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Assessment of the Immunomodulatory Properties of Human Mesenchymal Stem Cells (MSCs)
Published on: December 24, 2015
[Comparative study on activated immunocytes of human bone marrow and peripheral blood by cytokines]
Wen-Rong Huang1, Bo-Long Zhang, Hai-Jie Jin
1Department of Hematology, The General Hospital of PLA, Beijing 100853, China. Huangwr301@yahoo.com.cn
Insights
Combined cytokines significantly enhance immune cell numbers and cytotoxicity in both bone marrow and peripheral blood, supporting their use in cell immunotherapy.
Area of Science:
- Immunology
- Cell Biology
Background:
- Immunocytes in bone marrow and peripheral blood are crucial for immune responses.
- Understanding their activation and function is key for developing effective immunotherapies.
Purpose of the Study:
- To investigate the immunophenotype and cytotoxicity of bone marrow and peripheral blood mononuclear cells (MNC) after activation with combined cytokines.
- To assess the impact of interferon-gamma (IFN-γ), interleukin-1 (IL-1), interleukin-2 (IL-2), and anti-CD3 monoclonal antibody (McAb-CD3) on MNCs.
Main Methods:
- In vitro activation of bone marrow and peripheral blood MNCs using IFN-γ, IL-1, IL-2, and McAb-CD3.
- Observation of cell count and morphology during culture.
- Cytochemical staining and immunophenotype analysis (CD3, CD56, CD38) before and after culture.
- Cytotoxicity assessment using the MTT method.
Main Results:
- Combined cytokines significantly increased MNC numbers in both bone marrow and peripheral blood (P < 0.05), with a more pronounced increase in peripheral blood MNCs.
- Cytochemical staining revealed decreased POX and increased PAS activity.
- Positive ratios of CD3+, CD56+, and CD38+ cells increased significantly post-culture (P < 0.05).
- Peripheral blood MNCs showed a significant increase in CD3+ CD56+ cells, unlike bone marrow MNCs.
- No significant difference in cytotoxicity was observed between the two groups.
Conclusions:
- IFN-γ, IL-1, IL-2, and McAb-CD3 enhance the cell number and cytotoxicity of both bone marrow and peripheral blood MNCs.
- Activated MNCs demonstrate potential for use in cell-based immunotherapies.
Abstract:
To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
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