Dataset Information

Comprehensive Analysis of Protein Expression Levels and Phosphorylation Levels in Host Skin in Response to Tick (Haemaphysalis longicornis) Bite

ABSTRACT: Ticks are widely distributed ectoparasitic arthropods that suck blood from the body surface of livestock, wild animals, and humans. Ticks not only transmit a variety of pathogens but also cause various degrees of damage to their hosts' skin during blood feeding. To explore the molecular regulatory mechanism employed by the host skin to withstand tick bites, larval, nymphal, and adult Haemaphysalis longicornis, which is distributed in East Asia, were used to bite the skin tissues of healthy rabbits in the present study. The quantitative proteomic technology data-independent acquisition was then employed to investigate in depth the changes in protein expression and phosphorylation in rabbit skin after tick bite. The results showed that among the 4034 proteins and 1795 phosphorylated proteins identified, a total of 202 proteins and 435 phosphorylation sites were changed at all time points after H. longicornis bite. Regulated host proteins such as coronin, mannose receptor C-type 1, dendritic cell-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN), ezrin, and integrin play important roles in immune defence against tick bite. Fibrinogen, fibronectin, integrin-linked kinase (ILK), and tenascin C jointly regulate the coagulation and wound healing processes in the host. Trichohyalin and peptidyl arginine deiminase 3 were downregulated to induce hair loss in the host. In addition, the changes in the phosphorylation of interleukin-4 receptor, microtubule-actin crosslinking factor 1, and Nck adaptor protein 1 would play important roles in host immunity and vascular repair.

ORGANISM(S): Haemaphysalis Longicornis

SUBMITTER: Hui Wang

PROVIDER: PXD017694 | iProX | Mon Feb 24 00:00:00 GMT 2020

REPOSITORIES: iProX

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Comprehensive analysis of protein expression levels and phosphorylation levels in host skin in response to tick (Haemaphysalis longicornis) bite.

Zhang Xiaoli X Zhang Baowen B Masoudi Abolfazl A Wang Xiaoshuang X Xue Xiaomin X Li Mengxue M Xiao Qi Q Wang Minjing M Liu Jingze J Wang Hui H

Journal of proteomics 20200716

Ticks are parasitic arthropods that suck blood from the surface of most vertebrates. They can transmit a variety of pathogens. The blood sucking of ticks causes varying degrees of damage to the skin of the host. Proteins related to immune regulation, vascular repair, and wound healing in mammalian skin respond to tick bites by regulating their expression and post-translational modifications to protect the skin from injury. Phosphorylation of proteins, as the most common post-translational modifi ...[more]

PMID: 32682108

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Project description:Tick saliva contains many bioactive molecules that are involved in attachment to the host, blood feeding and transmission of pathogens. MicroRNAs (miRNAs) are a class of short non-coding RNAs with a length of 19-24 nucleotides. They act as regulators of gene expression by binding to their target mRNA at the post-transcriptional level and control a variety of cellular functions, including regulation of growth, metabolism, and development. The detection and characterizations of miRNAs from tick saliva may help explain the molecular mechanisms involved in the interaction between ticks, pathogens, and hosts. They may also contribute to the discovery of vaccines, which can control ticks and the pathogens they transmit. An RNA library was generated from the saliva of fed adult Haemaphysalis longicornis ticks, and it contained 17.4 million clean reads of 18-30 nucleotides. Overall, 319 known miRNAs and 1 novel miRNA were found. The 10 most abundantly expressed miRNAs present in tick saliva were miR-100_2, miR-315, miR-184_1, miR-100-5p_2, miR-5307, miR-184-3p_3, Let-7-5p_6, miR-71_5, miR-1-3p_6, and miR-10-5p_2. The miR-375, one of the abundantly expressed, was subjected to Quantitative Real-Time PCR analysis (qRT-PCR) in various tick developmental stages, as well as in different tissues isolated from adult ticks. The expression of miR-375 in different tick development stages was highest in unfed nymphs and lowest in the egg stage. In the tissues of adult ticks, miR-375 was most highly expressed in the salivary gland. To investigate the possible role of miR-375, Ant-375 was used to inhibit the miR-375. Treated group (Ant-375) had a reduced number of eggs (t(10)= 2.652, P=0.0242), eggs that were partially desiccated, and reduced egg hatchability (t(10)=2.272, P=0.044) compared to Ms-Ant and the non-injected control. This is the first study to investigate the miRNAs profile in tick saliva and the role of miR-375 in H. longicornis. The identification and characterization of miRNA in tick saliva may help to reveal the molecular mechanisms of interactions among ticks, pathogens, and hosts and suggest new vaccine strategies to control tick borne diseases.

Project description:Introduction: Ixodes 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. To address this issue, we measured 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. Results: Primary 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. Conclusions: Our 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 host response at the tick bite site and suggests both a potential model of the host cutaneous response and candidate genes for further description and investigation. Ear biopsies from BALB/cJ mice infested with Ixodes scapularis nymphs were assayed at 12, 48, 72, and 96 hours after infestation during a primary and secondary exposure. 3 mice were measured at each time point. Controls were 3 similarly housed but tick-free mice.

Dataset Information

Comprehensive Analysis of Protein Expression Levels and Phosphorylation Levels in Host Skin in Response to Tick (Haemaphysalis longicornis) Bite

Publications

Comprehensive analysis of protein expression levels and phosphorylation levels in host skin in response to tick (Haemaphysalis longicornis) bite.

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