Generation, Phenotyping and Functional Analysis of Dendritic Cells (DC) Derived from Human Monocytes and Acute Myeloid Leukaemia (AML) Cells
Dendritic cells (DC) are efficient and potent antigen-presenting cells in our immune system. The ability of DC to present antigens and stimulate T cells has prompted their application as therapeutic cancer vaccines. The objective of this study was to generate DC from two resources: monocytes and...
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Format: | Thesis |
Language: | English English |
Published: |
2004
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Subjects: | |
Online Access: | http://psasir.upm.edu.my/id/eprint/6283/1/FPSK%28M%29_2004_2.pdf |
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Summary: | Dendritic cells (DC) are efficient and potent antigen-presenting cells in our immune
system. The ability of DC to present antigens and stimulate T cells has prompted
their application as therapeutic cancer vaccines. The objective of this study was to
generate DC from two resources: monocytes and AML blasts. The generated DC
were evaluated for their morphology by phase contrast microscopy and May
Grunwald Giemsa staining. Viability of cells was determined by trypan blue dye
exclusion. Percentage of yields and immunophenotypes were carried out with flow
cytometry. The functional capability of DC was also tested in Mixed Leukocyte
Reactions and anti-leukaemia cytotoxicity assay. As a result, the generated DC
shown typical morphology of those reported and expressed DC surface markers
including CDl a, CD83, CD86, CD80 and HLA-DR. Down regulation of CD14 was
also observed for cultured monocytes. In MLR assay, both generated DC elicited
strong allo-stimulatory response up to more than 100 fold compared to preculture
cells. Mild anti-leukaemia cytotoxicity effect (1 5%) was also observed fiom primed
effector cells with AML antigen pulsed DC generated fiom monocytes. These data
indicate that DC were successfully generated fiom the two resources and they were
capable of eliciting immune response. |
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