[Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine].
Zhejiang Da Xue Xue Bao Yi Xue Ban
; 35(1): 39-44, 2006 01.
Article
em Zh
| MEDLINE
| ID: mdl-16470919
ABSTRACT
OBJECTIVE:
To detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.METHODS:
The 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.RESULTS:
A 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.CONCLUSION:
It 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.
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Base de dados:
MEDLINE
Assunto principal:
Leucócitos Mononucleares
/
Linfócitos T
/
Citocinas
/
Interferon gama
/
Doença Enxerto-Hospedeiro
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
Zh
Ano de publicação:
2006
Tipo de documento:
Article