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SAP-controlled T–B cell interactions underlie germinal centre formation


Generation of long-term antibody-mediated immunity depends on the germinal centre reaction, which requires cooperation between antigen-specific T and B lymphocytes. In human X-linked lymphoproliferative disease and its gene-targeted mouse model, loss-of-function mutations in signalling lymphocyte activation molecule-associated protein (SAP, encoded by SH2D1a) cause a profound defect in germinal centre formation by an as yet unknown mechanism. Here, using two-photon intravital imaging, we show that SAP deficiency selectively impairs the ability of CD4+ T cells to stably interact with cognate B cells but not antigen-presenting dendritic cells. This selective defect results in a failure of antigen-specific B cells to receive adequate levels of contact-dependent T-cell help to expand normally, despite Sap-/- T cells exhibiting the known characteristics of otherwise competent helper T cells. Furthermore, the lack of stable interactions with B cells renders Sap-/- T cells unable to be efficiently recruited to and retained in a nascent germinal centre to sustain the germinal centre reaction. These results offer an explanation for the germinal centre defect due to SAP deficiency and provide new insights into the bi-directional communication between cognate T and B cells in vivo.

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Figure 1: Sap-/- T cells normally interact with and are activated by DCs in vivo.
Figure 2: Sap -/- T cells are defective in adhesion to cognate B cells.
Figure 3: B cells fail to receive contact-dependent help from SAP-deficient T cells.
Figure 4: Defective GC recruitment and retention of Sap -/- T cells due to lack of efficient cognate interactions with B cells.

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H.Q. is in debt to Y. Hong for support, encouragement and inspiration. This work was funded by the intramural research programs of the National Institute of Allergy and Infectious Disease and National Human Genome Research Institute, National Institutes of Health, USA.

Author Contributions H.Q. and J.C. conducted the experiments. R.N.G. and P.L.S. contributed equally to this study. All authors contributed collectively to designing the experiments, interpreting the data and writing the paper.

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Correspondence to Pamela L. Schwartzberg or Ronald N. Germain.

Supplementary information

Supplementary Information 1

This files contains Supplementary Figures 1-10 with Legends and Supplementary Movie Legends 1-8 (PDF 18150 kb)

Supplementary Movie 1

Supplementary Movie 1, which shows sap+/+ and sap-/- T cell interactions with antigen-bearing DCs in the same lymph node. (MOV 2694 kb)

Supplementary Movie 2

Supplementary Movie 2, which shows sap+/+ and sap-/- T cell interactions with antigen-specific B cells in the same lymph node. (MOV 4422 kb)

Supplementary Movie 3

Supplementary Movie 3, which shows another example of sap+/+ and sap-/- T cell interactions with antigen-specific B cells in the same lymph node. (MOV 2852 kb)

Supplementary Movie 4

Supplementary Movie 4, which shows sap+/+ and sap-/- T cell interactions with antigen-specific B cells under non-competitive conditions. (MOV 10061 kb)

Supplementary Movie 5

Supplementary Movie 5, which shows examples of tracking B cells for their interaction histories with T cells. (MOV 11059 kb)

Supplementary Movie 6

Supplementary Movie 6, which shows further examples of tracking B cells for their interaction histories with T cells. (MOV 10038 kb)

Supplementary Movie 7

Supplementary Movie 7, which shows migration patterns of sap+/+ and sap-/- T cells within follicles containing non-cognate GCs. (QuickTime 9.0MB) (MOV 9244 kb)

Supplementary Movie 8

Supplementary Movie 8, which shows another example of migration patterns of sap+/+ and sap-/- T cells within follicles containing non-cognate GCs. (MOV 9621 kb)

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Qi, H., Cannons, J., Klauschen, F. et al. SAP-controlled T–B cell interactions underlie germinal centre formation. Nature 455, 764–769 (2008).

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