Document Detail


Generation of highly pure fusions of colorectal carcinoma and antigen-presenting cells.
MedLine Citation:
PMID:  20309577     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: The induction of potent T cell responses against tumors is the goal of tumor immunotherapy. One approach is the fusion of antigen-presenting cells (APCs) with tumor cells. Hybrid cells combine the antigenicity of tumors with the immunostimulatory capacity of APCs. However, contaminating unfused cells present in the fusion reaction may prevent the induction of antitumoral immune responses. Here, we present a simple and effective protocol to substantially elevate the purity of hybrid cells. METHODS: Colorectal tumor cell lines and CD40-activated B cells as APCs were fused using polyethylene glycol. Important parameters including cell numbers, concentrations, and handling and detection procedures were optimized. Combination of these optimized fusion conditions with both magnetic cell sorting and selective adherence delivered very pure preparations of APC/tumor cell hybrids. The T cell stimulatory capacity of these hybrids was tested using ELISpot. RESULTS: The optimization of the fusion resulted in maximal fusion efficiencies of 31.6% (n = 10, range 13.5-46.6%). Prelabeling of APCs with magnetic beads allowed for easy elimination of up to 94.3% of unfused tumor cells from the cell mixture by magnetic separation. Hybrid cell capacity to firmly adhere to plastic was then used to remove unfused B cells from the remaining cell mixture by simple washing. The obtained 85.0% pure hybrids cells readily induced antitumoral T cell responses. CONCLUSIONS: Our protocol delivers pure hybrid cell preparations with strong immunostimulatory potential. In subsequent experiments, the ability of hybrid cells to stimulate specific antitumoral T cell responses must be tested in vivo.
Authors:
Ulrike Klier; Claudia Maletzki; Ernst Klar; Michael Linnebacher
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-03-23
Journal Detail:
Title:  Langenbeck's archives of surgery / Deutsche Gesellschaft für Chirurgie     Volume:  395     ISSN:  1435-2451     ISO Abbreviation:  Langenbecks Arch Surg     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-04-02     Completed Date:  2010-07-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9808285     Medline TA:  Langenbecks Arch Surg     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  365-71     Citation Subset:  IM    
Affiliation:
Division of Molecular Oncology and Immunotherapy, Department of General Surgery, University of Rostock, Schillingallee 35, 18055, Rostock, Germany.
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MeSH Terms
Descriptor/Qualifier:
Aged, 80 and over
Antigen-Presenting Cells / immunology*
Antigens, CD40 / immunology
B-Lymphocytes / immunology*
Cell Line, Tumor
Colorectal Neoplasms
Humans
Hybrid Cells / immunology*
Male
Chemical
Reg. No./Substance:
0/Antigens, CD40

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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