Characterization of MHC class II-presented peptides generated from an antigen targeted to different endocytic compartments

Fernandes, Dancella M.; Vidard, Laurent; Rock, Kenneth L.

doi:10.1002/1521-4141(2000)30:8<2333::aid-immu2333>3.0.co;2-f

Cited by 34 publications

(16 citation statements)

References 30 publications

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“…[28][29][30] In addition, it was shown that TfR colocalizes with Ii in early endosomes which contain the proteolytic enzymes cathepsin B and D. 31 Thus it is very likely that peptide loading of MHC class II molecules occurs in this compartment. Further support for this conclusion stems from a recent report by Fernandes et al, 32 who demonstrate that in B cell lines TfR-OVA fusion protein is indeed found in early endosomes. Additionally, in this study more OVA peptides were generated and presented on MHC class II from the TfR-OVA fusion than from an OVA version that was targeted to the secretory pathway.…”

Section: Discussionmentioning

confidence: 83%

MHC class II presentation of endogenously expressed antigens by transfected dendritic cells

et al. 2001

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Dendritic cells (DC) present immunogenic epitopes of antigens in the context of MHC class I and class II molecules in association with costimulatory molecules, and efficiently activate both cytotoxic T cells and T helper cells. Gene modified DC expressing antigen encoding cDNA represent a particularly attractive approach for the immunotherapy of disease. We previously described a gene delivery system for DC based on receptor-mediated endocytosis of ligand/polyethylenimine (PEI) DNA transfer complexes that target cell surface receptors which are abundantly expressed on DC. Employing this gene delivery system, DC were generated that express chicken ovalbumin (OVA) cDNA as a model antigen and introduce antigen into the

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Section: Discussionmentioning

confidence: 83%

MHC class II presentation of endogenously expressed antigens by transfected dendritic cells

et al. 2001

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“…In this construct, ovalbumin is fused in frame with the transferrin receptor to produce a membrane-bound form of OVA. This protein is transported into the ER and exocytic compartments of cells and then localizes to the plasma membrane and early endosomes (31). In addition, to visualize their intracellular location and for the ease of selection and quantitation by flow cytometry, we also fused these forms of ovalbumin to EGFP.…”

Section: Resultsmentioning

confidence: 99%

“…The PCR product was directly cloned into pCR3.1 plasmid by using TA cloning kit (Invitrogen) and later also subcloned into pcDNA3.1. TfR-OVA was described (31) and subcloned into pcDNA3.1.zeocin. We also generated enhanced GFP (EGFP) fusion constructs with Cyto-OVA, FL-OVA, and TfR-OVA attached to the C terminus of EGFP by PCR.…”

Section: Methodsmentioning

confidence: 96%

Cellular protein is the source of cross-priming antigenin vivo

Shen

Rock

2004

Proc. Natl. Acad. Sci. U.S.A.

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176

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Cross-priming is essential for generating cytotoxic T lymphocytes to viral, tumor, and tissue antigens that are expressed exclusively in parenchymal cells. In this process, the antigen-bearing parenchymal cells must somehow transfer their antigens to bone marrow-derived professional antigen-presenting cells. Although intact proteins, small peptides, or peptide-heat shock protein complexes can all be acquired and presented by antigen-presenting cells, the physiologically relevant form of antigen that is actually transferred from parenchymal cells and cross-presented in vivo is unknown and controversial. To address this issue we have investigated the ability of fibroblasts stably expressing chicken ovalbumin constructs targeted to different subcellular compartments to crossprime cytotoxic T lymphocytes. Although these transfectants generated similar amounts of the immunogenic ovalbumin peptide, their cross-priming activity differed markedly. Instead, the cells cross-priming ability correlated with their steady-state levels of ovalbumin protein and͞or the physical form͞location of the protein. Moreover, in subcellular fractionation experiments, the crosspriming activity colocalized with antigenic protein. In addition, depletion of intact protein antigen from these cell fractions eliminated their cross-priming activity. In contrast, the major heat shock protein candidates for cross-presentation were separable from the cell's main sources of cross-priming antigen. Therefore, cellular proteins, rather than peptides or heat shock protein͞ peptide complexes, are the major source of antigen that is transferred from antigen-bearing cells and cross-presented in vivo.

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“…Several studies have demonstrated that targeting of an Ag to different cellular subcompartments can quantitatively and qualitatively influence the repertoire of CD4 ϩ and CD8 ϩ epitope-specific immune responses elicited by that Ag (5,(22)(23)(24)(25)(26)(27), and it has been postulated that cleavage by the enzymes of a specific cellular compartment can determine the repertoire of epitopes generated by the Ag's processing. Some of these studies have addressed the impact of different targeting mechanisms on a limited number of known CD4 ϩ and CD8 ϩ epitopes; however, the impact of these strategies on the in vivo epitope-specific T cell responses and on the modulation of the whole repertoire of activated cells induced by a protein Ag has not been systematically investigated.…”

Section: T He Establishment Of Mhc-restricted Cd4mentioning

confidence: 99%

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

Arruda

Sim²,

Chikhlikar

et al. 2006

The Journal of Immunology

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Ag processing is a critical step in defining the repertoire of epitope-specific immune responses. In the present study, HIV-1 p55Gag Ag was synthesized as a DNA plasmid with either lysosomal-associated membrane protein-1 (LAMP/gag) or human dendritic cell-LAMP (DC-LAMP/gag) and used to immunize mice. Analysis of the cellular trafficking of these two chimeras demonstrated that both molecules colocalized with MHC class II molecules but differed in their overall trafficking to endosomal/lysosomal compartments. Following DNA immunization, both chimeras elicited potent Gag-specific T and B cell immune responses in mice but differ markedly in their IL-4 and IgG1/IgG2a responses. The DC-LAMP chimera induced a stronger Th type 1 response. ELISPOT analysis of T cell responses to 122 individual peptides encompassing the entire p55gag sequence (15-aa peptides overlapping by 11 residues) showed that DNA immunization with native gag, LAMP/gag, or DC-LAMP/gag induced responses to identical immunodominant CD4+ and CD8+ peptides. However, LAMP/gag and DC-LAMP/gag plasmids also elicited significant responses to 23 additional cryptic epitopes that were not recognized after immunization with native gag DNA. The three plasmids induced T cell responses to a total of 39 distinct peptide sequences, 13 of which were induced by all three DNA constructs. Individually, DC-LAMP/gag elicited the most diverse response, with a specific T cell response against 35 peptides. In addition, immunization with LAMP/gag and DC-LAMP/gag chimeras also promoted Ab secretion to an increased number of epitopes. These data indicate that LAMP-1 and DC-LAMP Ag chimeras follow different trafficking pathways, induce distinct modulatory immune responses, and are able to present cryptic epitopes.

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Characterization of MHC class II-presented peptides generated from an antigen targeted to different endocytic compartments

Cited by 34 publications

References 30 publications

MHC class II presentation of endogenously expressed antigens by transfected dendritic cells

MHC class II presentation of endogenously expressed antigens by transfected dendritic cells

Cellular protein is the source of cross-priming antigenin vivo

Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes

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