Flow cytometry based identification of simian immunodeficiency virus Env-specific B lymphocytes
Ismael Ben F Fofana1, Arnaud D Colantonio, R Keith Reeves
1Microbiology Division, New England Primate Research Center, Harvard Medical School, One Pine Hill Drive, Southborough, MA 01772, USA.
Journal of Immunological Methods
|June 22, 2011
Summary
Researchers developed soluble Simian Immunodeficiency Virus (SIV) envelope proteins to detect and quantify SIV-specific B lymphocytes in macaques, aiding AIDS vaccine research. These reagents help analyze B cell responses in preclinical studies.
Area of Science:
- Virology
- Immunology
- Vaccinology
Background:
- The macaque model is crucial for preclinical AIDS vaccine and pathogenesis research.
- High-titer antibodies are induced by SIV infection, driving viral escape variants.
- Potent neutralizing antibodies (Nabs) against SIVmac239/251 are difficult to detect or isolate.
Purpose of the Study:
- To generate reagents for detecting and quantifying SIVmac Env-specific B lymphocytes.
- To overcome challenges in identifying and cloning potent Nabs from infected macaques.
- To facilitate analysis of B cell responses in SIV preclinical studies.
Main Methods:
- Generated recombinant, secreted, soluble SIVmac envelope (Env) proteins (gp120 and gp140).
- Utilized advances in labeling HIV-specific B lymphocytes for SIV studies.
- Tested soluble gp140 for stable oligomerization and reactivity with antibodies.
Main Results:
- Soluble SIVmac239 gp140 forms stable oligomers without additional modifications.
- Oligomeric gp140 reacted with anti-SIV Env monoclonal antibodies and depleted neutralizing antibodies from plasma.
- Soluble gp120 and gp140 bound to SIV-specific B cells in immunized macaques.
Conclusions:
- Developed novel soluble SIV Env proteins (gp120, gp140) as valuable reagents.
- These reagents enable detection and quantification of SIV Env-specific B lymphocytes.
- Useful for analyzing B cell responses in SIV-infected or vaccinated macaques for AIDS vaccine development.
