Depletion of conventional mature B cells and compromised specific antibody response in bovine immunoglobulin μ heavy-chain transgenic mice

Abstract

In this study, we introduced the bovine immunoglobulin μ heavy-chain gene (the orphaned gene on BTA11) into mouse germline cells. Bovine IgM was highly expressed in selected transgenic lines, and it largely inhibited rearrangements of the endogenous immunoglobulin heavy chain (IgH) genes in these lines. The forced expression of bovine IgM resulted in reduced numbers of pro- and pre-B cells but increased the number of immature B cells in the transgenic mice. Bovine IgM-expressing B cells can migrate from the bone marrow to the spleen, but most of the cells are arrested at the T1 transitional B cell stage, leading to a significantly lower number of T2 transitional and mature B cells in the spleen. Although the serum concentrations of endogenous IgM and IgG in the transgenic mice were significantly decreased, the IgA levels were slightly increased compared to the WT mice. The bovine IgM level in the serum was only one-tenth to one-fifth of that of endogenous mouse IgM, suggesting that most of the serum immunoglobulin were contributed by endogenous IgH gene-expressing B cells. These transgenic mice also exhibited a lower frequency of unique complementarity determining region 3 (CDR3) sequences in their VH repertoire and Vκ repertoire but exhibited an increased frequency of unique CDR3 in their Vλ repertoire. Compared to the WT mice, the transgenic mice had a significantly higher percentage of mouse IgM-expressing B cells that expressed λ chains. Finally, we showed that the transgenic mice were deficient in a specific antibody response to antigen stimulation.

Keywords

bovine Ig μ heavy-chain / transgenic mice / B cell development / allelic exclusion / immune response / Ig repertoire

References

[1] Flajnik M F. Comparative analyses of immunoglobulin genes: surprises and portents. Nature Reviews-Immunology, 2002, 2(9): 688–698

[2] Flajnik M F, Kasahara M. Origin and evolution of the adaptive immune system: genetic events and selective pressures. Nature Reviews-Genetics, 2010, 11(1): 47–59

[3] Matthias P, Rolink A G. Transcriptional networks in developing and mature B cells. Nature Reviews-Immunology, 2005, 5(6): 497–508

[4] Yancopoulos G D, Blackwell T K, Wood C, Thomas E, Boss M, Coffman R, Rosenberg N, Tonegawa S, Baltimore D. EMBO Journal, 1984, 3(6): 1209–1219

[5] ten Boekel E, Melchers F, Rolink A G. Precursor B cells showing H chain allelic inclusion display allelic exclusion at the level of pre-B cell receptor surface expression. Immunity, 1998, 8(2): 199–207

[6] Keyna U, Beck-Engeser G B, Jongstra J, Applequist S E, Jäck H M. Surrogate light chain-dependent selection of Ig heavy chain V regions. Journal of Immunology, 1995, 155(12): 5536–5542

[7] Kline G H, Hartwell L, Beck-Engeser G B, Keyna U, Zaharevitz S, Klinman N R, Jäck H M. Pre-B cell receptor-mediated selection of pre-B cells synthesizing functional mu heavy chains. Journal of Immunology, 1998, 161(4): 1608–1618

[8] Melchers F. The pre-B-cell receptor: selector of fitting immunoglobulin heavy chains for the B-cell repertoire. Nature Reviews-Immunology, 2005, 5(7): 578–584

[9] Pernis B, Chiappino G, Kelus A S, Gell P G. Cellular localization of immunoglobulins with different allotypic specificities in rabbit lymphoid tissues. The Journal of Experimental Medicine, 1965, 122(5): 853–876

[10] Weiler E. Differential activity of allelic gamma-globulin genes in antibody-producing cells. Proceedings of the National Academy of Sciences of the United States of America, 1965, 54(6): 1765–1772

[11] Vettermann C, Schlissel M S. Allelic exclusion of immunoglobulin genes: models and mechanisms. Immunological Reviews, 2010, 237(1): 22–42

