Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine.
- Author:
Xiao-jian MENG
1
;
Mao-fang LIN
;
Zhen CAI
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; pharmacology; CD28 Antigens; immunology; Cell Proliferation; Cells, Cultured; Cytokines; secretion; Graft vs Host Disease; immunology; Humans; Interferon-gamma; secretion; Interleukin-2; secretion; Leukocytes, Mononuclear; cytology; immunology; Lymphocyte Culture Test, Mixed; Muromonab-CD3; pharmacology; T-Lymphocytes; cytology; immunology
- From: Journal of Zhejiang University. Medical sciences 2006;35(1):39-44
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.
METHODSThe novel cytokine secretion assay (CKSA) was applied to detect T lymphocytes secreting IFN-gamma at single cell level in human mixed lymphocytes reaction. IFN-gamma secreting T cells were enriched by means of magnetic sorting system and expanded with OKT(3), anti-CD(3)mAb and IL-2 combination. Antigen specificity of the expanded cells was confirmed using enzyme linked immunospot assay.
RESULTSA sizable proportion of IFN-gamma secreting T lymphocytes could be detected [(1.12 +/-0.13)% compared with (0.23 +/-0.07)%] and be further enriched to (67.3 +/-10.5)%, or (93.8 +/-22.1) fold. T lymphocytes could be expanded up to 600-fold within 21-28 days and the specific IFN-gamma response of expanded cells was confirmed with stimulation of the relevant allogeneic PBMNC, which was significantly higher than the irrelevant PBMNC control.
CONCLUSIONIt is feasible to detect significantly increased IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation based on the CKSA technique at single cell level and these cells can be efficiently enriched and expanded for further research.
