Original Article
Zahra Faghih; Somayeh Rezaeifard; Akbar Safaei; Abbas Ghaderi; Nasrollah Erfani
Volume 10, Issue 4 , December 2013, Pages 193-204
Abstract
Background: CD8+ cytotoxic T lymphocytes have been recently divided based on their cytokine expression profile. Objective: To evaluate the percentages of CD8+ lymphocytes and their effector subsets including Tc1, Tc2 and Tc17 in the tumor draining lymph nodes (TDLNs) of patients with breast cancer. Methods: ...
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Background: CD8+ cytotoxic T lymphocytes have been recently divided based on their cytokine expression profile. Objective: To evaluate the percentages of CD8+ lymphocytes and their effector subsets including Tc1, Tc2 and Tc17 in the tumor draining lymph nodes (TDLNs) of patients with breast cancer. Methods: Single cell suspensions were obtained from TDLNs of 42 patients with breast cancer. Staining of the cell surface markers and intracellular cytokines was performed using appropriate fluorochrome-conjugated antibodies. The data was acquired on a four-color flow cytometer and was analyzed by CellQuestPro software package. The percentages of different CD8+ cell subtypes (Tc1, Tc2 and Tc17) were quantified in CD8+ T lymphocytes. The comparison was made between LN+ versus LN- patients, as well as patients in different clinico-pathological status. Results: The percentage of Tc1, Tc2 and Tc17 subsets were not significantly different between LN+ and LN- patients. Despite no difference in the percentages of Tc1 cells in LN+ patients with infiltrative ductal carcinoma (IDC), the mean expression of IFN-γ by Tc1 cells decreased significantly in comparison to LN- patients. On the other hand, the percentages of Tc2 and Tc17 effector subsets were increased in advanced stages (p=0.018 and p=0.009, respectively). Conclusion: As the first study to investigate various effector subtypes of CD8+ lymphocytes in TDLNs of patients with breast cancer, our data collectively suggests a positive association between IL-17- and IL-4-producing CD8+ T cell percentages (Tc2 and Tc17) in TDLNs with breast cancer progression. Although the number of Tc1 cells seems not to be affected by cancer progression, down-regulation of IFN-γ by these cells seems to be associated with tumor metastasis to TDLNs. These findings may have implications in cancer immunotherapy based on CD8+ effector subsets.
Original Article
Azam Torabi; Mojtaba Tahmoorespur; Fatemeh Vahedi; Nader Mosavari; Mohammadreza Nassiri
Volume 10, Issue 4 , December 2013, Pages 205-215
Abstract
Background: Tuberculosis is a disease with high morbidity, caused mainly by Mycobaterium tuberculosis (M.tb.). DNA vaccines show a promising future due to their unique advantages over conventional methods. The early-secreted antigen target (ESAT)-6 and culture filtrate protein (CFP)-10 of M.tb. antigens ...
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Background: Tuberculosis is a disease with high morbidity, caused mainly by Mycobaterium tuberculosis (M.tb.). DNA vaccines show a promising future due to their unique advantages over conventional methods. The early-secreted antigen target (ESAT)-6 and culture filtrate protein (CFP)-10 of M.tb. antigens have been identified as vaccine candidates against Mycobacteria and used as subunit vaccines, DNA or protein, in different studies. Objective: To investigate the potential of pcDNA3.1+ plasmid containing CFP-10 and ESAT-6 genes in induction of local immune responses after intramuscular injection in BALB/c mice. Methods: pcDNA 3.1+ CFP-10 and pcDNA3.1+ ESAT-6 plasmids were prepared and defined groups of mice were injected intramuscularly with the plasmids both separately and in combination. The RNA was extracted from muscles after one month and cDNA was made using RT-PCR. The expressions of IL-4, IL-10 and IFN-γ genes cytokines were evaluated using comparative real time PCR. Results: Expression of IL-4 and IL-10 increased in the injection site of the mice groups which received plasmids encoding ESAT-6 and CFP-10 individually or together. More than 10-fold increase in IFN-γ expression was found in samples taken from mice groups inoculated by plasmids encoding ESAT-6 and CFP-10 individually or together. Conclusion: pcDNA 3.1+ESAT-6 and pcDNA3.1+CFP-10 plasmids can increase the expression of IFN-γ in mice after immunization.
