Fatemeh Roodbari; Farzaneh Sabahi; Mohamad Nabi Sarbolouki; Farzaneh Barkhordari; Ahmad Adeli; Amel Jamedar; Fereidoun Mahboudi
Volume 9, Issue 2 , June 2012, , Pages 86-97
Abstract
Background: Development of an effective vaccine is highly needed in order to restrict the AIDS pandemic. DNA vaccines initiate both arms of immunity without the potential of causing disease. HIV-1 p24 and gp41 (gag and env) proteins play important roles in viral pathogenesis and are effective candidates ...
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Background: Development of an effective vaccine is highly needed in order to restrict the AIDS pandemic. DNA vaccines initiate both arms of immunity without the potential of causing disease. HIV-1 p24 and gp41 (gag and env) proteins play important roles in viral pathogenesis and are effective candidates for immune induction and vaccine design. Objective: In this study, new DNA vaccine candidates constructed from HIV-1 fused p24- gp41 or gp41 alone were evaluated in Balb/c mice for induction of cellular and humoral immune responses. Methods: Recombinant plasmids, pcDNA3.1/Hygro expression vector containing immunogenic sequences of fused p24-gp41 or gp41alone were produced. Dendrosome used as a system for carrying vectors in laboratory animals, and an IL-12 containing vector (pCAGGS-IL-12) was co-immunized with the p24-gp41 vector as a genetic adjuvant. Induction of effective immune responses against the designed vectors as DNA vaccine candidates in Balb/c mice was evaluated. Levels of total antibodies, IgG isotypes (IgG2a and IgG1); IFN-γ and IL-4 were measured by ELISA. MTT assay was used to evaluate lymphoproliferation. Results: The results confirmed that the immunogenic epitopes of both p24 and gp41 genes are highly effective inducers of immune responses, and administration of fused p24-gp41 alone or along with IL-12 resulted in further enhancement of immune responses. Group 4 that received fused fragments (p24-gp41) along with an IL-12 expressing vector demonstrated a significantly higher Stimulation Index (SI) and IFN- production (p<0.0001) with a significant increase in IgG2a/IgG1 ratio, indicating the stimulation of CMI towards Th1. Although gp41 containing vector (group 6) also showed significant increases in both proliferation and IFN- production, the responses were persistently lower than that of p24-gp41 containing vectors. Total antibody production was highest in group 6 as expected. Conclusion: Dendrosome proved to be an efficient carrier of recombinant plasmids constructed in this study. Further studies are necessary to evaluate these constructs as HIV vaccine candidates.
Sayeed Bayanolhagh; Mahtab Alinezhad; Kooroosh Kamali; Maryam Foroughi; Hamid Reza Khorram Khorshid; Minoo Mohraz; Fereidoun Mahboudi; Ali Akbar Pourfathollah
Volume 7, Issue 3 , September 2010, , Pages 162-176
Abstract
Background: Numerous evidences indicate that in some HIV-1 positive patients, the humoral and cellular immune responses are induced against HIV-1 proteins and this is inversely related to the progress of infection. Objective: The aim of this study was the evaluation of the Adenovectors containing HIV ...
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Background: Numerous evidences indicate that in some HIV-1 positive patients, the humoral and cellular immune responses are induced against HIV-1 proteins and this is inversely related to the progress of infection. Objective: The aim of this study was the evaluation of the Adenovectors containing HIV genes in induction of immune responses in mice. Methods: The HIV-1 genes including gag p24, rev, nef and exon-1 of tat were amplified from HIV-1 RNA (clade-A). The cDNA of each gene was cloned into a transfer vector. The transfer vector was then co-transformed into E. coli strain BJ5183 together with pAdenovector ΔE1/E3. The recombinant adenoviral construct was transfected into QBI-293A cells. Recombinant viruses were purified and titrated on 293 cell plates. Expression of transgenes was evaluated using western blotting. Then 1012 viral particles were injected into 15 groups of 5 mice and all patterns of combination of these 4 HIV-1 genes were evaluated. After 2 weeks, humoral and cellular immune responses were evaluated using ELISA, cell proliferation and ELISpot (IL-2, IL-4 and IFN-γ) assays, consecutively. Results: It was demonstrated that each gene was expressed. The response targets were mostly toward Th1, though several Th2 responses were also observed. Single injection in our study induced a good cellular response but the humoral responses were not as strong as the cellular ones. Conclusion: Considering and comparing all results and evaluating the various possible interactions revealed that simultaneous injection of tat and gag has enhanced the humoral and cellular responses.
