Integrated multivariate analysis of transcriptomic data reveals immunological mechanisms in mice after Leishmania infantum infection

Transcriptional analysis of complex biological scenarios has been extensively used, even though sometimes results may prove imprecise or difficult-to interpret due to an overwhelming amount of information. In this study, a large-scale Real-time qPCR experiment was coupled to multivariate statistical analysis to describe the main immunological events underlying the early L. infantum infection in livers of BALB/c mice. High-throughput qPCR was used to evaluate the expression of 223 genes related to immunometabolism 1-, 3-, 5- and 10-days post infection. This integrative analysis showed strikingly different gene signatures at 1- and 10-days post infection, revealing progression of infection in the experimental model based on the upregulation of particular immunological response patterns and mediators. This approach addresses the challenge of integrating large collections of transcriptional data for the identification of candidate biomarkers in experimental models.

[1]  M. Teixeira,et al.  CXCL10 treatment promotes reduction of IL-10+ regulatory T (Foxp3+ and Tr1) cells in the spleen of BALB/c mice infected by Leishmania infantum. , 2019, Experimental parasitology.

[2]  S. Prakash,et al.  Parasitic load determination by differential expression of 5-Lipoxygenase and PGE2 Synthases in Visceral Leishmaniasis. , 2019, Prostaglandins & other lipid mediators.

[3]  Gary D Bader,et al.  Pathway enrichment analysis and visualization of omics data using g:Profiler, GSEA, Cytoscape and EnrichmentMap , 2019, Nature Protocols.

[4]  A. González-García,et al.  Transcriptional Profiling of Immune-Related Genes in Leishmania infantum-Infected Mice: Identification of Potential Biomarkers of Infection and Progression of Disease , 2018, Front. Cell. Infect. Microbiol..

[5]  Y. Miyazaki,et al.  Functions of CD1d-Restricted Invariant Natural Killer T Cells in Antimicrobial Immunity and Potential Applications for Infection Control , 2018, Front. Immunol..

[6]  A. Al-Hendy,et al.  PDL-1 Blockade Prevents T Cell Exhaustion, Inhibits Autophagy, and Promotes Clearance of Leishmania donovani , 2018, Infection and Immunity.

[7]  M. Teixeira,et al.  Protection mediated by chemokine CXCL10 in BALB/c mice infected by Leishmania infantum , 2017, Memorias do Instituto Oswaldo Cruz.

[8]  Haley R Pipkins,et al.  Polyamine transporter potABCD is required for virulence of encapsulated but not nonencapsulated Streptococcus pneumoniae , 2017, PloS one.

[9]  Ron Diskin,et al.  Mapping of the Lassa virus LAMP1 binding site reveals unique determinants not shared by other old world arenaviruses , 2017, PLoS pathogens.

[10]  H. Spratt,et al.  Splenic CD4+ T Cells in Progressive Visceral Leishmaniasis Show a Mixed Effector-Regulatory Phenotype and Impair Macrophage Effector Function through Inhibitory Receptor Expression , 2017, PloS one.

[11]  B. Luxon,et al.  Transcriptional Profiling in Experimental Visceral Leishmaniasis Reveals a Broad Splenic Inflammatory Environment that Conditions Macrophages toward a Disease-Promoting Phenotype , 2017, PLoS pathogens.

[12]  Damian Szklarczyk,et al.  The STRING database in 2017: quality-controlled protein–protein association networks, made broadly accessible , 2016, Nucleic Acids Res..

[13]  A. González-García,et al.  The Challenge of Stability in High-Throughput Gene Expression Analysis: Comprehensive Selection and Evaluation of Reference Genes for BALB/c Mice Spleen Samples in the Leishmania infantum Infection Model , 2016, PloS one.

[14]  P. Mermelstein,et al.  Opposite Effects of mGluR1a and mGluR5 Activation on Nucleus Accumbens Medium Spiny Neuron Dendritic Spine Density , 2016, PloS one.

[15]  F. Kirchhoff,et al.  Vpu-Mediated Counteraction of Tetherin Is a Major Determinant of HIV-1 Interferon Resistance , 2016, mBio.

[16]  H. Bravo,et al.  Dual Transcriptome Profiling of Leishmania-Infected Human Macrophages Reveals Distinct Reprogramming Signatures , 2016, mBio.

