Cell-free Kshv
Cell-free Kshv - Related Videos
07:02
Identifying Dysregulated Genes Induced by Kaposi's Sarcoma-associated Herpesvirus (KSHV)
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Cited by 6 •
2010
Host cell factors play a critical role in the establishment and maintenance of Kaposi's sarcoma (KS). We outline methods to identify host cell factors altered in KSHV-infected DMVEC cells, and in KS tumor tissue. Cellular genes altered by virus will serve as potential target(s) for novel therapeutics.
00:06
Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected Cells
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Cited by 4 •
2019
We describe a protocol utilizing fluorescence in situ hybridization (FISH) to visualize multiple herpesviral RNAs within lytically infected human cells, either in suspension or adherent. This protocol includes quantification of fluorescence producing a nucleocytoplasmic ratio and can be extended for simultaneous visualization of host and viral proteins with immunofluorescence (IF).
07:10
Establishment and Quantification of De Novo Lytic Infection by Cell-free Kaposi's Sarcoma-Associated Herpesvirus
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2025
This protocol describes an in vitro system to model de novo lytic infection of Kaposi's sarcoma-associated herpesvirus using BAC16-derived virions. The method enables investigation of early viral replication and dissemination. Infectious virus production is quantified by GFP-based infectious units assays and real-time PCR of encapsidated viral genomes.
07:20
Bioenergetic Profile Experiment using C2C12 Myoblast Cells
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Cited by 190 •
2010
A description of a method for profiling mitochondrial function in cells is provided. The mitochondrial profile generated provides four parameters of mitochondrial function that can be measured in one experiment: basal respiration rate, ATP-linked respiration, proton leak, and reserve capacity.
12:16
Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma
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Cited by 22 •
2013
This article describes the preparation of freshly obtained melanoma tissue into primary cell cultures, and how to remove contaminations of erythrocytes and fibroblasts from the tumor cells. Finally, we describe how CD133+ putative melanoma stem cells are sorted from the CD133- bulk using Magnetic Activated Cell Sorting (MACS).
FAQs
Viral glycoproteins first interact with receptors on a susceptible cell, creating the molecular conditions needed for entry. These interactions can support membrane fusion or promote uptake through an endocytic route. Because receptor availability differs among cell types, this step helps explain cellular tropism, or the preference of KSHV for particular target cells, and provides a basis for testing host determinants of infection.
Cell-free KSHV may enter through direct fusion with the cell membrane or through endocytosis, in which the cell internalizes the particle. Although both routes deliver the viral genome toward the nucleus, they represent distinct entry mechanisms that can be examined separately. Comparing them helps investigators determine how cellular conditions and host factors influence the efficiency and outcome of infection.
Once the viral genome reaches the nucleus, KSHV can establish latency or proceed into lytic replication. These alternatives represent different infection outcomes and are central to understanding how viral persistence and active replication are regulated. A defined cell-free infection system allows researchers to examine how susceptible-cell properties, host factors, or experimental conditions influence which outcome follows entry.
Researchers can expose different susceptible cell types to comparable cell-free KSHV preparations and examine whether infection is initiated and how it develops. Differences among target cells can then be related to receptor interactions, entry capacity, and downstream viral outcomes. This approach separates effects caused by the incoming virus from effects that depend on the cellular environment, supporting controlled studies of host range and susceptibility.
Cell-free KSHV provides a defined target for testing whether antibodies can interfere with infection. Antibody-containing conditions can be compared with conditions lacking the antibody, while entry or subsequent infection is examined in susceptible cells. Reduced infection indicates neutralizing activity and helps connect immune recognition with disruption of viral attachment, entry, or another early stage of the infection process.
The system supports controlled testing of antiviral activity, immune recognition, and host factors that shape infection. Investigators can compare infection outcomes under different experimental conditions without relying on intact infected cells as the immediate experimental variable. These studies can clarify how KSHV enters cells, how immune components affect that process, and how such interactions relate to KSHV-associated disease.