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Elife. 2018 Aug 1;7. pii: e33051. doi: 10.7554/eLife.33051.

Capturing change in clonal composition amongst single mouse germinal centers.

Author information

1
Cleveland Clinic Lerner College of Medicine, Cleveland, United States.
2
Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, United States.
3
Howard Hughes Medical Institute, Maryland, United States.
4
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
5
Edwin L. Steele Laboratories for Tumor Biology, Department of Radiation Oncology, Massachusetts General Hospital, Boston, United States.
6
Department of Pediatrics, Harvard Medical School, Boston, United States.

Abstract

Understanding cellular processes occurring in vivo on time scales of days to weeks requires repeatedly interrogating the same tissue without perturbing homeostasis. We describe a novel setup for longitudinal intravital imaging of murine peripheral lymph nodes (LNs). The formation and evolution of single germinal centers (GCs) was visualized over days to weeks. Naïve B cells encounter antigen and form primary foci, which subsequently seed GCs. These experience widely varying rates of homogenizing selection, even within closely confined spatial proximity. The fluidity of GCs is greater than previously observed with large shifts in clonality over short time scales; and loss of GCs is a rare, observable event. The observation of contemporaneous, congruent shifts in clonal composition between GCs within the same animal suggests inter-GC trafficking of memory B cells. This tool refines approaches to resolving immune dynamics in peripheral LNs with high temporospatial resolution and minimal perturbation of homeostasis.

KEYWORDS:

B cell clonal development; germinal centers; immunology; inflammation; intravital imaging; mouse

PMID:
30066671
PMCID:
PMC6070335
DOI:
10.7554/eLife.33051
[Indexed for MEDLINE]
Free PMC Article

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