[17]  J. Estaquier,et al.  Regulation of immunity during visceral Leishmania infection , 2016, Parasites & Vectors.

[18]  R. Kumar,et al.  Combined Immune Therapy for the Treatment of Visceral Leishmaniasis , 2016, PLoS neglected tropical diseases.

[19]  Laura A. L. Dillon,et al.  Simultaneous transcriptional profiling of Leishmania major and its murine macrophage host cell reveals insights into host-pathogen interactions , 2015, BMC Genomics.

[20]  M. Thon,et al.  Identification of horizontally transferred genes in the genus Colletotrichum reveals a steady tempo of bacterial to fungal gene transfer , 2015, BMC Genomics.

[21]  R. Kumar,et al.  Immune Regulation during Chronic Visceral Leishmaniasis , 2014, PLoS neglected tropical diseases.

[22]  P. Greenberg,et al.  Tolerance and exhaustion: defining mechanisms of T cell dysfunction. , 2014, Trends in immunology.

[23]  P. Kaye,et al.  A Transcriptomic Network Identified in Uninfected Macrophages Responding to Inflammation Controls Intracellular Pathogen Survival , 2013, Cell host & microbe.

[24]  Yan Zhang,et al.  Regulatory Actions of Toll-Like Receptor 2 (TLR2) and TLR4 in Leishmania donovani Infection in the Liver , 2013, Infection and Immunity.

[25]  R. Bhadra,et al.  T cell exhaustion in protozoan disease. , 2012, Trends in parasitology.

[26]  M. Wilson,et al.  Receptor-mediated phagocytosis of Leishmania: implications for intracellular survival. , 2012, Trends in parasitology.

[27]  H. Gascan,et al.  Invariant NKT Cells Drive Hepatic Cytokinic Microenvironment Favoring Efficient Granuloma Formation and Early Control of Leishmania donovani Infection , 2012, PloS one.

[28]  B. Sharrack,et al.  CCL2 binding is CCR2 independent in primary adult human astrocytes , 2012, Brain Research.

[29]  N. Reiling,et al.  Leishmania major parasite stage‐dependent host cell invasion and immune evasion , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[30]  Gary D. Bader,et al.  clusterMaker: a multi-algorithm clustering plugin for Cytoscape , 2011, BMC Bioinformatics.

[31]  J. Donelson,et al.  The effects of macrophage source on the mechanism of phagocytosis and intracellular survival of Leishmania. , 2011, Microbes and infection.

[32]  E. Assier,et al.  Interleukin-23: A key cytokine in inflammatory diseases , 2011, Annals of medicine.

[33]  J. Polettini,et al.  Analysis of the expression of toll-like receptors 2 and 4 and cytokine production during experimental Leishmania chagasi infection. , 2011, Memorias do Instituto Oswaldo Cruz.

[34]  A. Satoskar,et al.  Role of chemokines in regulation of immunity against leishmaniasis. , 2010, Experimental parasitology.

[35]  A. Kariminia,et al.  Leishmania major lipophosphoglycan: discrepancy in Toll-like receptor signaling. , 2010, Experimental parasitology.

[36]  A. Kariminia,et al.  The involvement of TLR2 in cytokine and reactive oxygen species (ROS) production by PBMCs in response to Leishmania major phosphoglycans (PGs) , 2009, Parasitology.

[37]  Antonio Polley,et al.  Coregulation of CD8+ T cell exhaustion during chronic viral infection by multiple inhibitory receptors , 2008, Nature immunology.

[38]  C. Bogdan,et al.  TLR9 signaling is essential for the innate NK cell response in murine cutaneous leishmaniasis , 2007, European journal of immunology.

[39]  C. Engwerda,et al.  Balancing immunity and pathology in visceral leishmaniasis , 2007, Immunology and cell biology.

[40]  L. Hennighausen,et al.  Interleukin 27 negatively regulates the development of interleukin 17–producing T helper cells during chronic inflammation of the central nervous system , 2006, Nature Immunology.

[41]  J. Schwartzman,et al.  CCR5 Is Essential for NK Cell Trafficking and Host Survival following Toxoplasma gondii Infection , 2006, PLoS pathogens.

