Generation of highly pure fusions of colorectal carcinoma and antigen-presenting cells

Klier, Ulrike; Maletzki, Claudia; Klar, E.; Linnebacher, Michael

doi:10.1007/s00423-010-0598-1

Cited by 6 publications

(8 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Data presented here confirm our previous findings that cell hybrids of human MSI + tumor cells and CD40 Bs as APC can easily be generated [21]. Fusion cells retain desired characteristics of both fusion partners; in particular the expression of MSI-induced FSPs and of functional antigen processing and presentation machinery together with potent immunostimulatory capacity.…”

Section: Discussionsupporting

confidence: 86%

“…Semiallogenic cellular fusions were performed in the presence of polyethylene glycol. The fusion efficiencies were between 10 and 15% [data not shown] and thus very similar to published data [21]. However, additionally to fresh fusions we decided to obtain fusion cell clones as this allowed us to select variants expressing mutations in cMS giving rise to FSPs with high immunogenic potential and additionally high amounts of major histocompatibility complex (MHC) and of the costimulatory molecules CD40, CD80 and CD86.…”

Section: Resultssupporting

confidence: 74%

“…Fusion cells retain desired characteristics of both fusion partners; in particular the expression of MSI-induced FSPs and of functional antigen processing and presentation machinery together with potent immunostimulatory capacity. In the present study we worked with stable fusion cell clones instead of highly purified fresh fusion cells we described recently [21]. This approach was chosen to ensure a uniform fusion cell population concerning the immunostimulatory phenotype and the expression of FSPs.…”

Section: Discussionmentioning

confidence: 99%

“…Good arguments in favour of CD40 Bs as APC are the possibilities to easily isolate them even out of minimal amounts of peripheral blood, to propagate them for longer time periods and lastly the outstanding T cell stimulatory capacity of fully activated B cells [8,21,22]. …”

Section: Discussionmentioning

confidence: 99%

“…Very recently, we optimized the generation of cellular fusions consisting of CD40 Bs and MSI + CRC cells [21]. In the present study, we have evaluated the potency of T cell induction by semiallogenic cell fusions of a MSI + tumor cell line and CD40 Bs.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Semiallogenic fusions of MSI+tumor cells and activated B cells induce MSI-specific T cell responses

2011

Self Cite

View full text Add to dashboard Cite

BackgroundVarious strategies have been developed to transfer tumor-specific antigens into antigen presenting cells in order to induce cytotoxic T cell responses against tumor cells. One approach uses cellular vaccines based on fusions of autologous antigen presenting cells and allogeneic tumor cells. The fusion cells combine antigenicity of the tumor cell with optimal immunostimulatory capacity of the antigen presenting cells.Microsatellite instability caused by mutational inactivation of DNA mismatch repair genes results in translational frameshifts when affecting coding regions. It has been shown by us and others that these mutant proteins lead to the presentation of immunogenic frameshift peptides that are - in principle - recognized by a multiplicity of effector T cells.MethodsWe chose microsatellite instability-induced frameshift antigens as ideal to test for induction of tumor specific T cell responses by semiallogenic fusions of microsatellite instable carcinoma cells with CD40-activated B cells. Two fusion clones of HCT116 with activated B cells were selected for stimulation of T cells autologous to the B cell fusion partner. Outgrowing T cells were phenotyped and tested in functional assays.ResultsThe fusion clones expressed frameshift antigens as well as high amounts of MHC and costimulatory molecules. Autologous T cells stimulated with these fusions were predominantly CD4+, activated, and reacted specifically against the fusion clones and also against the tumor cell fusion partner. Interestingly, a response toward 6 frameshift-derived peptides (of 14 tested) could be observed.ConclusionCellular fusions of MSI+ carcinoma cells and activated B cells combine the antigen-presenting capacity of the B cell with the antigenic repertoire of the carcinoma cell. They present frameshift-derived peptides and can induce specific and fully functional T cells recognizing not only fusion cells but also the carcinoma cells. These hybrid cells may have great potential for cellular immunotherapy and this approach should be further analyzed in preclinical as well as clinical trials. Moreover, this is the first report on the induction of frameshift-specific T cell responses without the use of synthetic peptides.

show abstract

Section: Discussionsupporting

confidence: 86%

Section: Resultssupporting

confidence: 74%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Semiallogenic fusions of MSI+tumor cells and activated B cells induce MSI-specific T cell responses

2011

Self Cite

View full text Add to dashboard Cite

show abstract

Avitalized bacteria mediate tumor growth control via activation of innate immunity

Klier

Maletzki

Göttmann

et al. 2011

Cellular Immunology

View full text Add to dashboard Cite

Establishment, Characterization and Chemosensitivity of Three Mismatch Repair Deficient Cell Lines from Sporadic and Inherited Colorectal Carcinomas

et al. 2012

Self Cite

View full text Add to dashboard Cite

BackgroundColorectal cancer (CRC) represents a morphologic and molecular heterogenic disease. This heterogeneity substantially impairs drug effectiveness and prognosis. The subtype of mismatch repair deficient (MMR-D) CRCs, accounting for about 15% of all cases, shows particular differential responses up to resistance towards currently approved cytostatic drugs. Pre-clinical in vitro models representing molecular features of MMR-D tumors are thus mandatory for identifying biomarkers that finally help to predict responses towards new cytostatic drugs. Here, we describe the successful establishment and characterization of three patient-derived MMR-D cell lines (HROC24, HROC87, and HROC113) along with their corresponding xenografts.MethodologyMMR-D cell lines (HROC24, HROC87, and HROC113) were established from a total of ten clinicopathological well-defined MMR-D cases (120 CRC cases in total). Cells were comprehensively characterized by phenotype, morphology, growth kinetics, invasiveness, and molecular profile. Additionally, response to clinically relevant chemotherapeutics was examined in vitro and in vivo.Principal FindingsTwo MMR-D lines showing CIMP-H derived from sporadic CRC (HROC24: K-raswt, B-rafmut, HROC87: K-raswt, B-rafmut), whereas the HROC113 cell line (K-rasmut, B-rafwt) was HNPCC-associated. A diploid DNA-status could be verified by flow cytometry and SNP Array analysis. All cell lines were characterized as epithelial (EpCAM+) tumor cells, showing surface tumor marker expression (CEACAM+). MHC-class II was inducible by Interferon-γ stimulation. Growth kinetics as well as invasive potential was quite heterogeneous between individual lines. Besides, MMR-D cell lines exhibited distinct responsiveness towards chemotherapeutics, even when comparing in vitro and in vivo sensitivity.ConclusionsThese newly established and well-characterized, low-passage MMR-D cell lines provide a useful tool for future investigations on the biological characteristics of MMR-D CRCs, both of sporadic and hereditary origin. Additionally, matched patient-derived immune cells allow for comparative genetic studies.

show abstract

Generation of highly pure fusions of colorectal carcinoma and antigen-presenting cells

Abstract: 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.

Cited by 6 publications

References 19 publications

Semiallogenic fusions of MSI+tumor cells and activated B cells induce MSI-specific T cell responses

Semiallogenic fusions of MSI+tumor cells and activated B cells induce MSI-specific T cell responses

Avitalized bacteria mediate tumor growth control via activation of innate immunity

Establishment, Characterization and Chemosensitivity of Three Mismatch Repair Deficient Cell Lines from Sporadic and Inherited Colorectal Carcinomas

Contact Info

Product

Resources

About