Characterization of the microtranscriptome of macrophages infected with virulent, attenuated and saprophyte strains of Leptospira spp.

Garcia, Leandro Encarnação; Araújo, Erivelto Corrêa de; Melo, Larissa Martins; Bragato, Jaqueline Poleto; Peiró, Juliana Regina; Lima, Valéria Marçal Felix de; Marinho, Márcia; Arnold, Daniel; Lopes, Flávia L.

doi:10.1371/journal.pntd.0006621

Cited by 10 publications

(10 citation statements)

References 64 publications

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“…A recent work that examined the microtranscriptome of murine macrophages J774A.1 stimulated with virulent, attenuated, and saprophyte strains of Leptospira showed differential expression of 29 miRNAs. Of these 29 miRNAs, the highest fold change was observed for miR155-5p in cells stimulated with pathogenic L. interrogans ( 42 ). We did not see an upregulation of this miR155-5p, which could be due to different factors, including but not limited to the difference in the assay method, the cell type, and the culture conditions.…”

Section: Discussionmentioning

confidence: 99%

MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis

Mercy

Muthukumaran

Velusamy

et al. 2020

mSphere

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Leptospirosis remains a significant human health issue due to its systemic complications. Therefore, biomarkers that are more effective are urgently needed for the early diagnosis of leptospirosis. MicroRNAs (miRNAs) are evolutionarily conserved regulatory RNAs that have shown the potential to be used as biomarkers for diagnosis, prognosis, and therapy of infectious diseases. In this study, we performed an unbiased screen using the miRNome miRNA array to identify circulating miRNAs with the potential to serve as authentic biomarkers for early diagnosis of leptospirosis. Because leptospiral lipopolysaccharide (LPS) is the predominant leptospiral antigen and plays a vital role in immunological and biological activities, we used LPS treated and untreated in vitro (THP1 cells) and in vivo (BALB/c mice) surrogate models to identify the LPS-specific miRNAs. Differential expression analysis revealed 18 miRNAs to be associated strongly with LPS stimulation in THP1 cells. Of these, three (miR-let-7b-5p, miR-144-3p, and miR-21-5p) were observed to be present at increased levels in vivo. The identified miRNAs were validated for their biomarker potential using serum samples from leptospirosis-negative patients and patients with confirmed cases of leptospirosis. Identified miRNAs were able to discriminate the acute leptospiral infection from other febrile diseases with a test sensitivity and specificity of 93.2% and 88.19%, respectively. Gene functional enrichment and protein-protein interaction (PPI) network analysis revealed that the identified miRNAs play important roles in disease signal transduction, signaling by interleukins, the stress-activated protein kinase signaling cascade, the mitogen-activated protein kinase (MAPK) signaling pathway, and the cellular response to a transforming growth factor β (TGF-β) stimulus with a notable interconnection between these biological processes. IMPORTANCE Here, we used miRNAs that are differentially regulated by the LPS/TLR2 immune axis to devise a miRNA-based diagnosis for leptospirosis. The study established the role of the circulating stable miRNAs (miR-21-5p, miR-144-3p, and miR-let-7b-5p) as an early diagnostic marker for leptospirosis. These miRNAs can be used to diagnose acute leptospirosis and also to differentiate leptospiral infection from other bacterial and spirochetal infections, as proved by the use of human clinical samples. Thus, our findings indicate that miRNAs can play a crucial role in the diagnosis of infectious diseases, like leptospirosis, that are generally misdiagnosed.

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Section: Discussionmentioning

confidence: 99%

MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis

Mercy

Muthukumaran

Velusamy

et al. 2020

mSphere

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“…A previous study from our group has shown that BCL2 is a potentially down-regulated by mmu-mir-7667-3p following infection with L . interrogans , suggesting that cell survival could be compromissed after macrophages infection by the spirochete [29].…”

Section: Discussionmentioning

confidence: 99%

“…Murine monocyte-macrophage cells ( Mus musculus monocyte-macrophage cell line J774A.1), provided by the Paul Ehrlich cell bank (Rio de Janeiro, Brazil), was used as described [26]. Cells were maintained at 37° C, 5% CO2 in RPMI-1640 media (Sigma, USA) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, USA), 100 ug/mL streptomycin (Sigma Chemical Co St.Louis, MO), 0.03% L-glutamine solution (Sigma) and 100 UI/mL of penicillin, in 6-well cell culture plates (3cm/well) until confluency [22,29].…”

Section: Methodsmentioning

confidence: 99%

Gene expression is associated with virulence in murine macrophages infected with Leptospira spp

Garcia

et al. 2019

PLoS ONE

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Leptospira genus contains species that affect human health with varying degrees of pathogenicity. In this context, we aimed to evaluate the differences in the modulation of host gene expression by strains of Leptospira varying in virulence. Our data showed a high number of differentially expressed transcripts in murine macrophages following 6h of infection. Leptospira infection modulated a set of genes independently of their degree of virulence. However, pathway analysis indicated that Apoptosis, ATM Signaling, and Cell Cycle: G2/M DNA Damage Checkpoint Regulation were exclusively regulated following infection with the virulent strain. Taken together, results demonstrated that species and virulence play a role during host response to Leptospira spp in murine macrophages, which could contribute to understanding the pathogenesis of leptospirosis.

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“…In fact, Schulte et al [35] concluded that increased TNF-α, IL-1β and IL-6 can activate the coagulation system in endotoxemic models. The high concentration of IL-6 is an indicator of septic shock and correlates with the severity of leptospirosis [36].…”

Section: Discussionmentioning

confidence: 99%

Gene expression is associated with virulence in murine macrophages infected withLeptospiraspp

Júnior

Garcia

Araújo

et al. 2019

Preprint

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18Leptospira genus contains species that affect human health with varying degrees of 19 pathogenicity. In this context, we aimed to evaluate the differences in modulation of host 20 gene expression by strains of Leptospira with varied virulence degrees. Our data showed 21 a high number of differentilly expressed transcripts in murine macrophages following 6h 22 3 40 showed a high number of differentially expressed genes after 6h of infection by virulent 41 and attenuated L. interrogans, and to a lesser extent with L. biflexa saprophytic lineage. 42 This suggests that RNAs are modulated after infection by Leptospira in macrophages, in 43 a species and virulence related manner. It is hoped that the data produced will contribute 44 to further our understanding on the pathogenesis of leptospirosis. 45 46 47 48 49 50 51 52 53 4 54

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Characterization of the microtranscriptome of macrophages infected with virulent, attenuated and saprophyte strains of Leptospira spp.

Cited by 10 publications

References 64 publications

MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis

MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis

Gene expression is associated with virulence in murine macrophages infected with Leptospira spp

Gene expression is associated with virulence in murine macrophages infected withLeptospiraspp

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