Isolation of human immunodeficiency virus type 2 biological clones from peripheral blood lymphocytes

Area of Science:

• Virology
• Immunology
• Infectious Diseases

Background:

• Human immunodeficiency virus (HIV) isolation from peripheral blood mononuclear cells (PBMC) is crucial for research and diagnostics.
• Standard HIV type 1 (HIV-1) isolation protocols are effective for HIV-1 but not HIV-2, especially in asymptomatic individuals.
• Existing methods are inadequate for isolating HIV-2 from PBMC of asymptomatically infected individuals with high CD4+ T-cell counts.

Purpose of the Study:

• To describe a modified HIV isolation procedure specifically adapted for human immunodeficiency virus type 2 (HIV-2).
• To enable the isolation of HIV-2 from PBMC of asymptomatically infected individuals.
• To facilitate the study of HIV-2 variants in early-stage infections.

Main Methods:

• A variant HIV isolation procedure using adapted culture conditions for HIV-2.
• Utilizing a limiting dilution format to obtain individual HIV variants.
• Propagation of virus in co-cultures with healthy donor PBMC.

Main Results:

• The described procedure is effective for isolating HIV-2 from PBMC of "healthy" HIV-2-infected individuals.
• This method allows for the isolation of HIV-2 from individuals with high CD4+ T-cell counts and undetectable plasma viral RNA.
• The limiting dilution format enables the recovery of distinct biological clones of HIV-2.

Conclusions:

• The developed protocol significantly improves HIV-2 isolation from asymptomatic individuals.
• This advancement aids in the characterization of HIV-2 variants and understanding early-stage disease.
• The method provides a means to estimate the frequency of infectious HIV-2 variants in PBMC.