Immunogenicity of a New HIV-1 DNA Construct in a BALB/c Mouse Model
Mehdi
Mahdavi
Departments of Immunology
author
Masoumeh
Ebtekar
Departments of Immunology
author
Fereidoun
Mahboudi
Departments of
Biotechnology
author
Hamidreza
Korram Khorshid
Genetics Research Centre, University
of Social Welfare and Rehabilitation Sciences
author
Fatemeh
Rahbarizadeh
Medical Biotechnology, Tarbiat Modares University
author
Kayhan
Azadmanesh
Virology
author
Haydeh
Darabi
immunology Pasteur Institute of Iran
author
Farzaneh
Pourasgari
Department of Molecular Biology and Genetic Engineering, Stem Cell Technology
Company, Tehran, Iran
author
Zuhair Mohammad
Hassan
Departments of Immunology
author
text
article
2009
eng
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.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
163
173
https://iji.sums.ac.ir/article_17089_e96e7c79ed20a1afba9a052935017b8c.pdf
Heterologous Immunization, a Way out of the Problem of Monoclonal Antibody Production against Carcinoma Antigen 125
Sorour
Shojaeian
Department of Biochemistry, Tarbiat Modares University
author
Amir Hassan
Zamani
Nanobiotechnology
author
Mahmood
Jeddi-Tehrani
Monoclonal Antibody
author
Forouzandeh
Fereidooni
Cancer Institute, Imam Khomeini Medical Center
author
Ebrahim
Torkabadi
Reproductive Biotechnology Research Centers, Avicenna Research Institute, ACECR
author
Mohammad Mehdi
Akhondi
Reproductive Biotechnology Research Centers, Avicenna Research Institute, ACECR
author
Akram Sadat
Tabatabaei-Panah
Science and Research Branch, Islamic Azad University, Tehran, Iran
author
Abdolamir
Allameh
Department of Biochemistry, Tarbiat Modares University
author
text
article
2009
eng
Background: Monoclonal antibodies (mAbs) are essential tools for many molecular im-munology investigations, epitope mapping and molecular modelling, clinical laboratory di-agnostic tests and immunotherapy. Humoral immune response of immunized animals largely depends on the nature of antigen and the immunization technique. Polysaccharides and heavily-glycosylated proteins are very elusive targets incapable of mounting long-lasting, high affinity antibody responses. Carcinoma antigen 125 (CA 125), a well known tumor marker of ovarian cancer, is a mucin type antigen consisting of repetitive units of heavily glycosylated moieties which render production of mAbs very difficult. Objective: To evaluate the efficacy of heterologous antigen preparations as a way of mouse immuniza-tion in the production of anti-CA 125 mAb. Methods: Two different protocols of immuni-zation were used for priming of NMRI mice. In the first method, mice conventionally im-munized by three intraperitoneal injections of purified CA 125 and boosted by the antigen three days before fusion. In the second approach, mice were primed by three intraperitoneal injections of living CA 125 positive cells of OVCAR-3 cell line, and boosted by intrave-nous injection of the purified extracellular domain of CA 125. Production of mAb was per-formed by standard hybridoma technology and mAbs were characterized by different im-munoassays. Results: The first method failed to produce stable clones despite six time fu-sion. A total of ten stable clones, however, were produced in the second approach. Some of the clones were characterized and found to have excellent immunoreactivity when tested by ELISA assay, western blotting, intracellular and surface immunofluorescent staining of OVCAR-3 cell line and immunohistochemical staining of ovarian cancer tissues. Conclusion: Altogether the results of the present study clearly showed that heterologous antigen preparation is the method of choice for immunization when production of mono-clonal antibody against highly glycosylated poorly immunogenic antigens is concerned.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
174
185
https://iji.sums.ac.ir/article_17090_3e752bb38506d7d92fc3b6a590d47855.pdf
Mesenchymal Stem Cells Do Not Suppress Lymphoblastic Leukemic Cell Line Proliferation
Neda
Mousavi Niri
Department of Immunology
author
Mansooreh
Jaberipour
Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences,
Shiraz, Iran
author
Mahboobeh
Razmkhah
Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences,
Shiraz, Iran
author
Abbas
Ghaderi
Department of Immunology
author
Mojtaba
Habibagahi
Department of Immunology
author
text
article
2009
eng
