Iranian Journal of Immunology最新文献

Background: Gastroesophageal reflux disease (GERD) is a prevalent clinical condition that affects millions ‎of individuals worldwide.‎.

Objective‎: To assess the level of soluble HLA-E (sHLA-E) as a biomarker in the diagnosis and ‎immunopathogenesis of GERD patients.‎.

Methods‎: The case-control prospective study included 40 GERD patients who were consulted at the ‎Gastroenterology Unit of Al-Kindy Teaching Hospital, as along with 40 healthy control ‎subjects. The study period extended from January 2023 to May 2024. Blood was drawn from ‎both groups and serum was separated to assesss HLA-E using a sandwich enzyme-linked ‎immunosorbent assay (ELISA) kit.‎.

Results: There was a statistically significant difference in sHLA-E levels between GERD patients and ‎healthy controls (p = 0.021). The median sHLA-E level was significantly higher in GERD ‎patients (0.370 ng/mL) compared to controls (0.148 ng/mL). A receiver operating characteristic ‎‎(ROC) curve was generated to evaluate the diagnostic performance of soluble HLA-E (sHLA-E) ‎in predicting GERD. The area under the ROC curve (AUC) was calculated to assess the ‎discriminatory ability of sHLA-E, with a value of 0.649 (95% CI: 0.534-0.752, p = 0.021). The ‎optimal cutoff value for sHLA-E was determined to be ≤0.65 ng/mL, with a sensitivity of ‎‎85.1%, specificity of 27.3%, positive predictive value of 65.9%, negative predictive value of ‎‎69.4%, and accuracy of 35.0%.‎.

Conclusion‎: The study provides evidence of an association between elevated sHLA-E levels and GERD. It ‎also suggests that sHLA-E has a moderate discriminatory ability as a biomarker in predicting ‎GERD.‎.

Background: Studies have demonstrated that transmembrane tumor necrosis factor-α (tmTNF-α) plays an anti-inflammatory role. tmTNF-α has a dual function, acting as both a signaling ligand and a receptor that transmits reverse signaling to cells expressing tmTNF-α. However, the role and mechanisms of tmTNF-α reverse signaling in sepsis are not fully understood.

Objective‎: To explore the potential role and mechanisms of tmTNF-α reverse signaling in lipopolysaccharide (LPS)-induced inflammation.

Methods‎: The expression levels of tmTNF-α and TNF-α mRNA were evaluated using flow cytometry and real-time PCR, respectively. We employed the anti-TNF-α drug infliximab to stimulate tmTNF-α reverse signaling and measured interleukin-6 (IL-6) and monocyte chemoattractant protein (MCP)-1 production through real-time PCR and ELISA in THP-1-derived macrophages. The location of p65 was determined through immunofluorescence assay. The phosphorylation and acetylation of p65, as well as the expression levels of MCP-induced protein 1 (MCPIP1) and Sirtuin 1 (SIRT1), were evaluated using western blotting.

Results: Our findings revealed that tmTNF-α reverse signaling reduced the expression of IL-6 and MCP-1 triggered by LPS. tmTNF-α reverse signaling inhibited the nuclear translocation of p65, suppressed p65 phosphorylation and acetylation, and upregulated the expression of negative regulatory molecules MCPIP1 and SIRT1 in the LPS/ toll-like receptor 4 (TLR4) signaling pathway.

Conclusion‎: This study demonstrates that tmTNF-α reverse signaling plays a negative regulatory role in inflammation triggered by LPS by inhibiting the TLR4/ nuclear factor kappa B (NF-κB) signaling pathway. This study helps to further understand the function of tmTNF-α reverse signaling and offers new therapeutic possibilities for sepsis and other inflammatory disease conditions.

Background: Clinical features of SARS-CoV-2 infection vary, ranging from asymptomatic cases to pneumonia, and other serious complications. Some populations have been observed to be at higher risk for severe disease and death compared to other ethnical groups.

Objective: To evaluate two parameters of the innate immune system, that play a significant role in viral immunity.

Methods: In samples of peripheral blood from sixteen patients with severe COVID-19, ten with asymptomatic to mild illness, and fifteen healthy subjects, the percentage of NK and NKT cells, the expression of different NK cell receptors and the blood levels of pro-inflammatory cytokines were tested.

