Biomedical subjects

Federica Moschella

Publications and source records attributed to Federica Moschella.

4 recordsLinked to original sources

PubMed2004

Identification of tissue-restricted transcripts in human islets.

The purpose of our study was to identify transcripts specific for tissue-restricted, membrane-associated proteins in human islets that, in turn, might serve as markers of healthy or diseased islet cell masses. Using oligonucleotide chips, we obtained gene expression profiles of human islets for comparison with the profiles of exocrine pancreas, liver, and kidney tissue. As periislet presence of type 1 interferon is associated with the development of type 1 diabetes, the expression profile of human islets treated ex vivo with interferon-alpha2beta (IFNalpha2beta) was also determined. A set of genes encoding transmembrane- or membrane-associated proteins with novel islet-restricted expression was resolved by determining the intersection of the islet set with the complement of datasets obtained from other tissues. Under the influence of IFNalpha2beta, the expression levels of transcripts for several of the identified gene products were up- or down-regulated. One of the islet-restricted gene products identified in this study, vesicular monoamine transporter type 2, was shown to bind [3H]dihydrotetrabenazine, a ligand with derivatives suitable for positron emission tomography imaging. We report here the first comparison of gene expression profiles of human islets with other tissues and the identification of a target molecule with possible use in determining islet cell masses.

Culture Techniques↗

PubMed2003

Administration of different antigenic forms of altered peptide ligands derived from HIV-1 RTase influences their effects on T helper cell activation.

Genetic hypervariability of viruses such as HIV-1 facilitates appearance of escape mutants for immune response. HIV-1 isolates display variant epitopes, which may fail to stimulate T-lymphocyte responses or act as natural T-cell receptor antagonists, contributing to viral persistence. We evaluated the effect on epitope specific T-cell reactions of different amino acid substitutions in a residue of the 248-262 sequence of HIV-1 reverse transcriptase (peptide 23), showing variability in different viral isolates. Responses against such a determinant have been detected in long-term nonprogressive patients. The modified antigenic determinant was administered either as synthetic peptide or as recombinant protein. Our results show that certain amino acid substitutions abolished peptide binding to major histocompatibility complex (MHC); other modifications, although not affecting the formation of the MHC/peptide complex, either abrogated T-cell proliferation or exhibited an antagonistic effect. The results suggest that residue 11 of peptide 23 exhibits a double function; its alteration affects both the peptide affinity for the MHC and the MHC/peptide complex affinity for the T-cell receptor. Furthermore, we demonstrated that synthetic ligands and recombinant proteins may produce distinct functional effects, providing evidence that synthetic peptides, compared with corresponding epitopes generated by intracellular processing of recombinant proteins, may bind to the MHC groove in a different conformation.

Epitopes↗

PubMed2003

Shifting gene expression profiles during ex vivo culture of renal tumor cells: implications for cancer immunotherapy.

The use of cultured tumor cells rather than original tumor tissue for the preparation of therapeutic cancer vaccines represents an obvious solution to the problem of availability of adequate quantities of autologous tumor. In this study we investigated possible changes in gene expression accompanying the transition of renal cell carcinoma cells from the original tissue to cell populations in culture. In our study we employed cDNA microarray technology to compare the gene expression pattern of ex vivo cultured renal carcinoma cells to that of the original solid tumor tissue from which the cells were derived. Using this approach we detected changes in the expression of many genes mostly related to the cell lines' physiological properties. Some of the products of those genes showing differential expression between tumor-derived cell line and original tumor are known human autoantigens or tumor-associated antigens. Furthermore, analysis of overexpressed genes revealed the presence of several transcripts with restricted normal tissue distribution, representing self-antigens with potential to elicit autoimmunity. Our results suggest that adapting tumor tissue to culture can result in changes in the level of transcripts specific for known antigens and that more information regarding the composition of tumor cells and their byproducts used in vaccine trials is needed before the efficacy and safety of such procedures can truly be determined.

Adaptation, Physiological↗

PubMed2003

Gene expression profiling and functional activity of human dendritic cells induced with IFN-alpha-2b: implications for cancer immunotherapy.

PURPOSE: In this study, we have compared patterns of gene expression and functional activity of human dendritic cells (DCs) cultured under defined conditions in IFN-alpha-2b and recombinant human granulocyte macrophage colony-stimulating factor (DCA) with cells grown in granulocyte macrophage colony-stimulating factor and IL-4 (DC4) as an initial step in evaluating the clinical utility of DCA in cancer immunotherapy. EXPERIMENTAL DESIGN AND RESULTS: Comparison of mRNA transcript profiles between DCA and DC4 revealed different expression patterns for cytokines, chemokines, chemokine receptors, costimulatory molecules, and adhesion proteins. Many genes involved in antigen (Ag) processing were equally expressed in both populations; however, expression of transcripts involved in Ag presentation was increased in DCA. DCA also showed up-regulation of Toll-like receptor 2 and 3, as well as several tumor necrosis factor family ligands. Consistent with expression profiling, functional assays demonstrated that DCAs were more potent stimulators of naive T-cell responses than DC4 in an interleukin 15 and interleukin 1beta-dependent manner. DCA-mediated tumor cell-directed cytotoxicity induced apoptosis in different human tumor cell lines and internalized apoptotic bodies to a greater extent than DC4. Lastly, in vitro priming experiments, using apoptotic cells or peptide as sources of Ag, showed that DCA drove the expansion of tumor peptide Ag-specific autologous CD8+ T cells to a greater extent than DC4. CONCLUSIONS: The unique phenotype conferred by culturing DCs in IFN-alpha-2b may be useful in adoptive transfer regimens where the destruction of tumor cells in situ, initiation of T-cell responses toward tumor tissue with unknown Ags, and/or enhancement of pre-existing Ag-specific memory responses are desired outcomes.

Apoptosis↗