Scabies mite peritrophins are potential targets of human host innate immunity

Angela Mika, Priscilla Goh, Deborah Holt, David J Kemp, K FISCHER

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    Abstract

    Background: Pruritic scabies lesions caused by Sarcoptes scabiei burrowing in the stratum corneum of human skin facilitate opportunistic bacterial infections. Emerging resistance to current therapeutics emphasizes the need to identify novel targets for protective intervention. We have characterized several protein families located in the mite gut as crucial factors for host-parasite interactions. Among these multiple proteins inhibit human complement, presumably to avoid complement-mediated damage of gut epithelial cells. Peritrophins are major components of the peritrophic matrix often found in the gut of arthropods. We hypothesized that a peritrophin, if abundant in the scabies mite gut, could be an activator of complement.

    Methodology/Principal Findings: A novel full length scabies mite peritrophin (SsPTP1) was identified in a cDNA library from scabies mites. The amino acid sequence revealed four putative chitin binding domains (CBD). Recombinant expression of one CBD of the highly repetitive SsPTP1 sequence as TSP-hexaHis-fusion protein resulted in soluble protein, which demonstrated chitin binding activity in affinity chromatography assays. Antibodies against a recombinant SsPTP1 fragment were used to immunohistochemically localize native SsPTP1 in the mite gut and in fecal pellets within the upper epidermis, co-localizing with serum components such as host IgG and complement. Enzymatic deglycosylation confirmed strong N- and O-glycosylation of the native peritrophin. Serum incubation followed by immunoblotting with a monoclonal antibody against mannan binding lectin (MBL), the recognition molecule of the lectin pathway of human complement activation, indicated that MBL may specifically bind to glycosylated SsPTP1.

    Conclusions/Significance: This study adds a new aspect to the accumulating evidence that complement plays a major role in scabies mite biology. It identifies a novel peritrophin localized in the mite gut as a potential target of the lectin pathway of the complement cascade. These initial findings indicate a novel role of scabies mite peritrophins in triggering a host innate immune response within the mite gut.
    Original languageEnglish
    Article numbere1331
    Pages (from-to)1-11
    Number of pages11
    JournalPLoS Neglected Tropical Diseases
    Volume5
    Issue number9
    DOIs
    Publication statusPublished - 2011

    Fingerprint

    Scabies
    Mites
    Innate Immunity
    Chitin
    Mannose-Binding Lectin
    Proteins
    Mannose-Binding Lectin Complement Pathway
    Sarcoptes scabiei
    Host-Parasite Interactions
    Arthropods
    Nucleic Acid Repetitive Sequences
    Complement Activation
    Opportunistic Infections
    Serum
    Gene Library
    Glycosylation
    Affinity Chromatography
    Bacterial Infections
    Immunoblotting
    Lectins

    Cite this

    Mika, Angela ; Goh, Priscilla ; Holt, Deborah ; Kemp, David J ; FISCHER, K. / Scabies mite peritrophins are potential targets of human host innate immunity. In: PLoS Neglected Tropical Diseases. 2011 ; Vol. 5, No. 9. pp. 1-11.
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    title = "Scabies mite peritrophins are potential targets of human host innate immunity",
    abstract = "Background: Pruritic scabies lesions caused by Sarcoptes scabiei burrowing in the stratum corneum of human skin facilitate opportunistic bacterial infections. Emerging resistance to current therapeutics emphasizes the need to identify novel targets for protective intervention. We have characterized several protein families located in the mite gut as crucial factors for host-parasite interactions. Among these multiple proteins inhibit human complement, presumably to avoid complement-mediated damage of gut epithelial cells. Peritrophins are major components of the peritrophic matrix often found in the gut of arthropods. We hypothesized that a peritrophin, if abundant in the scabies mite gut, could be an activator of complement. Methodology/Principal Findings: A novel full length scabies mite peritrophin (SsPTP1) was identified in a cDNA library from scabies mites. The amino acid sequence revealed four putative chitin binding domains (CBD). Recombinant expression of one CBD of the highly repetitive SsPTP1 sequence as TSP-hexaHis-fusion protein resulted in soluble protein, which demonstrated chitin binding activity in affinity chromatography assays. Antibodies against a recombinant SsPTP1 fragment were used to immunohistochemically localize native SsPTP1 in the mite gut and in fecal pellets within the upper epidermis, co-localizing with serum components such as host IgG and complement. Enzymatic deglycosylation confirmed strong N- and O-glycosylation of the native peritrophin. Serum incubation followed by immunoblotting with a monoclonal antibody against mannan binding lectin (MBL), the recognition molecule of the lectin pathway of human complement activation, indicated that MBL may specifically bind to glycosylated SsPTP1. Conclusions/Significance: This study adds a new aspect to the accumulating evidence that complement plays a major role in scabies mite biology. It identifies a novel peritrophin localized in the mite gut as a potential target of the lectin pathway of the complement cascade. These initial findings indicate a novel role of scabies mite peritrophins in triggering a host innate immune response within the mite gut.",
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    author = "Angela Mika and Priscilla Goh and Deborah Holt and Kemp, {David J} and K FISCHER",
    year = "2011",
    doi = "10.1371/journal.pntd.0001331",
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    Scabies mite peritrophins are potential targets of human host innate immunity. / Mika, Angela; Goh, Priscilla; Holt, Deborah; Kemp, David J; FISCHER, K.

