ABSTRACT
Background: Psoriasis is a noncontagious auto-inflammatory chronic skin disease. So far, some of the inflammatory genes were upregulated in mouse model of psoriasis. This study examined changes in skin mRNA expression of L-kynureninase (Kynu), cathelicidin antimicrobial peptide (Camp), beta-defensin 2 (Defb2), and proenkephalin (Penk) in a mouse model of imiquimod-induced psoriasis. Materials and Methods: Tree groups of C57BL/6 female mice were allocated. The imiquimod (IMQ) cream was administered to the mice dorsal skin of the two groups to induce psoriatic inflammation. In the treatment group, IMQ was administered 10 min after hydrogel-containing M7 anti-IL-17A aptamer treatment. Vaseline (Vas) was administered to the negative control group. The psoriatic skin lesions were evaluated based on the psoriasis area severity index (PASI) score, histopathology, and mRNA expression levels of Kynu, Camp, Defb2, and Penk using real-time PCR. In order to assess the systemic response, the spleen and lymph node indexes were also evaluated. Results: The PASI and epidermal thickness scores were 6.01 and 1.96, respectively, in the IMQ group, and they significantly decreased after aptamer administration to 1.15 and 0.90, respectively (P < 0.05). Spleen and lymph node indexes showed an increase in the IMQ group, followed by a slight decrease after aptamer treatment (P > 0.05). Additionally, the mRNA expression levels of Kynu, Defb2, Camp, and Penk genes in the IMQ-treated region showed a significant 2.70, 4.56, 3.29, and 2.61-fold increase relative to the Vas mice, respectively (P < 0.05). The aptamer-treated region exhibited a significant decrease in these gene expression levels (P < 0.05). A positive correlation was found between Kynu, Penk, and Camp expression levels and erythema, as well as Camp expression with PASI, scaling, and thickness (P < 0.05). Conclusion: According to our results, it seems that Kynu, Camp, and Penk can be considered appropriate markers for the evaluation of psoriasis in IMQ-induced psoriasis. Also, the anti-IL-17 aptamer downregulated these important genes in this mouse model.
Subject(s)
Cathelicidins , Disease Models, Animal , Enkephalins , Imiquimod , Mice, Inbred C57BL , Protein Precursors , Psoriasis , beta-Defensins , Psoriasis/chemically induced , Psoriasis/metabolism , Animals , Mice , Female , beta-Defensins/metabolism , beta-Defensins/genetics , Protein Precursors/metabolism , Protein Precursors/genetics , Enkephalins/metabolism , Enkephalins/genetics , Antimicrobial Cationic Peptides/metabolism , Skin/metabolism , Skin/pathology , Skin/drug effects , Biomarkers/metabolismABSTRACT
Chimeric antigen receptor (CAR) T-cell therapy is an immunotherapy approach that has played a tremendous role in the battle against cancer for years. Since the CAR T lymphocytes are unrestricted-major histocompatibility complex T lymphocytes, they could identify more targets than natural T cells, resulting in practical and widespread functions. The good prospects of CAR T-cell therapy in oncology can be additionally applied to treat other diseases such as autoimmune and infectious diseases. CAR-T cell-derived immunotherapy for autoimmune disorders can be allocated to CAR-Tregs and chimeric autoantibody receptor T cells. Other generations of CARs target human immunodeficiency virus (HIV) proteins. In this review, we summarize CAR-T cell therapies in autoimmune disorders and HIV infection.
