Seminario Biología Molecular, Luisa Maradey
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This document presents research on the role of the NLRP12 protein during SARS-CoV-2 infection. The study found elevated expression of NLRP12 in human lung tissue and monocytes during COVID-19 infection. NLRP12 was found to directly interact with the viral M protein through its leucine-rich repeat domain. Methods used in the study included quantitative PCR, western blot, immunofluorescence, and immunohistochemistry to analyze protein expression and interactions between NLRP12 and the viral M protein. The results provide insights into the molecular mechanisms underlying inflammation during SARS-CoV-2 infection.
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Seminario Biología Molecular, Luisa Maradey
- 1. Luisa Fernanda Maradey Arbelaez III semestre Universidad Pontificia Bolivariana
- 2. COVID-19 Infectious disease caused by the SARS-CoV-2 virus which spreads by air and causes severe respiratory symptoms such as cough, fever, chills and sore throat. People with previous illnesses or other risk factors may contract the disease more easily and have complications. https://images.app.goo.gl/FJBMVCb3nJsELiFF8
- 3. What is inflammatory response? Process that is triggered by aggressions of any kind. Inmune reaction in which cells attack invading microorganisms and prevent tissue damage and repair them in case of destruction. Nucleotide-binding leucine-rich repeat– containing receptor 12 It’s an intracellular pattern recognition receptor that detects intracellular microbial pathogens. NLRP12
- 4. To observe elevated expression levels of NLRP12 in human peripheral monocytes and lung tissue during infection with SARS-CoV- 2. Analysis revealed that NLRP12 directly interacted with the M protein through its leucine-rich repeat domain. GENERAL OBJECTIVE
- 5. METHODS Quantitative Polymerase Chain Reacttion (PCRQ) Its objective is the direct enzymatic amplification of a gene or a DNA fragments, either directly or indirectly through RNA Laboratory technique used to detect a specific protein in a biological sample. Western Blot https://es.slideshare.net/EdgarHernandez71/western-blot-44994357
- 6. METHODS Immunofluorescence Immunohistochemistry A diagnostic technique that uses fluorescent substances to detect the presence of an antigen or antibody in cells or tissues. Is the identification of a specific molecule in a tissue by the interaction between an antigen and a labeled antibody. https://www.labclinics.com/2019/04/30/inmunofluorescencia-campo-amplio-versus-confocal/ https://labpap.com.mx/sitio/2023/01/13/inmunohistoquimica-tecnica-esencial-para-el-diagnostico-y-la-investigacion-en-patologia/
- 7. RESULTS Representative immunofluorescence staining of NLRP12 in the lung tissue from healthy donors and patients with COVID-19 A549 cells and dTHP-1 cells were transfected with plasmids containing FLAG-tagged M protein The exogenous interaction between NLRP12 and the structural proteins was analyzed by co- immunoprecipitation 1 3 2
- 8. RESULTS Bone marrow–derived macrophages from wild type C57BL/6J and NLRP12 knockout mice were transfected with siTRAF3 and infected with VSV-M. Hematoxylin and eosin staining was performed on lung tissue sections 4 5
- 9. DISCUSSION
- 10. Molecular biology, thanks to its advances, helps to identify the molecules involved in inflammatory processes, how they act and their implications in the organism in cases where health is affected. It is important to know how viruses and other pathogens carry out their functions in the organism in order to find solutions to these diseases that affect an entire population. CONCLUSIONS
- 11. MAPA
- 12. THANKS!