CD45 Depletion from Jurkat T Cell Vesicles Shows Exosomes Contain CD45
Author Information
Author(s): Coren Lori V, Shatzer Teresa, Ott David E
Primary Institution: AIDS and Cancer Virus Program, SAIC-Frederick, Inc., National Cancer Institute at Frederick, Frederick, Maryland, USA
Hypothesis
Do exosomes from Jurkat T cells contain CD45, and can they be removed from HIV-1 preparations using CD45 immunoaffinity depletion?
Conclusion
The study demonstrates that exosomes from Jurkat and SupT1/CCR5 cells contain CD45 and can be effectively removed from HIV-1 preparations.
Supporting Evidence
- CD45 is excluded from HIV-1 but present in vesicles from hematopoietic cells.
- Immunoblotting showed that CD45 was removed from vesicle preparations after depletion.
- Depletion effectively removed 95% of vesicular proteins from Jurkat E6-1 and 96% from SupT1/CCR5.
Takeaway
The researchers found that tiny bubbles called exosomes from certain immune cells have a protein called CD45, which can be taken out to make studying viruses easier.
Methodology
The study used CD45 immunoaffinity depletion to remove vesicles from HIV-1 preparations and analyzed the samples using immunoblotting and SDS-PAGE.
Limitations
Absolute biochemical purity of virion preparations may not be practically attainable.
Participant Demographics
The study involved Jurkat and SupT1/CCR5 cell lines, which are commonly used for HIV-1 production.
Digital Object Identifier (DOI)
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