Monitoring of tumor cell purging after highly efficient immunomagnetic selection of CD34 cells from leukapheresis products in breast cancer patients: Comparison of immunocytochemical tumor cell staining and reverse transcriptase-polymerase chain reaction
Monitoring of tumor cell purging after highly efficient immunomagnetic selection of CD34 cells from leukapheresis products in breast cancer patients: Comparison of immunocytochemical tumor cell staining and reverse transcriptase-polymerase chain reaction
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DOI：
10.1182/blood.v89.1.337.337_337_344
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发表时间：
1997-01-01
期刊：
影响因子：
20.3
通讯作者：
Dorken, B
中科院分区：
文献类型：
--
作者：
Mapara, MY;Korner, IJ;Dorken, B
我们研究了基于CD 34（+）细胞的免疫磁性选择，通过从白细胞去除产物（LP）中富集CD 34（+）造血祖细胞来间接净化乳腺癌患者的肿瘤细胞的效率。免疫细胞化学染色检测肿瘤细胞。此外，我们评估了细胞角蛋白19（CK 19）和一种新的表皮生长因子受体（EGF-R）特异性逆转录聚合酶链反应（RT-PCR）监测肿瘤细胞耗竭的能力。分析了13例乳腺癌患者的LP。进行了23次CD 34选择程序。将中位1.4 x 10（10）总有核细胞（[TNC]范围，0.88 - 3.5 x 10（10））和中位CD 34纯度2.5%（范围，0.4%-6.3%）输入选择程序。免疫磁性CD 34富集导致中位纯度为83.3%（范围：45%-95.4%），中位回收率为73.2%（范围：22%-95%）。在高剂量化疗后回输CD 34选择的细胞导致快速和持续的血液学恢复，在第+10天达到绝对中性粒细胞计数500/μ L，在第+11天达到血小板计数20，000/μ L。通过免疫细胞化学检测CK 19阳性细胞定量肿瘤细胞耗竭。通过该方法，可以证明1.9 log（范围，0.7至>3 log）的中值肿瘤细胞消除。肿瘤细胞的免疫细胞化学检测比RT-PCR更敏感，在87%的LP（17 - 21）中产生阳性结果，而阳性LP为58%（10/17）。然而，基于EGF-R的RT-PCR比基于CK 19的RT-PCR敏感得多（17个中的10个对17个中的1个）。尽管CD 34选择效率很高，但使用免疫细胞化学和EGF-R特异性RT-PCR进行CD 34富集后仍可检测到肿瘤细胞。因此，这种新的EGF-R特异性RT-PCR似乎是一种额外的方法来检测LP内污染的乳腺癌细胞的价值。(C)1997年，美国血液学会。
We studied the efficiency of indirect tumor cell purging via enrichment of CD34(+) hematopoietic progenitor cells from leukapheresis products (LP) in breast cancer patients based on immunomagnetic selection of CD34(+) cells. Detection of tumor cells was made by immunocytochemical staining. In addition, we evaluated the capacity of cytokeratin 19 (CK19)-and a novel epidermal growth factor receptor (EGF-R)-specific reverse transcriptase-polymerase chain reaction (RT-PCR) for monitoring tumor cell depletion. LP from 13 breast cancer patients were analyzed. Twenty-three CD34 selection procedures were performed. A median of 1.4 x 10(10) total nucleated cells ([TNC] range, 0.88 to 3.5 x 10(10)) with a median CD34 purity of 2.5% (range, 0.4% to 6.3%) were entered into the selection procedure. Immunomagnetic CD34 enrichment resulted in a median purity of 83.3% (range, 45% to 95.4%) and a median recovery of 73.2% (range, 22% to 95%). Retransfusion of CD34-selected cells after high-dose chemotherapy resulted in a rapid and sustained hematologic recovery, reaching an absolute neutrophil count of 500/mu L at day +10 and platelet count of 20,000/mu L at day +11. Tumor cell depletion was quantified by immunocytochemical detection of CK19-positive cells. By this method, a median tumor cell depletion of 1.9 log (range, 0.7 to >3 log) could be demonstrated. Immunocytochemical detection of tumor cells was more sensitive than RT-PCR, yielding positive results in 87% of LP (17 to 21) versus 58% positive LP (10 of 17). However, EGF-R-based RT-PCR was much more sensitive than CK19-based RT-PCR (10 of 17 v 1 of 17). Despite highly efficient CD34 selection, tumor cells were still detectable after CD34 enrichment using immunocytochemistry and EGF-R-specific RT-PCR. Thus, this novel EGF-R-specific RT-PCR appears to be of value as an additional method to detect contaminating breast cancer cells within LP. (C) 1997 by The American Society of Hematology.