Germinal center dynamics revealed by multiphoton microscopy with a photoactivatable fluorescent reporter

Cell. 2010 Nov 12;143(4):592-605. doi: 10.1016/j.cell.2010.10.032.

Abstract

The germinal center (GC) reaction produces high-affinity antibodies by random mutation and selective clonal expansion of B cells with high-affinity receptors. The mechanism by which B cells are selected remains unclear, as does the role of the two anatomically defined areas of the GC, light zone (LZ) and dark zone (DZ). We combined a transgenic photoactivatable fluorescent protein tracer with multiphoton laser-scanning microscopy and flow cytometry to examine anatomically defined LZ and DZ B cells and GC selection. We find that B cell division is restricted to the DZ, with a net vector of B cell movement from the DZ to the LZ. The decision to return to the DZ and undergo clonal expansion is controlled by T helper cells in the GC LZ, which discern between LZ B cells based on the amount of antigen captured and presented. Thus, T cell help, and not direct competition for antigen, is the limiting factor in GC selection.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens / immunology
  • B-Lymphocytes / cytology
  • Female
  • Germinal Center / cytology*
  • Germinal Center / immunology*
  • Humans
  • Immunity, Humoral
  • Lymph Nodes / cytology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Spleen / cytology
  • T-Lymphocytes / cytology

Substances

  • Antigens

Associated data

  • GEO/GSM589872