases are capable of increasing their immunofunctional activity under the influence of immune modifiers in vitro reactions; - the most active of the group of immunotropic immune response modifiers are T-activin and Thymogen; - it can be considered the optimal diagnostic technique to carry out a "stress" test on the tonsil tissue using nonspecific (alternating magnetic field of industrial (low) frequency and low-frequency ultrasound) and specific — a microbial vaccine when applied to the tonsil tissue; - a non-invasive method for recording changes in the tonsils after stimulation may be fluctuations in the level of secretory immunoglobulin A and α-interferon in saliva after the test; - an increase in the level of secretory immunoglobulin A and α-interferon by more than 1/3 of the initial level after the test is a clinically favorable sign and a recommendation for medical treatment to the managing of patients with CT.

COVID-19 infection in the palatine tonsil tissue and detritus: the detection of the virus compartment with RT-PCR

Two patients suffering from chronic recurrent tonsillitis were reported. The first patient was confirmed infected with COVID-19, 3 weeks prior to tonsillectomy. The detritus and tonsil specimen were further analysed through real-time PCR (RT-PCR) and revealed amplification of the fragment N and ORF1ab genes of SARS-CoV-2. The second patient had a negative IgM and positive IgG antibody for COVID-19; however, the nasopharyngeal swab indicated negative for SARS-CoV-2. Tonsillectomy was performed 2 weeks after the swab; the tonsil specimen was analysed through RT-PCR and revealed amplification of the N2 and RdRp gene of SARS-CoV-2. According to both results, the presence of the SARS-CoV-2 gene remains to be detected in tonsil and/or detritus after 2–3 weeks after recovery. Hence, it is suggested that it is necessary to use adequate protection when performing tonsillectomy on early recovered patients with COVID-19. Furthermore, tonsillectomy would be more advisable to be performed after the fourth week after recovery from COVID-19.

Overview of Helicobacter pylori Detection Using Rapid Urease Test and Giemsa Modification Staining in Chronic Tonsillitis Patients

BACKGROUND: Helicobacter pylori (H. pylori) is a Gram-negative bacteria and has been known for its role in causing gastric infection aused diseases such as gastric ulcer. H. pylori also implied to play a role in chronic tonsillitis, but this theory remains controversial. Many researches have different and contradictory results due to difficulty to accurately detect H. pylori in tonsillar tissue. There is still no appropriate method that able to detect H. pylori in tonsil tissue. AIM: The aim of the study was to detect H. pylori colonization in chronic tonsillitis and understand some of the methods of examination that can be done to detect H. pylori in tonsillar tissue. METHODS: This study is a descriptive study conducted on 25 respondents. Each sample was taken from patients with chronic tonsillitis who underwent tonsillectomy. Then, the rapid urease test (RUT) and the Giemsa modification staining were carried out to determine the presence of H. pylori. RESULTS: There were 19 people (76%) positive and 6 people (24%) negative for H. pylori using RUT. On examination with Giemsa modification staining obtained 19 people (76%) positive and 6 people (24%) negative for H. pylori. CONCLUSION: H. pylori can be found in most of chronic tonsillitis. Combination RUT and Giemsa modification staining examination can be a good option in detecting H. pylori in chronic tonsillitis.

Generation of tonsil organoids as an ex vivo model for SARS-CoV-2 infection

SUMMARYPalatine tonsil (hereinafter referred to as “tonsil”) plays role in the immune system’s first line of defense against foreign pathogens. Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a worldwide pandemic since the infection was first reported in China in December 2019. The aim of this study was to establish tonsil epithelial cell-derived organoids and to examine their feasibility as an ex vivo model for SARS-CoV-2 infection. Using an optimized protocol, we achieved 3D tonsil organoid culture from human tonsil tissue that reflects the distinctive characteristics of the tonsil epithelium, such as its cellular composition, histologic properties, and molecular biological features. Notably, we verified that SARS-CoV-2 can infect tonsil organoids with a robust replication efficiency. Furthermore, treatment with remdesivir, an antiviral agent, effectively protected them from viral infection. Therefore, tonsil organoids could be available for investigation of SARS-CoV-2 infection-mediated pathology and for preclinical screening of novel antiviral drug candidates.One-sentence SummaryThis study established tonsil epithelial cell-derived organoids and demonstrated their feasibility as an ex vivo model for SARS-CoV-2 infection.

Rhinovirus species and tonsillar immune responses

Background Rhinovirus A and C infections are important contributors to asthma induction and exacerbations. No data exist on the interaction of local immune responses in rhinovirus infection. Therefore, we aimed to determine the tonsillar immune responses according to rhinovirus A, B and C infections. Methods We collected tonsillar samples, nasopharyngeal aspirates and peripheral blood from 42 rhinovirus positive tonsillectomy patients. Fifteen respiratory viruses or their types were investigated from nasopharynx and tonsil tissue, and rhinovirus species were typed. The expression of 10 cytokines and 4 transcription factors (IFN-α, IFN-β, IFN-γ, IL-10, IL-13, IL-17, IL-28, IL-29, IL-37, TGF-β, FOXP3, GATA3, RORC2 and Tbet) were studied from tonsil tissue by quantitative PCR. A standard questionnaire of respiratory symptoms and health was filled by the patient or his/her guardian. The patients were divided into three groups by the determination of rhinovirus species. Results Overall, 16 patients had rhinovirus A, 12 rhinovirus B and 14 rhinovirus C infection. In rhinovirus B positive group there were significantly less men (P = 0.0072), less operated in spring (P = 0.0096) and more operated in fall (P = 0.030) than in rhinovirus A or C groups. Rhinovirus A positive patients had more respiratory symptoms (P = 0.0074) and particularly rhinitis (P = 0.036) on the operation day. There were no significant differences between the groups in virus codetection. In adjusted analysis, rhinovirus C infections were associated with increased IFN-α (P = 0.045) and decreased RORC2 expression (P = 0.025). Conclusions Rhinovirus species associated differently with clinical characteristics and tonsillar cytokine responses.