TACI-Fc和BCMA-Fc融合蛋白的表达、纯化以及生物功能鉴定
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摘要
B淋巴细胞刺激因子(B lymphocyte stimulator, BLyS),又称肿瘤坏死因子家族的B细胞活化分子(B cell-activating factor belonged to TNF family, BAFF)是一种重要的肿瘤坏死因子超家族成员。穿膜蛋白活化物(transmembrane activator and CAML interactor, TACI)和B细胞成熟抗原(B cell maturation antigen, BCMA)与BAFF-R (BAFF Receptor)一起成为BLyS的三个受体。本文中我们进行了人TACI全长及胞外区基因的钓取,与人IgG1的Fc段的连接,构建其原核表达载体。并进行了TACI-Fc和BCMA-Fc重组蛋白的诱导表达、纯化及生物学活性的鉴定。旨在为进一步研究BLyS与其受体的相互作用模式和开发用于治疗自身免疫疾病的BLyS拮抗剂奠定基础。该项研究取得的主要结果如下:
(1)经RT-PCR法成功克隆了编码人TACI的全长基因,大小约为900bp,测序结果与GenBank结果一致。
(2)扩增的TACI胞外区与人IgG1的Fc段基因连接后插入原核表达质粒pET28a,成功地构建了重组原核表达质粒pET28a-TACI-Fc。
(3)上述质粒和实验室已构建好的pET28a-BCMA-Fc重组质粒转化大肠杆菌BL21 (DE3),经IPTG诱导,并优化诱导条件,获得了较高水平的表达。SDS-PAGE分析可见,分别在大约40kDa和35kDa处有目的条带。Western Blotting和ELISA实验也显阳性。Protein A亲和层析柱进行纯化,得到高纯度的融合蛋白TACI-Fc和BCMA-Fc。
(4)已纯化的融合蛋白送北京进行MTT检测,结果表明TACI-Fc和BCMA-Fc融合蛋白可以阻断BLyS对Nalm-6细胞系的增殖作用,并且TACI-Fc较BCMA-Fc的阻断作用强。
B lymphocyte stimulator (BLyS), also known as B cell-activating factor belonged to TNF family (BAFF), is a member of tumor necrosis factor (TNF) family. Transmembrane activator and CAML interactor (TACI) and B cell maturation antigen (BCMA), which along with BAFF-R, are the three receptors of BLyS. In this project, we cloned the full length of TACI and ligated the extracellular fragment soluble TACI (sTACI) with human IgG1 Fc fragment, and constructed the prokaryotic expression vectors. Then pET28a-TACI-Fc and pET28a-BCMA-Fc recombinant plasmids were induced by IPTG to express the corresponding proteins. Next, the proteins were purified by Protein A chromatography, and their bioactivity was identified by MTT. We provide experimental base for further study on the action mode of TACI/BCMA with BLyS.
In summary, the results of this study are as follows:
(1)Clone the full length of TACI. The 900bp cDNA was amplified by RT-PCR, which was consistent with the sequence reported in GenBank.
(2)The extracellular fragment soluble TACI (sTACI) was ligated with human IgG1 Fc fragment. sTACI-Fc was subcloned into expression vector pET28a. The prokaryotic expression plasmid pET28a-TACI-Fc was successfully constructed.
(3)Recombinant plasmids pET28a-TACI-Fc and pET28a-BCMA-Fc constructed previously were transformed into E.coli BL21 (DE3) respectively. The expression was induced by IPTG and optimal conditions of the induction were achieved. SDS-PAGE analysis showed that about 40kDa and 35kDa fusion proteins were highly expressed. The expressed proteins were identified by Western Blotting and ELISA. The expressed proteins were purified by Protein A chromatography. SDS-PAGE and Western Blotting analysis showed that the target proteins were highly purified.
(4)The bioactivity of the purified proteins was indentified by MTT. The fusion proteins could inhibit the proliferation of BLyS to Nalm-6 cell lines. And the inhibitory effect of TACI-Fc fusion protein was more potent than that of BCMA-Fc.
