Project description:This project is focused on isolation and characterization of polar tube proteins (PTPs) of Anncaliia algerae using diverse bioinformatic and experimental methodologies - from mass spectrometry to heterologous expression to antibody production and immunolocalization. A novel protein was isolated from A. algerae polar tube extracts named PTP6 which could participates in recognition of host cell receptors as well.Moreover the obtained data on immunolabeling of the polar tubes in the process of extrusion, strongly support one of three hypothetical mechanisms for spore firing, namely the “everting-like-a-glove finger” model. The practical result of this fundamental research is the elaboration of the novel workflow that allows identification and isolation of PTPs in a given microsporidian species wich unites different approaches, such as homology search, sporal protein extractions by thiol-reducing agents, antibody-based approaches, as well as a protocol for purification of extruded polar tubes combined with mass spectrometry to identify new polar tube components.
Project description:We describe a biopsy proven case of microsporidial infection of the false vocal cords in a 69-yr-old male with a history of chronic lymphocytic leukemia. The patient had hoarseness for several weeks before his admission to the hospital for shortness of breath. He had received chemotherapy with fludarabine 6 wk before this hospital admission. A biopsy of vocal cord nodules demonstrated an organism that was identified as Anncaliia algerae by electron microscopy. Molecular analysis of the small subunit RNA gene amplified by polymerase chain reaction further confirmed the identification of this organism as A. algerae. This case illustrates the ability of this insect pathogen to cause disease in immune-compromised mammalian hosts.
Project description:We report a case of Anncaliia algerae microsporidia infection in an immunosuppressed kidney transplant recipient in China. Light microscopy and transmission electron microscopy initially failed to identify A. algerae, which eventually was detected by metagenomic next-generation sequencing. Our case highlights the supporting role of metagenomic sequencing in early identification of uncommon pathogens.
Project description:All microsporidia share a unique, extracellular spore stage, containing the infective sporoplasm and the apparatus for initiating infection. The polar filament/polar tube when exiting the spore transports the sporoplasm through it into a host cell. While universal, these structures and processes have been enigmatic. This study utilized several types of microscopy, describing and extending our understanding of these structures and their functions. Cryogenically preserved polar tubes vary in diameter from 155 to over 200 nm, noticeably larger than fixed-sectioned or negatively stained samples. The polar tube surface is pleated and covered with fine fibrillar material that projects from the surface and is organized in clusters or tufts. These fibrils may be the sites of glycoproteins providing protection and aiding infectivity. The polar tube surface is ridged with 5-6 nm spacing between ridges, enabling the polar tube to rapidly increase its diameter to facilitate the passage of the various cargo including cylinders, sacs or vesicles filled with particulate material and the intact sporoplasm containing a diplokaryon. The lumen of the tube is lined with a membrane that facilitates this passage. Careful examination of the terminus of the tube indicates that it has a closed tip where the membranes for the terminal sac are located.