Subject(s)
Autoimmune Diseases , HIV Infections , Receptors, Chimeric Antigen , Humans , T-Lymphocytes , HIV Infections/therapy , Immunotherapy, Adoptive/methods , Autoimmune Diseases/metabolism , Receptors, Antigen, T-Cell/metabolismABSTRACT
Interleukin (IL)-23 inhibitor monoclonal antibodies shown significant efficacy in treating autoimmune diseases. DNA or RNA aptamers exhibit comparable specificity to antibodies, are cost-effective, non-immunogenic, and do not have batch to batch variation. This study aimed to characterize a single-stranded DNA (ssDNA) aptamer targeting human IL-23. The alpha subunit of IL-23 (P19) and intact IL-23 were cloned, expressed, and the proteins finally were purified through Ni2+-iminodiacetic acid affinity chromatography. The selection and characterization of ssDNA aptamer against P19 were conducted using the protein-systematic evolution of ligands by exponential enrichment (SELEX). Dot blot assay was carried out to monitor binding of the aptamer output of SELEX rounds, to P19 protein. The dissociation constant (Kd) of aptamers with positive results in dot blot assay, determined based on their binding to IL-23 using an ELISA method. Recombinant P19 and IL-23 proteins were 26 and 72â¯kDa, respectively, observed on SDS-PAGE .12â¯%. The aptamers output from 7, 8, 9, 10, 11, and 12 rounds of the SELEX was monitored by dot blot assay, revealing that the aptamer from the round 8 has stronger luminescent signal and was selected for TA-cloning. After analyzing the biotinylated aptamers from clones, positive clones in dot blot assay and ELISA were sequenced. Finally, the Kd calculation revealed three aptamers with high affinity, named A23P3, A23P6, and A23P15 with Kd values of 1.37, 2.139, and 2.88â¯nM, respectively. Results of this study introduced three specific anti-IL-23 ssDNA aptamers with high affinity, which could be utilized for therapeutic and diagnostic purposes.
Subject(s)
Aptamers, Nucleotide , DNA, Single-Stranded , SELEX Aptamer Technique , Aptamers, Nucleotide/chemistry , SELEX Aptamer Technique/methods , Humans , Interleukin-23/antagonists & inhibitors , Recombinant Proteins , Interleukin-23 Subunit p19/antagonists & inhibitors , Chromatography, Affinity/methodsABSTRACT
BACKGROUND: Despite antitumor properties, chemotherapy medication can create conditions in tumor cells that work in favor of the tumor. Doxorubicin, commonly prescribed chemotherapy agents, can increase the risk of migration and invasion of tumor cells through overexpression of the CXCR4 gene by affecting downstream signaling pathways. The regulatory role of CXCR7 on CXCR4 function has been demonstrated. Therefore, it is hypothesized that combining doxorubicin with another anticancer drug could be a promising approach. METHODS: In this research, we evaluated the anti-invasive property of pioglitazone along with antitumor effects of doxorubicin on MDA-MB-231 breast cancer cell lines. RESULTS: There was no significant difference between two treatment groups in neither the expression nor changes in the expression of CXCR7 and CXCR4 genes (P < 0.05). Pioglitazone-doxorubicin combination reduced cell migration in tumor cells to a significantly higher extent compared to doxorubicin alone (P < 0.05). CONCLUSIONS: Co-administration of pioglitazone and doxorubicin might reduce cell migration in breast cancer tumor cells, and that cell migration function is independent of some specific proteins.
Subject(s)
Breast Neoplasms , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Cell Line, Tumor , Doxorubicin/pharmacology , Doxorubicin/therapeutic use , Female , Humans , Neoplasm Invasiveness/genetics , Pioglitazone/pharmacology , Pioglitazone/therapeutic useABSTRACT
BACKGROUND: Mesenchymal stem cells have restorative effects on premature ovarian failure (POF). The aim of this study was to investigate the effects of human umbilical cord vein MSCs (hUCV-MSCs) on follicular quantitative parameters and histological changes of ovaries in the cyclophosphamide (CTX)-induced POF in mice. MATERIALS AND METHODS: C57BL/6 mice were divided into three groups (10 mice in each group). In the control group, phosphate-buffered saline (PBS) was injected via tail vein following 15 days injection of PBS intraperitoneally (IP). In the CTX group, CTX was administered IP for 15 days and then PBS was injected via tail vein. In the CTX + hUCV-MSCs group, following CTX administration, single dose of the 1 × 106 of hUCV-MSCs were injected into tail vein. H&E, trichrome and PAS staining as well as TUNEL assay were performed on the ovaries tissue sections. The number of follicles, follicular quantitative parameters and apoptotic index were obtained. The serum levels of estradiol and FSH were measured in the mice. RESULTS: In the CTX + hUCV-MSCs group, degenerative changes were decreased and follicular quantitative parameters increased in the ovarian follicles compared to the CTX group. In this group number of follicles was increased, apoptotic index was decreased, estradiol and FSH levels were decreased and increased, respectively, all of them improved compared to the CTX group. The mean percentage areas of collagen fibers content were decreased compared to the CTX group. CONCLUSION: Results showed that, hUCV-MSCs administration increases follicular quantitative parameters and improve degenerative changes in the follicles following CTX injury.
