Development of imaging-based techniques to examine signalling properties of channel catfish (Ictalurus punctatus) leukocyte immune-type receptors (IpLITRs)

• Author / Creator

  Jones, Lena O.

• Cells of the innate immune system have the ability to rapidly detect and eliminate pathogens using surface-expressed immunoregulatory receptors that translate extracellular binding into effector functions such as degranulation, cytokine secretion, cell-mediated cytotoxicity, and phagocytosis. Teleost leukocyte immune-type receptors (LITRs) are a family of immunoregulatory receptors capable of inducing effector responses when heterologously expressed in mammalian immune cells. The mechanisms by which these receptors induce effector responses remain unknown; however, these effector responses are all dependent on cytoskeletal rearrangements. Many of the intracellular signaling components that facilitate cytoskeleton remodelling are conserved through evolution due to the ubiquitous requirement for cells to change shape or move. The main objective of my thesis was to develop imaging-based assays to examine LITR-induced signaling activities that culminate in effector responses such as phagocytosis. Using the phagocytic process as a bioassay, I developed experimental strategies employing cellular transfections, imaging flow cytometry, and confocal microscopy to monitor the LITR-mediated phagocytic process and associated signaling. I showed that two LITR-types induced very distinct cytoskeletal remodelling patterns to induce a phagocytic response. Additionally, cytoskeletal remodelling was mirrored by the distribution of a phospholipid species known to participate in the induction of filamentous actin (the main cytoskeletal component) polymerization. Overall, my work set the stage for testing potential signaling activities that IpLITRs are believed to induce.

• Subjects / Keywords

  • Leukocyte immune-type receptors
  • Imaging
  • Phagocytosis
  • Flow cytometry

• Graduation date

  Spring 2017

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R3RJ49661

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.