[en] 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.
Disciplines :
Immunology & infectious disease Biotechnology
Author, co-author :
Jacobs, Nathalie ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Giannini, Sandra ; Université de Liège - ULiège > Département des sciences cliniques > Département des sciences cliniques
Al-Saleh, Walid
Hubert, Pascale ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Boniver, Jacques ; Centre Hospitalier Universitaire de Liège - CHU > Anatomie pathologique
Delvenne, Philippe ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Language :
English
Title :
Generation of T lymphocytes from the epithelium and stroma of squamous pre-neoplastic lesions of the uterine cervix.
Publication date :
1999
Journal title :
Journal of Immunological Methods
ISSN :
0022-1759
eISSN :
1872-7905
Publisher :
Elsevier Science
Volume :
223
Issue :
1
Pages :
123-9
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
Centre de Recherche Interuniversitaire en Vaccinologie with a grant from the Walloon Region and SmithKline Beecham Bio- logicals Belgian Fund for Medical Scientific Research B104-CT98-0097 EU contract Oeuvre Belge du Cancer F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
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