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Related Experiment Video

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Improved flow-based method for HIV/SIV envelope-specific memory B-cell evaluation in rhesus macaques.

Venkatramanan Mohanram1, Thorsten Demberg1, Iskra Tuero1

  • 1Section on Immune Biology of Retroviral Infection, Vaccine Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, United States.

Journal of Immunological Methods
|June 24, 2014
PubMed
Summary

Developing a new flow cytometry method rapidly identifies HIV envelope-specific memory B cells. This technique aids HIV vaccine development by efficiently monitoring immune responses in preclinical and clinical trials.

Keywords:
Envelope-specific memory B cellsHIVSIV

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Area of Science:

  • Immunology
  • Vaccinology
  • Biotechnology

Background:

  • Eliciting potent, long-lasting broadly neutralizing HIV envelope (Env)-specific antibodies is crucial for effective HIV vaccine development.
  • Rapid and efficient monitoring of memory B cell development is critical for advancing vaccine design in preclinical and clinical trials.

Purpose of the Study:

  • To develop an improved, rapid, and efficient flow cytometry-based method for identifying gp120-specific memory B cells.
  • To enable direct staining of memory B cells without prior cell sorting or enrichment in various tissues.

Main Methods:

  • Utilized an improved flow cytometry technique for direct staining of memory B cells.
  • Applied the method to peripheral blood, bone marrow, and mucosal tissues (rectal tissue) in rhesus macaques.
  • Demonstrated staining of both HIV and Simian Immunodeficiency Virus (SIV) Env-specific memory B cells.

Main Results:

  • Successfully demonstrated the identification of Env-specific memory B cells in multiple tissue types.
  • Validated the method through statistically significant correlations with ELISPOT assay results for memory B cell quantification.
  • Showed significant correlations with serum binding antibody titers determined by ELISA.

Conclusions:

  • The developed flow cytometry method offers a rapid and efficient means to identify and quantify Env-specific memory B cells.
  • This technique facilitates the study of homing and trafficking of Env-specific memory B cells.
  • The method is a valuable tool for monitoring immune responses in HIV vaccine development and research.