Generation of T lymphocytes from the epithelium and stroma of squamous pre-neoplastic lesions of the uterine cervix.
In this study, we have developed a simple and efficient technique for the isolation of viable lymphocytes from the epithelium and stroma of small pre-neoplastic squamous intraepithelial lesions (SIL) of the uterine cervix. Following the separation of the epithelium from the stroma using dispase II, both biopsy fragments were used to generate T lymphocytes. The stroma-derived lymphocytes were obtained by collecting and culturing the cells migrating out of the biopsy in the presence of IL2 (50 U/ml). An average of 0.7 x 10(6) and 1.4 x 10(6) lymphocytes could be obtained after 20 and 30 days of culture, respectively. For the expansion of lymphocytes derived from the pre-neoplastic epithelium (SIL) it was necessary to use a combination of irradiated peripheral blood mononuclear cells (PBMC) as a feeder layer with PHA (0.1%), in addition to IL2 (50 U/ml). Interestingly, these lymphocytes could be obtained using either allogeneic or syngeneic PBMCs. With this protocol, we were able to generate up to 100 x 10(6) lymphocytes from the epithelium, the majority of which were T lymphocytes.
University of Liège, Department of Pathology, Belgium. n.jacobs@ulg.ac.be
N
N
Jacobs
S L
SL
Giannini
W
W
Al-Saleh
P
P
Hubert
J
J
Boniver
P
P
Delvenne
D002278:Carcinoma in Situ / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000276:immunology / Q000473:pathology; D002469:Cell Separation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000276:immunology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000494:pharmacology; D008213:Lymphocyte Activation; D011230:Precancerous Conditions / Q000276:immunology*; D017154:Stromal Cells / Q000276:immunology / Q000473:pathology; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000473:pathology; D002583:Uterine Cervical Neoplasms / Q000276:immunology*; D002578:Uterine Cervical Dysplasia / Q000276:immunology*
D007376:Interleukin-2