TY - JOUR
T1 - Germinal center dynamics revealed by multiphoton microscopy with a photoactivatable fluorescent reporter
AU - Victora, Gabriel D.
AU - Schwickert, Tanja A.
AU - Fooksman, David R.
AU - Kamphorst, Alice O.
AU - Meyer-Hermann, Michael
AU - Dustin, Michael L.
AU - Nussenzweig, Michel C.
N1 - Funding Information:
We would like to thank J. Lippincott-Schwartz (NIH), R. Tsien (UCSD), and B. Schaefer (USU) for the PA-GFP, tdTomato, and UbC constructs, respectively; the MSKCC Genomics Facility for performing the microarray experiments; J. Zavadil (NYU) for guidance in microarray analysis; A. North and S. Bhuvanendran (Rockefeller University Bio-Imaging Resource Center) for help with multiphoton imaging; R. Scheuerman (University of Texas Southwestern Medical Center) for ACS gene lists for GSEA analysis; and T. Cameron for helpful discussion. This work was supported in part by NIH grants R01 AI072529 (M.C.N. and M.L.D.) and P01 AI071195 (M.L.D.). Support for the Rockefeller University multiphoton microscope and cell sorter was granted by the Empire State Stem Cell Fund through NYSDOH Contract #C023046. D.R.F. was supported by NIH training grant CA009161-34. M.M.-H. was supported by the EU-NEST project MAMOCELL in FP6. M.C.N. is an HHMI Investigator.
PY - 2010/11/12
Y1 - 2010/11/12
N2 - 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.
AB - 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.
KW - Molimmuno
UR - https://www.scopus.com/pages/publications/78149477766
UR - https://www.scopus.com/pages/publications/78149477766#tab=citedBy
U2 - 10.1016/j.cell.2010.10.032
DO - 10.1016/j.cell.2010.10.032
M3 - Article
C2 - 21074050
AN - SCOPUS:78149477766
SN - 0092-8674
VL - 143
SP - 592
EP - 605
JO - Cell
JF - Cell
IS - 4
ER -