[en] The immunodeficiency disorder, X-linked agammaglobulinemia (XLA), results from mutations in the gene encoding Bruton tyrosine kinase (Btk). Btk is required for pre-B cell clonal expansion and B-cell antigen receptor signaling. XLA patients lack mature B cells and immunoglobulin and experience recurrent bacterial infections only partially mitigated by life-long antibody replacement therapy. In pursuit of definitive therapy for XLA, we tested ex vivo gene therapy using a lentiviral vector (LV) containing the immunoglobulin enhancer (Eµ) and IgB (B29) minimal promoter to drive B lineage-specific human Btk expression in Btk/Teck-/- mice, a strain that reproduces the features of human XLA. After transplantation of EµB29-Btk-LV-transduced stem cells, treated mice showed significant, albeit incomplete, rescue of mature B cells in the bone marrow, peripheral blood, spleen, and peritoneal cavity, and improved responses to T-independent and T-dependent antigens. LV-teated B cells exhibited enhanced B-cell antigen receptor signaling and an in vivo selective advantage in the peripheral versus central B-cell compartment. Secondary transplantation showed sustained Btk expression, viral integration, and partial functional responses, consistent with long-term stem cell marking; and serial transplantation revealed no evidence for cellular or systemic toxicity. These findings strongly support pursuit of B lineage-targeted LV gene therapy in human XLA.
Rawlings DJ, Witte ON. Bruton's tyrosine kinase is a key regulator in B-cell development. Immunol Rev. 1994;138:105-119.
Rawlings DJ, Witte ON. The Btk subfamily of cytoplasmic tyrosine kinases: structure, regulation and function. Semin Immunol. 1995;7(4):237-246.
Tsukada S, Saffran DC, Rawlings DJ, et al. Defi- cient expression of a B cell cytoplasmic tyrosine kinase in human X-linked agammaglobulinemia. Cell. 1993;72(2):279-290. (Pubitemid 23044942)
Conley ME, Brown P, Pickard AR, et al. Expression of the gene defect in X-linked agammaglobulinemia. N Engl J Med. 1986;315(9):564-567.
Hendriks RW, de Bruijn MF, Maas A, Dingjan GM, Karis A, Grosveld F. Inactivation of Btk by insertion of lacZ reveals defects in B cell development only past the pre-B cell stage. EMBO J. 1996;15(18):4862-4872. (Pubitemid 26315797)
Quintans J, McKearn JP, Kaplan D. B cell heterogeneity: I. A study of B cell subpopulations involved in the reconstitution of an X-linked immune defect of B cell differentiation. J Immunol. 1979;122(5):1750-1756.
Quan ZS, Dick RF, Regueiro B, Quintans J. B cell heterogeneity: II. Transplantation resistance in xid mice which affects the ontogeny of B cell subpopulations. Eur J Immunol. 1981;11(8):643-649.
Rohrer J, Conley ME. Correction of X-linked immunodeficient mice by competitive reconstitution with limiting numbers of normal bone marrow cells. Blood. 1999;94(10):3358-3365. (Pubitemid 29536129)
Yu PW, Tabuchi RS, Kato RM, et al. Sustained correction of B-cell development and function in a murine model of X-linked agammaglobulinemia (XLA) using retroviral-mediated gene transfer. Blood. 2004;104(5):1281-1290.
Santilli G, Thornhill SI, Kinnon C, Thrasher AJ. Gene therapy of inherited immunodeficiencies. Expert Opin Biol Ther. 2008;8(4):397-407.
Cavazzana-Calvo M, Fischer A. Gene therapy for severe combined immunodeficiency: are we there yet? J Clin Invest. 2007;117(6):1456-1465. (Pubitemid 46871328)
Hacein-Bey-Abina S, von Kalle C, Schmidt M, et al. A serious adverse event after successful gene therapy for X-linked severe combined immunodeficiency. N Engl J Med. 2003;348(3):255-256. (Pubitemid 36077924)
Ott MG, Schmidt M, Schwarzwaelder K, et al. Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1. Nat Med. 2006;12(4):401-409.
