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[Detection, enrichment and expansion of T lymphocytes mediating alloresponse based on cytokine].
Meng, Xiao-jian; Lin, Mao-fang; Cai, Zhen.
Afiliação
  • Meng XJ; Department of Hematology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
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.
Assuntos
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Coleções: 01-internacional 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
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Coleções: 01-internacional 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