(1)经RT-PCR法成功克隆了编码人TACI的全长基因,大小约为900bp,测序结果与GenBank结果一致。
(2)扩增的TACI胞外区与人IgG1的Fc段基因连接后插入原核表达质粒pET28a,成功地构建了重组原核表达质粒pET28a-TACI-Fc。
(3)上述质粒和实验室已构建好的pET28a-BCMA-Fc重组质粒转化大肠杆菌BL21 (DE3),经IPTG诱导,并优化诱导条件,获得了较高水平的表达。SDS-PAGE分析可见,分别在大约40kDa和35kDa处有目的条带。Western Blotting和ELISA实验也显阳性。Protein A亲和层析柱进行纯化,得到高纯度的融合蛋白TACI-Fc和BCMA-Fc。
(4)已纯化的融合蛋白送北京进行MTT检测,结果表明TACI-Fc和BCMA-Fc融合蛋白可以阻断BLyS对Nalm-6细胞系的增殖作用,并且TACI-Fc较BCMA-Fc的阻断作用强。
B lymphocyte stimulator (BLyS), also known as B cell-activating factor belonged to TNF family (BAFF), is a member of tumor necrosis factor (TNF) family. Transmembrane activator and CAML interactor (TACI) and B cell maturation antigen (BCMA), which along with BAFF-R, are the three receptors of BLyS. In this project, we cloned the full length of TACI and ligated the extracellular fragment soluble TACI (sTACI) with human IgG1 Fc fragment, and constructed the prokaryotic expression vectors. Then pET28a-TACI-Fc and pET28a-BCMA-Fc recombinant plasmids were induced by IPTG to express the corresponding proteins. Next, the proteins were purified by Protein A chromatography, and their bioactivity was identified by MTT. We provide experimental base for further study on the action mode of TACI/BCMA with BLyS.
In summary, the results of this study are as follows:
(1)Clone the full length of TACI. The 900bp cDNA was amplified by RT-PCR, which was consistent with the sequence reported in GenBank.
(2)The extracellular fragment soluble TACI (sTACI) was ligated with human IgG1 Fc fragment. sTACI-Fc was subcloned into expression vector pET28a. The prokaryotic expression plasmid pET28a-TACI-Fc was successfully constructed.
(3)Recombinant plasmids pET28a-TACI-Fc and pET28a-BCMA-Fc constructed previously were transformed into E.coli BL21 (DE3) respectively. The expression was induced by IPTG and optimal conditions of the induction were achieved. SDS-PAGE analysis showed that about 40kDa and 35kDa fusion proteins were highly expressed. The expressed proteins were identified by Western Blotting and ELISA. The expressed proteins were purified by Protein A chromatography. SDS-PAGE and Western Blotting analysis showed that the target proteins were highly purified.
(4)The bioactivity of the purified proteins was indentified by MTT. The fusion proteins could inhibit the proliferation of BLyS to Nalm-6 cell lines. And the inhibitory effect of TACI-Fc fusion protein was more potent than that of BCMA-Fc.
