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Application of immunomagnetic screening strategy for separation of CD4+ and CD8+ T cell subpopulations of peripheral blood / 中国实验血液学杂志
Journal of Experimental Hematology ; (6): 205-209, 2005.
Article in Chinese | WPRIM | ID: wpr-356592
ABSTRACT
To evaluate the separation of T lymphocyte subsets by immunomagnetic beads and to find optimization of strategy for specific binding of antibody-coated beads to cells, two strategies to isolate enriched T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from small volumes (< 5 ml) of peripheral blood by using immunomagnetic beads or complement cytotoxicity method were compared. The purity and activity of CD4+ T cells and CD8+ T cells were measured by using flow cytometry, trypan-blue dye exclusion test, etc. The results showed that the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using immunomagnetic beads were (94.2 +/- 1.4)% and (93.8 +/- 3.0)% respectively, higher than those of control group and the group of using completement cytotoxicity method (P < 0.05). At the same time, the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using complement cytotoxicity method were (76.0 +/- 2.8)% and (77.0 +/- 3.0)% respectively, higher than those of unenriched group (P < 0.05). The trypan-blue dye exclusion test confirmed that there were no influences on activity of CD4+ T cells and CD8+ T cells when immunomagnetic beads were used for separation of these cells from peripheral blood. It is concluded that the immunomagnetic bead method has a higher efficiency for separation of CD4+ T cells and CD8+ T cells from peripheral blood than complement cytotoxic method, especially for small sample. This method has no influence on activity and proliferation of T lymphocyte subpopulations, and would be expected to establish conditions for research of biological characteristics of CD4+ T cells and CD8+ T cells in future.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Leukocytes, Mononuclear / CD4-Positive T-Lymphocytes / Immunomagnetic Separation / CD8-Positive T-Lymphocytes / Cell Biology / Allergy and Immunology / Flow Cytometry / Methods Type of study: Diagnostic study / Screening study Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2005 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Leukocytes, Mononuclear / CD4-Positive T-Lymphocytes / Immunomagnetic Separation / CD8-Positive T-Lymphocytes / Cell Biology / Allergy and Immunology / Flow Cytometry / Methods Type of study: Diagnostic study / Screening study Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2005 Type: Article