Performing Department
(N/A)
Non Technical Summary
Honey bees are ecologically vital in maintaining a balanced ecosystem. Honey bee colonies face many biotic and abiotic stressors, including parasites, pathogens, and chemical pesticides, which all have led to significant losses during the last two decades. The innate immune system, including hemocytes (immune cells), represents the first line of defense against invading pathogens. The innate immune system of bees is divided into cellular and humoral components, which protect against a wide range of invading microorganisms. Hemocytes, professional immune cells equivalent to vertebrate leukocytes, represent the cellular arm. Following microbial infection, hemocytes execute cell-mediated responses, including phagocytosis, encapsulation, and nodulation, clearing pathogens. The humoral component of the immune system produces soluble effector molecules that activate the complement pathway, prophenoloxidase pathway, and melanization cascade and produce reactive oxygen and nitrogen species. Due to the complexity of hemocyte typing in the honey bee, the immunological compartments and mechanisms of cellular immunity have not been investigated in detail. A singular objective of this seed grant is to gain a comprehensive understanding of the complex interactions involving the honey bee (Apis mellifera) hemocytes, and intracellular microsporidian parasites (Nosema apis & Nosema ceranae), with the ultimate goal of developing a hemocyte atlas to advance our knowledge of parasitic infection dynamics. Building upon our preliminary unpublished data, we hypothesize that intracellular microsporidian parasites manipulate the honey bee immune cells to facilitate systemic dissemination within the host bee. Objective 1 will generate single-cell RNA sequencing from uninfected and parasite-infected honey bee hemocytes for downstream analyses. Objective 2 will investigate the role of phagocytic and non-phagocytic hemocytes on honey bee survival against bacterial and parasitic infection. The outcome of data generated in this aim will constitute a valuable resource for the honey bee research community as a reference for gene function studies at single-cell resolution. The deep insights into bee hemocyte-Nosema interactions will contribute to our fundamental knowledge of bee infection dynamics and potentially uncover novel targets for intervention strategies. Additionally, the outcome of this project could be extrapolated to other solitary bee species of importance to U. S. agriculture and is expected to provide a critical comparative understanding of bee immunology.
Animal Health Component
100%
Research Effort Categories
Basic
85%
Applied
10%
Developmental
5%
Goals / Objectives
This project seeks to fill the knowledge gap of Honey Bee's innate immune system by gaining a comprehensive understanding of the complex interaction involving the honey bee hemocytes, and intracellular microsporidian parasites, with the ultimate goal of developing a hemocyte atlas to advance our knowledge of parasitic infection dynamics. This innovative project will integrate a multidisciplinary approach combining hemocyte biology, single-cell genomics, and translational functional genomics by persuing following objectives.Objective #1:To generate single-cell RNA sequencing from uninfected and parasite-infected honey bee hemocytes for downstream analyses.Objective #2:To investigate the role of phagocytic and non-phagocytic hemocytes on honey bee survival against bacterial and parasitic infection.
Project Methods
The proposed experiments will be conducted using adult honey bees. Hemolymph will be collected, and hemocytes will be isolated.Samples with >90% cell viability will be selected for 10X genomic libraries. Single-cell RNA Seq data will be processed andanalyzed, and a Clodronate-mediated depletion of phagocytic hemocytes will be conducted.