Evaluation of the cervical cytobrush sampling technique for the preparation of CD45+ mononuclear cells from the human cervix.
A cytobrush technique developed to prepare mononuclear cells from the intraepithelial layer of the endocervix has been evaluated. Specimens yielded approximately 4-6x10(6) cells, of which 10-15% were CD45+. Between 10% and 15% of these CD45+ cells were mononuclear leukocytes. The non-leukocyte cell fraction exhibited high levels of autofluorescence and for flow cytometry analysis, it was necessary to exclude these cells by gating. Macrophages constituted approximately 60% and T lymphocytes, 40% of the mononuclear cells in cytobrush samples. The CD4/CD8 T-cell ratio was similar to that observed in blood. In 9 of 13 specimens, B lymphocytes constituted less than 1% of the mononuclear cell fraction suggesting that the mononuclear cells were derived from the intraepithelial compartment rather then the deeper lamina propria. Lack of B lymphocytes also indicates minimal blood contamination in these samples, a conclusion supported by labelling for the red blood cell (RBC) glycoprotein glycophorin A. However, the need to monitor all samples for possible blood contamination was indicated by 4 of 13 samples in which B lymphocytes accounted for 2-8% of the mononuclear cells.
Prakash
M
M
Department of Immunology, Imperial College of Science Technology and Medicine at Chelsea and Westminster Hospital, 369 Fulham Road, London, UK. m.prakash@ic.ac.uk
Patterson
S
S
Kapembwa
M S
MS
D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002584:Cervix Uteri / Q000166:cytology* / Q000276:immunology*; D004847:Epithelial Cells / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008264:Macrophages / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology
D017493:Leukocyte Common Antigens