Krajewska M, Wang H G, Krajewski S, Zapata J M, Shabaik A, Gascoyne R, Reed J C
Apoptosis & Cell Death Research Program, The Burnham Institute, La Jolla, California 92037, USA.
Cancer Res. 1997 Apr 15;57(8):1605-13.
The in vivo patterns of CPP32 (Caspase-3) gene expression were determined using an immunohistochemical approach and paraffin-embedded normal human tissues. A rabbit polyclonal antiserum was generated against recombinant human CPP32 protein and shown to be specific by immunoblot analysis of various human tissues and cell lines. CPP32 immunoreactivity was selectively found in certain cell types and was typically present within the cytosol, although occasional cells also contained nuclear immunostaining. CPP32 immunostaining was easily detected, for example, in epidermal keratinocyes, cartilage chondrocytes, bone osteocytes, heart myocardiocytes, vascular smooth muscle cells, bronchial epithelium, hepatocytes, thymocytes, plasma cells, renal tubule epithelium, spermatogonia, prostatic secretory epithelial cells, uterine endometrium and myometrium, mammary ductal epithelial cells, and the gastrointestinal epithelium of the stomach, intestine, and colon. In contrast, little or no CPP32 immunoreactivity was observed in endothelial cells, alveolar pneumocytes, kidney glomeruli, mammary myoepithelial cells, Schwann cells, and most types of brain and spinal cord neurons. Consistent with a role for CPP32 in apoptotic cell death, clear differences in the relative intensity of CPP32 immunostaining were noted in some shorter-lived types of cells compared to longer-lived, including (a) germinal center (high) versus mantle zone (low) B lymphocytes within the secondary follicles of lymph nodes, spleen, and tonsils; (b) mature neutrophils (high) versus myeloid progenitor cells (low) in bone marrow; (c) corpus luteal cells (high) versus follicular granulosa cells (low) in the ovary; and (d) prostate secretory epithelial cells (high) versus basal cells (low). These findings establish for the first time the cell type- and differentiation-specific patterns of expression of an interleukin-1beta converting enzyme/CED-3 (Caspase) family protease.
采用免疫组化方法和石蜡包埋的正常人组织，确定了CPP32（半胱天冬酶-3）基因在体内的表达模式。制备了针对重组人CPP32蛋白的兔多克隆抗血清，并通过对各种人体组织和细胞系的免疫印迹分析表明其具有特异性。CPP32免疫反应性选择性地存在于某些细胞类型中，通常存在于细胞质中，不过偶尔也有细胞核免疫染色的细胞。例如，在表皮角质形成细胞、软骨细胞、骨细胞、心肌细胞、血管平滑肌细胞、支气管上皮细胞、肝细胞、胸腺细胞、浆细胞、肾小管上皮细胞、精原细胞、前列腺分泌上皮细胞、子宫内膜和子宫肌层、乳腺导管上皮细胞以及胃、小肠和结肠的胃肠道上皮细胞中很容易检测到CPP32免疫染色。相比之下，在内皮细胞、肺泡上皮细胞、肾小球、乳腺肌上皮细胞、施万细胞以及大多数类型的脑和脊髓神经元中，观察到很少或没有CPP32免疫反应性。与CPP32在凋亡性细胞死亡中的作用一致，与寿命较长的细胞相比，在一些寿命较短的细胞类型中，CPP32免疫染色的相对强度存在明显差异，包括：（a）淋巴结、脾脏和扁桃体次级滤泡内生发中心（高）与套区（低）B淋巴细胞；（b）骨髓中成熟中性粒细胞（高）与髓系祖细胞（低）；（c）卵巢中黄体细胞（高）与卵泡颗粒细胞（低）；以及（d）前列腺分泌上皮细胞（高）与基底细胞（低）。这些发现首次确立了白细胞介素-1β转化酶/CED-3（半胱天冬酶）家族蛋白酶的细胞类型和分化特异性表达模式。