引文
[1] Moore PA, Belvedere O, Orr A, et al., BLyS: Member of the tumor necrosis factor family and B lymphocyte stimulator, Science, 1999, 285(5425): 260~263
[2] Shu HB, Hu WH, Johnson H, TALL-1 is a novel member of the TNF family that is down-regulated by mitogens, J. Lenkoc. Biol., 1999, 65(5): 680~683
[3] Schneider P, Mackay F, Steiner V, et al., BAFF, a novel ligand of the tumor necroses factor family, stimulates B cell growth, J. Exp. Med., 1999, 189(11): 1747~1756
[4] Mukhopadhyay A, Ni J, Zhai Y, et al., Identification and characterization of a novel cytokine, THANK, a TNF homologue that activates apoptosis, nuclear factor-kappaB, and c-Jun NH2-terminal kinase, J. Biol. Chem., 1999, 274(23): 15978~15981
[5] Kem C, Cornuel JF, Billard C, et a1., Involvement of BAFF and APRIL in the resistance to apoptosis of B-Cell through an autocrine pathway, Blood, 2004, 103(2): 679~688
[6] Roschke V, Sosnovtseva S, Ward CD, et a1., BLyS and APRIL form biologically active heterotrimers that are expressed in patients with systemic immune-based rheumatic diseases, J. Immunol., 2002, 169(8): 4314~4321
[7] Vallerskog T, Heimburger M, Gunnarsson I, et al., Differential effects on BAFF and APRIL levels in rituximab-treated patients with systemic lupus erythematosus and theumatoid arthritis, Arthritis. Res. Ther., 2006, 8(6): R167
[8] Mackay F, Woodcock SA, Lawton P, Mice transgenic for BAFF develop lymphocytic disorders along with autoimmune manifestations, J. Exp. Med., 1999, 190(11): 1697~1710
[9] Thompson JS, Bixler SA, Qian F, et a1., BAFF-R, a newly identified TNF receptor that specifically interacts with BAFF, Science, 2001, 293: 2108~2111
[10] Yan M, Identification of a novel receptor for B lymphocyte stimulator that is mutated in a mouse strain with severe B cell deficiency, Curr. Biol., 2001, 11(19): 1547~1552
[11] Shulga-Morskaya S, B cell-activating factor belonging to the TNF family acts through separate receptors to support B cell survival and T cell-independent antibody formation, J. Immunol., 2004, 173(4): 2331~2341
[12] Sasaki Y, Casola S, Kutok JL, et al., TNF family member B cells activating factor (BAFF) receptor-dependent and independent roles for BAFF in B cell physiology, J. Immunol., 2004, 173: 2245~2252
[13] Novak AJ, Bram RJ, Kay NE, et al., Aberrant expresion of B-1ymphocyte stimulator by B chronic lymphocytic leukemia cells: a mechanism for survival, Blood, 2002, 100(8): 2973~2979
[14] Zhang X, Park CS, Yoon SO, et al., BAFF supports human B cell differentiation in the lymphoid follicles through distinct receptors, Int. Immunol., 2005, 17(6): 779~788
[15] Xu LG, Shu HB, TNFR-associated factor-3 is associated with BAFF-R and negatively regulates BAFF-R-mediated NF-kappa B activation and IL-10 production, J. Immunol., 2002, 169: 6883~6889
[16] Ni CZ, Oqanesyan G, Welsh K, et al., Key molecular contacts promote recognition of the BAFF receptors by TNF receptor-associated factor 3: implications for intracellular signaling regulation, J. Immunol., 2004, 173(12): 7394~9400
[17] Shu HB, Johnson H, B cell maturation protein is a receptor for the tumor necrosis factor family member TALL-1, Proc. Natl. Acad. Sci. USA, 2000, 97: 9156~9l61
[18] O’Connor BP, Raman VS, Erickson LD, et al., BCMA is essential for the survival of long-lived bone marrow plasma cells, J. Exp. Med., 2004, 199(1): 91~98
[19] von Bülow GU, Bram RJ, NF-AT activation induced a CAML-interacting member of the rumor necrosis factor receptor superfamily, Science, 1997, 278(5335): 138~141
[20] Treml LS, Carlesso G, Hoek KL, et al., TLR stimulation modifies BLyS receptor expression in follicular and marginal zone B cells, J. Immunol., 2007, 178(12): 7531~7539
[21] Hahne M, Kataoka T, Schroter M, et a1., APRIL, a new ligand of the tumor necrosis factor family, stimulates tumor cell growth. J. Exp. Med., 1998, 188(6): 1185~1190
[22] Bischof D, Elsawa SF, Mantchev G, et al., Selective activation of TACI by syndecan-2, Blood, 2006, 107(8): 3235~3242
[23] Schiemann B, Gommerman JL, Vora K, et al., An essential role for BAFF in the normal development of B cells through a BCMA-independent pathway, Science, 2001, 293(5537): 2111~2114
[24] Woodland RT, Fox CJ, Schmidt MR, et al., Multiple signaling pathways promote B lymphocytes stimulato dependent B-cell growth and survival, Blood, 2008, 111(2): 750~760
[25] Yan M, Marsters SA, Grewal IS, et al., Identification of a receptor for BLyS demonstrates a crucial role in humoral immunity, Nat. Immunol., 2000, 1: 37~41
[26] Laabi Y, Strasser A, Lymphocyte survival-ignorance is BLyS, Science, 2000, 289(5481): 883~884
[27] Do RK, Hatada E, Lee H, et al., Attenuation of apoptosis underlies B lymphocyte stimulator enhancement of humoral immune response, J. Exp. Med., 2000, 192(7): 953~964
[28] Rodriguez B, Shneider P, Mauri D, et al., T cell costimulation by the TNF ligand BAFF, J. Immunol., 2001, 167(11): 6225~6231.
