Transcriptional profiling of the murine cutaneous response during initial and subsequent infestations with Ixodes scapularis nymphs

Abstract

BackgroundIxodes scapularis ticks are hematophagous arthropods capable of transmitting many infectious agents to humans. The process of blood feeding is an extended and continuous interplay between tick and host responses. While this process has been studied extensively in vitro, no global understanding of the host response to ticks has emerged.MethodsTo address this issue, we used PCR-arrays to measure skin-specific expression of 233 discrete genes at 8 time points during primary and secondary infestations of mice with pathogen-free I. scapularis nymphs. Selected results were then validated at the mRNA and protein levels by additional real-time PCR and bioplex assay.ResultsPrimary infestation was characterized by the late induction of an innate immune response. Lectin pattern recognition receptors, cytokines, and chemokines were upregulated consistent with increased neutrophil and macrophage migration. Gene ontology and pathway analyses of downregulated genes suggested inhibition of gene transcription and Th17 immunity. During the secondary infestation, additional genes were modulated suggesting a broader involvement of immune cells including CD8 and CD4 positive T lymphocytes. The cytokine response showed a mixed Th1/Th2 profile with a potential for T regulatory cell activity. Key gene ontology clusters observed during the secondary infestation were cell migration and activation. Matrix metalloproteinases were upregulated, apoptosis-related genes were differentially modulated, and immunoreceptor signaling molecules were upregulated. In contrast, transcripts related to mitogenic, WNT, Hedgehog, and stress pathways were downregulated.ConclusionsOur results support a model of tick feeding where lectin pattern recognition receptors orchestrate an innate inflammatory response during primary infestation that primes a mixed Th1/Th2 response upon secondary exposure. Tick feeding inhibits gene transcription and Th17 immunity. Salivary molecules may also inhibit upregulation of mitogenic, WNT, Hedgehog, and stress pathways and enhance the activity of T regulatory cells, production of IL-10, and suppressors of cytokine signaling molecules (SOCS). This study provides the first comprehensive transcriptional analysis of the murine host response at the I. scapularis bite site and suggests both a potential model of the host cutaneous response and candidate genes for further description and investigation.

Characterization of the early local immune response to Ixodes ricinus tick bites in human skin

“…In BALB/c mice, very early (6 – 12 h post infestation) tick-bite lesions are characterized histologically by a strong innate immune reaction with a predominance of neutrophils and mononuclear cells, intermingled with CD4+ and CD8+ T cells in a ratio of 2.2:1 (24). Consistent with these results, molecular investigations of early lesions in mice found upregulation of the chemoattractants for monocytes, neutrophils, and dendritic cells CXCL1, CXCL5, CCL2, and CXCL14 (25, 26). These early alterations in tick bite lesions in murine skin are comparable with our findings in human skin, which showed an initial strong, transient innate immune response.…”

The putative role of Rhipicephalus microplus salivary serpins in the tick-host relationship

“…Neutrophils play an essential part as triggers of an immune response against tick feeding (Heinze et al, 2012a; Heinze et al, 2012b). athepsin G, neutrophil elastase and proteinase 3 are the most abundant molecules present in neutrophil azurophil granules (Korkmaz et al, 2008; Pham, 2006; Pham, 2008).…”

Partial pathogen protection by tick-bite sensitization and epitope recognition in peptide-immunized HLA DR3 transgenic mice

“…Though first identified in Guinea pigs, the Th2 bias of repeated tick infestations was best represented by gene expression and cytokine profile studies in BALB/c, C3H/HeN and C3H/HeJ murine strains. 29-32 However, in those mouse models, repeat tick exposure induced immune response changes but neither BALB/c nor C3H/HeN mice developed resistance 16,33-35 …”