Results: We observed that patients with severe COVID-19 showed significantly lower frequencies of both CD56dim and CD56bright NK cells compared to patients with mild illness or healthy controls. Furthermore, patients with severe manifestation of COVID-19 exhibited an aberrant expression of the natural cytotoxicity receptors NKp30, NKp44 and NKp46. Similarly, NK cells from these patients showed statistically significant differences in the expression of various killer immunoglobulin-like receptors (KIRs) in the two main cell subsets (CD56bright, CD56dim) compared to controls or patients with mild disease. Moreover, patients with severe illness displayed decreased frequency of NKT cells (defined as CD3+CD56+) and elevated blood levels of the cytokines IL-6 and IL-8.

Conclusion: This study suggests that the abnormal features of NK and NKT cells observed in patients with severe SARS-CoV-2 infection may play an important role in the outcome of this infectious disease in various population groups.

Background: Patients with thin endometrium undergoing frozen-thawed embryo transfer often encounter challenges with pregnancy outcomes. Enhancing endometrial receptivity and immune tolerance may improve these outcomes.

Objective: To investigate the effects of intrauterine perfusion of granulocyte colony-stimulating factor (G-CSF) and human chorionic gonadotropin (HCG) on regulatory T cells (Tregs) and pregnancy outcomes in patients with thin endometrium undergoing frozen-thawed embryo transfer.

Methods: 150 patients with thin endometrium were randomly assigned to three groups: a control group that received no intervention, an HCG group, and a G-CSF group. The effectiveness of the treatments was assessed by comparing uterine parameters, Treg levels, and pregnancy outcomes across the groups.

Results: The HCG and G-CSF groups exhibited significant improvements compared to the control group, including increased endometrial thickness, enhanced blood flow, higher expression of endometrial receptivity markers (integrin αvβ3, osteopontin), and elevated Treg levels. Notably, the G-CSF group demonstrated even greater enhancements compared to the HCG group, with significantly higher endometrial thickness, better blood flow, increased receptivity markers, and elevated Treg levels. Additionally, the G-CSF group achieved significantly higher biochemical and clinical pregnancy rates compared to both the HCG and control groups. This highlights the potential of G-CSF in improving pregnancy outcomes for patients with a thin endometrium.

Conclusion: The intrauterine perfusion of G-CSF significantly enhanced pregnancy outcomes in patients with thin endometrium by improving endometrial blood flow, immune tolerance, thickness, Treg induction, and embryo implantation. These findings suggest that G-CSF could be a promising therapeutic option for this patient population.

Background: Gamma-delta (γδ) T cells are a distinct subset of T cells with a receptor composed of γ and δ chains. Their ability to directly recognize stress-induced molecules and non-peptide antigens expressed by cancer cells, along with their capacity to produce cytokines and interact with other immune cells, makes them potentially significant contributors to immune-based treatments.

Objective: To investigate the presence and frequency of Tγδ cells in tumor-draining lymph nodes of patients with bladder cancer (BC), and to assess their association with prognostic parameters.

Methods: Forty-nine fresh tumor-draining lymph nodes from untreated patients with BC were minced to obtain single cells. The cells were surface-stained with anti-CD3, anti-TCRγδ, and anti-HLA-DR antibodies, then acquired on a four-color FACSCalibur flow cytometer, and analyzed by FlowJo software.

Results: On average, 2.07% ± 1.99% of CD3+ lymphocytes in regional nodes of BC exhibited a γδ T phenotype. A considerable percentage of these cells (37.90% ± 24.42%) expressed HLA-DR. Statistical analysis revealed that while the frequency of γδ T cells showed no variation among patients with different prognoses, the HLA-DR+ subset was higher in T4 patients than in T2 patients (p=0.031). These cells also tended to be increased in stage III compared to stage II (p=0.077).

Conclusion: The data collectively indicated an association of HLA-DR expressing γδ T cells with prognostic factors related to tumor progression (higher T-group and stage), suggesting their potential involvement in disease progression. However, future research, including longitudinal studies with larger cohorts, needs to validate these findings and elucidate the functional roles of γδ T cells in the immune response against BC.

Background: Developing effective targeted treatment approaches to overcome drug resistance remains a crucial goal in cancer research. Immunotoxins have dual functionality in cancer detection and targeted therapy.

