Antigen Trafficking
Antigen Trafficking - Related Videos
11:27
Analysis of SCAP N-glycosylation and Trafficking in Human Cells
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Cited by 17 •
2016
We describe a modified method for membrane fraction isolation from human cells and sample preparation for the detection of SCAP N-glycosylation and total protein by using western blot. We further introduce a GFP-labeling method to monitor SCAP trafficking using confocal microscopy. This protocol can be used in regular biology laboratories.
07:55
Imaging the Intracellular Trafficking of APP with Photoactivatable GFP
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Cited by 6 •
2015
While the transport of cell surface proteins is relatively easily studied, visualizing the trafficking of intracellular proteins is much more difficult. Here, we use constructs incorporating photoactivatable GFP and demonstrate a method to accurately follow the amyloid precursor protein from the Golgi apparatus to down-stream compartments and follow its clearance.
06:57
Development of an Antigen-driven Colitis Model to Study Presentation of Antigens by Antigen Presenting Cells to T Cells
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Cited by 9 •
2016
In this antigen-driven colitis model, OT-II CD4+ T cells expressing a red fluorescent protein were adoptively transferred into RAG-/- mice that express a green fluorescent protein in mononuclear phagocytes (MPs). The hosts were challenged with Escherichia coli (E.coli) expressing the ovalbumin protein (OVA) fused to a cyan fluorescent protein (CFP).
07:48
Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis
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Cited by 1 •
2015
Receptor trafficking modulates signaling and cell responsiveness to ligands and is, itself, responsive to cell conditions, including ligand-induced signaling. Here, we describe a powerful and flexible technique for quantitatively assessing drug-induced receptor trafficking using immunolabeling and colocalizational analysis.
09:59
Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
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Cited by 3 •
2017
Labeling the extracellular domain of a membrane protein with a pH sensitive fluorophore, superecliptic pHluorin (SEP), allows subcellular localization, expression, and trafficking to be determined. Imaging SEP-labeled proteins with total internal reflection fluorescence microscopy (TIRFM) enables the quantification of protein levels in the peripheral ER and plasma membrane.
FAQs
Within antigen-presenting cells, extracellular proteins first undergo endocytosis, then move through vesicular transport to endosomal and lysosomal compartments. There, proteolysis breaks proteins into peptides, creating material that can be loaded onto MHC class II. Thus, trafficking controls not only location but also the processing stage at which antigen becomes available for immune recognition.
Selected intracellular antigens can access MHC class I presentation pathways, showing that cellular origin and intracellular routing influence which presentation system receives antigen-derived material. This parallel route is important because it broadens the types of antigenic information that immune cells can display. In infection research, it helps connect intracellular pathogen-associated material with T-cell activation.
Endosomal and lysosomal compartments do more than provide storage sites. Their involvement places antigen in environments where proteolysis can generate peptides suitable for MHC class II loading. The resulting compartmental control links intracellular transport to presentation efficiency, allowing trafficking decisions to influence how strongly immune cells detect a given antigen.
Because trafficking determines which peptides reach MHC molecules, it can affect whether antigen exposure supports T-cell activation or immune tolerance. The same cellular transport principle therefore has consequences beyond infection detection: altered routing may change the immune response to self or pathogen-derived material. This makes trafficking relevant to autoimmune disease research and host-pathogen studies.
Researchers can evaluate antigen trafficking by following its consequences at successive stages: delivery to endosomal or lysosomal compartments, proteolytic peptide generation, loading onto MHC molecules, and downstream effects on T-cell activation or tolerance. These linked outcomes help connect intracellular transport with immune detection rather than treating presentation as an isolated event.
In vaccine research, antigen trafficking is relevant because the route taken by vaccine-associated antigen can influence whether peptides enter MHC class II presentation or, for selected intracellular antigens, MHC class I pathways. Studying these routes helps researchers consider how antigen delivery relates to T-cell responses and the design of immune-stimulating strategies.
Therapeutic research can target antigen trafficking when the goal is to modify antigen presentation. Because transport controls access to processing compartments and MHC pathways, changing this system could alter immune detection. The topic therefore provides a mechanistic framework for investigating interventions in conditions where antigen presentation contributes to disease, including autoimmune disorders.