Localized skin inflammation during cutaneous leishmaniasis drives a chronic, systemic IFN-γ signature

Abstract

Cutaneous leishmaniasis is a localized infection controlled by CD4+ T cells that produce IFN-γ within lesions. Phagocytic cells recruited to lesions, such as monocytes, are then exposed to IFN-γ which triggers their ability to kill the intracellular parasites. Consistent with this, a transcriptional analysis of lesions from patients identified the presence of a strong interferon stimulated gene (ISG) signature. To determine what systemic responses are occurring that might influence the disease, we performed RNA sequencing (RNA-seq) on the blood of L. braziliensis-infected patients, as well as healthy controls. Functional enrichment analysis identified a transcriptional ISG signature as the dominant response in the blood of patients. An increase in monocytes and macrophages in the blood, estimated from our RNA-seq dataset, was positively correlated with this ISG signature. Consistent with this result, patients had circulating IFN-γ in their serum. A cytotoxicity signature, which is a dominant feature in the lesions, was also found in the blood and correlated with an increased abundance of cytolytic cells. Thus, two transcriptional signatures present in lesions were found systemically, although with a substantially reduced number of differentially expressed genes (DEGs). Finally, we found that the number of DEGs and ISGs in leishmaniasis was similar to tuberculosis – another localized infection – but significantly less than observed in malaria. In contrast, the cytolytic signature and increased cytolytic cell abundance was not found in tuberculosis or malaria. Our results indicate that systemic signatures can reflect what is occurring in leishmanial lesions. Furthermore, the presence of an ISG signature in blood monocytes and macrophages suggests a mechanism to limit systemic spread of the parasite, as well as enhance parasite control by pre-activating cells prior to lesion entry.Author summaryCutaneous leishmaniasis caused by the protozoan Leishmania braziliensis exhibits two dominant inflammatory responses in cutaneous lesions: Interferon-γ (IFN-γ)-mediated signaling, which promotes parasite control, and cytolysis mediated by cytotoxic CD8+ T and NK cells, which promotes increased pathology. To determine if these responses were limited to cutaneous lesions, we performed RNA-seq on the blood of cutaneous leishmaniasis (CL) patients, and detected both transcriptional signatures in the peripheral blood. The presence of interferon stimulated genes, as well as circulating IFN-γ, suggests that protective immune responses are not limited to the lesion site, but are occurring systemically. This may be one mechanism to ensure optimal control of the parasites, both by limiting their systemic spread and by pre-activating cells to kill the parasites prior to entry into the lesions. The cytolytic transcriptional signature was uniquely detectable in the blood of L. braziliensis patients when compared to the blood of patients with tuberculosis (TB) or malaria, further emphasizing the importance of this pathway in cutaneous leishmaniasis. Taken together, these data suggest that this localized infection has a systemic component that may have an impact the development of the disease.

Pathogenic CD8 T cell responses are driven by neutrophil-mediated hypoxia in cutaneous leishmaniasis

“…To determine whether hypoxia is a feature of human disease, we compared transcriptional signatures in blood and skin from healthy subjects (HS) and cutaneous leishmaniasis (CL) patients using previously published datasets (51,76). To estimate levels of hypoxia-gene expression in clinical samples, we performed single-sample GSEA using the Harris et al hypoxia gene signature (58), and referred to this variable as the "hypoxia score" (Supp.…”

“…Data in Fig. 6 are derived from published transcriptional profiling (51,76). The analysis carried out for this current study was performed from the filtered, normalized gene expression matrix available for download as a text file on NCBI GEO accession #GSE162760 and #GSE214397.…”

Advances in Understanding Leishmania Pathobiology: What Does RNA-Seq Tell Us?

“…The blood of L. braziliensis infected patients was also recently analyzed using RNA-seq to determine systemic responses that might be influencing the disease ( Amorim et al, 2020 ). A strong interferon stimulated gene (ISG) signature has been identified which was correlated with an increase in circulating monocytes and macrophages ( Amorim et al, 2020 ). A cytotoxicity signature was correlated with an increase in cytolytic cells ( Amorim et al, 2020 ).…”

“…A strong interferon stimulated gene (ISG) signature has been identified which was correlated with an increase in circulating monocytes and macrophages ( Amorim et al, 2020 ). A cytotoxicity signature was correlated with an increase in cytolytic cells ( Amorim et al, 2020 ).…”