Original Article
Ghasem Mosayebi; Shaban Ali Alizadeh; Ali Alasti; Alireza Amouzandeh Nobaveh; Ali Ghazavi; Mahsa Okhovat; Mohammad Rafiei
Volume 10, Issue 4 , December 2013, Pages 216-228
Abstract
Background: The appendix is considered as part of the gut-associated lymphoid tissue; however, lymphocyte subsets in this tissue are not fully defined. Objective: To investigate and compare the function and phenotype of lymphocyte subsets in peripheral blood and appendix of patients with normal and inflamed ...
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Background: The appendix is considered as part of the gut-associated lymphoid tissue; however, lymphocyte subsets in this tissue are not fully defined. Objective: To investigate and compare the function and phenotype of lymphocyte subsets in peripheral blood and appendix of patients with normal and inflamed appendix tissues. Methods: Peripheral blood samples and appendiceal mononuclear cells were obtained from 81 patients (mean age; 23 ± 10.5 years), clinically suspected of having appendicitis. The phenotypic characteristics of lymphocyte subsets in peripheral blood (before and 48-72 hrs after appendectomy) and in appendix tissue were analyzed by three color-flow cytometry. The proliferative response of mononuclear cells was assessed by MTT method. Results: The frequency of CD19+DR+, HLA-DR+ and CD19+ cells in the appendix tissue were significantly higher than that of the peripheral blood in all the groups (p<0.001). The percentage of CD19+ cells and HLA-DR+CD19+ cells significantly decreased after appendectomy in the peripheral blood of the patients with acute appendicitis (p=0.047 and p=0.03, respectively). CD19 and HLA-DR plus CD19 had better diagnostic efficiency compared with T cell markers (area under the ROC curve [AUC]= 0.76 and 0.73, respectively). Conclusion: These results indicate a significant difference in CD19+ and HLA-DR+ lymphocytes between peripheral blood and the appendix tissue.
Original Article
Saeed Solooki; Arash Khozaei; Seyedeh Azra Shamsdin; Mohammad Jafar Emami; Farnaz Khademolhosseini
Volume 10, Issue 4 , December 2013, Pages 229-237
Abstract
Background: Primary malignant bone tumors are heterogeneous groups of neoplasms, which affect mainly children and adolescents. The most common types are Osteosarcoma, Ewing sarcoma and chondrosarcoma. Elevation of sCD30 and sCD40L has been observed in lymphoma, leukemia and autoimmune disorders. Objective: ...
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Background: Primary malignant bone tumors are heterogeneous groups of neoplasms, which affect mainly children and adolescents. The most common types are Osteosarcoma, Ewing sarcoma and chondrosarcoma. Elevation of sCD30 and sCD40L has been observed in lymphoma, leukemia and autoimmune disorders. Objective: To evaluate serum concentrations of sCD30 and sCD40L in patients with primary malignant bone tumors. Method: Fifty-four cases (31 Osteosarcomas, 14 Ewing sarcomas, and 9 Chondrosarcomas) and 54 healthy controls enrolled in this study. Cases with the history of prior treatment (surgery, chemotherapy and radiotherapy) were excluded from the study. Serum levels of sCD30 and sCD40L were detected by an enzyme linked immunosorbent assay (ELISA). Results: Mean serum concentration of sCD30 in Ewing sarcoma was significantly higher than that of the control groups (p=0.007), but mean serum concentrations of sCD30 in osteosarcoma and chondrosarcoma groups were not significantly different, compared to the controls (p=0.41 and p=0.11, respectively). Mean serum concentrations of sCD40L in osteosarcoma, Ewing sarcoma and chondrosarcoma groups were significantly higher than that of the control group (p<0.0001). In addition, the mean serum level of sCD40L in chondrosarcoma patients was higher than that of both Ewing sarcoma and osteosarcoma groups (p<0.001). Conclusion: sCD30 and sCD40L increase in primary bone tumors; however the significant of these findings for diagnosis or prognosis of these tumors needs further investigation.
Original Article
Leila Rezanezhad; Jaleh Zolghadri; Behrouz Gharesi-Fard
Volume 10, Issue 4 , December 2013, Pages 238-246
Abstract
Background: Preeclampsia (PE) is a pregnancy specific syndrome that is associated with high maternal and fetal morbidity and mortality. Glucose regulated protein78 (GRP78) is an Endoplasmic Reticulum (ER) protein which is expressed on the cell surfaces of trophoblast cells under stress or hypoxic condition. ...