Subject(s)
Cyclophosphamide/adverse effects , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/metabolism , Ovarian Follicle/metabolism , Primary Ovarian Insufficiency , Umbilical Cord/metabolism , Animals , Cyclophosphamide/pharmacology , Disease Models, Animal , Female , Heterografts , Humans , Mesenchymal Stem Cells/pathology , Mice , Ovarian Follicle/pathology , Primary Ovarian Insufficiency/chemically induced , Primary Ovarian Insufficiency/metabolism , Primary Ovarian Insufficiency/pathology , Primary Ovarian Insufficiency/therapy , Umbilical Cord/pathologyABSTRACT
Gluten sensitivity contributes to various degrees of neurological manifestations and neurodegenerative immunological changes. We investigated the experimental features of anti-gliadin immune responses in the central nervous system (CNS) of mice. Female C57BL6 mice were divided into three groups. Mice immunized with complete Freund's adjuvant (CFA) or gliadin emulsified in CFA, and the control group received phosphate-buffered saline (PBS). Immunohistochemistry, hematoxylin-eosin, and Luxol fast blue staining were performed on the sections. The serum levels of interleukin (IL)-17 and interferon-gamma (IFN-γ) were measured using enzyme-linked immunosorbent assay (ELISA). Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assess the mRNA levels of chemokine (C-X-C motif) ligand-2 (CXCL-2), C-C motif chemokine ligand-2 (CCL-2), and CXCL-10. In gliadin+CFA immunized mice, the microscopic lesions included perivascular edema, focal-microgliosis, and acute neuronal necrosis in the cortex, subcortical, Purkinje cell layer, and ventral horn of the spinal cord. While extravasation of anti-IgG antibodies and selective targeting of Purkinje cells were observed in gliadin+CFA immunized mice. A significant increase in serum IL-17 and IFN-g levels (p<0.05), as well as expression of CXCL-2, CCL-2, and CXCL-10 in mice immunized with gliadin+CFA, were monitored versus controls. Our findings indicated that the immune responses directed against gliadin peptides might contribute to blood-brain barrier breakdown, extravasation of serum anti-IgG, gliosis, and acute neuronal necrosis in the cortex and cerebellar Purkinje cells. Anti-IgG antibodies may cause extravasation of blood-born anti-gliadin antibodies and selective targeting of Purkinje cells observed in mice immunized with peptide tryptic (pt) -gliadin in CFA.
Subject(s)
Brain/drug effects , Freund's Adjuvant/administration & dosage , Gliadin/administration & dosage , Spinal Cord/drug effects , Animals , Autoimmunity/drug effects , Brain/immunology , Brain/pathology , Cytokines/blood , Cytokines/genetics , Female , Immunization , Immunoglobulin G/immunology , Mice, Inbred C57BL , Spinal Cord/pathologyABSTRACT
Co-inhibitory molecules modulate immune responses. Immunomodulatory properties of mesenchymal stem cells (MSCs) turn them into ideal candidates for cell therapy. This study was designed to investigate the immunomodulatory effect of adipose-derived stem cells (ASCs) on inflammatory environment of a co-culture of allogenic peripheral blood mononuclear cells (PBMCs) in a two-way mixed leukocyte reaction (twMLR) setting. ASCs were co-cultured with allogenic PBMCs in twMLR setting for four days. The proliferation of peripheral blood mononuclear cells (PBMCs), levels of interleukin (IL)-10, and expression of interferon-gamma (IFN-γ), B7-1, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed death-ligand 1 (PD-L1), +, and CD200R1 genes, as well as cell surface expression of CD200 and CD200R1, were measured in twMLR as control, and co-culture groups on days 0, 2 and 4 of the experiment. The proliferation of PBMCs was suppressed on days 2 and 4 of co-culture. The expression of CD200 (p=0.014), CD200R1, CTLA-4, and PD1 genes increased on days 2 and 4 of the co-culture compared to twMLR. CD200 expressing PBMCs decreased by 1.75% on day 2 of the co-culture but increased by 6.23% on day 4 of the co-culture (p=0.013) compared to the same days of twMLR. IL-10 levels increased in the co-culture supernatants on days 2 and 4 compared to twMLR (p<0.05). Our results showed that ASCs upregulate the CD200/CD200R1 axis more than PD-1/PD-L1 and CTLA-4/B7-1 pathways in the twMLR. Also, elevated expression of CD200R1 in the final day of co-culture was similar to PD-1 expression pattern. This finding suggests a role for the CD200/CD200R1 axis in later modulation of the immune response.
