Junwei Chen; Meng Wu; Jinhua Yang; Jing Wang; Yue Qiao; Xiaofeng Li
Volume 14, Issue 2 , June 2017, Pages 90-98
Fateme Sadri-Ardalani; Moslem Ahmadi; Azam Hemmati; Shaghayegh Emami; Samira Farid; Mohammad Mehdi Amiri; Mahmood Jeddi-Tehrani; Mahdi Shabani; Fazel Shokri
Volume 14, Issue 2 , June 2017, Pages 99-110
Abstract
Background: In addition to passive immunotherapy using anti-HER2 monoclonal antibodies, active immunotherapy via HER2 targeting is an interesting approach to inducing specific anti-tumor immune responses. We have recently reported the immunogenicity of HER2 subdomains following DNA immunization and HER2 ...
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Background: In addition to passive immunotherapy using anti-HER2 monoclonal antibodies, active immunotherapy via HER2 targeting is an interesting approach to inducing specific anti-tumor immune responses. We have recently reported the immunogenicity of HER2 subdomains following DNA immunization and HER2 protein boosting. In the present study, we evaluated the immunogenicity of different HER2 extracellular subdomains for the induction of anti-HER2 antibody response in BALB/c mice. Objective: To investigate and characterize antibody responses to human recombinant proteins of HER2 extracellular subdomains in immunized mice. Methods: Four subdomains of HER2 extracellular domain were expressed in E.coli; subsequently, purified recombinant proteins were intraperitoneally injected in BALB/c mice with Freund's adjuvant. The anti-HER2 antibody response was detected by ELISA, immunoblotting and flow cytometry. Results: All the four HER2 subdomains along with the full extracellular domain (fECD) were able to induce specific anti-HER2 antibodies. Although anti-HER2 subdomains antibodies could not react with eukaryotic recombinant fECD protein by ELISA, they were able to recognize this protein by immunoblotting under both reduced and non-reduced conditions. Furthermore, only the sera of mice immunized with fECD protein could recognize native HER2 on HER2 overexpressing tumor cells (>99%) by flow cytometry. Moreover, fECD immunized mice sera inhibited the proliferation of tumor cells by XTT assay. Conclusion: The prokaryotic recombinant proteins of HER2 extracellular subdomains are immunogenic, yet the induced specific antibodies do not react with the native HER2 protein due to the paucity of post-translation modifications and /or distortion of the native conformation of isolated HER2 extracellular subdomains which might be potentially effective for induction of cell mediated immune response against HER2.
Amir Savardashtaki; Bahador Sarkari; Farzane Arianfar; Zohreh Mostafavi-Pour
Volume 14, Issue 2 , June 2017, Pages 111-122
Abstract
Background: Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. Objective: The current study aimed at sub-cloning a gene, encoding the B8/1 subunit ...
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Background: Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. Objective: The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodiagnosis of human CE. Methods: The coding sequence for AgB8/1 subunit of Echinococcus granulosus was selected from GenBank and was gene-optimized. The sequence was synthesized and inserted into pGEX-4T-1 vector. Purification was performed with GST tag affinity column. Diagnostic performance of the produced recombinant antigen, native antigen B and a commercial ELISA kit were further evaluated in an ELISA system, using a panel of sera from CE patients and controls. Results: SDS-PAGE demonstrated that the protein of interest had a high expression level and purity after GST tag affinity purification. Western blotting verified the immunoreactivity of the produced recombinant antigen with the sera of CE patients. In an ELISA system, the sensitivity and specificity (for human CE diagnosis) of the recombinant antigen, native antigen B and commercial kit were respectively 93% and 92%, 87% and 90% and 97% and 95%. Conclusion: The produced recombinant antigen showed a high diagnostic value which can be recommended for serodiagnosis of CE in Iran and other CE-endemic areas. Utilizing the combination of other subunits of AgB8 would improve the performance value of the introduced ELISA system.
Hamid Reza Rahimi; Tahereh Mohammadzadeh; Seyed Mahmoud Sadjjadi; Bahador Sarkari; Farzaneh Zahabiun
Volume 14, Issue 2 , June 2017, Pages 123-133
Abstract
Background: Echinococcosis is a zoonotic parasitic disease caused by the larval stage of Echinococcus granulosus. Several native and recombinant antigens, derived from different stages of E. granulosus life cycle, have been used for vaccine trials. In vitro reared adult worms are good candidates for ...
