In-vitro activation of cytotoxic T lymphocytes by fusion of mouse hepatocellular carcinoma cells and lymphotactin gene-modified dendritic cells.

Abstract

AIM:

To investigate the in-vitro activation of cytotoxic T lymphocytes (CTLs) by fusion of mouse hepatocellular carcinoma (HCC) cells and lymphotactin gene-modified dendritic cells (DCs).

METHODS:

Lymphotactin gene modified DCs (DCLptn) were prepared by lymphotactin recombinant adenovirus transduction of mature DCs which differentiated from mouse bone marrow cells by stimulation with granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-alpha). DCLptn and H22 fusion was prepared using 50% PEG. Lymphotactin gene and protein expression levels were measured by RT-PCR and ELISA, respectively. Lymphotactin chemotactic responses were examined by in-vitro chemotaxis assay. In-vitro activation of CTLs by DCLptn/H22 fusion was measured by detecting CD25 expression and cytokine production after autologous T cell stimulation. Cytotoxic function of activated T lymphocytes stimulated with DCLptn/H22 cells was determined by LDH cytotoxicity assay.

RESULTS:

Lymphotactin gene could be efficiently transduced to DCs by adenovirus vector and showed an effective biological activity. After fusion, the hybrid DCLptn/H22 cells acquired the phenotypes of both DCLptn and H22 cells. In T cell proliferation assay, flow cytometry showed a very high CD25 expression, and cytokine release assay showed a significantly higher concentration of IFN-gamma and IL-2 in DCLptn/H22 group than in DCLptn, DCLptn+H22, DC/H22 or H22 groups. Cytotoxicity assay revealed that T cells derived from DCLptn/H22 group had much higher anti-tumor activity than those derived from DCLptn, H22, DCLptn+H22, DC/H22 groups.

CONCLUSION:

Lymphotactin gene-modified dendritoma induces T-cell proliferation and strong CTL reaction against allogenic HCC cells. Immunization-engineered fusion hybrid vaccine is an attractive strategy in prevention and treatment of HCC metastases.

• Authors: Xi-Ling Sheng, Hao Zhang
• Journal: World journal of gastroenterology (World J Gastroenterol) Vol. 13 Issue 44 Pg. 5944-50 (Nov 28 2007) ISSN: 1007-9327 [Print] United States
• PMID: 17990361 (Publication Type: Journal Article)

Chemical References

• Chemokines, C
• Cytokines
• Interleukin-2 Receptor alpha Subunit
• Xcl1 protein, mouse

Topics

• Adenoviridae
• Animals
• Carcinoma, Hepatocellular (immunology, metabolism, pathology)
• Cell Fusion
• Cell Line, Tumor
• Cell Proliferation
• Cells, Cultured
• Chemokines, C (genetics, metabolism)
• Cytokines (metabolism)
• Dendritic Cells (immunology, metabolism, pathology)
• Female
• Interleukin-2 Receptor alpha Subunit (metabolism)
• Liver Neoplasms, Experimental (immunology, metabolism, pathology)
• Lymphocyte Activation
• Mice
• Mice, Inbred BALB C
• T-Lymphocytes, Cytotoxic (immunology, pathology, physiology)
• Transduction, Genetic