[29] Stohl W, SLE-systemic lupus erythematosus: a BLySful, yet BAFFling, disorder, Arthritis. Res. Ther., 2003, 5 (3): 136~138
[30] Khare SD, Sarosi L, Severe B cell hyperplasia and autoimmune disease in TALL-1 transgenic mice, Proc. Natl. Acad. Sci. USA, 2000, 97(7): 3370~3375
[31] Gross A, Johnston J, Mudn S, et al., TACI and BCMA are receptors for a TNF homokgue implicated in B-cell autoimmune disease, Nature, 2000, 404(678): 995~999
[32] Zhang J, Roschke V, Baker KP, et a1., Cutting edge: a role for B lymphocyte stimulator in systemic lupus erythematosus, J. Immunol., 2001, 166(1): 6~10
[33] Stohl W, Metyas S, Tan SM, et a1., B lymphocyte stimulator overexpression in patients with systemic lupus erythematosus longitudinal observations, Arthritis Rheum., 2003, 48(12): 3475~3486
[34] Cheema GS, Roschke V, Hilbert DM, et a1., Elevated serum B lymphocyte stimulator levels in patients with systemic immune-based rheumatic diseases, Arithritis Rheum., 2001, 44(6): 1313~1319
[35] Davis JR, Gross J, Gescak B, et a1., zTNF4 and soluble TACI receptor levels in serum and urine may reflect disease activity in patients with SLE, Arthritis Rheum., 2001, 44: S99
[36] Tan SM, Xu D, Roschke V, et a1., Local production of B lymphocyte stimulator protein and APRIL in arthritic joints of patients with inflammatory arthritis, Arthritis Rheum., 2003, 48(4): 982~992
[37] Groom J, Kalled SL, Culter AH, et a1., Association of BAFF/BLyS over- expression and altered B cell diferentiation with Sjogren’s syndrome, J. Clin. Invest., 2002, 109(1): 59~68
[38] Lavie F, Miceli-Richard C, Quillard J, et a1., Expression of BAFF (BLyS) in T cell infiltrating labial salivary glands from patients with Sjogren’s syndrome. J. Pathol., 2004, 202(4): 496~502
[39] Voutsadakis IA, Apoptosis and the pathogenesis of lymphoma, Acta. Oncol., 2000, 39(2): 151~156
[40] Batten M, Fletcher C, Ng L, et a1., TNF deficiency fails to protect BAFF transgenic mice against autoimmunity and reveals a predisposition to B cell lymphomas, J. Immunol., 2004, 172(2): 812~822
[41] Briones J, Timmerman JM, Hilbert DM, et al., BLyS and BLyS receptor expression in non-Hodgkin’s lymphoma, Exp. Hematol., 2002, 30(2): 135~141
[42] Novak AJ, Grote DM, Stenson M, et a1., Expression of BLys and its receptors in B-cell non-Hodgkin lymphoma: correlation with disease activity and patient outcome, Blood, 2004, 104(8): 2247~2253
[43] Novak AJ, Darce JR, Arendt BK, et a1., Expression of BCMA, TACI, and BAFF-R in multiple myeloma: a mechanism for growth and survival, Blood, 2004, 103(2): 689~694
[44] Moreaux J, Legouffe E, Jourdan E, et a1., BAFF and APRIL protect myeloma cells from apoptosis induced by interleukin 6 deprivation and dexamethasone, Blood, 2004, 103(8): 3148~3157
[45] Drachman DB, Myasthenia gravis, N. Engl. J. Med, 1994, 330(25): 1797~1810
