Scabies mite peritrophins are potential targets of human host innate immunity

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

Research output: Contribution to journalArticleResearchpeer-review

4 Downloads (Pure)

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.
@article{823833774fcd46ce83cf661449b8bb61,
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.",
keywords = "chitin, complementary DNA, immunoglobulin G, mannan binding lectin, scabies mite peritrophin 1, secretory protein, unclassified drug, complement, insect protein, amino acid sequence, article, binding affinity, binding site, complement activation, controlled study, deglycosylation, DNA library, human, human tissue, immune response, immunohistochemistry, innate immunity, mite, nonhuman, nucleotide sequence, protein binding, protein expression, protein glycosylation, protein localization, protein targeting, scabies, affinity chromatography, animal, chemistry, DNA sequence, gastrointestinal tract, genetics, host parasite interaction, immunology, metabolism, molecular genetics, protein tertiary structure, Sarcoptes scabiei, Animals, Chitin, Chromatography, Affinity, Complement System Proteins, DNA, Complementary, Gastrointestinal Tract, Host-Parasite Interactions, Humans, Immunity, Innate, Insect Proteins, Molecular Sequence Data, Protein Binding, Protein Structure, Tertiary, Scabies, Sequence Analysis, DNA",
author = "Angela Mika and Priscilla Goh and Deborah Holt and Kemp, {David J} and K FISCHER",
year = "2011",
doi = "10.1371/journal.pntd.0001331",
language = "English",
volume = "5",
pages = "1--11",
journal = "PLoS Neglected Tropical Diseases",
issn = "1935-2727",
publisher = "Public Library of Science (PLoS)",
number = "9",

}

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 -