Subject(s)
Adipose Tissue/immunology , Antigens, CD/immunology , Immunologic Factors/immunology , Leukocytes, Mononuclear/immunology , Mesenchymal Stem Cells/immunology , Orexin Receptors/immunology , Up-Regulation/immunology , Adipocytes/immunology , Adult , Cells, Cultured , Coculture Techniques/methods , Humans , Immunomodulation/immunology , Interferon-gamma/immunology , Interleukin-10/immunology , Young AdultABSTRACT
Purpose: Idiopathic pulmonary fibrosis (IPF) is a progressive lung disorder with few available treatments. Mesenchymal stem cell therapy (MSCT), an innovative approach, has high therapeutic potential when used to treat IPF. According to recent data, preconditioning of MSCs can improve their therapeutic effects. Our research focuses on investigating the anti-inflammatory and antifibrotic effects of H2 O2 -preconditioned MSCs (p-MSCs) on mice with bleomycin-induced pulmonary fibrosis (PF). Methods: Eight-week-old male C57BL/6 mice were induced with PF by intratracheal (IT) instillation of bleomycin (4 U/kg). Human umbilical cord vein-derived MSCs (hUCV-MSCs) were isolated and exposed to a sub-lethal concentration (15 µM for 24 h) of H2 O2 in vitro. One week following the injection of bleomycin, 2×105 MSCs or p-MSCs were injected (IT) into the experimental PF. The survival rate and weight of mice were recorded, and 14 days after MSCs injection, all mice were sacrificed. Lung tissue was removed from these mice to examine the myeloperoxidase (MPO) activity, histopathological changes (hematoxylin-eosin and Masson's trichrome) and expression of transforming growth factor beta 1 (TGF-ß1) and alpha-smooth muscle actin (α-SMA) through immunohistochemistry (IHC) staining. Results: Compared to the PF+MSC group, p-MSCs transplantation results in significantly decreased connective tissue (P<0.05) and collagen deposition. Additionally, it is determined that lung tissue in the PF+pMSC group has increased alveolar space (P<0.05) and diminished expression of TGF-ß1 and α-SMA. Conclusion: The results demonstrate that MSCT using p-MSCs decreases inflammatory and fibrotic factors in bleomycin-induced PF, while also able to increase the therapeutic potency of MSCT in IPF.
ABSTRACT
Background: Mesenchymal stem cells (MSCs) can be used to treat premature ovarian failure (POF). Different methods have already been applied to detect MSCs in tissues. This study aimed to investigate the quantitative distribution of CM-DiI-labeled human umbilical cord vein MSCs (hUCV-MSCs) in different regions of the ovarian tissue of the cyclophosphamide (CTX)-induced POF in mice. Methods: Adult female C57BL/6 mice (n = 40) were divided into four groups: (1) Mice receiving PBS as control (Ctrl) group; (2) mice receiving hUCV-MSCs intravenously as Ctrl + hUCV-MSCs group; (3) mice receiving CTX intraperitoneally (i.p.) as CTX group; (4) mice receiving CM-DiI-labeled hUCV-MSCs after CTX injection as CTX + hUCV-MSCs group. Histological changes and CM-DiI-labeled hUCV-MSCs distribution were analyzed in the ovarian tissues. Quantitative real-time PCR was performed to detect human mitochondrial cytochrome b (MTCYB) gene in the ovarian tissues of the mice. Results: The mean number of the fluorescent hUCV-MSCs was 20 ± 2.5 (57.1%) in the medulla, 11.3 ± 2.8 (32.2%) in the cortex, and 5.5 ± 1 (15%) in the germinal epithelium of the ovarian tissue (p < 0.05). Moreover, MTCYB gene was detected in the mice ovaries of the CTX + hUCV-MSCs group, but not in other groups. Conclusion: Our findings suggest that the distribution of the transplanted hUCV-MSCs in different regions of the ovarian tissue is not equal, and it is greater in the medulla than the cortex and germinal epithelium. This is the first report of quantitative distribution of MSCs in different regions of ovarian tissue in the POF model.