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Background: Echinococcosis is a zoonotic parasitic disease caused by the larval stage of Echinococcus granulosus. Several native and recombinant antigens, derived from different stages of E. granulosus life cycle, have been used for vaccine trials. In vitro reared adult worms are good candidates for vaccination as they do not produce fertile egg/s and do not have any risk of contamination for researchers. Objective: To evaluate different antigens derived from in vitro reared E. granulosus adult worms for the immunization of BALB/c mice against secondary hydatidosis. Methods: Viable protoscoleces (PCSs) of sheep hydatid cyst were cultivated in S.10E.H media. Excretory secretory (E/S) and crude antigens were prepared from reared adult worms. A total of fifty BALB/c mice, each 8-weeks-old, were divided into 5 groups of 10 mice. Three groups were subcutaneously immunized with crude, E/S and immunodominant antigens on days 1 and 28. The fourth group received only PBS and the fifth group had no injection. Three weeks following the second immunization, all groups were challenged, intraperitoneal, with viable PSCs. After the autopsy of the mice and opening their abdominal wall, cysts were counted and measured followed by histopathological observations. Results: The highest protective immunity (98.7%) against hydatidosis was induced by crude antigen, followed by E/S and immunodominant antigens. Conclusion: Antigens (crude antigens in particular) derived from in vitro reared E. granulosus adult worms, and their different protein components are suitable candidates for the vaccination of intermediate hosts against hydatidosis.
Haideh Namdari; Maryam Izad; Zahra Amirghofran
Volume 14, Issue 2 , June 2017, Pages 134-150
Abstract
Background: Euphorbia plants are traditionally used in folk medicine for infections, inflammation, and cancer. Objectives: To investigate the effects of the butanolic extracts of Euphorbia micorociadia and Euphorbia osyridea on specific transcription factors and cytokines expression of T cell subsets. ...
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Background: Euphorbia plants are traditionally used in folk medicine for infections, inflammation, and cancer. Objectives: To investigate the effects of the butanolic extracts of Euphorbia micorociadia and Euphorbia osyridea on specific transcription factors and cytokines expression of T cell subsets. Methods: Activated mouse splenocytes were cultured in the presence of non-cytotoxic concentrations of the extracts. Cells were evaluated for the gene expressions of T cell transcription factors and cytokines of T helper (Th)1 [T-bet and interferon gamma (IFNγ)], Th17 [retinoic acid receptor related orphan receptor (RORγt) and interleukin (IL)-17], and T regulatory (Treg) cells [forkhead box P3(Foxp3), IL-10, and Transforming growth factor (TGF)-β] using real-time PCR. The cytokine secretions were evaluated by ELISA and Foxp3 protein expression by flow cytometry. Results: Both E. osyridea and E. microciadia extracts at 0.1 μg/ml increased T-bet expression [>1.73 relative fold change (RFC), p<0.05] and IFNγ production (>1195 pg/ml, p<0.001). Both decreased Foxp3 (<0.41 RFC, p<0.05) expression. At the higher concentration both extracts significantly reduced T-bet mRNA as well as IFNγ, IL-17, IL-10, and TGF-β cytokines and Foxp3 at the mRNA and protein levels. Conclusion: These data showed the immunomodulatory effects of E. osyridea and E. micorociadia extracts on T cell-mediated responses. The extracts caused upregulation of Th1 and downregulation of Treg cells at a low concentration which suggested their possible therapeutic value in tumor models and infectious diseases. The observed immunosuppressive effects at the higher concentration potentially make these plants candidates for identification of active components and studying their mechanisms of action.
Kiandokht Borhani; Taravat Bamdad; Tayebeh Hashempour
Volume 14, Issue 2 , June 2017, Pages 151-158
Abstract
Background: Lenalidomide, a synthetic immunomodulatory drug, has a wide range of features including anti-angiogenic and anti-proliferative properties. To date, researchers have shown that lenalidomide is capable of ameliorating the immune system factors and antitumor responses. Most researchers have ...
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Background: Lenalidomide, a synthetic immunomodulatory drug, has a wide range of features including anti-angiogenic and anti-proliferative properties. To date, researchers have shown that lenalidomide is capable of ameliorating the immune system factors and antitumor responses. Most researchers have reported that lenalidomide enhances the immune response in certain cancer patients through several pathways including the stimulation of Natural Killer cells; notwithstanding, it is still crucial to investigate the effect of lenalidomide on the activity of NK cell cytotoxicity both in vitro and in vivo. Objective: To evaluate the in vitro impact of lenalidomide, of different doses, on NK cytotoxicity activity and an in vivo investigation to find the adjuvant behavior of lenalidomide. Methods: NK cytotoxocity was measured with the lactate dehydrogenase (LDH) release assay via K562 cells. Lenalidomide was prepared at 1 mM, 2 mM, 4 mM and 8 mM for in vitro study. In addition, the adjuvant properties of lenalidomide were assessed in ten mice groups using NS3 HCV DNA vaccine model of antigen pcDNA3.1(+)/NS3. Results: The results showed that, comparisons to other doses, 4 mMol of lenalidomide was able to noticeably increase NK cytotoxicity activity. Furthermore, the animal model indicated that lenalidomide stimulated NK cytotoxicity in vivo, augmenting it from 16.67% ± 2.07% for the control group to 38.17% ± 2.87% for the lenalidomide-treated. Conclusion: Treatment by lenalidomide and pcDNA3.1(+)/NS3 improves NK cytotoxicity up to 66.80% suggesting that lenalidomide can be used in parallel with such therapeutic vaccines as cancer vaccine or virus vaccines.
