Effective isolation of GALT cells: Insights into the intestine immune response of rainbow trout (Oncorhynchus mykiss) to different bacterin vaccine preparations

Ahmed Attaya* (Corresponding Author), Christopher J. Secombes* (Corresponding Author), Tiehui Wang* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

The teleost gut is a multifunction complex structure that plays a pivotal immunological role in homeostasis and the maintenance of health, in addition to digestion of food and/or nutrient absorption. In vitro examination of the intestine leucocyte repertoire has the potential to aid our understanding of gut immune competence and allows a rapid screen of host-microorganism interactions in different immunological contexts. To explore this possibility, in the present study we investigated the response of isolated gut leucocytes to 4 bacterins of Aeromonas salmonicida, prepared from different strains, combinations and strains grown in different environments, in comparison to a Yersinia ruckeri bacterin for which a commercial/effective oral booster vaccine has been developed. To aid this study we also optimized further our method of GALT cell isolation from rainbow trout, so as to avoid mechanical clearance of the intestine contents. This drastically increased the cell yield from ∼12 × 106 to ∼210 × 106/fish with no change in the percent cell viability over time or presence of transcripts typical of the key leucocyte types needed for the study of immune modulation (i.e. T- and B-cells, dendritic cells and macrophages). A wide array of immune transcripts were modulated by the bacterins, demonstrating the diversity of GALT cell responses to bacterial stimulation. Indeed, the GALT leucocyte responses were sensitive enough to distinguish the different bacterial species, strains and membrane proteins, as seen by distinct kinetics of immune gene expression. However, the response of the GALT cells was often relatively slow and of a low magnitude compared to those of PBL. These results enhance our knowledge of the gut biocapacity and help validate the use of this model for screening of oral vaccine candidates.
Original languageEnglish
Pages (from-to)378-392
Number of pages14
JournalFish & Shellfish Immunology
Volume105
Early online date29 Jun 2020
DOIs
Publication statusE-pub ahead of print - 29 Jun 2020

Keywords

  • Rainbow trout
  • GALT cells
  • Cell yield
  • Leucocyte types
  • Aeromonas salmonicida
  • Yersinia ruckeri
  • Bacterin
  • Immune modulation
  • Gene expression

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