Korean J Hematol.  2006 Sep;41(3):186-193. 10.5045/kjh.2006.41.3.186.

Induction of Myeloma Cell Line-specific Cytotoxic T Lymphocytes using Monocyte-derived Dendritic Cells Pulsed with Myeloma Cell Line Lysates

• Park MS
• Park JS
• Kang HK
• Kim SK
• Won JH
• Choi BH
• Shin SW
• Zhu XW
• Jin CJ
• Pham TN
• Cho D
• Nam JH
• Kim YJ
• Kim YK
• Yang DH
• Chung IJ
• Kim HJ
• Lee JJ

• KMID: 2252300
• DOI: http://doi.org/10.5045/kjh.2006.41.3.186

Abstract

BACKGROUND: In multiple myeloma (MM), the idiotype (ID) determinant of the paraprotein has been used for immunotherapy using dendritic cells (DCs). However, ID-specific immune responses showed limited clinical responses after the Id vaccination. Therefore, an alternative approach using DCs pulsed with other tumor antigens is required.
METHODS
We investigated the possibility of immunotherapy for MM using myeloma cell line-specific cytotoxic T lymphocytes (CTLs), that were stimulated in vitro by monocyte-derived DCs pulsed with the myeloma cell line ysates. CD14+ cells isolated from the peripheral blood of HLA-A0201+ healthy donors were cultured in the presence of GM-CSF and IL-4. On day 6, the immature DCs were pulsed with the myeloma cell line lysates (IM-9: HLA0201+ and ARH-77: HLA0201+), and then maturation of DCs was induced by the addition of TNF- alpha for 2 days. CTL lines were generated by a 2 time stimulation with DCs to the autologous CD3+ T cells.
RESULTS
DCs pulsed with myeloma cell lysates showed the production of IL-12p70, but less than that of unpulsed DCs. CTLs lines stimulated with the DCs pulsing, for the myeloma cell line lysates, showed potent cytotoxic activities against autologous target cells, but not against HLA-A2-cell lines (RPMI-8226). Mature DCs pulsed with the myeloma cell line lysates showed a higher stimulatory capacity for autologous CTL when compared with mature non-pulsed DCs.
CONCLUSION
These results suggest that DCs pulsed with the myeloma cell line lysates can generate potent myeloma cell line-specific CTLs for the myeloma cell-based immunotherapeutic approach in MM.

Keyword

Dendritic cells; Myeloma; Lysate; Cell line

MeSH Terms

Antigens, Neoplasm
Cell Line*
Dendritic Cells*
Granulocyte-Macrophage Colony-Stimulating Factor
Humans
Immunotherapy
Interleukin-4
Multiple Myeloma
T-Lymphocytes
T-Lymphocytes, Cytotoxic*
Tissue Donors
Vaccination
Antigens, Neoplasm
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-4

Figure

• 

  Fig. 1 Phenotypic analysis of monocytes (top panel), immature dendritic cells (DCs) (middle panel), or mature DCs (bottom panel). CD14+ cells isolated from the MNCs of normal donor by MACS were cultured with a combination of GM-CSF and IL-4. On day 6, the immature DCs (middle panel) were pulsed with tumor antigens, and then maturation was induced by the addition of TNF-α (bottom panel).

• 

  Fig. 2 Production of IL-12 by DCs pulsed with or without myeloma cell line lysates (A: ARH-77, B: IM-9) after the stimulation with CD40L-transfected J558 cells for 24 hr. IL-12p70 concentrations in 24 hr supernatants were determined by ELISA. Results, expressed as mean±SD of duplicate culture, are from one representative experiment of two.

• 

  Fig. 3 ELISPOT data showing the mean number of spots and standard deviation per 1～4×105 T cells. Comparison of IFN-γ secreting cells of CTL generated from stimulation with DCs pulsed with ARH-77 lysates (A) or IM-9 lysates (B) against autologous myeloma cell line lysates or RPMI-8226. The data are shown as the mean IFN-γ secreting cell (±SD) of duplicate culture from one representative experiment of two.

• 

  Fig. 4 T cell-stimulatory capacity of DCs with or without myeloma cell line lysates (ARH-77: A, IM-9: B) for CTLs that were generated from stimulation with DCs with myeloma cell line lysates. The data are shown as the mean cpm (±SD) of triplicate cultures from one representative experiment of two.

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