Biology subjects

Gottschalk, E.

Publications and source records attributed to Gottschalk, E..

2 recordsLinked to original sources

bioRxiv2021

Discovering tumor-reactive T-cell receptors through single-cell sequencing of tumor-infiltrating lymphocytes

We evaluated the utility of single-cell sequencing of tumor-infiltrating lymphocytes (TIL) for tumor-reactive T-cell receptor (TCR) discovery. Using the MC38 cell line as our tumor model in mice, we show that expression of exogenous TCRs via mRNA electroporation in human T cells provides an easy and quick path to validating tumor-specific candidate TCRs. We detail the identification and validation of four novel MC38-reactive mouse TCRs with varying levels of reactivity to the target cells. Validating our process, one of the MC38 TCRs is specific against a previously reported neoantigen (ASMTNMELM in the Adpgk gene). Consideration of these methodologies may aid in the development of rapid TCR-based therapies for the treatment of cancer and human disease. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=197 SRC="FIGDIR/small/470597v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@792d44org.highwire.dtl.DTLVardef@18af6aforg.highwire.dtl.DTLVardef@498c2aorg.highwire.dtl.DTLVardef@9127e8_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2019

Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy

Three-dimensional (3D) cell culture systems with tumor spheroids are being adopted for research on the antitumor activity of drug treatments and cytotoxic T cells. Analysis of the cytotoxic effect on 3D tumor cultures within a 3D scaffold, such as collagen, is challenging. Image-based approaches often use confocal microscopy, which greatly limits the sample size of tumor spheroids that can be assayed. We explored a system where tumor spheroids growing in a collagen gel within a microfluidics chip can be treated with drugs or co-cultured with T cells. We attempted to adapt the system to measure the death of cells in the tumor spheroids directly in the microfluidics chip via automated widefield fluorescence microscopy. We were able to successfully measure drug-induced cytotoxicity in tumor spheroids, but had difficulties extending the system to measure T cell-mediated tumor killing. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=193 SRC="FIGDIR/small/842039v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@c95725org.highwire.dtl.DTLVardef@785728org.highwire.dtl.DTLVardef@a23dceorg.highwire.dtl.DTLVardef@187bd74_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