[42]  H. Weiner,et al.  Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells , 2006, Nature.

[43]  Y. Iwakura,et al.  The IL-23/IL-17 axis in inflammation. , 2006, The Journal of clinical investigation.

[44]  P. Kaye,et al.  Invariant NKT Cells Are Essential for the Regulation of Hepatic CXCL10 Gene Expression during Leishmania donovani Infection , 2005, Infection and Immunity.

[45]  R. D. Hatton,et al.  Interleukin 17–producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages , 2005, Nature Immunology.

[46]  J. Mesirov,et al.  From the Cover: Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005 .

[47]  T. Yoshimoto,et al.  A Role for IL-27 in Early Regulation of Th1 Differentiation1 , 2005, The Journal of Immunology.

[48]  A. Gebert,et al.  Cutting Edge: Neutrophil Granulocyte Serves as a Vector for Leishmania Entry into Macrophages1 , 2004, The Journal of Immunology.

[49]  P. Kaye,et al.  The immunopathology of experimental visceral leishmaniasis , 2004, Immunological reviews.

[50]  P. Shannon,et al.  Cytoscape: a software environment for integrated models of biomolecular interaction networks. , 2003, Genome research.

[51]  P. Scott,et al.  Interleukin-12 Regulates Chemokine Gene Expression during the Early Immune Response to Leishmania major , 2003, Infection and Immunity.

[52]  T. Mcclanahan,et al.  IL-27, a heterodimeric cytokine composed of EBI3 and p28 protein, induces proliferation of naive CD4+ T cells. , 2002, Immunity.

[53]  M. Olivier,et al.  Leishmania-induced cellular recruitment during the early inflammatory response: modulation of proinflammatory mediators. , 2002, The Journal of infectious diseases.

[54]  H. Murray Tissue granuloma structure‐function in experimental visceral leishmaniasis , 2001, International journal of experimental pathology.

[55]  T. Mak,et al.  WSX-1 is required for the initiation of Th1 responses and resistance to L. major infection. , 2001, Immunity.

[56]  F.J. Sauvage,et al.  Development of Th1-type immune responses requires the type I cytokine receptor TCCR , 2000, Nature.

[57]  C. Nathan,et al.  Macrophage Microbicidal Mechanisms In Vivo: Reactive Nitrogen versus Oxygen Intermediates in the Killing of Intracellular Visceral Leishmania donovani , 1999, The Journal of experimental medicine.

[58]  P. Kaye,et al.  Leishmania donovani infection initiates T cell‐independent chemokine responses, which are subsequently amplified in a T cell‐dependent manner , 1999, European journal of immunology.

[59]  A. Zlotnik,et al.  T-cell subsets: Chemokine receptors guide the way , 1998, Current Biology.

[60]  G. Rainaldi,et al.  IL-12 induces IFN-gamma expression and secretion in mouse peritoneal macrophages. , 1997, Journal of immunology.

[61]  S. Beverley,et al.  Leishmania major: promastigotes induce expression of a subset of chemokine genes in murine macrophages. , 1997, Experimental parasitology.

[62]  F. Sutterwala,et al.  Leishmania major-human macrophage interactions: cooperation between Mac-1 (CD11b/CD18) and complement receptor type 1 (CD35) in promastigote adhesion , 1996, Infection and immunity.

[63]  F. Derouin,et al.  Culture microtitration: a sensitive method for quantifying Leishmania infantum in tissues of infected mice , 1995, Antimicrobial agents and chemotherapy.

[64]  A. Minty,et al.  Monocyte chemotactic protein 3 is a most effective basophil- and eosinophil-activating chemokine , 1994, The Journal of experimental medicine.

[65]  K. Squires,et al.  Acquired resistance and granuloma formation in experimental visceral leishmaniasis. Differential T cell and lymphokine roles in initial versus established immunity. , 1992, Journal of immunology.

[66]  Antonio Polley,et al.  Coregulation of CD8+ T cell exhaustion by multiple inhibitory receptors during chronic viral infection , 2009, Nature Immunology.

[67]  J. Carl,et al.  IL27: its roles in the induction and inhibition of inflammation. , 2008, International journal of clinical and experimental pathology.