CAR-modified marrow infiltrating lymphocytes efficiently target malignant plasma cells with very low antigen density

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The delivery of antigens on degradable nanoparticles, directly to dendritic cells residing in lymph nodes, offers great opportunities for vaccine design. This master thesis investigates the issues related to the development of vaccines based on ultrasmall (25nm)nanoparticles, capable of draining to lymph nodes via lymphatic flow and to spontaneously activate complement factor C3 as an adjuvant. The first part investigates a Mycobacterium tuberculosis preventive vaccine in collaboration with Northwestern (Ja

Isolated primary human cells from different donors vary in their permissiveness-the ability of cells to be infected and sustain the replication of human immunodeficiency virus type 1 (HIV-1). We used replicating HIV-1 and single-cycle lentivirus vectors in a population approach to identify polymorphic steps during viral replication. We found that phytohemagglutinin-stimulated CD4(+) CD45RO(+) CD57(-) T cells from healthy blood donors (n = 128) exhibited a 5.2-log-unit range in virus production. For 20 selec

The ionotropic 5HT3 receptor was expressed in transiently transfected mammalian cells, yielding an unprecedented high concn. of up to 12 million receptors per cell. Receptor traffic in the plasma membrane of live cells was obsd. continuously over 24 h by fluorescence scanning confocal microscopy. This was possible by using 5HT3 receptor-specific fluorescent ligands with high binding affinity and low off-rate to pulse label receptors at any time after appearance on the cell surface, and label subsequently th

A novel in situ product removal (ISPR) method that uses microcapsules to ext. inhibitory products from the reaction suspension is introduced into fermn. technol. More specifically, L-phenylalanine (L-phe) was transformed by Saccharomyces cerevisiae to 2-phenylethanol (PEA), which is inhibitory toward the yeast. In order to continuously remove PEA from the vicinity of the cells, the reaction suspension was brought into contact with capsules of 2.2-mm diam. that had a hydrophobic core of di-Bu sebacate and an

The metabolic and cell cycle status of primary T lymphocytes conditions their susceptibility to human immunodeficiency virus (HIV) and HIV-derived vectors. While in fully quiescent T lymphocytes the reverse transcription and nuclear import of these retroelements are impaired, leading to an abortive infection, various stimuli can induce a state of virus permissiveness. Here, we studied the modalities by which interleukin-7 (IL-7), an important controller of T-cell homeostasis, exerts this effect. IL-7-expose

Fluorescent proteins expressed in mammalian cells can be quantified quickly and noninvasively with a standard fluorescence plate reader. We have previously exploited this quality in cell growth assessment (Hunt et al., 1999b). In this work, different CHO cell lines constitutively expressing fluorescent proteins were evaluated as model systems for process development and optimization. Our results demonstrate that the fluorescence of these cell lines quickly reveals conditions that might improve the overall p

Various methods exist to transfect mammalian cells in culture. It is generally accepted that individual methods have to be optimized for each of the cell lines or cell types used. Despitethe use of optimized protocols, significant day-to-day variationsin transfection efficiency regularly occur. We postulate that the;status' of cell populations prior to transfection is involved insuch variability. This study evaluates standardized transfectionsdone at different phases of the cell cycle. Cell synchronizationw

We have recently reported that Notch 1, a member of the Notch multigene family, is essential for the development of murine T cells. Using a mouse model in which Notch 1 is inactivated in bone marrow (BM) precursors we have shown that B cells instead of T cells are found in the thymus of BM chimeras. However, it is not clear whether these B cells develop by default from a common lymphoid precursor due to the absence of Notch 1 signaling, or whether they arise as a result of perturbed migration of BM-derived

The ability of organisms, or individual cells, to react to external chemical signals, which are detected and transduced by cell-surface receptors, is crucial for their survival. These receptors are the targets of the majority of clinically used medicines. Combinatorial genetics can provide almost unlimited numbers of mutant receptor proteins and combinatorial chemistry can produce large libraries of potential therapeutic compounds that act on these membrane receptors. What is missing for the fundamental und

The Nef protein of primate lentiviruses triggers the accelerated endocytosis of CD4 and of class I major histocompatibility complex (MHC-I), thereby down-modulating the cell surface expression of these receptors. Nef acts as a connector between the CD4 cytoplasmic tail and intracellular sorting pathways both in the Golgi and at the plasma membrane, triggering the de novo formation of CD4-specific clathrin-coated pits (CCP). The downstream partners of Nef in this event are the adapter protein complex (AP) of

A review with 6 refs. of the authors' work in developing a biophys. technique to investigate the structure, function and dynamics of membrane receptors. A fluorescence-based approach has been developed and applied to the prototypic G protein-coupled receptor NK2. A fluorescent unnatural amino acid was introduced at known sites into NK2 by suppression of nonsense codons with the aid of a chem. misacylated synthetic tRNA specifically designed for the incorporation of unnatural amino acids during expression in

CAR-modified marrow infiltrating lymphocytes efficiently target malignant plasma cells with very low antigen density

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Evaluating the use of nanoparticles as an antigen delivery system

Entry and transcription as key determinants of differences in CD4 T-cell permissiveness to human immunodeficiency virus type 1 infection

Monitoring expression and clustering of the ionotropic 5HT3 receptor in plasma membranes of live biological cells

xxvel type of in situ extraction: Use of solvent containing microcapsules for the bioconversion of 2-phenylethanol from L-phenylalanine by Saccharomyces cerevisiae

Modalities of interleukin-7-induced human immunodeficiency virus permissiveness in quiescent T lymphocytes

Fluorescent proteins in animal cells for process development: optimization of sodium butyrate treatment as an example

S-phase synchronized CHO cells show elevated transfection efficiency and expression using CaPi

Notch 1-deficient common lymphoid precursors adopt a B cell fate in the thymus

Fluorescence techniques: shedding light on ligand-receptor interactions

Nef-induced CD4 and major histocompatibility complex class I (MHC-I) down-regulation are governed by distinct determinants: N-terminal alpha helix and proline repeat of Nef selectively regulate MHC-I trafficking

Probing ligand-receptor recognition in G protein-coupled receptors through biosynthetic incorporation of fluorescent amino acids at specific sites and measurement of distances by fluorescence energy transfer