[46] Matusevicius D, Navikas V, Palasik W, et a1., Tumor necrosis factor alpha, lymphotoxin, interleukin (IL)-6, IL-10, IL-12 and perforin mRNA expression in mononuclear cells in response to acetylcholine receptor is augmented in myasthenia gravis, J. Neuroimmunol., 1996, 71(1~2): 191~198
[47] Gu JH, Ju SQ, Xu MY, Measurement of B lymphocyte stimulator mRNA expression in children with infectious mononucleosis by real-time fluorescence quantitative method, Zhongguo Dang Dai Er Ke Za Zhi, 2007, 9(6): 553~556
[48] Hever A, Roth RB, Hevezi P, et al., Human endometriosis is associated with plama cells and overexpression of B lymphocyte stimulator, Proc. Natl. Acad. Sci. USA, 2007, 104(30): 12451~12456
[49] Sidhu SS, Fairbrother WJ, Deshayes K, Exploring protein-protein interactions with phage display, Chembiochem., 2003, 4(1): 14~25
[2] Shu HB, Hu WH, Johnson H, TALL-1 is a novel member of the TNF family that is down-regulated by mitogens, J. Lenkoc. Biol., 1999, 65(5): 680~683
[3] Schneider P, Mackay F, Steiner V, et al., BAFF, a novel ligand of the tumor necroses factor family, stimulates B cell growth, J. Exp. Med., 1999, 189(11): 1747~1756
[4] Mukhopadhyay A, Ni J, Zhai Y, et al., Identification and characterization of a novel cytokine, THANK, a TNF homologue that activates apoptosis, nuclear factor-kappaB, and c-Jun NH2-terminal kinase, J. Biol. Chem., 1999, 274(23): 15978~15981
[5] Kem C, Cornuel JF, Billard C, et a1., Involvement of BAFF and APRIL in the resistance to apoptosis of B-Cell through an autocrine pathway, Blood, 2004, 103(2): 679~688
[6] Roschke V, Sosnovtseva S, Ward CD, et a1., BLyS and APRIL form biologically active heterotrimers that are expressed in patients with systemic immune-based rheumatic diseases, J. Immunol., 2002, 169(8): 4314~4321
[7] Vallerskog T, Heimburger M, Gunnarsson I, et al., Differential effects on BAFF and APRIL levels in rituximab-treated patients with systemic lupus erythematosus and theumatoid arthritis, Arthritis. Res. Ther., 2006, 8(6): R167
[8] Mackay F, Woodcock SA, Lawton P, Mice transgenic for BAFF develop lymphocytic disorders along with autoimmune manifestations, J. Exp. Med., 1999, 190(11): 1697~1710
[9] Thompson JS, Bixler SA, Qian F, et a1., BAFF-R, a newly identified TNF receptor that specifically interacts with BAFF, Science, 2001, 293: 2108~2111
[10] Yan M, Identification of a novel receptor for B lymphocyte stimulator that is mutated in a mouse strain with severe B cell deficiency, Curr. Biol., 2001, 11(19): 1547~1552
[11] Shulga-Morskaya S, B cell-activating factor belonging to the TNF family acts through separate receptors to support B cell survival and T cell-independent antibody formation, J. Immunol., 2004, 173(4): 2331~2341
