IgT, a primitive immunoglobulin class specialized in mucosal immunity

Abstract

Teleost fish are the most primitive bony vertebrates that contain immunoglobulins. In contrast to mammals and birds, these species are devoid of immunoglobulin A (IgA) or a functional equivalent. This observation suggests that specialization of immunoglobulin isotypes into mucosal and systemic responses took place during tetrapod evolution. Challenging that paradigm, here we show that IgT, an immunoglobulin isotype of unknown function, acts like a mucosal antibody. We detected responses of rainbow trout IgT to an intestinal parasite only in the gut, whereas IgM responses were confined to the serum. IgT coated most intestinal bacteria. As IgT and IgA are phylogenetically distant immunoglobulins, their specialization into mucosal responses probably occurred independently by a process of convergent evolution.

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Figure 1: Purification and structural characterization of IgT.
Figure 2: Identification of a previously unknown B cell lineage that expresses only surface IgT.
Figure 3: Phagocytic and intracellular killing capacities of IgT+ B cells.
Figure 4: Proliferative and immunoglobulin-secreting capacities of IgT+ and IgM+ B cells in response to microbial stimulation.
Figure 5: Accumulation of IgT+ B cells in the GALT of fish that survived infection with C. shasta.
Figure 6: Immune responses in the gut of trout infected with C. shasta are mediated by the IgT system.
Figure 7: Most trout gut luminal bacteria are predominantly coated with IgT.
Figure 8: Gut mucus IgT and IgM associate with a trout pIgR.

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Acknowledgements

We thank G. Warr (National Science Foundation) for anti–trout IgM; D. Artis and P. Boudinot for critical reading of the manuscript; the Morphology Core of the Center for Molecular Studies in Digestive and Liver Diseases of the University of Pennsylvania, especially G. Swain, for help and advice on immunohistochemistry; C. Pletcher and the staff of the Flow Cytometry and Cell Sorting Facility of the University of Pennsylvania for cell sorting; and Y. Liu, L. Zhang and C. Zhou for technical assistance in cell transfection and immunofluorescence microscopy analyses. Supported by the National Science Foundation (NSF-MCB-0719599 to J.O.S.), the US National Institutes of Health (R01GM085207-01 to J.O.S.) and the United States Department of Agriculture (USDA-NRI 2006-01619 and USDA-NRI 2007-01719 to J.O.S.).

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Y.-A.Z. purified trout IgT, developed polyclonal and monoclonal antibodies to IgT and a polyclonal antibody to trout pIgR, analyzed the biochemical features of IgT and the gene-expression profiles of IgT+ and IgM+ cells, measured specific IgT and IgM titers to C. shasta and did all experiments involved in cloning and functional studies of trout pIgR; I.S. developed the protocols to obtain leukocytes and bacteria from trout GALT and the immunohistochemistry protocols to detect IgT+ and IgM+ cells and C. shasta in trout lymphoid tissues and did immunohistochemistry studies and flow cytometry of IgT+ and IgM+ cells in trout GALT and gut luminal bacteria; J.L. did the flow cytometry of IgT+ and IgM+ cells of trout lymphoid organs, measured the phagocytotic capacity and intracellular bacterial killing of B cells and contributed to the evaluation of total IgT and IgM concentrations in serum and gut mucus, together with Y.-A.Z.; D.P. analyzed IgT and IgM coating on gut bacteria, did the B cell proliferation studies and tested the production of IgT and IgM after stimulation with microbial products; S.B. infected fish with C. shasta and provided samples from survivor and control fish; Z.X. contributed to the evaluation of the production of IgT and IgM by cultured trout leukocytes and sorted B cells; S.E.L. and J.B. contributed to the experimental design and discussions related to C. shasta infection; J.O.S. designed the overall study, contributed to data analysis and wrote the main body of the paper; and all authors read and commented on the paper. J.L. and D.P. contributed equally to this work.

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Correspondence to J Oriol Sunyer.

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Supplementary Figures 1–9, Supplementary Table 1 and Supplementary Methods (PDF 1247 kb)

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Zhang, YA., Salinas, I., Li, J. et al. IgT, a primitive immunoglobulin class specialized in mucosal immunity. Nat Immunol 11, 827–835 (2010). https://doi.org/10.1038/ni.1913

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