Cytokines are mediators for polarization of immune response in vaccines. Studies show that co-immunization of DNA vaccines with granulocyte-macrophage colony-stimulating factor (GM-CSF) can increase immune responses. Here, experimental mice were immunized with HIV-1 DNA vaccine with GM-CSF and boosted with recombinant vaccine. Lymphocyte proliferation with Brdu and CTL activity, IL-4, IFN-γ, IL-17 cytokines, total antibody, and IgG1 and IgG2a isotypes were assessed with ELISA. Results show that GM-CSF as adjuvant in DNA immunization significantly increased lymphocyte proliferation and IFN-γ cytokines, but CTL response was tiny increased. Also GM-CSF as adjuvant decreased IL-4 cytokine vs mere vaccine group. IL-17 in the group that immunized with mixture of DNA vaccine/GM-CSF was significantly increased vs DNA vaccine group. Result of total antibody shows that GM-CSF increased antibody response in which both IgG1 and IgG2a increased. Overall, results confirmed the beneficial effect of GM-CSF as adjuvant to increase vaccine immunogenicity. The hallmark result of this study was to increase IL-17 cytokine with DNA vaccine/GM-CSF immunized group. This study for the first time provides the evidence of the potency of GM-CSF in the induction of IL-17 in response to a vaccine, which is important for control of infection such as HIV-1.

Journal homepage: www.zums.ac.ir/jhehp Background: Neutrophils are the most abundant white blood cells in humans. Recently, a novel strategy called the formation neutrophil extracellular traps (NETs) was described. NETs is a new strategy for pathogen response. This study focused on whether LPS induced NETs release in vitro in the HL60 cell line. Methods: In this study, the HL60 cell line was used for culture and DMSO for induction and differentiation. Flow cytometry was used to evaluate CD11b in the differentiated cells, and the NBT assay was used to evaluate the functionality of the differentiated HL-60 cells. Neutrophil-like cells were incubated with LPS (200 ng/ml) for 45 min, followed by incubation for 25 min with 100 ng/ml Hoechst 33342. Trypan blue as vital staining was used for viability. The statistical significance of the difference between the control and treated groups was evaluated using a one-way ANOVA. Results: Our results showed that 75% NETs was produced by HL-60 differentiated neutrophil cells exposed to 200 ng/ml LPS in 45 minues. Conclusion: Consequently, the LPS-induced infection and lethality may occur through various mechanisms. Thus, understanding the molecular mechanisms regulating NET formation in LPS-induced neutrophil-like cells would support the development of new therapeutic methods.