[12] Sasaki Y, Casola S, Kutok JL, et al., TNF family member B cells activating factor (BAFF) receptor-dependent and independent roles for BAFF in B cell physiology, J. Immunol., 2004, 173: 2245~2252
[13] Novak AJ, Bram RJ, Kay NE, et al., Aberrant expresion of B-1ymphocyte stimulator by B chronic lymphocytic leukemia cells: a mechanism for survival, Blood, 2002, 100(8): 2973~2979
[14] Zhang X, Park CS, Yoon SO, et al., BAFF supports human B cell differentiation in the lymphoid follicles through distinct receptors, Int. Immunol., 2005, 17(6): 779~788
[15] Xu LG, Shu HB, TNFR-associated factor-3 is associated with BAFF-R and negatively regulates BAFF-R-mediated NF-kappa B activation and IL-10 production, J. Immunol., 2002, 169: 6883~6889
[16] Ni CZ, Oqanesyan G, Welsh K, et al., Key molecular contacts promote recognition of the BAFF receptors by TNF receptor-associated factor 3: implications for intracellular signaling regulation, J. Immunol., 2004, 173(12): 7394~9400
[17] Shu HB, Johnson H, B cell maturation protein is a receptor for the tumor necrosis factor family member TALL-1, Proc. Natl. Acad. Sci. USA, 2000, 97: 9156~9l61
[18] O’Connor BP, Raman VS, Erickson LD, et al., BCMA is essential for the survival of long-lived bone marrow plasma cells, J. Exp. Med., 2004, 199(1): 91~98
[19] von Bülow GU, Bram RJ, NF-AT activation induced a CAML-interacting member of the rumor necrosis factor receptor superfamily, Science, 1997, 278(5335): 138~141
[20] Treml LS, Carlesso G, Hoek KL, et al., TLR stimulation modifies BLyS receptor expression in follicular and marginal zone B cells, J. Immunol., 2007, 178(12): 7531~7539
[21] Hahne M, Kataoka T, Schroter M, et a1., APRIL, a new ligand of the tumor necrosis factor family, stimulates tumor cell growth. J. Exp. Med., 1998, 188(6): 1185~1190
[22] Bischof D, Elsawa SF, Mantchev G, et al., Selective activation of TACI by syndecan-2, Blood, 2006, 107(8): 3235~3242
[23] Schiemann B, Gommerman JL, Vora K, et al., An essential role for BAFF in the normal development of B cells through a BCMA-independent pathway, Science, 2001, 293(5537): 2111~2114
[24] Woodland RT, Fox CJ, Schmidt MR, et al., Multiple signaling pathways promote B lymphocytes stimulato dependent B-cell growth and survival, Blood, 2008, 111(2): 750~760
[25] Yan M, Marsters SA, Grewal IS, et al., Identification of a receptor for BLyS demonstrates a crucial role in humoral immunity, Nat. Immunol., 2000, 1: 37~41
[26] Laabi Y, Strasser A, Lymphocyte survival-ignorance is BLyS, Science, 2000, 289(5481): 883~884
[27] Do RK, Hatada E, Lee H, et al., Attenuation of apoptosis underlies B lymphocyte stimulator enhancement of humoral immune response, J. Exp. Med., 2000, 192(7): 953~964
[28] Rodriguez B, Shneider P, Mauri D, et al., T cell costimulation by the TNF ligand BAFF, J. Immunol., 2001, 167(11): 6225~6231.
