A comparison of flow cytometry and immunohistochemistry in human colorectal cancers.
In human colorectal cancer it has been reported that some tumours lack the HLA-ABC antigens. This has been interpreted as reflecting tumour escape from the immune system. Earlier data have been obtained by immunohistochemistry. In this study, we compared the expression of HLA-ABC, HLA-DR, CD80 (B7-1) and CD54 (ICAM-1) in 20 tumours using both a conventional immunohistochemistry two-layer technique and multiparameter flow cytometry, gating on an epithelial cell marker. Colorectal cancer tissue used in flow cytometry was dissociated with collagenase, deoxyribonuclease and hyaluronidase. The intensity of expression of HLA-ABC, HLA-DR and CD80 was unaffected by the enzymes, but CD54 was decreased by 30%. The reproducibility of flow cytometry was good. Microscopy of sections revealed that about 5% of each tumour sample consisted of normal epithelium, but even after correction for this, flow cytometry was superior to immunohistochemistry in 33 out of 80 cases, and showed that tumours described as HLA-ABC negative by immunohistochemistry were in fact weakly positive for HLA-ABC. We conclude that flow cytometry and immunohistochemistry are complementary, and that flow cytometry is superior to immunohistochemistry for detecting antigens/epitopes present in low amounts.
Department of Clinical Immunology, Odense University Hospital, Denmark.
A C
AC
Diederichsen
T P
TP
Hansen
O
O
Nielsen
C
C
Fenger
J C
JC
Jensenius
P B
PB
Christensen
T
T
Kristensen
J
J
Zeuthen
D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D018122:B7-1 Antigen / Q000302:isolation & purification; D015179:Colorectal Neoplasms / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006680:HLA Antigens / Q000302:isolation & purification*; D015234:HLA-A Antigens / Q000302:isolation & purification; D015235:HLA-B Antigens / Q000302:isolation & purification; D015236:HLA-C Antigens / Q000302:isolation & purification; D006684:HLA-DR Antigens / Q000302:isolation & purification; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D018799:Intercellular Adhesion Molecule-1 / Q000302:isolation & purification; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results
D000951:Antigens, Neoplasm; D018122:B7-1 Antigen; D006680:HLA Antigens; D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D006684:HLA-DR Antigens; D018799:Intercellular Adhesion Molecule-1