Subject(s)
Carbocyanines/metabolism , Cell Movement , Mesenchymal Stem Cells/cytology , Ovary/injuries , Ovary/pathology , Staining and Labeling , Umbilical Cord/cytology , Animals , Cyclophosphamide , Cytochromes b/genetics , Cytochromes b/metabolism , Female , Humans , Mice, Inbred C57BL , Primary Ovarian Insufficiency/pathologyABSTRACT
The immunomodulatory properties of type 2 diabetic patients' adipose-derived mesenchymal stem cells (D-ASCs) has not been extensively studied. In this study, we compared the immunomodulatory properties of D-ASCs and non-diabetic subjects mesenchymal stem cells (ND-ASCs) in co-culture with mixed leukocyte reaction (MLR). ASCs were isolated from adipose tissue samples of type 2 diabetic and non-diabetic subjects (age: 40-55). D-ASCs and ND-ASCs were co-cultured with two-way MLR. Peripheral blood mononuclear cells (PBMCs) proliferation ratio, protein levels of IFN-γ and IL-10, mRNA expression of COX-2, TNF-α, TGF-ß1 and IL-6 genes in MLR, D-ASCs and ND-ASCs co-cultures were assessed using XTT, ELISA and Real-time qRT-PCR, respectively. PBMCs proliferation on days 2 and 4 of D-ASCs co-culture was higher than ND-ASCs co-culture of the same days (p < 0.001). IFN-γ level decreased on day 4 compared to day 2 of ND-ASCs co-culture, but its level had not changed in D-ASCs co-culture. COX-2 expression on days 2 and 4 of D-ASCs co-culture was lower than ND-ASCs co-culture of the same days (p < 0.05). The expression of TNF-α and IL-6 on days 2 and 4 of D-ASCs co-culture were higher than ND-ASCs co-culture of the same days (p < 0.001). TGF-ß1 on day 4 of ND-ASCs co-culture showed a slightly higher expression than D-ASCs co-culture of the same day. Lower suppression of PBMCs proliferation, declined expression of anti-inflammatory and upregulated expression of pro-inflammatory factors in D-ASCs co-culture, indicated an impairment of these cells in modulation of the inflammatory condition.
ABSTRACT
In vivo and in vitro aging of the mesenchymal stromal cells (MSCs) can affects their properties. We investigated the immunomodulatory properties of the term and preterm human umbilical cord vein MSCs (UCV-MSCs) at the passages (P) 2 and 5. Term and preterm UCV-MSCs at P2 and 5 were co-cultured with two-way mixed lymphocyte reaction. Proliferation, IFN-γ and IL-10 protein levels, mRNA levels of the COX-2, TGF-ß1, TNF-α, IL-4 and FoxP3 were assessed. The term UCV-MSCs and P5 of the term and preterm UCV-MSCs had stronger inhibitory effects on cell proliferation than the preterm UCV-MSC and P2, respectively (Pâ¯=â¯0.001). In supernatants of the co-cultures, IFN-γ was higher in the term UCV-MSC than the preterm UCV-MSC, while IL-10 was higher in the preterm UCV-MSCs than the term UCV-MSCs. Also in the co-cultures, COX-2 expression in the term UCV-MSCs and P2 was higher than the preterm UCV-MSCs and P5, respectively and TGF-ß1 expression in the term UCV-MSCs was higher than preterm. Conclusively it appears that the term UCV-MSCs, and P5 of the term and preterm UCV-MSCs showed a higher immunomodulatory ability than the preterm UCV-MSCs and P2, respectively.
