CpG DNA activation and plasma-cell differentiation of CD27- naive human B cells
CpG DNA activation and plasma-cell differentiation of CD27- naive human B cells
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DOI：
10.1182/blood-2006-03-008441
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发表时间：
2007-02-15
期刊：
影响因子：
20.3
通讯作者：
Zand, Martin S.
中科院分区：
文献类型：
--
作者：
Huggins, Jennifer;Pellegrin, Tina;Zand, Martin S.
据报道，初始 CD27(-) B 细胞的非甲基化 CpG DNA 激活需要 B 细胞受体 (BCR) 交联。我们描述了一种使用 CpG DNA 的培养系统，通过连续步骤对初始 CD19(+)CD27(-) 人外周血 B 细胞进行 T 细胞独立激活，从而诱导有效的 CD138(+) 浆细胞分化。 CD27(+)和CD27- B细胞在3步系统中培养：(1)第0天至第4天：CpG、IL-2/10/15； (2)第4至7天：IL-2/6110/15和抗CD40L； (3)第7至10天：IL-6/15、IFN-α、肝细胞生长因子和透明质酸。 CpG DNA 激活后，CD27+ 和 CD27- B 细胞均上调胞质内 TLR-9。 CD27- B 细胞激活需要细胞与细胞接触。幼稚 B 细胞和记忆 B 细胞均进展为浆细胞表型：CD19（低）CD20（低）CD27（+）CD38（+）HLA-DRlow。 70% 的 CD27 衍生的 CD138(+) 细胞表现出有效的 V 链重排，没有体细胞突变，证实了它们起源于幼稚前体。 CD27(+)B细胞来源的浆细胞主要为IgG(+)，而CD27(-)B细胞来源的浆细胞主要为IgM(+)。我们的结果表明，在某些条件下，初始 B 细胞会增加 TLR-9 表达并在没有 BCR 信号转导的情况下在 CpG DNA 刺激下增殖。除了其免疫学意义外，该系统应该是一种询问初始 B 细胞抗原特异性的有价值的方法。
Unmethylated CpG DNA activation of naive CD27(-) B cells has been reported to require B-cell-receptor (BCR) cross-linking. We describe a culture system using CpG DNA with sequential steps for T-cell-independent activation of naive CD19(+)CD27(-) human peripheral blood B cells that induces efficient CD138(+) plasma-cell differentiation. CD27(+) and CD27- B cells were cultured in a 3-step system: (1) days 0 to 4: CpG, IL-2/10/15; (2) days 4 to 7: IL-2/6110/15 and anti-CD40L; (3) days 7 to 10: IL-6/15, IFN-alpha, hepatocyte growth factor, and hyaluronic acid. Both CD27+ and CD27- B cells up-regulated intracytoplasmic TLR-9 following CpG DNA activation. CD27- B-cell activation required cell-cell contact. Both naive and memory B cells progressed to a plasma-cell phenotype: CD19(low)CD20(low)CD27(+)CD38(+)HLA-DRlow. Seventy percent of the CD27-derived CD138(+) cells demonstrated productive V chain rearrangements without somatic mutations, confirming their origin from naive precursors. Plasma cells derived from CD27(+) B cells were primarily IgG(+), while those from CD27(-) B cells were IgM(+). Our results indicate that under certain conditions, naive B cells increase TLR-9 expression and proliferate to CpG DNA stimulation without BCR signaling. In addition to its immunologic significance, this system should be a valuable method to interrogate the antigenic specificity of naive B cells.