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Updated: Aug 12, 2026
In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function
Published on: October 16, 2013
Increased expression of CD80 and CD86 in in vitro-infected CD3+ cells producing cytoplasmic HIV type 1 p24
1Division of AIDS, Sexually Transmitted Diseases, and Tuberculosis Laboratory Research (DASTLR); National Center for Infectious Diseases; Centers for Disease Control and Prevention, Department of Health and Human Services, Public H.
Insights
Productive human immunodeficiency virus type 1 (HIV) infection increases expression of CD80 and CD86 on lymphocytes. This finding, using multiparameter flow cytometry, impacts understanding of HIV pathogenesis and potential therapies.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Differentiating HIV-infected cells from uninfected ones is crucial for understanding infection effects.
- Previous studies lacked methods to distinguish productively infected cells.
Purpose of the Study:
- To investigate the impact of productive HIV infection on cell surface molecule expression.
- To differentiate effects on actively HIV-producing cells versus non-productively infected cells.
Main Methods:
- Used multiparameter flow cytometry to analyze HIV-infected and uninfected human peripheral blood mononuclear cells.
- Employed cell permeabilization and cytoplasmic fluorescent staining to detect HIV p24 antigen.
- Compared surface molecule expression (CD25, CD38, HLADR, CD28, CD80, CD86) between p24+ and p24- cells.
Main Results:
- Productively HIV-infected cells (p24+) showed increased expression of CD25, CD38, HLADR, and CD28 compared to p24- cells.
- A significant proportion of p24+ cells expressed CD80 or CD86, which are typically not found on CD3+ lymphocytes.
- This increased CD80/CD86 expression was observed in both resting and stimulated HIV-infected cells, independent of stimulation status.
Conclusions:
- Productive HIV infection is associated with increased expression of CD80 and CD86 on lymphocytes.
- This finding provides insights into HIV pathogenesis.
- The methodology offers a valuable tool for studying HIV's effects on cellular molecules.
Abstract:
Determining the effects of HIV infection on the expression of cell surface molecules has been limited by an inability to differentiate between productively infected cells and those without productive infection. We inoculated human peripheral blood mononuclear cells from healthy, human immunodeficiency virus type 1 (HIV) antibody-negative donors with HIV; noninoculated cells were also examined. Using multiparameter flow cytometry, we differentiated cells actively producing HIV cytoplasmic p24 antigen during acute, in vitro HIV infection from those not producing detectable cytoplasmic p24. For both resting and PHA-stimulated cells inoculated with HIV (R/H and P/H), a higher proportion of p24+ cells expressed CD25, compared with p24-cells (p = 0.031 and p = 0.008, respectively), consistent with either increased viral replication in stimulated cells or increased stimulation secondary to productive HIV infection. Findings were similar for the expression of CD38, HLADR, and CD28. A striking proportion of p24+ cells expressed CD80 or CD86, antigens not usually expressed by CD3+ lymphocytes. The increased expression appeared to be independent of stimulation status in that it occurred in both the R/H and P/H treatment groups but not in resting or PHA-stimulated uninfected cells. CD28 expression was generally comparable between CD3+ cells that did and did not express CD80 or CD86. Multiparameter flow cytometry, in association with improved techniques for cell permeabilization and cytoplasmic fluorescent staining, should prove useful in examining the effects of productive HIV infection on surface and cytoplasmic cellular molecules. Using this approach, we found an association between productive infection and increased expression of CD80 and CD86. This association has implications for HIV disease pathogenesis and, potentially, HIV therapy.