    In: PLoS Neglected Tropical Diseases, Vol. 5, No. 9, e1331, 2011, p. 1-11.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Scabies mite peritrophins are potential targets of human host innate immunity

    AU - Mika, Angela

    AU - Goh, Priscilla

    AU - Holt, Deborah

    AU - Kemp, David J

    AU - FISCHER, K

    PY - 2011

    Y1 - 2011

    N2 - Background: Pruritic scabies lesions caused by Sarcoptes scabiei burrowing in the stratum corneum of human skin facilitate opportunistic bacterial infections. Emerging resistance to current therapeutics emphasizes the need to identify novel targets for protective intervention. We have characterized several protein families located in the mite gut as crucial factors for host-parasite interactions. Among these multiple proteins inhibit human complement, presumably to avoid complement-mediated damage of gut epithelial cells. Peritrophins are major components of the peritrophic matrix often found in the gut of arthropods. We hypothesized that a peritrophin, if abundant in the scabies mite gut, could be an activator of complement. Methodology/Principal Findings: A novel full length scabies mite peritrophin (SsPTP1) was identified in a cDNA library from scabies mites. The amino acid sequence revealed four putative chitin binding domains (CBD). Recombinant expression of one CBD of the highly repetitive SsPTP1 sequence as TSP-hexaHis-fusion protein resulted in soluble protein, which demonstrated chitin binding activity in affinity chromatography assays. Antibodies against a recombinant SsPTP1 fragment were used to immunohistochemically localize native SsPTP1 in the mite gut and in fecal pellets within the upper epidermis, co-localizing with serum components such as host IgG and complement. Enzymatic deglycosylation confirmed strong N- and O-glycosylation of the native peritrophin. Serum incubation followed by immunoblotting with a monoclonal antibody against mannan binding lectin (MBL), the recognition molecule of the lectin pathway of human complement activation, indicated that MBL may specifically bind to glycosylated SsPTP1. Conclusions/Significance: This study adds a new aspect to the accumulating evidence that complement plays a major role in scabies mite biology. It identifies a novel peritrophin localized in the mite gut as a potential target of the lectin pathway of the complement cascade. These initial findings indicate a novel role of scabies mite peritrophins in triggering a host innate immune response within the mite gut.

    AB - Background: Pruritic scabies lesions caused by Sarcoptes scabiei burrowing in the stratum corneum of human skin facilitate opportunistic bacterial infections. Emerging resistance to current therapeutics emphasizes the need to identify novel targets for protective intervention. We have characterized several protein families located in the mite gut as crucial factors for host-parasite interactions. Among these multiple proteins inhibit human complement, presumably to avoid complement-mediated damage of gut epithelial cells. Peritrophins are major components of the peritrophic matrix often found in the gut of arthropods. We hypothesized that a peritrophin, if abundant in the scabies mite gut, could be an activator of complement. Methodology/Principal Findings: A novel full length scabies mite peritrophin (SsPTP1) was identified in a cDNA library from scabies mites. The amino acid sequence revealed four putative chitin binding domains (CBD). Recombinant expression of one CBD of the highly repetitive SsPTP1 sequence as TSP-hexaHis-fusion protein resulted in soluble protein, which demonstrated chitin binding activity in affinity chromatography assays. Antibodies against a recombinant SsPTP1 fragment were used to immunohistochemically localize native SsPTP1 in the mite gut and in fecal pellets within the upper epidermis, co-localizing with serum components such as host IgG and complement. Enzymatic deglycosylation confirmed strong N- and O-glycosylation of the native peritrophin. Serum incubation followed by immunoblotting with a monoclonal antibody against mannan binding lectin (MBL), the recognition molecule of the lectin pathway of human complement activation, indicated that MBL may specifically bind to glycosylated SsPTP1. Conclusions/Significance: This study adds a new aspect to the accumulating evidence that complement plays a major role in scabies mite biology. It identifies a novel peritrophin localized in the mite gut as a potential target of the lectin pathway of the complement cascade. These initial findings indicate a novel role of scabies mite peritrophins in triggering a host innate immune response within the mite gut.

    KW - chitin

    KW - complementary DNA

    KW - immunoglobulin G

    KW - mannan binding lectin

    KW - scabies mite peritrophin 1

    KW - secretory protein

    KW - unclassified drug

    KW - complement

    KW - insect protein

    KW - amino acid sequence

    KW - article

    KW - binding affinity

    KW - binding site

    KW - complement activation

    KW - controlled study

    KW - deglycosylation

    KW - DNA library

    KW - human

    KW - human tissue

    KW - immune response

    KW - immunohistochemistry

    KW - innate immunity

    KW - mite

    KW - nonhuman

    KW - nucleotide sequence

    KW - protein binding

    KW - protein expression

    KW - protein glycosylation

    KW - protein localization

    KW - protein targeting

    KW - scabies

    KW - affinity chromatography

    KW - animal

    KW - chemistry

    KW - DNA sequence

    KW - gastrointestinal tract

    KW - genetics

    KW - host parasite interaction

    KW - immunology

    KW - metabolism

    KW - molecular genetics

    KW - protein tertiary structure

    KW - Sarcoptes scabiei

    KW - Animals

    KW - Chitin

    KW - Chromatography, Affinity

    KW - Complement System Proteins

    KW - DNA, Complementary

    KW - Gastrointestinal Tract

    KW - Host-Parasite Interactions

    KW - Humans

    KW - Immunity, Innate

    KW - Insect Proteins

    KW - Molecular Sequence Data

    KW - Protein Binding

    KW - Protein Structure, Tertiary

    KW - Scabies

    KW - Sequence Analysis, DNA

    U2 - 10.1371/journal.pntd.0001331

    DO - 10.1371/journal.pntd.0001331

    M3 - Article

    VL - 5

    SP - 1

    EP - 11

    JO - PLoS Neglected Tropical Diseases

    JF - PLoS Neglected Tropical Diseases

    SN - 1935-2727

    IS - 9

    M1 - e1331

    ER -