Diversity and functional specialization of oyster immune cells uncovered by integrative single-cell level investigations

Abstract

Mollusks are a major component of animal biodiversity and play a critical role in ecosystems and global food security. The Pacific oyster, Crassostrea (Magallana) gigas , is the most farmed bivalve mollusk in the world and is becoming a model species for invertebrate biology. Despite the extensive research on hemocytes, the immune cells of bivalves, their characterization remains elusive. Here, we were able to extensively characterize the diverse hemocytes and identified at least seven functionally distinct cell types and three hematopoietic lineages. A combination of single-cell RNA sequencing, quantitative cytology, cell sorting, functional assays, and pseudo-time analyses was used to deliver a comprehensive view of the distinct hemocyte types. This integrative analysis enabled us to reconcile molecular and cellular data and identify distinct cell types performing specialized immune functions, such as phagocytosis, reactive oxygen species production, copper accumulation, and expression of antimicrobial peptides. This study emphasized the need for more in depth studies of cellular immunity in mollusks and non-model invertebrates and set the ground for further comparative immunology studies at the cellular level. Pacific oysters are vital to the ecosystem. They are also a popular seafood and are increasingly used in life science research as a model to represent animals without a backbone (known as invertebrates). However, these oysters are prone to deadly infections caused by bacteria and viruses.Like humans, oysters and other invertebrates need an immune system to fight infections. Immune cells called hemocytes – which travel through the oyster’s body in a blood-like fluid called hemolymph – help eliminate bacteria, viruses and other disease-causing microbes by absorbing them, releasing toxic molecules, and producing natural antibiotics called antimicrobial peptides. However, it is still unclear how many types of hemocytes Pacific oysters have or what each type does.De La Forest Divonne et al. used single-cell RNA sequencing and other cell biology techniques to study the genetic activity and anatomy of immune cells in the Pacific oyster. The experiments confirmed that the oysters have at least seven distinct types of hemocytes, each with a specialized immune role, for example, some eat microbes while others produce antimicrobial peptides. The team also mapped how immature hemocytes develop into mature hemocytes with these specialist roles.This work provides the first detailed atlas of oyster immune cells and reveals how their immune system is organized and operates. A deeper understanding of oyster immune cells may guide the development of new strategies to reduce disease outbreaks in farmed or wild oysters. Before these benefits can be realized, future studies must test how each type of hemocyte responds to actual infections and explore whether targeted treatments or breeding programs can enhance the immune systems of farmed oysters.