Subject(s)
Gestational Age , Immunomodulation , Mesenchymal Stem Cells/immunology , Umbilical Cord/immunology , Cyclooxygenase 2/immunology , Cytokines/immunology , Humans , Infant, Newborn , Infant, Premature/immunology , Mesenchymal Stem Cells/pathology , Umbilical Cord/pathologyABSTRACT
PURPOSE: Migration ability of mesenchymal stem cells (MSCs) towards chemotactic mediators is a determinant factor in cell therapy. MSCs derived from different sources show different properties. Here we compared the migration ability of the term and the pre-term human umbilical cord vein MSCs (hUCV-MSCs). MATERIALS/METHODS: MSCs were isolated from term and pre-term umbilical cord vein, and cultured to passage 3-4. Migration rate of both groups was assessed in the presence of 10% FBS using chemotaxis assay. Surface expression of CXCR4 was measured by flow cytometery. The relative gene expression of CXCR4, IGF1-R, PDGFRα, MMP-2, MMP-9, MT1-MMP and TIMP-2 were evaluated using real time PCR. RESULTS: The isolation rate of the pre-term hUCV-MSCs was higher than the term hUCV-MSCs. Phenotype characteristics and differentiation ability of the term and pre-term hUCV-MSCs were not different. The migration rate of the pre-term hUCV-MSCs was more than the term hUCV-MSCs. Gene and surface expressions of the CXCR4 were both significantly higher in the pre-term hUCV-MSCs (P≤0.05). The mRNA levels of PDGFRα, MMP-2, MMP-9, MT1-MMP and TIMP-2 showed no significant difference between the two groups. CONCLUSION: Our results showed that the gestational age can affect the migration ability of the hUCV-MSCs.
Subject(s)
Cell Movement , Gestational Age , Mesenchymal Stem Cells/cytology , Umbilical Cord/cytology , Umbilical Veins/cytology , Cell Differentiation , Cell Membrane/metabolism , Gene Expression Regulation , Humans , Mesenchymal Stem Cells/metabolism , Receptors, CXCR4/metabolismABSTRACT
In recent studies, mesenchymal stromal cells (MSCs) have been increasingly employed to treat various diseases like pulmonary fibrosis (PF). There are very few MSCs in tissues so in order to obtain their sufficient numbers for therapeutic applications, their in vitro expansion is necessary. The aim of this study was to investigate the effects of long-term culture of the human umbilical cord vein MSCs (hUCV-MSCs) on pulmonary fibrosis in mice. MSCs were first isolated from human umbilical cord vein and cultured up to 18 passages. In C57BL/6 mice, 15 min after belomycin instillation, UCV-MSCs at passages (P) 0, 4, 8, 12, and 18 (long-term culture) were transplanted intratracheally. Mice were weighted every 5 days and were euthanized on day 21. For histopathological examination, the lung sections were stained with hematoxylin-eosin (HE) and Masson's trichrome. The mRNA expression of TGF-ß1, alpha-smooth muscle actin (α-SMA), and collagen type I alpha 1 (COL1A1) in lung tissues were assessed using RT-PCR. For cell tracking, human cytochrome B DNA was detected in mice lung tissues by PCR. The weight of mice receiving long-term culture of UCV-MSCs increased compared to other mice (p=0.056). Also, transplantation of UCV-MSCs at P18 led to increased alveolar space and decreased connective tissue and collagen deposition of the lung tissues. The mRNA expression of TGF-ß1, α-SMA, and COL1A1 also decreased in this group. The results showed that intratracheally transplanted long-term culture of the UCV-MSCs attenuated lung fibrosis in mice.
Subject(s)
Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Pulmonary Fibrosis/therapy , Umbilical Veins/cytology , Animals , Biomarkers , Bleomycin/adverse effects , Cells, Cultured , Collagen Type I, alpha 1 Chain , Disease Models, Animal , Gene Expression Profiling , Heterografts , Humans , Immunohistochemistry , Immunophenotyping , Male , Mesenchymal Stem Cell Transplantation/methods , Mice , Mice, Inbred C57BL , Pulmonary Fibrosis/etiology , Pulmonary Fibrosis/metabolism , Pulmonary Fibrosis/pathology , TranscriptomeABSTRACT
BACKGROUND: Multiple etiologies have been hypothesized for prostate cancer, including genetic defects and infectious agents. A recently reported gamaretrovirus, xenotropic murine leukemia virus-related virus (XMRV) has been reported to be detected in prostate cancer. However, this virus has not been detected in similar groups of patients in other studies. Herein, we sought to detect XMRV in prostate cancers and benign controls in Sanandaj, west of Iran. MATERIALS AND METHODS: In a case-control study, genomic DNA was extracted from formalin fixed and paraffin embedded prostate tissues from a total of 163 Iranian patients. We developed a conventional and a nested PCR assay using primers targeting to an env specific sequence of XMRV. PCR assays were carried out on 63 prostate cancers and 100 benign prostate hyperplasias. RESULTS: Beta-actin sequences were successfully detected in the DNA extracts from all prostate tissues, confirming DNA extraction integrity. We did not detect XMRV in samples either from prostate cancers or benign prostate hyperplasias using XMRV specific primers. CONCLUSIONS: We conclude that in our population XMRV does not play a role in genesis of prostate cancer.
