Analysis of T cell proliferative responses to antigenic or mitogenic stimuli are based on e.g. the incorporation tritiated thymidine BrdU, CFSE labeling and detection secreted cytokines by ELISA ELISPOT assay. There inherent drawbacks these methods rather long ex vivo expansion, stimulation protocols inability distinguish specific populations assumption importance certain cytokines. Rapid ident...

The stable incorporation of the intracellular dye CSFE into lymphocytes is a powerful tool to monitor lymphocyte migration in vivo and to quantitatively analyze cell division both in vivo and in vitro. This unit describes methods for labeling high or low numbers of lymphocytes with CSFE. Protocols are provided to use CSFE-labeled cells in cell transfer studies or as cells to be cultured in vitr...

The spontaneous proliferation of peripheral blood mononuclear cells (PBMCs) is a hallmark of the human T-lymphotropic virus (HTLV) type-1. Cell proliferation is usually measured using a [(3)H]thymidine incorporation assay. This study aims to quantify the spontaneous proliferation of PBMCs using flow cytometry. PBMCs were cultured for 24 to 120 hours in the presence of 5,6-carboxyfluorescein dia...

We have recently shown that activin A, a member of TGF-beta superfamily, stimulates mouse B cells to express IgA isotype but other isotypes. In the present study, we further characterized effects of activin A on B cell growth and IgA expression. We found that activin A did not have effect on LPS-stimulated cell viability. In parallel, CFSE staining analysis revealed that activin A did not alter...

Mobilization of tumor-reactive CD8+ T cells remains the major challenge of cancer immunotherapy. Knowing how and when the T cell response expands and differentiates after antigen stimulation would make a significant contribution to the development of tumor vaccines. In the current study, we used CFSE-based cell sorting and cDNA microarray to identify the gene expression profile of adjacent gene...

Roy J. and Lucille A. Carver College of Medicine Medical Student Research Day September 4, 2015 Sponsored by: Medical Student Research Club and Medical Student Research Council Funds provided by University of Iowa Student Government Free to University of Iowa Students Neuroantigen-specific CD8+ T cells in experimental autoimmune encephalitis (EAE) Tina Arkee, B.S., Alexander Boyden, PhD, and Ni...

The activation, localization, phenotypic changes, and function of CFSE-labeled naive influenza-specific CD8(+) and CD4(+) T cells following influenza infection were examined. Response of adoptively transferred CD8(+) T cells was seen earliest in draining lymph node. Highly activated cells were found later in the lung, airways, and spleen, were cytolytic, and expressed IFN-gamma upon restimulati...

Data regarding the role of TGF-beta for the in vivo function of regulatory CD4(+)CD25(+) T cells (Treg) are controversial. A transgenic mouse model with impaired TGF-beta signaling specifically in T cells was used to assess the role of endogenous TGF-beta for the in vivo function of CD4(+)CD25(+) Treg in a murine model of colitis induced by dextran sulfate. Transfer of wild-type, but not transg...

Background. Due to their ability recruit immune cells kill tumor directly, bispecific T cell engager antibodies (BiTE) hold great potential in redirecting therapies. BiTE is able activate through CD3 and target them tumor-expressed antigens. However, there are many components the microenvironment (TME) such as mesenchymal stem (MSCs) that may interfere with function. Herein, we designed an anti...