[12] Lutz J, Heideman M R, Roth E, van den Berk P, Müller W, Raman C, Wabl M, Jacobs H, Jäck H M. Pro-B cells sense productive immunoglobulin heavy chain rearrangement irrespective of polypeptide production. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(26): 10644–10649

[13] Nussenzweig M C, Shaw A C, Sinn E, Danner D B, Holmes K L, Morse H C 3rd, Leder P. Allelic exclusion in transgenic mice that express the membrane form of immunoglobulin mu. Science, 1987, 236(4803): 816–819

[14] Rusconi S, Köhler G. Transmission and expression of a specific pair of rearranged immunoglobulin mu and kappa genes in a transgenic mouse line. Nature, 1985, 314(6009): 330–334

[15] Weaver D, Costantini F, Imanishi-Kari T, Baltimore D. A transgenic immunoglobulin mu gene prevents rearrangement of endogenous genes. Cell, 1985, 42(1): 117–127

[16] Roth P E, Doglio L, Manz J T, Kim J Y, Lo D, Storb U. Immunoglobulin gamma 2b transgenes inhibit heavy chain gene rearrangement, but cannot promote B cell development. The Journal of Experimental Medicine, 1993, 178(6): 2007–2021

[17] Nussenzweig M C, Shaw A C, Sinn E, Campos-Torres J, Leder P. Allelic exclusion in transgenic mice carrying mutant human IgM genes. The Journal of Experimental Medicine, 1988, 167(6): 1969–1974

[18] Herzenberg L A, Stall A M, Braun J, Weaver D, Baltimore D, Herzenberg L A, Grosschedl R. Depletion of the predominant B-cell population in immunoglobulin mu heavy-chain transgenic mice. Nature, 1987, 329(6134): 71–73

[19] Jasper P J, Rhee K J, Kalis S L, Sethupathi P, Yam P C, Zhai S K, Knight K L. B lymphocyte deficiency in IgH-transgenic rabbits. European Journal of Immunology, 2007, 37(8): 2290–2299

[20] Zhao Y, Kacskovics I, Pan Q, Liberles D A, Geli J, Davis S K, Rabbani H, Hammarstrom L. Artiodactyl IgD: the missing link. Journal of Immunology, 2002, 169(8): 4408–4416

[21] Tobin-Janzen T C, Womack J E. Comparative mapping of IGHG1, IGHM, FES, and FOS in domestic cattle. Immunogenetics, 1992, 36(3): 157–165

[22] Hayes H C, Petit E J. Mapping of the beta-lactoglobulin gene and of an immunoglobulin M heavy chain-like sequence to homoeologous cattle, sheep, and goat chromosomes. Mammalian Genome, 1993, 4(4): 207–210

[23] Zhao Y, Kacskovics I, Rabbani H, Hammarstrom L. Physical mapping of the bovine immunoglobulin heavy chain constant region gene locus. Journal of Biological Chemistry, 2003, 278(37): 35024–35032

[24] Kuroiwa Y, Kasinathan P, Sathiyaseelan T, Jiao J A, Matsushita H, Sathiyaseelan J, Wu H, Mellquist J, Hammitt M, Koster J, Kamoda S, Tachibana K, Ishida I, Robl J M. Antigen-specific human polyclonal antibodies from hyperimmunized cattle. Nature Biotechnology, 2009, 27(2): 173–181

[25] Eggen A, Gautier M, Billaut A, Petit E, Hayes H, Laurent P, Urban C, Pfister-Genskow M, Eilertsen K, Bishop M D. Construction and characterization of a bovine BAC library with four genome-equivalent coverage. Genetics Selection Evolution, 2001, 33(5): 543–548

[26] Hosseini A, Campbell G, Prorocic M, Aitken R. Duplicated copies of the bovine JH locus contribute to the Ig repertoire. International Immunology, 2004, 16(6): 843–852

