Biomedical subjects

Rik J Scheper

Publications and source records attributed to Rik J Scheper.

76 records · Page 5Linked to original sources

PubMedDate not supplied

A single-component CD40-targeted adenovirus vector displays highly efficient transduction and activation of dendritic cells in a human skin substrate system.

Dendritic cell (DC) based tumor vaccination usually involves the administration of ex vivo generated autologous DC. Transduction of DC by viral vectors in vivo has been proposed as a more standardized and easily clinically applicable approach. Previously, we have reported that an Ad5 vector targeted to CD40 via genetic capsid incorporation of CD40L achieves selective transduction of DC in vitro. In the present study, we evaluate the ability of this vector to deliver transgenes in a stringent human substrate system. We report the capacity of this CD40-targeted vector to infect, with high efficiency, cutaneous DC resident in human skin explants, while simultaneously inducing their activation and maturation. This latest generation of single-component, fully targeted vectors should make feasible the clinical testing of in vivo DC-targeted vaccines.

Adenoviridae↗

PubMedDate not supplied

The CXCR3 targeting chemokine CXCL11 has potent antitumor activity in vivo involving attraction of CD8+ T lymphocytes but not inhibition of angiogenesis.

The IFN-gamma-inducible and CXCR3-targeting human CXC chemokines CXCL9 (Mig) and CXCL10 (IP10) have potent antitumor activity through attraction of cytotoxic T lymphocytes and inhibition of angiogenesis. The more recently identified CXCR3-targeting chemokine CXCL11 (I-TAC/IP9) proved to be a more potent chemokine than CXCL9 and CXCL10 in vitro, both in chemotaxis assays with CXCR3+ T lymphocytes and in calcium mobilization experiments. However, its antitumor activity in vivo has not been shown so far. To investigate this, mice were challenged with EL4 T-cell lymphoma cells, genetically modified to produce murine CXCL11. Tumor growth curves showed complete rejection of CXCL11-producing tumors but not of control tumors. Tumor infiltrate analysis by flow cytometry showed a clear correlation between rejection of CXCL11-producing tumors and an increase of tumor-infiltrating CD8+CXCR3+ as well as CD8+CXCR3- T lymphocytes. In vivo CD8 T-cell depletion completely abrogated the antitumor effect. No difference in angiogenesis between control and CXCL11-producing tumors was observed. In survivors, rechallenge experiments with wild-type tumor cells suggested development of protective antitumor immunity involving tumor-specific IFN-gamma production by CD8+ T lymphocytes. These experiments show, for the first time, antitumor activity of CXCL11 in vivo, which warrants exploration for its potential role in anticancer immunotherapy.

Animals↗

PubMedDate not supplied

Constitutively active STAT5b induces cytokine-independent growth of the acute myeloid leukemia-derived MUTZ-3 cell line and accelerates its differentiation into mature dendritic cells.

The CD34(+) human acute myeloid leukemia-derived cell line MUTZ-3 is dependent on hematopoietic growth factors for its proliferation and is able to differentiate into dendritic cells (DCs) in response to the combination of granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor necrosis factor-alpha. This cell line carries human leukocyte antigen (HLA)-A2.1, HLA-A3, and HLA-B44, which cover most of the caucasian population, and it could therefore be used as an off-the-shelf allogeneic DC-based vaccine. Signal transduction and activation of transcription (STAT) 5b is involved in cytokine signal transduction, particularly of cytokines involved in DC precursor growth and differentiation. The constitutively active form of STAT5b induced cytokine-independent growth of MUTZ-3 cells. Furthermore, STAT5b-transduced cells differentiated into mature DCs in 3 to 4 days after stimulation with DC differentiation-inducing cytokines, reducing the culture period to obtain mature DCs with 5 days compared with unmodified MUTZ-3-derived mature DC cultures. Both DC types expressed DC maturation markers and were equally effective in inducing primary T-cell responses. DCs derived from the STAT5b-transduced cells had a more stable mature phenotype after cytokine deprivation, which was reflected in a better performance in functional assays. In conclusion, these results show that STAT5b-transduced MUTZ-3 can be propagated in cytokine-free medium and rapidly differentiated into functional mature DCs that sustain a mature phenotype over a period of 3 to 5 days in the absence of differentiation-inducing cytokines. The simplified propagation and rapid differentiation into mature DCs may facilitate clinical application of this cell line as an allogeneic DC-based vaccine.

Antigens, CD↗

PubMedDate not supplied

Assessment of the classical MDR phenotype in epithelial ovarian carcinoma using primary cultures: a feasibility study.

This in vitro feasibility study has assessed a number of techniques and their applicability when looking at the role of multidrug resistance (MDR) in solid tumours. Fresh tumour material was obtained from 34 patients, (11 previously treated, 23 untreated) with ovarian adenocarcinoma. Doxorubicin sensitivity was measured using the MTT assay +/- the cyclosporins, Pgp expression was assessed by immunocytochemistry with the MRK-16 MoAb and flow cytometry was used to assess intracellular drug accumulation +/- PSC 833. 85% of samples showed some evidence of modest chemosensitisation by the cyclosporins (median 1.74-fold). We saw a marked variation in the number of Pgp positive cells between patients (1-87%, median 31%). 63% of samples tested showed an enhancement of DNR accumulation in the presence of PSC 833, with a median increase of 7% (sample range 0-29%). The present study highlights some of the technical difficulties encountered when working with fresh tumour material ex vivo. We conclude that screening of patients for their suitability to enter clinical trials incorporating MDR modulating agents is technically demanding, but feasible.

ATP Binding Cassette Transporter, Subfamily B, Mem↗