Simultaneous transcriptional profiling of Leishmania major and its murine macrophage host cell reveals insights into host-pathogen interactions

Dillon, Laura A. L.; Suresh, Rahul; Okrah, Kwame; Bravo, Héctor Corrada; Mosser, David M.; El-Sayed, Najib M.

doi:10.1186/s12864-015-2237-2

Cited by 84 publications

(119 citation statements)

References 101 publications

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“…Moreover, LmPRL-1 was still detectable in macrophage lysates even at 72 h after infection with promastigotes, when the parasites had already transformed into amastigotes. This observation on the protein level corroborates and extends recent mRNA expression profiling experiments (59,60). The expression of the gene LmjF.…”

Section: Discussionsupporting

confidence: 75%

“…The expression of the gene LmjF. 16.230, coding for LmPRL-1, was upregulated (1.27-fold) during the transition of procyclic into metacyclic promastigotes (59) and after macrophage infection by promastigotes (1.49-fold) (60). However, LmPRL-1 was hardly detectable in macrophages directly infected with amastigotes purified from mouse skin lesions or in lysates of these amastigotes.…”

Section: Discussionmentioning

confidence: 97%

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Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

et al. 2017

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Similar to other intracellular pathogens, Leishmania parasites are known to evade the antimicrobial effector functions of host immune cells. To date, however, only a few virulence factors have been described for Leishmania major, one of the causative agents of cutaneous leishmaniasis. Here, we have characterized the expression and function of an L. major phosphatase, which we termed LmPRL-1. This enzyme shows a strong structural similarity to the human phosphatases of regenerating liver (PRL-1, -2, and -3) that regulate the proliferation, differentiation, and motility of cells. The biochemical characterization of the L. major phosphatase revealed that the enzyme is redox sensitive. When analyzing the subcellular localization of LmPRL-1 in promastigotes, amastigotes, and infected macrophages, we found that the phosphatase was predominantly expressed and secreted by promastigotes via the exosome route. Finally, we observed that ectopic expression of LmPRL-1 in L. major led to an increased number of parasites in macrophages. From these data, we conclude that the L. major phosphatase LmPRL-1 contributes to the intracellular survival of the parasites in macrophages.KEYWORDS Leishmania, exosome, macrophage, tyrosine phosphatase L eishmaniasis is an infectious disease prevalent worldwide that is caused by kinetoplastid protozoan parasites belonging to the genus Leishmania (1). In nature, this heteroxenous parasite is transmitted by the bites of sand flies. Following the inoculation of flagellated promastigotes into the dermis of the host, the parasites are rapidly endocytosed by phagocytic cells, in which they differentiate into amastigotes, multiply, and reach inner compartments and organs, such as draining lymph nodes, spleen, liver, and bone marrow. The life cycle of the parasite is completed after ingestion of amastigote-infected cells by sand flies during their blood meal. In the digestive tract of their vector, parasites transform back into extracellular promastigotes that develop into infectious metacyclics (2). Depending on the status of the host immune system and the Leishmania species, the infection of mammals leads either to mostly self-healing skin lesions (cutaneous leishmaniasis [CL]) or to a systemic disease, termed visceral leishmaniasis (VL) or kala-azar, that is lethal if untreated (3-5).To survive in their hosts, Leishmania parasites need to quickly adapt their growth, metabolism, and mechanisms of protection to their new environment. Interestingly, only a few genes are differentially expressed during this adaptive process, suggesting an important role for the regulation of protein translation (6). Leishmania speciesspecific gene diversity also participates in the adaptation of the parasite to its organ-

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Section: Discussionsupporting

confidence: 75%

Section: Discussionmentioning

confidence: 97%

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

et al. 2017

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“…In both species, a multitude of transcriptional-mediated cellular processes are fine-tuned: activated, maintained and repressed to ensure survival. The recently-described dual RNA-Seq approach allows simultaneous host-pathogen monitoring during their interactions (Baddal et al, 2015; Dillon et al, 2015; Westermann et al, 2016). In this study, we exploited the dual RNA-Seq approach by application to a pneumococcal infection model to human lung alveolar epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…In a thought experiment, Vogel and co-workers argued that simultaneous profiling of host and pathogen transcriptomics by dual RNA-Seq might provide valuable insights for infection biology (Westermann et al, 2012). Recent dual RNA-Seq studies were successful in elucidating the role of sRNAs in the intracellular pathogen Salmonella typhimurium (Westermann et al, 2016), cross-talk in the Gram-negative LRTI pathogen Haemophilus influenzae (Baddal et al, 2015) and transcription profiles in the protozoan Leishmania major (Dillon et al, 2015) during their respective infection.…”

