AAV2/8 was produced by transient transfection and purification from culture supernatant by PEG precipitation and cesium chloride ultracentrifugation. Virus was quantified by qPCR against CMV sequences and functionally validated in vitro to confirm gene expression prior to use in vivo. Mice were given single injections with purified vector in the gastrocnemius muscle. Antibody concentration in the serum was determined using an ELISA specific for either total human IgG or human IgG against HIV-gp120. Humanized mice expressing antibodies were produced by adoptive transfer of expanded huPBMCs into mice previously transduced with AAV vectors. HIV challenge was carried out via IP or IV injection and blood was sampled weekly to determine the ratio of CD4 to CD8 cells by flow cytometry.
Protocol detail
Humanized Mouse Model for HIV Study
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Adoptive transfer
Antibodies
Antibody
Blood
Cd8 cells
Cesium chloride
Elisa
Flow cytometry
Gastrocnemius muscle
Gene expression
Hiv gp120
Human
Mice
Serum
Transfection
Transient
Ultracentrifugation
Vectors
Virus
Corresponding Organization :
Other organizations : California Institute of Technology, University of California, Los Angeles
Top 5 similar protocols
Protocol cited in 39 other protocols
Variable analysis
independent variables
- Vector type (AAV2/8)
- Virus production method (transient transfection and purification from culture supernatant)
- Virus purification method (PEG precipitation and cesium chloride ultracentrifugation)
- Method of virus administration (single injections in the gastrocnemius muscle)
- HIV challenge method (IP or IV injection)
- Antibody concentration in serum (total human IgG or human IgG against HIV-gp120)
- Ratio of CD4 to CD8 cells in blood
- Gene expression (validated in vitro)
- Virus quantification method (qPCR against CMV sequences)
- Functional validation of gene expression in vitro
- Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!