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 intra epithelial layer of the endocervix has been evaluated. Specimens yielded ap proximately 4-6 X 10(6) cells, of which 10-15% were CD45 +. Between 10% and 15% of these CD45 + cells were mononuclear leukocytes. The non-leukocyte c ell fraction exhibited high levels of autofluorescence and for flow cytomet ry analysis, it was necessary to exclude these cells by gating. Macrophages constituted approximately 60% and T lymphocytes, 40% of the mononuclear ce lls in cytobrush samples. The CD4/CD8 T-cell ratio was similar to that obse rved in blood. In 9 of 13 specimens, B lymphocytes constituted less than 1% of the mononuclear cell fraction suggesting that the mononuclear cells wer e derived from the intraepithelial compartment rather then the deeper lamin a propria. Lack of B lymphocytes also indicates minimal blood contamination in these samples, a conclusion supported by labelling for the red blood ce ll (RBC) glycoprotein glycophorin A. However, the need to monitor all sampl es for possible blood contamination was indicated by 4 of 13 samples in whi ch B lymphocytes accounted for 2-8% of the mononuclear cells. (C) 2001 Else vier Science B.V. All rights reserved.