Event Abstract Back to Event Global transcriptome analysis of alveolar macrophage from tuberculosis patients and non-infected controls Lelia Lavalett1, 2, Héctor A. Rodríguez3, 4, Héctor Ortega5, Mauricio Rojas1 and Luis F. Barrera1* 1 Universidad de Antioquia (UdeA), Instituto de Investigaciones Médicas (IIM), Facultad de Medicina, Colombia 2 Universidad Nacional de Colombia, Sede Medellín, Facultad de Ciencias, Colombia 3 Universidad Nacional de Colombia, Sede Medellín, Facultad de Ciencias, Colombia 4 Corporación para Investigaciones Biológicas (CIB), Colombia 5 Clínica Cardiovascular Santa María, Colombia Introduction. Alveolar macrophages (AMs) are an important focus of tuberculosis (TB) research since they play an essential role in the immune response to infection with Mycobacterium tuberculosis (Mtb). During lung infection, Mtb subverts the bactericidal mechanisms of AMs. Therefore, understanding this host-pathogen relationship is fundamental to the development of new therapies to prevent and cure TB. We hypothesized that there was an effect of the disease (TB) on patterns of gene expression in AMs from patients with active TB (n=4) compared to AMs from non-infected control subjects (n=4). Methods. Total RNA from at least 3 replica wells was obtained with the RNeasy minikit (Qiagen). RNA (RIN>7.5) were used to prepare cRNAs. Biotinylated cRNAs were prepared from 0.55 µg of total RNA using the Illumina TotalPrep RNA Amplification kit (Ambion). Following fragmentation, 0.75 µg of cRNA were hybridized to the Illumina Expression Human HT-12v4 Expression Bead Chip, targeting 47,323 probes (more than 31,000 coding sequences) according to the protocols provided by the manufacturer. Array data export processing and analysis was performed using Illumina Genome Studio v2011.1. After log transformation of raw data, samples were Quantile normalized using the R software (v2.15.1) from BioConductor. Unsupervised methods with hierarchical clustering was calculated using complete Euclidean distance transformation. Functional annotation and clustering were done using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) v6.7. Results. We found altered gene expression for 46 genes in TB versus non-infected controls that were differentially expressed by ≥1.5 or ≤1,5-log2 fold (FDR=0.05); 29 genes were upregulated, and 17 genes were downregulated. Moreover, some of the downregulated (CAMP, TGFB1, MARCO, CD163, IL1B) and upregulated (CHIT1, MMP7, STAT1, CCL5) genes have been previously associated with regulation of the antimicrobial innate immune response, excessive inflammation, tissue damage, disease progression and susceptibility to TB. Analysis of functional annotation of these 46 genes showed significant enrichment in different functional categories including defense and inflammatory response, chemotaxis, antigen processing and presentation, and prostaglandin metabolic process. Conclusion. These results suggest different functional pathways altered in alveolar macrophages after challenge with Mtb and support the idea of functional correlations of pathogenesis-driven gene expression signatures in TB. Our findings may partially contribute to identifying critical macrophage response genes associated with different outcomes in active pulmonary TB, which could help to elucidate the pathogenesis of the disease Acknowledgements Funded by Colciencias, Bogotá, Grant 1115-519-29046 Keywords: human, Tuberculosis, alveolar macrophages, Microarrays, Functional Genomics Conference: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015. Presentation Type: Poster Presentation Topic: Infectious and parasitic diseases Citation: Lavalett L, Rodríguez HA, Ortega H, Rojas M and Barrera LF (2015). Global transcriptome analysis of alveolar macrophage from tuberculosis patients and non-infected controls. Front. Immunol. Conference Abstract: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología. doi: 10.3389/conf.fimmu.2015.05.00152 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 28 Apr 2015; Published Online: 14 Sep 2015. * Correspondence: Prof. Luis F Barrera, Universidad de Antioquia (UdeA), Instituto de Investigaciones Médicas (IIM), Facultad de Medicina, Medellin, Colombia, luis.barrera@udea.edu.co Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Lelia Lavalett Héctor A Rodríguez Héctor Ortega Mauricio Rojas Luis F Barrera Google Lelia Lavalett Héctor A Rodríguez Héctor Ortega Mauricio Rojas Luis F Barrera Google Scholar Lelia Lavalett Héctor A Rodríguez Héctor Ortega Mauricio Rojas Luis F Barrera PubMed Lelia Lavalett Héctor A Rodríguez Héctor Ortega Mauricio Rojas Luis F Barrera Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.