Ramina Fatemi; Ebrahim Mirzadegan; Zohreh Vahedian; Amir Hassan Zarnani; Mahmood Jeddi-Tehrani; Farah Idali
Volume 14, Issue 2 , June 2017, Pages 159-171
Abstract
Background: 17β-estradiol (E2) has been known to modulate immune response. Recent studies indicate that E2 at pregnancy level plays a role in regulating T cell response. Objective: To investigate the optimum dose of E2 (from 10-9 to 10-7 M) in mediating the generation of regulatory T cells (Tregs), ...
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Background: 17β-estradiol (E2) has been known to modulate immune response. Recent studies indicate that E2 at pregnancy level plays a role in regulating T cell response. Objective: To investigate the optimum dose of E2 (from 10-9 to 10-7 M) in mediating the generation of regulatory T cells (Tregs), using naïve human CD4+ T cells from healthy women. Methods: Naïve peripheral T cells were purified and conditioned with soluble anti-CD28 in anti-CD3-coated plates in the presence or absence of E2. Flow cytometry was employed to assess the expression pattern of forkhead boxP3 (FOXP3) and programmed death-1 (PD-1). Proliferation and cytokine secretions were analyzed, using XTT and ELISA assays. Results: In the presence of different doses of E2, the expression levels of anti-CD3/CD28 antibody-stimulated CD25/FOXP3 and FOXP3/PD-1 in conditioned T cells (cT) were peaked at 1 ng/ml (early pregnancy level, E2(1)) (47.14% (37.3-74.9) and 32% (27.7-52.5), respectively) and a slight, but not significant, increase after declining at 36 ng/ml (late pregnancy/pharmaceutical, E2(36)) (19.4% (15.2-24.5) and 15.8% (10.6-26.8), respectively). E2(1) cT showed a significantly reduced proliferation capacity (p<0.05) and secretion of IL-10 was enhanced in supernatants of E2(1 and 36) cT (p<0.05). In contrast to decreased TNF-a and IFN-g secretions in E2(1) cT supernatants, E2(36) stimulated TNF-a and IFN-g secretions (pConclusion: Our results indicate that the differential effect of E2 on generation of Tregs is consistent with the possibility that lower levels of pregnancy E2 are most efficient in induction of Tregs.
Tahereh Poordast; Fateme Sadat Najib; Rasoul Baharlou; Atena Bijani; Shaghayegh Moradi Alamdarloo; Alieh Poordast
Volume 14, Issue 2 , June 2017, Pages 172-179
Abstract
Background: Preeclampsia is a common pregnancy-specific disorder associated with significant maternal and fetal morbidity and mortality worldwide. It has been proposed that the imbalance between two CD4+ T cell subtypes, regulatory T cells (Treg) and T-helper 17 cells (Th17), is involved in the pathophysiology ...
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Background: Preeclampsia is a common pregnancy-specific disorder associated with significant maternal and fetal morbidity and mortality worldwide. It has been proposed that the imbalance between two CD4+ T cell subtypes, regulatory T cells (Treg) and T-helper 17 cells (Th17), is involved in the pathophysiology of preeclampsia. Objectives: To determine the serum levels of IL-17, IL-21, IL-23 and TGF-β in patients with preeclampsia. Methods: Blood samples were collected from 30 preeclampsia patients, 30 normotensive pregnant women and 30 healthy individuals with no history of malignancies or autoimmune disorders based on simple sampling. The serum levels of IL-17, IL-21, IL-23 and TGF-β were measured by the enzyme linked immunosorbent assay (ELISA). Results: The serum levels of IL-17 and TGF-β were significantly higher in preeclampsia patients compared to normal pregnant group and healthy individuals (p>0.0001) but interestingly, the opposite was the case for IL-23 (p=0.005). However, there were no significant differences in IL-21 between preeclampsia and normal pregnant group. Conclusions: Our results conclude that contrary to IL-21, serum levels of IL-17 and TGF-β significantly increased in preeclampsia compared to normal pregnant women, supporting an imbalance of cytokine profile in preeclamtic patients.
