Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 39-44, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-355148
ABSTRACT
<p><b>OBJECTIVE</b>To detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Leukocytes, Mononuclear
/
T-Lymphocytes
/
Cells, Cultured
/
Cytokines
/
Interferon-gamma
/
Interleukin-2
/
Muromonab-CD3
/
Lymphocyte Culture Test, Mixed
/
CD28 Antigens
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2006
Type:
Article
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