Journal Article

Characterization of Anopheles gambiae immune cells through genetic and functional immunophenotyping

Kwon, Hyeogsun, Smith, Ryan C., Samantsidis, George-Rafael

2025

Overview

Mosquito immune cells, or hemocytes, are integral components of the innate immune responses that define vector competence. To date, the characterization and functional classification of hemocytes has been hindered by the limited availability of genetic resources. Here, we map the composition of mosquito hemocytes by engineering five transgenic Anopheles gambiae lines that express fluorescent proteins under the control of candidate hemocyte promoters. We characterize these five transgenic lines through gene expression and microscopy-based approaches, and examine mosquito immune cell populations by leveraging advanced spectral imaging flow cytometry. We classify mosquito hemocytes into twelve distinct populations based on size, granularity, and ploidy, while defining these hemocyte subtypes based on their phagocytic capacity and the expression of genetic markers. By simultaneously analyzing these morphological and genetic properties, our work highlights the complexity and plasticity of mosquito hemocytes and provides the foundation for deeper investigations into their roles in immunity and pathogen transmission. Classification of mosquito hemocytes into subtypes has thus far relied on morphological properties. Here, the authors develop new genetic resources that facilitate the visualization and analysis of hemocyte subtypes in Anopheles gambiae . By combining the expression of hemocyte-specific genetic markers with the analysis of morphology, ploidy, and phagocytic capacity, the authors identify twelve distinct subtypes, revealing the complexity and plasticity of these immune cells with unprecedented resolution.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 14/63 / 38/1 / 38/35 / 38/39 / 45/71 / 631/208/200 / 631/250/2502/248 / 631/250/2503 / 64 / Adults / Animals / Animals, Genetically Modified / Anopheles - cytology / Anopheles - genetics / Anopheles - immunology / Anopheles gambiae / Aquatic insects / Cells / Classification / Complexity / Female / Females / Flow Cytometry / Fluorescence / Gene expression / Genetic analysis / Genetic engineering / Genetic markers / Genetic resources / Granulocytes / Hemocytes / Hemocytes - classification / Hemocytes - cytology / Hemocytes - immunology / Hemocytes - metabolism / Homeostasis / Humanities and Social Sciences / Immune response / Immune system / Immunity, Innate / Immunophenotyping - methods / Innate immunity / Insects / Microscopy / Morphology / Mosquitoes / multidisciplinary / Pathogens / Phagocytes / Phagocytosis / Plastic properties / Plasticity / Ploidy / Populations / Quorum sensing / Science / Science (multidisciplinary)