Golay J, Erba E, Bernasconi S, Peri G, Introna M
Institute of Pharmacological Research Mario Negri, Milan, Italy.
J Immunol. 1994 Jul 15;153(2):543-53.
The A-myb gene is structurally related to the c-mby proto-oncogene, a transcription factor involved in the regulation of hemopoietic proliferation and differentiation. Recent evidence has shown that A-myb also functions as a transcriptional activator. We have previously demonstrated that A-myb RNA is not expressed in most mature human leukocytes at rest or after mitogenic or functional activation. We show here, by using cell sorting, PCR, and Western analyses that A-myb is most highly expressed in the subsets of human tonsillar B lymphocytes with the phenotypes CD38+, CD39-, and SIgM-. The preferential expression of A-myb in these populations was seen at both the RNA and protein levels. CD38 was consistently best at separating high from low A-myb-expressing cells, whereas other markers (CD10, 22, 23, 77, 11a, and 49d) did not correlate with A-myb expression. The CD38+ population expressing the highest levels of A-myb was shown to contain mostly cycling cells inasmuch as more than 95% were in the late G1, S, G2, and M phases of the cell cycle. In addition, A-myb expression always correlated with the percentage of cells in S/G2/M in the populations sorted with either CD38, CD39, or sIgM. Small resting tonsillar B lymphocytes induced to proliferate in vitro by several different polyclonal B cell activators did not, however, express detectable levels of A-myb, although these cells were demonstrated to express CD38 and enter the S/G2/M phases of the cell cycle. These data suggest that A-myb is a marker of in vivo-activated but not in vitro-activated B lymphocytes. Finally, A-myb was also found to be highly expressed in five of seven Burkitt's lymphoma lines and in none of three EBV lymphoblastoid cell lines. This finding is in agreement with the phenotype of the normal B cells that express high levels of A-myb in vivo and suggests that A-myb may be specifically induced within germinal center B cells.
A-myb基因在结构上与c-myb原癌基因相关,c-myb是一种参与造血细胞增殖和分化调控的转录因子。最近的证据表明,A-myb也作为转录激活因子发挥作用。我们之前已经证明,在大多数静止或经有丝分裂原或功能激活后的成熟人类白细胞中,A-myb RNA不表达。我们在此通过细胞分选、PCR和蛋白质免疫印迹分析表明,A-myb在人扁桃体B淋巴细胞中表型为CD38+、CD39-和SIgM-的亚群中表达最高。在这些群体中,A-myb在RNA和蛋白质水平上均呈现优先表达。CD38始终是区分高表达和低表达A-myb细胞的最佳标志物,而其他标志物(CD10、22、23、77、11a和49d)与A-myb表达无关。表达最高水平A-myb的CD38+群体显示主要包含处于细胞周期的细胞,因为超过95%的细胞处于细胞周期的G1晚期、S期、G2期和M期。此外,在用CD38、CD39或sIgM分选的群体中,A-myb表达始终与处于S/G2/M期的细胞百分比相关。然而,几种不同的多克隆B细胞激活剂在体外诱导增殖的小的静止扁桃体B淋巴细胞,尽管这些细胞被证明表达CD38并进入细胞周期的S/G2/M期,但并未表达可检测水平的A-myb。这些数据表明,A-myb是体内激活而非体外激活的B淋巴细胞的标志物。最后,还发现A-myb在7个伯基特淋巴瘤细胞系中的5个中高表达,而在3个EBV淋巴母细胞系中均不表达。这一发现与体内高表达A-myb的正常B细胞表型一致,并表明A-myb可能在生发中心B细胞内被特异性诱导。