Naldini L, Verma IM. Lentiviral vectors. Adv Virus Res. 2000;55:599-609.
Modlich U, Navarro S, Zychlinski D, et al. Insertional transformation of hematopoietic cells by self-inactivating lentiviral and gammaretroviral vectors. Mol Ther. 2009;17(11):1919-1928.
Wu X, Li Y, Crise B, Burgess SM. Transcription start regions in the human genome are favored targets for MLV integration. Science. 2003;300(5626):1749- 1751. (Pubitemid 36712571)
Schroder AR, Shinn P, Chen H, Berry C, Ecker JR, Bushman F. HIV-1 integration in the human genome favors active genes and local hotspots. Cell. 2002;110(4):521-529. (Pubitemid 35232295)
Banerji J, Olson L, Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983;33(3):729-740.
Cockerill PN, Garrard WT. Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites. Cell. 1986;44(2):273-282.
Thompson AA, Wood WJ Jr, Gilly MJ, Damore MA, Omori SA, Wall R. The promoter and 5′ flanking sequences controlling human B29 gene expression. Blood. 1996;87(2):666-673. (Pubitemid 26030322)
Dull T, Zufferey R, Kelly M, et al.Athird-generation lentivirus vector with a conditional packaging system. J Virol. 1998;72(11):8463-8471. (Pubitemid 28471781)
Ellmeier W, Jung S, Sunshine MJ, et al. Severe B cell deficiency in mice lacking the tec kinase family members Tec and Btk. J Exp Med. 2000;192(11):1611-1624.
Andrews SF, Rawlings DJ. Transitional B cells exhibit a B cell receptor-specific nuclear defect in gene transcription. J Immunol. 2009;182(5):2868-2878.
Takata M, Kurosaki T. A role for Bruton's tyrosine kinase in B cell antigen receptor-mediated activation of phospholipase C-gamma 2. J Exp Med. 1996;184(1):31-40. (Pubitemid 26247212)
Scharenberg AM, Humphries LA, Rawlings DJ. Calcium signalling and cell-fate choice in B cells. Nat Rev Immunol. 2007;7(10):778-789.
Fluckiger AC, Li Z, Kato RM, et al. Btk/Tec kinases regulate sustained increases in intracellular Ca2+ following B-cell receptor activation. EMBO J. 1998;17(7):1973-1985. (Pubitemid 28154527)
Montecino-Rodriguez E, Dorshkind K. New perspectives in B-1 B cell development and function. Trends Immunol. 2006;27(9):428-433. (Pubitemid 44216193)
Plebani A, Soresina A, Rondelli R, et al. Clinical, immunological, and molecular analysis in a large cohort of patients with X-linked agammaglobulinemia: an Italian multicenter study. Clin Immunol. 2002;104(3):221-230. (Pubitemid 35191097)
Van der Hilst JC, Smits BW, van der Meer JW. Hypogammaglobulinaemia: cumulative experience in 49 patients in a tertiary care institution. Neth J Med. 2002;60(3):140-147.
Howard V, Greene JM, Pahwa S, et al. The health status and quality of life of adults with X-linked agammaglobulinemia. Clin Immunol. 2006;118(2):201-208. (Pubitemid 43199659)
Winkelstein JA, Marino MC, Lederman HM, et al. X-linked agammaglobulinemia: report on a United States registry of 201 patients. Medicine (Baltimore). 2006;85(4):193-202.
Winkelstein JA, Conley ME, James C, Howard V, Boyle J. Adults with X-linked agammaglobulinemia: impact of disease on daily lives, quality of life, educational and socioeconomic status, knowledge of inheritance, and reproductive attitudes. Medicine (Baltimore). 2008;87(5):253-258.