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Background: Preeclampsia (PE) is a pregnancy specific syndrome that is associated with high maternal and fetal morbidity and mortality. Glucose regulated protein78 (GRP78) is an Endoplasmic Reticulum (ER) protein which is expressed on the cell surfaces of trophoblast cells under stress or hypoxic condition. GRP78 has a role in aggressive behavior of invasive cells and may play a role in normal placentation. Objectives: To investigate the autoantibody against GRP78 in the sera of patients with PE and to assess the correlation between antibody and severity of the disease. Methods: We evaluated the anti-GRP78 antibody within the sera of fifty pre-eclamptic (12 severe and 38 mild PE) and fifty healthy pregnant women using a home-made ELISA assay. Furthermore, western blot technique was used to assess the expression of GRP78 in placenta of healthy and pre-eclamptic women in their third trimester. The presence of anti-GRP78 antibody in the serum samples from pre-eclamptic and healthy women was also assessed. Results: GRP78 was expressed by placenta, and both healthy and preeclamptic women produced anti-GRP78 antibody. Although no significant difference was found between the pre-eclamptic and healthy women regarding the level of anti-GRP78 antibody, the difference between severe pre-eclamptic and healthy control women was statistically significant (p<0.003). Conclusion: The findings of the present study indicated that measurement of anti-GRP78 antibody may provide a new marker for severe pre-eclampsia. Yet, future studies are required to confirm this notion.
Applied Immunology Series
Masoumeh Rajabibazl; Mohammad Javad Rasaee; Mehdi Forouzandeh; Azam Rahimpour
Volume 10, Issue 4 , December 2013, Pages 247-258
Abstract
Background: Single domain antibodies from camel heavy chain antibodies (VHH or nanobody), are advantages due to higher solubility, stability, high homology with human antibody, lower immunogenicity and low molecular weight. These criteria make them candidates for production of engineered antibody fragments ...
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Background: Single domain antibodies from camel heavy chain antibodies (VHH or nanobody), are advantages due to higher solubility, stability, high homology with human antibody, lower immunogenicity and low molecular weight. These criteria make them candidates for production of engineered antibody fragments particularly in transgenic animals. Objective: To study the development of transgenic chicken using a recombinant retrovirus containing fluonanobody. Methods: The retrovirus constructs containing nanobody genes along with secretory signals and GFP gene were established and packed. The virus particle containing the obtained fusion gene was injected into the eggs in stage X. Molecular detection and protein analysis was done in the G0 chickens. Results: The rate of hatched chicken after gene manipulation was estimated to be about 33%. Real-Time PCR assay showed that the nanobody along with GFP gene were integrated in cells of 1.2% of chickens. Conclusion: We conclude that although the rate of gene transfer by recombinant viruses in chickens is low, it would be possible to transfect the target camel immunoglobulin gene into chicken genome.
Applied Immunology Series
Mahmood Shams; Mahmood Jeddi-Tehrani; Farzaneh Notash Haghighat; Ali Ahmad Bayat; Jafar Mahmoudian; Mohammad Reza Rezvani
Volume 10, Issue 4 , December 2013, Pages 259-266
Abstract
Background: Human CD34 is a transmembrane glycoprotein which is expressed in human hematopoietic stem cells (HSCs) and the small- vessel endothelial cells of a variety of tissues. CD34 plays a critical role as a marker for diagnosis and classification of leukemia. Anti CD34 antibodies are used for isolation ...
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Background: Human CD34 is a transmembrane glycoprotein which is expressed in human hematopoietic stem cells (HSCs) and the small- vessel endothelial cells of a variety of tissues. CD34 plays a critical role as a marker for diagnosis and classification of leukemia. Anti CD34 antibodies are used for isolation and purification of HSCs from bone marrow, peripheral blood and cord blood. Objective: To characterize a newly produced monoclonal antibody against a human CD34 peptide. Methods: Anti CD34 monoclonal antibody (Clone 2C10-D3) was purified from mouse ascitic fluid and hybridoma cell culture supernatants by affinity chromatography and its immune reactivity was examined by ELISA. The purified antibody was further characterized using Western blot and flow cytometry on TF1 (Human Erythroblast) cell line. Results: ELISA experiment revealed that the antibody recognized CD34 peptide. Western blot analysis on TF1 cell lysate confirmed the reactivity of the antibody with a 42 KDa protein. Blocking the antibody with a saturating concentration of specific CD34 peptide resulted in loss of its activity with TF1 lysate in Western blot. The 2C10-D3 antibody reacted with TF1 cells in flow cytometry in a similar manner to a commercial anti CD34 monoclonal antibody. Conclusions: Our data suggest that the anti CD34 monoclonal antibody (Clone 2C10-D3) is an appropriate antibody to study the CD34+ cells by flow cytometry and Western blot.
Letter To The Editor
Hamid Nasri
Volume 10, Issue 4 , December 2013, Pages 267-269