Subject(s)
Adenocarcinoma/etiology , Prostatic Neoplasms/etiology , Retroviridae Infections/complications , Xenotropic murine leukemia virus-related virus/pathogenicity , Adult , Aged , Aged, 80 and over , DNA, Viral/genetics , Follow-Up Studies , Humans , Iran , Male , Middle Aged , Neoplasm Grading , Polymerase Chain Reaction , Prognosis , Prostate/virology , Retroviridae Infections/virology , Xenotropic murine leukemia virus-related virus/genetics , Xenotropic murine leukemia virus-related virus/isolation & purificationABSTRACT
BACKGROUND/PURPOSE: A T-helper cell type 1-specific response leads to the elimination of intracellular infection with Brucella. Studies have shown that naloxone (NLX) can promote a cellular immune response in this respect. The current study was carried out to evaluate the induction of protective immunity in mice against brucellosis by vaccination with a combination of NLX, alum, and heat-killed Brucella melitensis 16 M (HKB). METHODS: Mice were categorized into five groups and received intraperitoneal vaccination on Days 0 and 7. Then serum levels of interferon (IFN)-γ and interleukin (IL)-4, the bacterial load, and the survival rate were measured 2 weeks after the last vaccination. RESULTS: The serum levels of IFN-γ, IL-4, and immunoglobulin G in the NLX + alum + HKB group were shown to be significantly increased (p < 0.05). Furthermore, the lowest bacterial load was observed in this group. The survival rate in groups vaccinated with combinations containing adjuvants was 100%. CONCLUSION: The combination of NLX and alum enhanced the immunogenicity of HKB, which can be used in the vaccination of animals and humans at risk of the disease.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Alum Compounds/administration & dosage , Brucella Vaccine/immunology , Brucella melitensis/immunology , Brucellosis/prevention & control , Naloxone/administration & dosage , Vaccination/methods , Animals , Antibodies, Bacterial/blood , Bacterial Load , Brucella Vaccine/administration & dosage , Brucellosis/immunology , Disease Models, Animal , Immunoglobulin G/blood , Interferon-gamma/blood , Interleukin-4/blood , Male , Mice , Mice, Inbred BALB C , Serum/immunology , Serum/microbiology , Survival Analysis , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/immunologyABSTRACT
BACKGROUND: Malassezia furfur is a lipophilic yeast that causes skin disease. OBJECTIVE: To evaluate the level of IL-10, IFN-γ and IL-12P70 in co-incubation of peripheral blood mononuclear cells (PBMCs) with M. furfur grown in the presence of some different types of natural oils. METHODS: PBMCs were obtained from blood samples of normal volunteers. M. furfur was cultured in different culture media containing almond oil, fish oil, walnut oil, full-fat milk, and a fat-free medium; and the yeasts grown were harvested and used for co-incubation with PBMCs in vitro. The IFN-γ, IL-10, and IL-12P70 levels were measured at different time intervals using ELISA methods. RESULTS: Generally, IFN-γ and IL-10 levels in the co-incubation of yeasts with walnut oil group (WOG) and fish oil group (FOG) were higher than those in the almond oil group (AOG) and full-fat milk group (FFMG). Although the IL-12P70 was higher in groups such as AOG, FOG, and WOG; the increase was not statistically significant. CONCLUSION: The results demonstrated that the type of fat used by M. furfur in the culture media can influence the immune response and increasesIFN-γ and IL-10 levels in an early time point of the culture system.