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Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells
Published on: July 23, 2017
Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus)
Eliane Esteves1, Flavio A Lara, Daniel M Lorenzini
1Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo. Av. Prof. Lineu Prestes, 1374, CEP 05508-900, São Paulo, SP, Brazil.
Abstract:
The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors, antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens.
Insights
Researchers characterized the BME26 cell line from the cattle tick Rhipicephalus (Boophilus) microplus. These cells exhibit immune responses and can be used to study tick-borne pathogens.
Area of Science:
- Veterinary Entomology
- Cell Biology
- Molecular Biology
Background:
- The cattle tick Rhipicephalus (Boophilus) microplus is a significant agricultural pest.
- Understanding tick cellular and molecular mechanisms is crucial for developing control strategies.
Purpose of the Study:
- To characterize the BME26 cell line derived from R. (B.) microplus embryos.
- To investigate the cellular and molecular features relevant to tick immunity and physiology.
Main Methods:
- Electron microscopy to examine cellular structures.
- Gene expression analysis via cDNA library sequencing and annotation.
- Functional assays including phagocytosis and gene transfection.
Main Results:
- BME26 cells possess glycogen, mitochondria, and vesicles involved in endocytosis and autophagy.
- Cells phagocytosed yeast and expressed antimicrobial peptide genes (microplusin, defensin).
- Sequencing identified 898 unique mRNA sequences, including novel transcripts and those related to immune response (e.g., ferritin, NADPH oxidase).
- Transfected cells expressed a reporter gene for up to 5 weeks.
Conclusions:
- The BME26 cell line is a valuable tool for studying R. (B.) microplus biology.
- This cell line can be used to analyze innate immune responses and other biological processes in ticks.
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