Mehdi Mahdavi; Masoumeh Ebtekar; Fereidoun Mahboudi; Hamidreza Korram Khorshid; Fatemeh Rahbarizadeh; Kayhan Azadmanesh; Haydeh Darabi; Farzaneh Pourasgari; Zuhair Mohammad Hassan
Volume 6, Issue 4 , December 2009, , Pages 163-173
Abstract
Background: Cell mediated immunity, especially cytotoxic T cell responses against HIV-1 infection, plays a critical role in controlling viral replication and disease progres-sion. DNA vaccine is a novel technology which is known to stimulate strong cellular immune responses. Many DNA vaccines have been ...
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Background: Cell mediated immunity, especially cytotoxic T cell responses against HIV-1 infection, plays a critical role in controlling viral replication and disease progres-sion. DNA vaccine is a novel technology which is known to stimulate strong cellular immune responses. Many DNA vaccines have been tested for HIV infection but there is still no effective vaccine against this infection. Construction of a vaccine consisting of multiple conserved and immunogenic epitopes may increase vaccine efficacy. Objective: In the present study, a DNA vaccine candidate constructed from HIV-1 P24-Nef was evaluated and cellular immune responses were assessed in murine BALB/c model. Methods: HIV-1 P24-Nef gene was cloned in pCDNA3.1 expression vector. Mice were immunized with DNA construct and IL-4 and IFN-γ evaluation was per-formed using ELISPOT. Cytotoxicity response was evaluated with Granzyme B ELIS-POT assay and lymphocyte proliferation was evaluated with LTT assay. Results: Analysis of immune responses showed that, compared to control groups, the candidate vaccine induced production of higher levels of both IL-4 and IFN-γ (p<0.05). Cytotox-icity and lymphocyte proliferation responses of mice vaccinated with the candidate vac-cine were significantly increased compared to control groups (p<0.05). Conclusion: HIV-1 P24-Nef DNA construct displayed strong immunogenicity in a murine model.
Reza Mansouri; Firoozeh Akbari; Mohammad Vodjgani; Fereidoun Mahboudi; Fathollah Kalantar; Mahroo Mirahmadian
Volume 4, Issue 3 , December 2007, , Pages 179-185
Abstract
Background: Preeclampsia is a major cause of mortality and morbidity and is also a leading cause of preterm birth and intrauterine growth retardation. Several studies have reported abnormal levels of cytokines in women with preeclampsia. Objectives: To detect serum levels of various cytokines in pregnant ...
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Background: Preeclampsia is a major cause of mortality and morbidity and is also a leading cause of preterm birth and intrauterine growth retardation. Several studies have reported abnormal levels of cytokines in women with preeclampsia. Objectives: To detect serum levels of various cytokines in pregnant women with and without preeclampsia in the third trimester of pregnancy. Methods: Thirty patients with preeclampsia and thirty normal pregnant women were enrolled in the study. Blood samples were taken and serum levels of IFN γ, IL-12p70, IL-18, IL-15, IL-4 and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). Results: Preeclamptic women had significantly increased levels of circulating IL-12p70 (p < 0.05), IL-18 (p < 0.001), IL-4 (p < 0.001), IL-15 (p < 0.05) and IFN γ (p < 0.001). By contrast, circulating levels of IL-10 were not significantly different between the two groups. Conclusions: The present study supports the hypothesis of altered immune response in preeclampsia and suggests that dysregulation of cytokine expression occurs in preeclampsia with increased levels of IFN γ, IL-12p70, IL-15, IL-18 and IL-4.