Background: Several studies have demonstrated the immunosuppresive effects of mes-enchymal stem cells (MSCs) in allogeneic or mitogenic interactions. Cell-cell contact inhibition and secretion of suppressive soluble factors have been suggested in this re-gard. Objective: To investigate if adipose derived MSCs could inhibit Jurkat lym-phoblastic leukemia T cell proliferation during coculture. Methods: Adherent cells with the ability of cellular growth were isolated from normal adipose tissues. Initial charac-terization of growing cells by flow cytometry suggested their mesenchymal stem cell characteristics. Cells were maintained in culture and used during third to fifth culture passages. Jurkat or allogeneic peripheral blood mononuclear cells (PBMCs) were la-beled with carboxy fluorescein diacetate succinimidyl ester and cocultured with increas-ing doses of MSCs or MSC culture supernatant. Proliferation of PBMCs or Jurkat cells under these conditions was assessed by flow cytometry after 2 and 3 days of coculture, respectively. Results: Results showed the expression of CD105, CD166 and CD44, and the absence of CD45, CD34 and CD14 on the surface of MSC like cells. Moreover, ini-tial differentiation studies showed the potential of cell differentiation into hepatocytes. Comparison of Jurkat cell proliferation in the presence and absence of MSCs showed no significant difference, with 70% of cells displaying signs of at least one cell division. Similarly, the highest concentration of MSC culture supernatant (50% vol/vol) had no significant effect on Jurkat cell proliferation (p>0.6). The same MSC lots significantly suppressed the allogeneic PHA activated PBMCs under similar culture conditions. Conclusion: Using Jurkat cells as a model of leukemia T cells, our results indicated an uncertainty about the suppressive effect of MSCs and their inhibitory metabolites on tumor or leukemia cell proliferation. Additional systematic studies with MSCs of differ-ent sources are needed to fully characterize the immunological properties of MSCs be-fore planning clinical applications.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
186
194
https://iji.sums.ac.ir/article_17091_937e3c8e0c7dffc831adeefad1df9ca9.pdf
In Vitro Fertilized Embryos do not Secrete Detectable HLA-G on Day Two
Zahra
Alinejad
Departments of Immunology
author
Reza
Jafari Shakib
Departments of Immunology
author
Kambiz
Forghan-parast
Microbiology
author
Ziba
Zahiri
Gynaecology
author
Hossein
Sadri
Fertility Clinic, Family Hospital, Rasht, Iran
author
Farangis
Nagafi
Departments of Immunology
author
Zahra
Roushan
Statistics, Faculty of Medicine, Guilan
University of Medical Sciences, Rasht, Iran
author
text
article
2009
eng
Background: Pregnancy is a successful transplantation. The factors evading rejection of the fetus are poorly understood. Recently an interest has grown, in HLA-G molecules as one of these factors. Since these antigens are mainly expressed on the surface of cytotro-phoblasts that are in direct contact with maternal tissues, it has been suggested that these molecules may have a role in induction of immune tolerance in mothers. Objective: to find the association of soluble HLA-G (sHLA-G) and the success of pregnancy with intracyto-plasmic sperm injection (ICSI) procedure. Methods: In this study, the supernatant of 287 individually cultured embryos corresponding to 96 women under ICSI procedure were as-sayed for soluble HLA-G1 and G5 by a sandwich ELISA. Results: Clinical pregnancy suc-cessfully occurred in 30 of candidates. No differences in clinical parameters (age, infertility duration, stimulation regimen) were observed between pregnant and nonpregnant women under ICSI procedure. None of the embryo supernatants in either group showed any detect-able sHLA-G molecules. Conclusion: Our results showed that detectable level of sHLA-G is not produced by day 2 embryos and such a measurement may not provide reliable infor-mation for embryo selection and estimation of pregnancy success.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
195
201
https://iji.sums.ac.ir/article_17092_bf8a423eedf2e7c8a31405b695d7bbfb.pdf
Performance of Latex Agglutination Test (KAtex) In Diagnosis of Visceral Leishmaniasis in Iran
Mohammad Amin
Ghatei
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
Shiraz, Iran
author
Gholamreza
Hatam
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
Shiraz, Iran
author
Seyed Mohammad Hossein
Hosseini
Razi Vaccine and Serum Research Institute, South Branch, Shiraz, Iran
author
Bahador
Sarkari
Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
Shiraz, Iran
author
text
article
2009
eng
Background: Visceral leishmaniasis (VL) is an endemic disease in some parts of Iran. Many techniques have been used for diagnosis of VL, among which the urine based la-tex agglutination test (KAtex) is a promising one. Objective: To compare three diag-nostic tests of VL including KAtex, ELISA and Direct Agglutination Test (DAT) in VL patients and healthy controls in the south west of Iran. Methods: Serum (n = 29) and urine samples (n = 31) were collected from parasitologically confirmed VL patients. Control samples were obtained from healthy individuals (n = 61) and also from patients with infectious diseases other than VL. The collected serum samples were tested by DAT and ELISA using crude antigen from promastigotes of Leishmania infantum and the urine samples were tested by KAtex. Results: Sensitivity and specificity of KAtex for diagnosis of VL was found to be 83.9% and 100%, respectively. Sensitivities of DAT and ELISA were 93.1% and 86.2% and their specificities were 100% and 90.5%, respectively. Conclusion: KAtex yielded a satisfactory sensitivity and specificity in di-agnosis of VL in Iran and can be recommended as a rapid, field applicable and reliable test for diagnosis of VL in this region.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