Section: Introductionmentioning

confidence: 99%

Time-resolved dual RNA-Seq reveals extensive rewiring of lung epithelial and pneumococcal transcriptomes during early infection

Aprianto

Slager

Holsappel

et al. 2016

Preprint

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“…RNAseq analysis of host-pathogen interactions has already been successfully applied for eukaryotic pathogens [11][12][13][14][15] . Transcripts of eukaryotic pathogens, unlike bacteria, are polyadenylated, and occur with comparable abundance to the hosts.…”

Section: Comparison To Other Currently Available Methodsmentioning

confidence: 99%

A highly multiplexed and sensitive RNA-seq protocol for simultaneous analysis of host and pathogen transcriptomes

et al. 2016

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EDITORIAL SUMMARY:This protocol enables simultaneous analysis of host and bacterial transcripts by RNA-Seq. Including procedures for efficient host and bacterial cell lysis, barcoding of samples, and analysis of both mammalian and microbial reads from mixed host-pathogen RNA-Seq data.TWEET: Simultaneous analysis of host and pathogen transcriptomes by RNA-Seq. Abstract The ability to simultaneously characterize the bacterial and host expression programs during infection would facilitate a comprehensive understanding of pathogen-host interactions. While RNA-Seq has greatly advanced our ability to study the transcriptomes of prokaryote and eukaryotes separately, limitations in existing protocols for generating and analyzing RNA-Seq data have hindered simultaneous profiling of host and bacterial pathogen transcripts from the same sample. Here we provide a detailed protocol for simultaneous analysis of host and bacterial transcripts by RNA-Seq. Importantly, this protocol details the steps required for efficient host and bacteria lysis, barcoding of samples, technical advances in sample preparation for low yield sample inputs and a computational pipeline to analyze both mammalian and microbial reads from mixed hostpathogen RNA-Seq data. Sample preparation takes 3 d from cultured cells to pooled libraries. Data analysis takes an additional day. Compared with previous methods, the protocol detailed here provides a sensitive, facile and generalizable approach, suitable for large-scale studies, which will enable the field to obtain in-depth analysis of hostpathogen interactions in infection models. Introduction Intracellular bacterial pathogens, such as Mycobacterium tuberculosis, Salmonella enterica, Legionella pneumophilia, and Neisseria gonorrhea, spend a significant portion of their life-cycle surviving and replicating within host cells. The cellular interaction entails both a complex virulence program executed by the bacterial pathogen during infection 1 and activation of an orchestrated defense response by the host to counter the pathogen 2 . Genomic approaches have been employed in recent years to uncover substantial molecular details of this rich host-pathogen biology 3, 4 . However, technical constraints limit these studies to profiling either the host or the pathogen -while a comprehensive understanding of host-pathogen interactions requires simultaneous analysis of the associated gene expression changes in both the pathogen and the host 5 . The limitations of conventional protocols for simultaneous analysis of host and pathogen transcripts include 1) the inability to obtain high quality RNA with efficient lysis of both bacterial and mammalian host cells; 2) inefficient depletion of both microbial and mammalian ribosomal RNA species; 3) inability to simultaneously process polyadenylated and non-polyadenylated transcripts from low yield samples (>10ng of RNA); and 4) a lack of robust computational approaches for analyzing the often small subset of bacterial transcripts in infected cells or tissue. Recently, sever...

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Simultaneous transcriptional profiling of Leishmania major and its murine macrophage host cell reveals insights into host-pathogen interactions

Cited by 84 publications

References 101 publications

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

Time-resolved dual RNA-Seq reveals extensive rewiring of lung epithelial and pneumococcal transcriptomes during early infection

A highly multiplexed and sensitive RNA-seq protocol for simultaneous analysis of host and pathogen transcriptomes

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