[27] Loder F, Mutschler B, Ray R J, Paige C J, Sideras P, Torres R, Lamers M C, Carsetti R. B cell development in the spleen takes place in discrete steps and is determined by the quality of B cell receptor-derived signals. The Journal of Experimental Medicine, 1999, 190(1): 75–89

[28] Carsetti R, Rosado M M, Wardmann H. Peripheral development of B cells in mouse and man. Immunological Reviews, 2004, 197(1): 179–191

[29] Koti M, Kataeva G, Kaushik A K. Novel atypical nucleotide insertions specifically at VH-DH junction generate exceptionally long CDR3H in cattle antibodies. Molecular Immunology, 2010, 47(11–12): 2119–2128

[30] Kaushik A K, Kehrli M E Jr, Kurtz A, Ng S, Koti M, Shojaei F, Saini S S. Somatic hypermutations and isotype restricted exceptionally long CDR3H contribute to antibody diversification in cattle. Veterinary Immunology and Immunopathology, 2009, 127(1–2): 106–113

[31] Shimizu A, Nussenzweig M C, Mizuta T R, Leder P, Honjo T. Immunoglobulin double-isotype expression by trans-mRNA in a human immunoglobulin transgenic mouse. Proceedings of the National Academy of Sciences of the United States of America, 1989, 86(20): 8020–8023

[32] Shimizu A, Nussenzweig M C, Han H, Sanchez M, Honjo T. Trans-splicing as a possible molecular mechanism for the multiple isotype expression of the immunoglobulin gene. The Journal of Experimental Medicine, 1991, 173(6): 1385–1393

[33] Ekman A, Niku M, Liljavirta J, Iivanainen A. Bos taurus genome sequence reveals the assortment of immunoglobulin and surrogate light chain genes in domestic cattle. BMC Immunology, 2009, 10(1): 22–32

[34] Stall A M, Kroese F G, Gadus F T, Sieckmann D G, Herzenberg L A, Herzenberg L A. Rearrangement and expression of endogenous immunoglobulin genes occur in many murine B cells expressing transgenic membrane IgM. Proceedings of the National Academy of Sciences of the United States of America, 1988, 85(10): 3546–3550

[35] Forni L. Extensive splenic B cell activation in IgM-transgenic mice. European Journal of Immunology, 1990, 20(5): 983–989

[36] Barreto V, Cumano A. Frequency and characterization of phenotypic Ig heavy chain allelically included IgM-expressing B cells in mice. Journal of Immunology, 2000, 164(2): 893–899

[37] Roes J, Rajewsky K. Immunoglobulin D (IgD)-deficient mice reveal an auxiliary receptor function for IgD in antigen-mediated recruitment of B cells. The Journal of Experimental Medicine, 1993, 177(1): 45–55

[38] Nitschke L, Kosco M H, Köhler G, Lamers M C. Immunoglobulin D-deficient mice can mount normal immune responses to thymus-independent and -dependent antigens. Proceedings of the National Academy of Sciences of the United States of America, 1993, 90(5): 1887–1891

[39] Knott J, Bona C, Kaushik A. The primary antibody repertoire of κ-deficient mice is characterized by non-stochastic Vλ1 + VH gene family pairings and a higher degree of self-reactivity. Scandinavian Journal of Immunology, 1998, 48(1): 65–72

[40] Khan W N. B cell receptor and BAFF receptor signaling regulation of B cell homeostasis. Journal of Immunology, 2009, 183(6): 3561–3567

[41] Macpherson A J, Lamarre A, McCoy K, Harriman G R, Odermatt B, Dougan G, Hengartner H, Zinkernagel R M. IgA production without mu or delta chain expression in developing B cells. Nature Immunology, 2001, 2(7): 625–631

Acknowledgements

The authors want to thank Liying Du for technical help with flow cytometry, Dr. Xiaorong Gu for help with statistical analysis and Prof. Lennart Hammarstrom for critical reading of the manuscript. This work was supported by the National Science Fund for Distinguished Young Scholars (30725029), the National Basic Research Program of China (2010CB945300).

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg