Protocol to identify and sort antigen-specific B cells from Plasmodium-infected mice

Area of Science:

• Immunology
• Parasitology
• Molecular Biology

Background:

• Malaria remains a significant global health challenge, necessitating detailed understanding of host immune responses.
• Identifying antigen-specific B cells is crucial for developing effective vaccines and therapies.
• Current methods may lack the specificity or throughput for comprehensive B cell analysis.

Purpose of the Study:

• To establish a protocol for isolating antigen-specific B cells in a rodent malaria model.
• To enable downstream multi-omic single-cell analyses of these B cells.
• To provide a reproducible method for malaria immunology research.

Main Methods:

• Development of SpyCage reagents targeting the Plasmodium falciparum MSP_1-19 antigen.
• Preparation of splenic single-cell suspensions from infected rodents.
• B cell enrichment and fluorescent staining for cell sorting.
• Utilizing flow cytometry for isolation of live, antigen-specific B cells.

Main Results:

• Successful identification and isolation of MSP_1-19-specific B cell populations.
• Demonstration of the protocol's compatibility with single-cell RNA sequencing, V(D)J sequencing, and ATAC sequencing.
• Generation of high-quality data for downstream molecular analyses.

Conclusions:

• The described protocol provides a robust method for isolating antigen-specific B cells in malaria research.
• This technique facilitates in-depth investigation of B cell responses at the single-cell level.
• The protocol is adaptable for studying other antigen-specific immune responses.