[29] Stohl W, SLE-systemic lupus erythematosus: a BLySful, yet BAFFling, disorder, Arthritis. Res. Ther., 2003, 5 (3): 136~138
[30] Khare SD, Sarosi L, Severe B cell hyperplasia and autoimmune disease in TALL-1 transgenic mice, Proc. Natl. Acad. Sci. USA, 2000, 97(7): 3370~3375
[31] Gross A, Johnston J, Mudn S, et al., TACI and BCMA are receptors for a TNF homokgue implicated in B-cell autoimmune disease, Nature, 2000, 404(678): 995~999
[32] Zhang J, Roschke V, Baker KP, et a1., Cutting edge: a role for B lymphocyte stimulator in systemic lupus erythematosus, J. Immunol., 2001, 166(1): 6~10
[33] Stohl W, Metyas S, Tan SM, et a1., B lymphocyte stimulator overexpression in patients with systemic lupus erythematosus longitudinal observations, Arthritis Rheum., 2003, 48(12): 3475~3486
[34] Cheema GS, Roschke V, Hilbert DM, et a1., Elevated serum B lymphocyte stimulator levels in patients with systemic immune-based rheumatic diseases, Arithritis Rheum., 2001, 44(6): 1313~1319
[35] Davis JR, Gross J, Gescak B, et a1., zTNF4 and soluble TACI receptor levels in serum and urine may reflect disease activity in patients with SLE, Arthritis Rheum., 2001, 44: S99
[36] Tan SM, Xu D, Roschke V, et a1., Local production of B lymphocyte stimulator protein and APRIL in arthritic joints of patients with inflammatory arthritis, Arthritis Rheum., 2003, 48(4): 982~992
[37] Groom J, Kalled SL, Culter AH, et a1., Association of BAFF/BLyS over- expression and altered B cell diferentiation with Sjogren’s syndrome, J. Clin. Invest., 2002, 109(1): 59~68
[38] Lavie F, Miceli-Richard C, Quillard J, et a1., Expression of BAFF (BLyS) in T cell infiltrating labial salivary glands from patients with Sjogren’s syndrome. J. Pathol., 2004, 202(4): 496~502
[39] Voutsadakis IA, Apoptosis and the pathogenesis of lymphoma, Acta. Oncol., 2000, 39(2): 151~156
[40] Batten M, Fletcher C, Ng L, et a1., TNF deficiency fails to protect BAFF transgenic mice against autoimmunity and reveals a predisposition to B cell lymphomas, J. Immunol., 2004, 172(2): 812~822
[41] Briones J, Timmerman JM, Hilbert DM, et al., BLyS and BLyS receptor expression in non-Hodgkin’s lymphoma, Exp. Hematol., 2002, 30(2): 135~141
[42] Novak AJ, Grote DM, Stenson M, et a1., Expression of BLys and its receptors in B-cell non-Hodgkin lymphoma: correlation with disease activity and patient outcome, Blood, 2004, 104(8): 2247~2253
[43] Novak AJ, Darce JR, Arendt BK, et a1., Expression of BCMA, TACI, and BAFF-R in multiple myeloma: a mechanism for growth and survival, Blood, 2004, 103(2): 689~694
[44] Moreaux J, Legouffe E, Jourdan E, et a1., BAFF and APRIL protect myeloma cells from apoptosis induced by interleukin 6 deprivation and dexamethasone, Blood, 2004, 103(8): 3148~3157
[45] Drachman DB, Myasthenia gravis, N. Engl. J. Med, 1994, 330(25): 1797~1810
[46] Matusevicius D, Navikas V, Palasik W, et a1., Tumor necrosis factor alpha, lymphotoxin, interleukin (IL)-6, IL-10, IL-12 and perforin mRNA expression in mononuclear cells in response to acetylcholine receptor is augmented in myasthenia gravis, J. Neuroimmunol., 1996, 71(1~2): 191~198
[47] Gu JH, Ju SQ, Xu MY, Measurement of B lymphocyte stimulator mRNA expression in children with infectious mononucleosis by real-time fluorescence quantitative method, Zhongguo Dang Dai Er Ke Za Zhi, 2007, 9(6): 553~556
[48] Hever A, Roth RB, Hevezi P, et al., Human endometriosis is associated with plama cells and overexpression of B lymphocyte stimulator, Proc. Natl. Acad. Sci. USA, 2007, 104(30): 12451~12456
[49] Sidhu SS, Fairbrother WJ, Deshayes K, Exploring protein-protein interactions with phage display, Chembiochem., 2003, 4(1): 14~25
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