IMMUNOCYTOCHEMICAL DOUBLE STAINING OF CYTOKERATIN AND PROSTATE SPECIFIC ANTIGEN IN INDIVIDUAL PROSTATIC TUMOR-CELLS

被引:83
作者
RIESENBERG, R [1 ]
OBERNEDER, R [1 ]
KRIEGMAIR, M [1 ]
EPP, M [1 ]
BITZER, U [1 ]
HOFSTETTER, A [1 ]
BRAUN, S [1 ]
RIETHMULLER, G [1 ]
PANTEL, K [1 ]
机构
[1] UNIV MUNICH, INST IMMUNOL, W-8000 MUNICH 2, GERMANY
关键词
D O I
10.1007/BF00268022
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Early dissemination of malignant cells is the main cause for metastatic relapse in patients with solid tumours. By use of monoclonal antibodies (mAbs) specific for cytokeratins, disseminated individual epithelial tumour cells can now be identified in mesenchymal organs such as bone marrow. Further to characterize such cells in patients with prostate cancer, an immunocytochemical procedure was developed for simultaneous labelling of cytokeratin component no. 18 (CK18) and prostate specific antigen (PSA). In a first step, cells were incubated with mAb ER-PR8 against PSA and secondary gold-conjugated goat anti-mouse antibodies. In a second step, biotinylated mAb CK2 to CK18 was applied as primary antibody and subsequently incubated with complexes of streptavidin-conjugated alkaline phosphatase, which were developed with the Newfuchsin substrate. The binding of gold-labelled antibodies was visualized by silver enhancement. The sensitivity and specificity of the technique was demonstrated on cryostat sections of hyperplastic prostatic tissue, and cytological preparations of LNCaP prostatic tumour cells. Double staining was restricted to cells derived from the secretory epithelium of the prostate. Cross-reactivity between both detection systems was excluded by several controls, including the use of unrelated antibodies of the same isotype and the staining of CK18+/PSA- HT29 colon carcinoma cells. CK18+ cells co-expressing PSA were found in bone marrow aspirates from 5 out of 13 patients with carcinomas of the prostate, a finding that is consistent with the relative fraction of double-positive LNCaP cells. The specificity of CK18 for epithelial tumour cells in bone marrow was supported by negative staining of 12 control aspirates from patients with benign prostatic hypertrophy. In conclusion, the approach presented appears to be a reliable method to phenotype individual prostatic carcinoma cells.
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页码:61 / 66
页数:6
相关论文
共 32 条
[1]   MEMBRANE ULTRAFILTRATION - DIAFILTRATION TECHNIQUE AND ITS APPLICATION TO MICROSOLUTE EXCHANGE AND BINDING PHENOMENA [J].
BLATT, WF ;
ROBINSON, SM .
ANALYTICAL BIOCHEMISTRY, 1968, 26 (01) :151-&
[2]   EXPRESSION OF SIMPLE EPITHELIAL TYPE CYTOKERATINS IN STRATIFIED EPITHELIA AS DETECTED BY IMMUNOLOCALIZATION AND HYBRIDIZATION INSITU [J].
BOSCH, FX ;
LEUBE, RE ;
ACHTSTATTER, T ;
MOLL, R ;
FRANKE, WW .
JOURNAL OF CELL BIOLOGY, 1988, 106 (05) :1635-1648
[3]   IMMUNOENZYMATIC LABELING OF MONOCLONAL-ANTIBODIES USING IMMUNE-COMPLEXES OF ALKALINE-PHOSPHATASE AND MONOCLONAL ANTI-ALKALINE PHOSPHATASE (APAAP COMPLEXES) [J].
CORDELL, JL ;
FALINI, B ;
ERBER, WN ;
GHOSH, AK ;
ABDULAZIZ, Z ;
MACDONALD, S ;
PULFORD, KAF ;
STEIN, H ;
MASON, DY .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1984, 32 (02) :219-229
[4]   PREDICTION OF EARLY RELAPSE IN PATIENTS WITH OPERABLE BREAST-CANCER BY DETECTION OF OCCULT BONE-MARROW MICROMETASTASES [J].
COTE, RJ ;
ROSEN, PP ;
LESSER, ML ;
OLD, LJ ;
OSBORNE, MP .
JOURNAL OF CLINICAL ONCOLOGY, 1991, 9 (10) :1749-1756
[6]   MONOCLONAL CYTOKERATIN ANTIBODIES THAT DISTINGUISH SIMPLE FROM STRATIFIED SQUAMOUS EPITHELIA - CHARACTERIZATION ON HUMAN-TISSUES [J].
DEBUS, E ;
WEBER, K ;
OSBORN, M .
EMBO JOURNAL, 1982, 1 (12) :1641-1647
[7]  
DEBUS E, 1984, AM J PATHOL, V114, P121
[8]   LIGHT MICROSCOPIC DETECTION OF LEUKOCYTE CELL-SURFACE ANTIGENS WITH A ONE-NANOMETER GOLD PROBE [J].
DEVALCK, V ;
RENMANS, W ;
SEGERS, E ;
LEUNISSEN, J ;
DEWAELE, M .
HISTOCHEMISTRY, 1991, 95 (05) :483-490
[9]   MONOCLONAL-ANTIBODY THAT DEFINES THE PROSTATE SPECIFIC ANTIGEN [J].
DONN, F ;
BRUNS, T ;
VONMEYERINCK, L ;
SCHULZ, M ;
BECKER, WM ;
BECKER, H ;
KLOSTERHALFEN, H .
PROSTATE, 1989, 14 (03) :237-249
[10]   CYTOSKELETAL COMPONENTS OF LYMPHOID ORGANS .1. SYNTHESIS OF CYTOKERATIN-8 AND CYTOKERATIN-18 AND DESMIN IN SUBPOPULATIONS OF EXTRAFOLLICULAR RETICULUM CELLS OF HUMAN LYMPH-NODES, TONSILS, AND SPLEEN [J].
FRANKE, WW ;
MOLL, R .
DIFFERENTIATION, 1987, 36 (02) :145-163