202
207
https://iji.sums.ac.ir/article_17093_0beb28f664e650f7b9f526dc8e6b8c2b.pdf
Qualitative and Semi-Quantitative Comparison of an Rk39 Strip Test and Direct Agglutination Test for Detection of Anti-Leishmania Donovani Antibodies in the Sudan
Durria
Mansour
Laboratory of Biomedical Research, Ahfad University for Women, P.O. Box 167, Omdurman, Sudan
author
Elfadil
M. Abass
Laboratory of Biomedical Research, Ahfad University for Women, P.O. Box 167, Omdurman, Sudan
author
Abdelhafeiz
Mahamoud
Laboratory of Biomedical Research, Ahfad University for Women, P.O. Box 167, Omdurman, Sudan
author
Abdallah el
Harith
Retired Microbiologist, Wijngaard 155, 8212 CJ, Lelystad, The Netherlands
author
text
article
2009
eng
Background: Until now, the comparison of the rK39 strip test (RKT) and direct ag-glutination test (DAT) for detection of visceral leishmaniasis (VL) is exclusively based on either positive or negative qualification of the reaction outcome. Objective: In this study, we compared the diagnostic performance of RKT and DAT for VL both qualitatively and semi-quantitatively. Methods: For comparison based on semi-quantitative grounds, the execution of RKT and DAT was according to the standard procedures. For comparison on semi-qualitative grounds with DAT, the RKT was ap-plied to aliquots from positive samples that were two-fold serially diluted in saline to determine, as for the DAT, the end-point reaction in RKT. Results: While qualita-tively both RKT and DAT demonstrated comparable reliability for VL detection (sen-sitivity = 96% and specificity = 98.7% or 99.3%), no significant correlation (r = 0.13) could be established between intensities of their positive reactions in 25 cases studied. A negative correlation was further determined in those 25 VL cases between the posi-tive intensities of the RKT and antibody levels measured semi-quantitatively with the same procedure (r = -0.36) or the DAT (r = -0.30). Irrespective of the low, moderate or high antibody levels measured with RKT (<1:8 and 1:16-1:32 >1:256) or DAT (< 1:25,600 and 1:51,200- 1:409,600 > 1:3,276,800) in patients with confirmed or uncon-firmed VL infection, exclusively strong positive intensities were obtained with RKT. Conclusion: For further optimizing diagnosis and simultaneously assessing magni-tude of immune response to L. donovani infection in Sudanese patients, the combined application of RKT and DAT is recommended.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
208
215
https://iji.sums.ac.ir/article_17094_5b84205c195fba087076752a2775955b.pdf
Tehranolide Could Shift the Immune Response towards Th1 and Modulate the Intra-Tumor Infiltrated T Regulatory Cells
Shokoofe
Noori
Department of Biochemistry
author
Mohammad
Taghikhani
Department of Biochemistry
author
Zuhair
M. Hassan
Immunology, Tarbiat Modares University, Tehran, Iran
author
Abdolamir
Allameh
Department of Biochemistry
author
Ali
Mostafaei
Department of
Immunology, Kermanshah University of Medical Sciences, Kermanshah, Iran
author
text
article
2009
eng
Background: Artemisia diffusa contains a new type of sesquiterpene lactone with an endoperoxide group (Tehranolide). Objective: Due to the existing similarity between the structures of Tehranolide and Artemisinin, it was hypothesized that Tehranolide would have similar effects as Artemisinin. In this study, the immunotherapeutic effec-tiveness of Tehranolide was investigated by direct intra-tumoral injection. Methods: Tehranolide was purified from Artemisia diffusa, and its effect on the tumor volume was investigated. The splenocyte proliferation, shifting of cytokine profile, and the presence of naturally-occurring CD4+CD25+Foxp3+ Treg cells were assessed to describe the anti-tumor immune response. Results: Analysis of immune response showed that, intra-tumoral injection of Tehranolide decreased the rate of tumor growth compared to control group. Furthermore, the proliferative response of mice treated with Tehranolide was en-hanced. In comparison with the control group, production of both IL-4 and IFN-γ was in-duced (p<0.05). The results indicated a decrease in tumor CD4+CD25+Foxp3+ T lym-phocytes in the Tehranolide-treated group compared to the control group. Conclusion: Treatment of tumors with Tehranolide attenuated CD4+CD25+Foxp3+ Treg cell-mediated immune suppression and elicited a persistent anti-tumor immunity against can-cer.
Iranian Journal of Immunology
Shiraz Institute for Cancer Research
1735-1383
6
v.
4
no.
2009
216
224
https://iji.sums.ac.ir/article_17095_199d44eb75d408ca966b1a19d64464b0.pdf