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Density and phenotype of tumour-associated mononuclear cells in colonic carcinomas determined by computer-assisted video image analysis.
Authors:M N Norazmi   A W Hohmann   J M Skinner   L R Jarvis     J Bradley
Affiliation:Department of Clinical Immunology, Flinders Medical Centre, South Australia.
Abstract:The density and phenotypes of tumour-associated mononuclear cells (TAMC) in tissue sections of colonic carcinomas was determined by the technique of video image analysis (VIA). This technique allowed an accurate and objective enumeration of both total mononuclear cells (MC) in H&E stained sections and individual types of cells as revealed by immunoperoxidase staining with monoclonal antibodies in frozen sections. This enumeration allowed reliable statistical analysis of the differences between sample groups. Using this technique it was found that the density of MC in histiologically normal tissue was significantly higher than in tumour tissue. Tumours from patients with the best prognosis (stage A) had significantly higher numbers of TAMC than stage B (P less than 0.02), C (P less than 0.002) and D (P less than 0.002) tumours. The differences in the density of TAMC between tumours obtained from stage B and C and that between C and D were not significant, whereas stage B had a significantly higher TAMC density than stage D tumours (P less than 0.05). Comparing tumour differentiation, well differentiated adenocarcinomas had a significantly higher (P less than 0.05) TAMC density than poorly differentiated tumours but not moderately differentiated tumours. Moderately and poorly differentiated adenocarcinomas did not differ significantly in the density of TAMC. In examining the phenotype of these cells, it was found that T lymphocytes formed the majority of the TAMC with the CD4+ subset predominating in 28 of 29 cases. Similarly, all sections of normal colon (taken at least 4 cm away from the tumour) had more CD4+ than CD8+ cells. The proportion of the total leucocyte population that was CD3+ was comparable in normal and tumour tissue. Generally, few macrophages were present in either tumour or normal tissues. B cells (CD21%) and subset of NK cells (CD57+) were not detected in the tumours. There were no significant differences in the proportion of leucocytes which were CD4+, CD8+ and CD14+ (macrophages) between the normal colon and the tumour tissues. The types of cells in the TAMC population did not differ with tumour stage or differentiation or with the density of the TAMC itself.
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