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Lymphocyte subsets in Atlantic cod (Gadus morhua) interrogated by single-cell sequencing.

Naomi Croft Guslund1, Anders K Krabberød2,3, Simen F Nørstebø4

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Atlantic cod lack a key immune pathway but possess diverse T and B cells. This study reveals novel CD8-negative T cell subsets, crucial for understanding fish immunity.

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Area of Science:

  • Immunology
  • Aquatic Animal Health
  • Genomics

Background:

  • Atlantic cod (Gadus morhua) immune systems are unique due to the loss of the major histocompatibility complex class II presentation pathway.
  • Previous identification of CD8-positive T cells, B cells, and plasma cells highlights the need for deeper characterization of lymphocyte subsets to understand immune adaptations.
  • The absence of CD4-positive T lymphocytes in Atlantic cod necessitates further investigation into their immune cell repertoire.

Purpose of the Study:

  • To characterize lymphocyte heterogeneity in Atlantic cod spleen using single-cell RNA sequencing.
  • To identify and describe distinct T cell and B cell subsets.
  • To propose a differentiation trajectory for B cells in Atlantic cod.

Main Methods:

  • Single-cell RNA sequencing (scRNA-seq) was employed to analyze spleen lymphocytes from Atlantic cod.
  • Bioinformatic analyses were used to identify and classify different lymphocyte subpopulations.
  • Gene expression patterns were examined to define T and B cell subsets and their potential functions.

Main Results:

  • Five distinct T cell subsets and eight distinct B cell subsets were identified in the Atlantic cod spleen.
  • A B cell differentiation trajectory was proposed based on the observed subsets.
  • A novel subpopulation of CD8-negative T cells was discovered, with some subsets expressing monocyte chemotactic protein 1b homologue and others expressing scavenger receptor m130 homologue.

Conclusions:

  • The study uncovers significant lymphocyte heterogeneity in Atlantic cod, including novel CD8-negative T cell subsets.
  • These findings provide a foundational understanding of the diverse immune states within B and T cell populations in Atlantic cod.
  • Further research can build upon this detailed immune cell atlas to explore adaptive immunity in fish lacking MHC class II.