Replacing human peripheral blood leukocytes with LRS-separated leukocytes for scientific research / 中华微生物学和免疫学杂志

ABSTRACT

Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.