Drabek D, Raguz S, De Wit TP, et al. Correction of the X-linked immunodeficiency phenotype by transgenic expression of human Bruton tyrosine kinase under the control of the class II major histocompatibility complex Ea locus control region. Proc Natl Acad Sci U S A. 1997;94(2):610-615. (Pubitemid 27053681)
Maas A, Dingjan GM, Grosveld F, Hendriks RW. Early arrest in B cell development in transgenic mice that express the E41K Bruton's tyrosine kinase mutant under the control of the CD19 promoter region. J Immunol. 1999;162(11):6526-6533. (Pubitemid 29309375)
Kersseboom R, Middendorp S, Dingjan GM, et al. Bruton's tyrosine kinase cooperates with the B cell linker protein SLP-65 as a tumor suppressor in Pre-B cells. J Exp Med. 2003;198(1):91-98. (Pubitemid 36870189)
Feldhahn N, Rio P, Soh BN, et al. Deficiency of Bruton's tyrosine kinase in B cell precursor leukemia cells. Proc Natl Acad Sci U S A. 2005;102(37):13266-13271. (Pubitemid 41325012)
Saffran DC, Parolini O, Fitch-Hilgenberg ME, et al. Brief report: a point mutation in the SH2 domain of Bruton's tyrosine kinase in atypical X-linked agammaglobulinemia. N Engl J Med. 1994;330(21):1488-1491. (Pubitemid 24151406)
Kornfeld SJ, Haire RN, Strong SJ, et al. A novel mutation (Cys145-->Stop) in Bruton's tyrosine kinase is associated with newly diagnosed X-linked agammaglobulinemia in a 51-year-old male. Mol Med. 1996;2(5):619-623. (Pubitemid 26353110)
Satterthwaite AB, Cheroutre H, Khan WN, Sideras P, Witte ON. Btk dosage determines sensitivity to B cell antigen receptor cross-linking. Proc Natl Acad Sci U S A. 1997;94(24):13152-13157. (Pubitemid 27518491)
Pinschewer DD, Ochsenbein AF, Satterthwaite AB, Witte ON, Hengartner H, Zinkernagel RM. A Btk transgene restores the antiviral TI-2 antibody responses of xid mice in a dose-dependent fashion. Eur J Immunol. 1999;29(9):2981-2987. (Pubitemid 30099022)
Zhang F, Thornhill SI, Howe SJ, et al. Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells. Blood. 2007;110(5):1448-1457. (Pubitemid 47443959)
Himmelmann A, Thevenin C, Harrison K, Kehrl JH. Analysis of the Bruton's tyrosine kinase gene promoter reveals critical PU.1 and SP1 sites. Blood. 1996;87(3):1036-1044. (Pubitemid 26049874)
Rohrer J, Conley ME. Transcriptional regulatory elements within the first intron of Bruton's tyrosine kinase. Blood. 1998;91(1):214-221. (Pubitemid 28018747)
Jo EK, Kanegane H, Nonoyama S, et al. Characterization of mutations, including a novel regulatory defect in the first intron, in Bruton's tyrosine kinase gene from seven Korean X-linked agammaglobulinemia families. J Immunol. 2001;167(7):4038-4045. (Pubitemid 32910850)
Yu L, MohamedAJ, Simonson OE, et al. Proteasome-dependent autoregulation of Bruton tyrosine kinase (Btk) promoter via NF-kappaB. Blood. 2008;111(9):4617-4626.
Buckley RH, Schiff SE, Schiff RI, et al. Hematopoietic stem-cell transplantation for the treatment of severe combined immunodeficiency. N Engl J Med. 1999;340(7):508-516. (Pubitemid 29093286)
Haddad E, Le Deist F, Aucouturier P, et al. Longterm chimerism and B-cell function after bone marrow transplantation in patients with severe combined immunodeficiency with B cells: a single-center study of 22 patients. Blood. 1999;94(8):2923-2930. (Pubitemid 29477323)
Liu A, Vosshenrich CA, Lagresle-Peyrou C, et al. Competition within the early B-cell compartment conditions B-cell reconstitution after hematopoietic stem cell transplantation in nonirradiated recipients. Blood. 2006;108(4):1123-1128. (Pubitemid 44232004)
