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Molecular tracking devices quantify antigen distribution and archiving in the lymph node.

ABSTRACT: Molecular tracking devices quantify antigen distribution and archiving in the lymph node.

PROVIDER: PRJNA633370 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12478200_1.fastq.gz	Fastqsanger.gz
	SRR12478200_2.fastq.gz	Fastqsanger.gz
	SRR12478201_1.fastq.gz	Fastqsanger.gz
	SRR12478201_2.fastq.gz	Fastqsanger.gz
	SRR12478202_1.fastq.gz	Fastqsanger.gz

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Molecular tracking devices quantify antigen distribution and archiving in the lymph node.

Project description:Live, attenuated vaccines generate humoral and cellular immune memory, increasing the duration of protective immune memory. We previously found that antigens derived from vaccination or viral infection persist beyond infection clearance via acquisition and retention (“archiving”) in lymphatic endothelial cells (LECs). Archived antigen maintains an effector-like pool of antigen-specific memory T cells, enhancing clearance of subsequent infections. Technical limitations of fluorescent labeling have precluded a complete picture of antigen archiving across cell types in lymph tissue. We developed a “molecular tracking device” to follow the distribution, acquisition, and retention of antigen in the lymph node. We immunized mice with an antigen conjugated to a nuclease-resistant DNA tag and used single-cell mRNA sequencing to quantify its abundance across major cell types in the lymph node two weeks later, validating its acquisition by migratory dendritic cells and durable retention by LECs and unexpected stromal cell types. Variable antigen levels in LECs enabled the identification and validation of caveolar endocytosis as a mechanism of antigen acquisition. Molecular tracking devices enable new approaches to study dynamic tis-sue dissemination at cellular resolution in vivo. We developed a 'molecular tracking device' to follow the distribution, acquisition, and retention of antigen across cell types in the lymph node. We immunized mice with an antigen conjugated to a nuclease-resistant DNA tag and used single-cell mRNA sequencing to quantify its abundance in immune and stromal cell types two weeks later, validating its retention by LECs and transfer to migratory dendritic cells and defining new reservoirs of archived antigen in stromal cells. Molecular tracking devices broadly enable new approaches to study molecular dissemination and retention in live animals.

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