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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
Activation of interleukin 4- and interleukin 6-secreting cells by HIV-specific synthetic peptides
D M Klinman1, B F Haynes, J Conover
1Section of Retroviral Immunology, Food and Drug Administration, Bethesda, Maryland 20892, USA.
Peptides were synthesized in which the type-specific determinant of the V3 loop region of gp120 (SP10) was expressed C terminal to a conserved T helper epitope (T1) on the same molecule. These T1-SP10 peptides can stimulate both cell-mediated and humoral immune responses. The current work used a novel approach to study the nature and specificity of the response elicited by these peptides. Cytokine-specific ELIspot assays were used to examine the number, kinetics and fine specificity of cells induced to secrete IL-4 and IL-6 in mice immunized with T1-SP10 peptides. Results indicate that the peptides activated cytokine-secreting cells in a dose-dependent manner in vivo. In vitro restimulation experiments demonstrated that both the SP10 and T1 regions contributed to this activation. Consistent with previous studies, mice sequentially immunized with peptides expressing different V3 loop regions generated B cell responses that were larger and more cross-reactive than those induced by a single peptide. Sequential immunizations had less effect on the number or specificity of the cytokine-producing cells.
Peptides were synthesized in which the type-specific determinant of the V3 loop region of gp120 (SP10) was expressed C terminal to a conserved T helper epitope (T1) on the same molecule. These T1-SP10 peptides can stimulate both cell-mediated and humoral immune responses. The current work used a novel approach to study the nature and specificity of the response elicited by these peptides. Cytokine-specific ELIspot assays were used to examine the number, kinetics and fine specificity of cells induced to secrete IL-4 and IL-6 in mice immunized with T1-SP10 peptides. Results indicate that the peptides activated cytokine-secreting cells in a dose-dependent manner in vivo. In vitro restimulation experiments demonstrated that both the SP10 and T1 regions contributed to this activation. Consistent with previous studies, mice sequentially immunized with peptides expressing different V3 loop regions generated B cell responses that were larger and more cross-reactive than those induced by a single peptide. Sequential immunizations had less effect on the number or specificity of the cytokine-producing cells.
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