Objective: This study aimed to engineer a recombinant chimeric fusion protein by combining a nanobody-targeting domain with an exotoxin effector domain. The chimeric protein was designed to bind surface-expressed GRP78 on cancer cells, facilitating internalization and inducing apoptosis to inhibit proliferation and survival.

Methods: Using a flexible linker, we designed two constructs linking VHH nanobody domains to Pseudomonas exotoxin (PE) domains II, III, and Ib. These constructs were then optimized for expression in E. coli BL21 (DE3) using the pET28a vector. Following the expression of the recombinant proteins, we purified them and tested their binding capability, cytotoxicity, and ability to induce apoptosis in breast cancer cell lines MDA-MB-231 and MCF-7, as well as in control cell lines HEK-293 and MDA-MB-468. The binding affinity was measured using a cell-based ELISA, internalization was assessed through Western blotting, cytotoxicity was evaluated by an MTT assay, and apoptosis was determined using flow cytometry with an Annexin V kit.

Results: The immunotoxin specifically bound to cancer cells expressing csGRP78. The results of the cytotoxicity test showed that the cytotoxic effect of two constructs, I and II, depended on concentration and time. With an increase in both components, the effect of recombinant proteins also increased. Both constructs were able to penetrate and induce apoptosis in csGRP78+ cells.

Conclusion: These immunotoxin structures showed therapeutic potential against GRP78-expressing cancers, making them suitable candidates for targeted therapy pending in vivo studies.

Background: Interleukin-2 (IL-2) is a well-known cytokine that plays a crucial role in stimulating immune cells, including natural killer (NK) cells and cytotoxic T cells. It has been studied as an immunotherapy for a variety of diseases, including cancer. However, due to its short serum half-life, high doses of IL-2 are required which can result in systemic toxicities like capillary leak syndrome.

Objective: To demonstrate the enhanced antitumor efficacy of Albumin Binding Domain-conjugated IL-2 (ABD-IL-2) at a lower dose compared to IL-2.

Methods: IL-2 and ABD-IL-2 were purified using Ni-NTA resin with a histidine sequence added to their C-terminal region for purification purpose. Peripheral blood lymphocytes were stimulated with IL-2 and ABD-IL-2 to assess their function. 4T1 cells were injected into BALB/c mice to establish a breast cancer model with metastasis evaluated in the lungs.

Results: Both recombinant proteins significantly stimulated T lymphocyte proliferation compared to the negative control (P=0.000, P=0.001). Administration of both proteins reduced the size of isolated tumors in the breast cancer mouse model. The control group had more nodules and larger lung metastatic centers (P=0.000). Metastasis to secondary lymphoid organs occurred only in the control group.

Conclusion: By using ABD-IL-2 at a one-third concentration compared to IL-2, we aimed to reduce administration toxicity associated with high doses of IL-2 in immunotherapy. This approach shows potential for improving IL-2-based treatments while minimizing adverse effects.

Background: Breast cancer is the leading cause of cancer-related deaths in women. Cytokines have been linked to various cancers, and both benign and malignant breast diseases are associated with inflammation. However, there is limited understanding of how the immune system's cytokine response varies among different subtypes of breast cancer.

Objective: To assess cytokine levels in breast cancer patients according to their subtypes and investigate the potential role of these cytokines in treatment.

Methods: Patients with stage 1-2 breast cancer and healthy volunteers were included in the study. The breast cancer patients were classified as luminal A, luminal B, and triple negative based on ER, PR, HER2 receptor status, and Ki67 score of trucut biopsy results. Multiplex assay and flow cytometry were used to quantify the concentrations of IL-1β, IFN-α2, IFN-γ, TNF-α, MCP-1, IL-6, IL-8, IL-10, IL-12p (p70), IL-17A, IL-18, IL-23, and IL-33 in serum samples collected from all participants. Age, menopausal status, and hematologic parameters were also compared between groups.

Results: The study involved 19 luminal A, 20 luminal B, 18 triple-negative patients and 21 healthy volunteers. TNF-α, IL-6, IL-8, IL-10, IL-12p (p70), IL-18, and IL-23 cytokines were significantly higher in breast cancer patients than in healthy volunteers. Significant differences in IFN-γ, IL-6, IL-8, IL-10, IL-12p (p70), IL-17A, IL-18, and IL-23 were observed between subtypes, with triple-negative patients showing lower cytokine levels, except for MCP-1.

