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Updated: Aug 17, 2026
Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
Published on: September 14, 2014
HIV-1 MN Env 15-mer peptides better detect HIV-1 specific CD8 T cell responses compared with consensus subtypes B and
Alleluiah Rutebemberwa1, Jeffrey R Currier, Linda Jagodzinski
1Henry M. Jackson Foundation and the US Military HIV Research Program, Rockville, Maryland 20850, USA. arutebemberwa@hivresearch.org
Objective:
To compare the ability of three Env (15-mer) peptide sets derived from the HIV-1 MN, the subtype B consensus, and the group M consensus to detect HIV-1 specific interferon (IFN)-gamma responses in HIV-1 subtype B infected subjects.
Methods:
Peripheral blood mononuclear cells were obtained from 17 HIV-1 subtype B seropositive and 5 HIV-1 seronegative subjects. Peptide matrices comprising each peptide set were used in IFN-gamma Elispot assays to screen for T cell epitopes. Following matrix deconvolution, individual peptides were analyzed by IFN-gamma intracellular cytokine-staining to confirm and characterize the responding cells.
Results:
HIV specific IFN-gamma responses were detected in 17 of 17 HIV-1 seropositive and none of 5 HIV-1 seronegative subjects by Elispot. Within the 17 HIV-1 seropositives, 16, 14, and 11 subjects responded to MN, B consensus, and group M env peptides, respectively. Responses were confirmed by intracellular cytokine analysis in 14 subjects and were in the CD3CD8 compartment. Cross-recognition of 'equivalent' peptides (i.e., peptides mapping to the same sequence region from the three peptide sets) was observed in 9 of 17 subjects. Peptide set specific responses to individual peptides were also observed; 11, 1, and 1 subjects demonstrated peptide set specific responses to MN, B consensus, and consensus group M, respectively.
Conclusion:
MN derived Env peptides were better able to detect HIV-1 specific CD8 T cell responses, many of which were not detectable by the equivalent clade or group consensus peptides. No single peptide set detected all the IFN-gamma responses within an individual. These results demonstrate the importance of reagent selection for monitoring of HIV responses in HIV-1 infected individuals and subsequently vaccine recipients.
Insights
The HIV-1 MN Env peptide set detected more HIV-1 specific T cell responses than consensus peptides. Peptide selection is crucial for monitoring HIV and guiding vaccine development.
Area of Science:
- Immunology
- Virology
- Vaccinology
Background:
- Monitoring immune responses to Human Immunodeficiency Virus (HIV) is critical for understanding viral control and vaccine efficacy.
- Interferon-gamma (IFN-γ) producing T cells are key indicators of cellular immunity against viral infections.
Purpose of the Study:
- To evaluate three different 15-mer Env peptide sets derived from HIV-1 MN, subtype B consensus, and group M consensus for their ability to detect HIV-1 specific IFN-γ responses.
- To compare the sensitivity and specificity of these peptide sets in individuals infected with HIV-1 subtype B.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) were collected from HIV-1 subtype B infected and seronegative individuals.
- IFN-γ Elispot assays were performed using peptide matrices to screen for T cell epitopes.
- Intracellular cytokine staining was used to confirm and characterize responding T cells, specifically CD8+ T cells.
Main Results:
- All 17 HIV-1 seropositive subjects showed IFN-γ responses via Elispot, while none of the 5 seronegative subjects did.
- The HIV-1 MN Env peptide set identified responses in 16 subjects, compared to 14 for the subtype B consensus and 11 for the group M consensus.
- CD8+ T cell responses were confirmed, and cross-recognition of equivalent peptides was observed in 9 subjects, with some peptide set-specific responses also noted.
Conclusions:
- HIV-1 MN Env peptides demonstrated superior ability in detecting HIV-1 specific CD8+ T cell responses compared to consensus peptide sets.
- No single peptide set was sufficient to detect all IFN-γ responses within an individual.
- The choice of peptide reagent is critical for accurate monitoring of HIV-specific immune responses and for informing the design of effective HIV vaccines.
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