Genetic analysis of basophil function in vivo

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Abstract

Contributions by basophils to allergic and helminth immunity remain incompletely defined. Using sensitive interleukin 4 (Il4) reporter alleles, we demonstrate here that basophil IL-4 production occurs by a CD4+ T cell–dependent process restricted to the peripheral tissues affected. We genetically marked and achieved specific deletion of basophils and found that basophils did not mediate T helper type 2 (TH2) priming in vivo. Two-photon imaging confirmed that basophils did not interact with antigen-specific T cells in lymph nodes but engaged in prolonged serial interactions with T cells in lung tissues. Although targeted deletion of IL-4 and IL-13 in either CD4+ T cells or basophils had a minimal effect on worm clearance, deletion from both lineages demonstrated a nonredundant role for basophil cytokines in primary helminth immunity.

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Figure 1: IL-4 production by basophils after parasite infection is restricted to affected tissues.
Figure 2: CD4+ T cell activation induces basophil IL-4 production in vitro.
Figure 3: Basophil lineage tracking and deletion in vivo.
Figure 4: Priming of CD4+ T cells in the absence of basophils.
Figure 5: Basophils and CD4+ T cells interact in the lungs but not in the lymph nodes.
Figure 6: Basophil-derived cytokines contribute to anti-helminth immunity.

Change history

  • 26 May 2011

    In the version of this article initially published, the label ‘DEL-OV’ in the bottom left graph in Figure 5c is incorrect. The correct label is 'DEL-OVA'. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank K.C. Lim and N. Flores for technical support; E. Thornton and M. Krummel for training in lung-slice preparation; D. Kioussis (Medical Research Council National Institute for Medical Research) for huCD2-DsRed transgenic mice; and J. Cyster (University of California, San Francisco) for mice and two-photon microscope use. Supported by the US National Institutes of Health (AI026918 and AI077439), the Howard Hughes Medical Institute and the Sandler Asthma Basic Research Center at the University of California San Francisco, San Francisco.

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Authors

Contributions

B.M.S., H.-E.L., C.D.C.A. and R.M.L. conceived of the work; H.-E.L. generated Basoph8 reporter mice; B.M.S. designed and did most experiments; C.D.C.A. contributed two-photon imaging data; J.K.B. and B.M.S. analyzed basophils in the small intestine; D.W. and B.M.S. generated data from mice infected with S. mansoni cercariae; L.E.C. analyzed mast cells in the skin in Basoph8 mice; J.K.M. provided S. mansoni cercariae and eggs; and B.M.S., C.D.C.A. and R.M.L. wrote the manuscript.

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Correspondence to Christopher D C Allen or Richard M Locksley.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Figures 1–11 (PDF 1074 kb)

Supplementary Video 1

Basophils and antigen-specific CD4+ T cells do not interact in draining lymph nodes after primary immunization with S. mansoni eggs and OVA. A time-lapse sequence of 24 μm z-projection images from an inguinal lymph node explant is shown approximately three days after subcutaneous immunization. Elapsed time is indicated as hh:mm:ss. (MOV 7192 kb)

Supplementary Video 2

Basophils and antigen-specific CD4+ T cells do not interact in draining lymph nodes after primary immunization with papain and OVA. A time-lapse sequence of 32 μm z-projection images from an inguinal lymph node explant is shown approximately 2.25 days after subcutaneous immunization. Elapsed time is indicated as hh:mm:ss. (MOV 5385 kb)

Supplementary Video 3

Antigen-specific CD4+ T cells interact with cognate antigen-specific B cells, but not basophils, in draining lymph nodes after primary immunization. A time-lapse sequence of 33 μm z-projection images from an inguinal lymph node explant is shown approximately 3.7 days after subcutaneous immunization with a mixture of papain and DEL-OVA. Elapsed time is indicated as hh:mm:ss. (MOV 10866 kb)

Supplementary Video 4

Basophils and antigen-specific CD4+ T cells engage in multiple serial interactions in the lung after primary N. brasiliensis infection. A time-lapse sequence of 29 μm z-projection images from a lung slice is shown approximately 7.5 days after N. brasiliensis infection, with intranasal OVA administered on days 1 and 6. Elapsed time is indicated as hh:mm:ss. The video corresponds to the still images shown in Fig. 5a and the characteristics of the serial encounters are quantified in Fig. 5d,e and Supplementary Fig. 8. (MOV 12493 kb)

Supplementary Video 5

Basophils and polyclonal T cells engage in multiple serial interactions in the lung after primary N. brasiliensis infection. A time-lapse sequence of 26 μm z-projection images from a lung slice is shown approximately 7.25 days after N. brasiliensis infection of a Basoph8 mouse carrying a hCD2-dsRed transgene. This transgene is highly expressed in T cells and weakly expressed in subsets of dendritic cells, macrophages, NK cells, and other undefined cell types. In this image sequence, a single green basophil and bright red T cell are shown interacting in the center, with a weakly fluorescent macrophage off to the right side. Elapsed time is indicated as hh:mm:ss. (MOV 4431 kb)

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Sullivan, B., Liang, HE., Bando, J. et al. Genetic analysis of basophil function in vivo. Nat Immunol 12, 527–535 (2011). https://doi.org/10.1038/ni.2036

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