Host Factor Analysis
Host Factor Analysis - Related Videos
08:20
RNAi Screening for Host Factors Involved in Vaccinia Virus Infection using Drosophila Cells
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Cited by 10 •
2010
Novel host factors involved in viral infection can be identified through cell-based genome-wide loss of function RNAi screening. A Drosophila cell culture model is particularly amenable to this approach due to the ease and efficiency of RNAi. Here we demonstrate this technique using vaccinia virus as an example.
15:29
Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells
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Cited by 6 •
2016
Two assays for microscopy-based high-throughput screening of host factors involved in Brucella infection are described. The entry assay detects host factors required for Brucella entry and the endpoint assay those required for intracellular replication. While applicable for alternative approaches, siRNA screening in HeLa cells is used to illustrate the protocols.
03:35
Analysis of Virus-Induced Host Transcriptional Suppression Using Flow Cytometry
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2026
Source: Kalveram, B., et al. Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis. J. Vis. Exp. (2013)This video demonstrates the use of click chemistry and flow cytometry to measure virus-induced suppression of host-cell transcription. Mammalian cells are infected with a virus that expresses a nuclear virulence protein disrupting the host transcription factor complex, while control cells remain transcriptionally active. Incorporation of 5-ethynyl uridine...
10:08
Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets
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Cited by 2 •
2024
This article outlines the protocol for quantifying the percentage of Tissue Factor (TF)-positive platelets using whole blood flow cytometry, assessing the protein: (1) intracellularly in resting conditions, and (2) on the cell surface, in both resting and activated conditions. Guidance is also provided for evaluating TF-positive platelets in platelet-rich plasma.
12:23
Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions
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Cited by 5 •
2015
Effector translocation into host cells via a type III secretion system is a common virulence strategy among gram-negative bacteria. A beta-lactamase effector fusion based assay for quantitative analysis of translocation was applied. In Yersinia infected cells, conversion of a FRET reporter by the beta-lactamase is monitored using laser scanning microscopy.
FAQs
The key interpretation is whether changing a host factor increases or decreases infection-related outcomes. A factor that supports infection may be required for entry or replication, whereas a factor that restricts infection may contribute to immune defense or pathogen clearance. Comparing perturbed hosts with appropriate comparison groups helps connect the factor to susceptibility rather than merely to infection-associated changes.
Researchers can perturb candidate host genes or proteins and then measure whether pathogen entry changes. A strong reduction after altering a particular factor can identify a component associated with entry, while broader pathway measurements can show how that component relates to cellular processes. These findings help map receptor-mediated routes that pathogens use to gain access to host cells.
Innate immune defenses can restrict infection before adaptive responses are considered, so host factor analysis examines how cellular pathways influence pathogen replication, persistence, or clearance. Measuring immune signaling alongside infection outcomes helps separate factors that directly support pathogen growth from those that activate protective responses, clarifying how host susceptibility and resistance arise.
Genetic screens, RNA interference, and CRISPR-based editing perturb host factors to test their functional contribution to infection. Transcriptomics can reveal broader changes in gene activity, while protein-interaction assays examine molecular associations. Used together, these approaches connect causal perturbations with pathway-level responses and interaction information, strengthening interpretation of candidate host factors.
A typical workflow begins by selecting host genes, proteins, or cellular pathways for comparison or perturbation. Researchers then apply a genetic, RNA-based, transcriptomic, or protein-interaction approach and measure infection, replication, persistence, clearance, or immune signaling. The resulting changes are interpreted to identify factors associated with pathogen entry, host defense, or disease susceptibility.
The measurements can indicate whether a host factor is associated with pathogen entry, replication, persistence, or clearance. They can also identify changes in immune signaling and reveal relationships among host proteins or pathways. Together, these outcomes help organize candidate factors into mechanisms that support infection or contribute to protective host responses.
This approach is useful when researchers need to explain why infection succeeds in some host contexts or how immune defenses limit it. Findings can guide therapeutic target discovery, vaccine design, and host-directed treatments. Because these strategies act on host mechanisms rather than only on the pathogen, they may be less vulnerable to pathogen resistance.