Levitt L, Kipps T J, Engleman E G, Greenberg P L
Blood. 1985 Mar;65(3):663-79.
The efficacy of four separate methods of human bone marrow T lymphocyte depletion was assessed, and the effect of T cells and monocytes on in vitro growth of marrow (CFU-GEMM, BFU-E, and CFU-GM) and peripheral blood (BFU-E) hematopoietic progenitors was determined. Extent of T cell depletion was assessed by multiparameter fluorescent cell sorter (FACS) analysis and by functional studies. Cells staining positively by FACS analysis for one or more of three separate fluorescent pan-T cell monoclonal antibodies (MCAbs) comprised 8.4% to 9.5% of control marrow mononuclear cells (MNCs). T cells constituted 3.2% to 5.1% of marrow following single, sequential, or combination treatment with two different pan-T cell MCAbs (Leu 1 and TM1) plus complement, 1.5% to 2.2% of marrow following solid-phase immunoabsorption ("panning"), 0.2% of marrow after sheep cell rosetting, and only 0.05% of marrow after FACS selective cell sorting and gated separation. T cells made up 59% to 73% of control peripheral blood MNCs and 0.8% to 2.8% of peripheral MNCs following sheep cell rosetting plus treatment with Leu 1 MCAb and complement. Mitogen (PHA, Con A) and allogeneic MLC-induced blastogenic responses (stimulation indices, experimental/control or E/C) revealed a concordant decrement in marrow T cell function after MCAb plus complement (E/C of 3.9 to 9.0), after panning (E/C of 1.6 to 3.5) and after sheep cell rosetting (E/C of 0.7 to 1.3), compared with control marrow (E/C of 5.3 to 15.7). After T cell depletion, marrow BFU-E growth was 95% to 120% of control, CFU-GM growth was 90% to 108% of control, and CFU-GEMM growth was 89% to 111% of control. Marrow T cell and/or monocyte depletion did not alter erythropoietin-dependent BFU-E growth in the absence of Mo-conditioned medium (81% to 95% of control), and the addition of as many as 50 to 100 X 10(3) purified marrow monocytes or T cells to 10(5) autologous nonadherent T cell-depleted marrow target cells had a negligible (P greater than .1) effect on marrow BFU-E growth in vitro. Peripheral blood (PB) BFU-E/10(5) T-depleted target cells were 106% +/- 19% of expected; PB BFU-E growth was significantly diminished after monocyte depletion alone (7% +/- 6% of expected) or after monocyte plus T cell depletion (8% +/- 4% of expected).(ABSTRACT TRUNCATED AT 400 WORDS)
评估了四种不同的人骨髓T淋巴细胞清除方法的疗效,并确定了T细胞和单核细胞对骨髓(CFU - GEMM、BFU - E和CFU - GM)以及外周血(BFU - E)造血祖细胞体外生长的影响。通过多参数荧光细胞分选仪(FACS)分析和功能研究评估T细胞清除的程度。通过FACS分析对三种不同的荧光泛T细胞单克隆抗体(mAb)中的一种或多种呈阳性染色的细胞占对照骨髓单个核细胞(MNC)的8.4%至9.5%。用两种不同的泛T细胞mAb(Leu 1和TM1)加补体进行单次、序贯或联合处理后,T细胞占骨髓的3.2%至5.1%;固相免疫吸附(“淘选”)后占骨髓的1.5%至2.2%;绵羊细胞玫瑰花结形成后占骨髓的0.2%;FACS选择性细胞分选和门控分离后仅占骨髓的0.05%。T细胞占对照外周血MNC的59%至73%,绵羊细胞玫瑰花结形成加Leu 1 mAb和补体处理后占外周MNC的0.8%至2.8%。丝裂原(PHA、Con A)和同种异体混合淋巴细胞培养诱导的增殖反应(刺激指数,实验/对照或E/C)显示,与对照骨髓(E/C为5.3至15.7)相比,mAb加补体(E/C为3.9至9.0)、淘选(E/C为1.6至3.5)和绵羊细胞玫瑰花结形成(E/C为0.7至1.3)后骨髓T细胞功能一致下降。T细胞清除后,骨髓BFU - E生长为对照的95%至120%,CFU - GM生长为对照的90%至108%,CFU - GEMM生长为对照的89%至111%。在没有单核细胞条件培养基的情况下,骨髓T细胞和/或单核细胞清除不会改变促红细胞生成素依赖的BFU - E生长(为对照的81%至95%),并且向10⁵个自体非贴壁T细胞清除的骨髓靶细胞中添加多达50至100×10³个纯化的骨髓单核细胞或T细胞对体外骨髓BFU - E生长的影响可忽略不计(P>0.1)。外周血(PB)BFU - E/10⁵个T细胞清除的靶细胞为预期值的106%±19%;单独单核细胞清除后(为预期值的7%±6%)或单核细胞加T细胞清除后(为预期值的8%±4%),PB BFU - E生长显著降低。(摘要截短于400字)