Immune Netw.  2007 Jun;7(2):80-86. 10.4110/in.2007.7.2.80.

Enhancement of Proliferation and Antigen Presentation of Human B Cells in Vitro by K562 Cells Expressing CD40L

• Park JY
• Yoon SH
• Kim EK
• Yun SO
• Sohn HJ
• Kim TG

• KMID: 2150630
• DOI: http://doi.org/10.4110/in.2007.7.2.80

Abstract

BACKGROUND: CD40-activated B (CD40-B) cells might be an attractive source of autologous antigen-presenting cells (APCs) for immunotherapy due to the convenience to obtain from peripheral blood and expand in vitro. Moreover, CD40-B cells were found to be comparable with DCs in their capacity to raise antigen-specific CD8+ T cells. Here, we have established K562 cells expressing CD40L to expand CD40-activated B cells used for APCs.
METHODS
After activation of B cell by K562/CD40L, CD40-B cells were examined by counting B cell numbers. Surface expression of CD54, CD80, CD86 and HLA class II was measured by flow cytometry. The CD40-B cells were tested for its function as APC by mixed lymphocyte reactions (MLR) and by induction of T cell responses specific for pp65 peptide in vitro.
RESULTS
The expansion of B cells by K562/CD40L increased about 6-folds compared with anti-CD40 or K562. Furthermore, the expression of CD54, CD80, CD86 and HLA class II was up-regulated by K562/CD40L. B cells by K562/CD40L showed comparable antigen presentation activity with mature DCs as shown in MLR, INF-gamma ELISPOT assay.
CONCLUSION
These results suggest that K562/CD40L could be used to generate activated B cells as potent APCs which could be useful for cellular vaccination and adoptive immunotherapy.

Keyword

Antigen-presenting cell; B cell; CD40L; K562; immunotherapy

MeSH Terms

Antigen Presentation*
Antigen-Presenting Cells
B-Lymphocytes*
CD40 Ligand*
Cell Count
Enzyme-Linked Immunospot Assay
Flow Cytometry
Humans*
Immunotherapy
Immunotherapy, Adoptive
K562 Cells*
Lymphocyte Culture Test, Mixed
T-Lymphocytes
Vaccination
CD40 Ligand