Conclusion: The decreased levels of cytokines in triple-negative breast cancer indicate lower immunogenicity leading to more aggressive tumor progression as a result of an insufficient immune response.

Background: Rosemary (Ros) is a member of the Lamiaceae family known for its antitumor properties. However, its low water solubility and impaired bioavailability are limiting factors when using rosemary extract. Liposomes are synthetic vesicles that offer permeability, improved bioavailability, and lack of immunogenicity and toxicity, making them ideal for delivering various drugs.

Objective: To prepare liposomes (HSPC/Chol/mPEG2000-DSPE) containing rosemary alcoholic extract (LipRos) and evaluate its antitumor properties in a mouse model of colorectal cancer (CRC).

Methods: LipRos were prepared and characterized. CRC was induced in Balb/c mice by subcutaneous injection of C26 cells, and tumor size was monitored continuously. The MTT assay was performed to evaluate cytotoxicity, and liver and kidney function tests were conducted to assess safety. The expression of the pro-apoptotic gene B-cell-lymphoma-2 (Bcl-2), the anti-apoptotic gene Bcl-2-associated-X-protein (Bax), and the expression of cytokines Tumor-necrosis-factor-alpha (TNF-α), Transforming-growth-factor-beta (TGF-β), and Interferon-gamma (IFN-γ) were investigated using real-time PCR. Flow cytometry was used to evaluate the count of cytotoxic (CTL) and regulatory T lymphocytes (Tregs) in spleen and tumor tissue.

Results: The results showed that the size of liposomal formulations and their encapsulation efficiency were 113.4 nm and 85%, respectively. The MTT assay demonstrated insignificant cytotoxicity of LipRos on splenocytes, and the tumor size was significantly reduced in the LipRos group (P=0.00045). LipRos also significantly decreased Bcl-2 gene expression (P=0.0086), increased Bax and IFN-γ gene expression (P=0.031), and enhanced the infiltration of CTLs in tumor tissue (P=0.023).

Conclusion: This study showed that PEGylated (Poly-Ethylene-Glycol) liposomes containing rosemary extract exhibit an antitumor effect on C26 colorectal cancer cells through multiple mechanisms. These findings can be utilized in future studies.

Background: CD8+ T cells have been found to accumulate in atherosclerotic plaques. However, the specific role of CD8+ T cell subsets in the development of atherosclerosis is still not fully understood.

Objective: To investigate the presence and functions of type 1 CD8+ T (Tc1) cells and interleukin-17 (IL-17)-producing CD8+ T (Tc17) cells.

Methods: Apolipoprotein E-deficient mice were fed a high-fat diet to induce atherosclerosis. Flow cytometry was used to identify and isolate aortic CD8+ T cell subsets, which were then cultured in vitro to assess their pro-inflammatory activities. The cholesterol content of the CD8+ T cell subsets was quantified.

Results: T-box expressed in T cells (T-bet)+ Tc1 cells and retinoic acid-related orphan receptor gamma t (RORγt)+ Tc17 cells were found in the atherosclerotic aorta. Aortic CD8+ T cells showed lower pro-inflammatory activity compared to splenic counterparts, with less interferon-gamma (IFN-γ) (P <0.01) and tumor necrosis factor-alpha (TNF-α) production (P <0.01). Surprisingly, aortic CD8+ T cells expressed little IL-17 and interleukin-21 (IL-21) despite the presence of Tc17 cells. Aortic Tc1 and Tc17 cells expressed high levels of 2B4 and programmed cell death protein 1 (PD-1). Furthermore, aortic Tc1 and Tc17 cells had higher cholesterol contents than splenic CD8+ T cells (P <0.05, respectively). Cholesterol treatment decreased IFN-γ expression in Tc1 cells (P <0.001) and reduced IL-17 expression in Tc17 cells (P <0.001). Additionally, cholesterol up-regulated 2B4 and PD-1 on Tc1 (P <0.001) and Tc17 cells (P <0.001).

Conclusion: Aortic CD8+ T cells, particularly aortic Tc17 cells, are functionally exhausted in atherosclerosis, possibly due to the influence of cholesterol.