Journal of immunoassay & immunochemistry最新文献

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system. Inflammasomes, particularly NLRP3 and AIM2, have been implicated in MS pathogenesis. Interferon-β (IFN-β) and Rituximab (RTX) are treatment agents for relapsing-remitting MS (RRMS), but their impact on inflammasome regulation remains unclear. This study evaluates the expression of NLRP3 and AIM2 inflammasomes in MS patients responding to IFN-β and RTX, alongside newly diagnosed cases and healthy controls. Blood samples from IFN-β-treated patients (n = 23), RTX-treated patients (n = 23), newly diagnosed people (n = 20), and healthy controls (n = 12) were analysed. mRNA levels of NLRP3 and AIM2 were measured by RT-PCR, and plasma IL-1β was assessed using ELISA. Results revealed AIM2 expression was significantly higher in RTX-treated patients compared to other groups, while NLRP3 showed no significant differences. IL-1β levels were elevated in all patient groups, but no correlations were found between disease/treatment duration and inflammasome or IL-1β levels. RTX significantly increases AIM2 expression, suggesting its potential as a biomarker or therapeutic target in MS. Further research is needed to clarify AIM2's role in disease processes.

Considering the high tendency of hepatitis B virus (HBV) to mutate and escape diagnosis, and the high prevalence of its chronic infection in Nigeria, it is expedient to wholistically study the various genes to understand the changes and to prepare for such to enhance the diagnosis, management, and control of the disease. This study was designed to detect the four genes of HBV in different categories of participants. Venous blood samples were collected from sick and apparently healthy individuals and screened for HBsAg using the ELISA method. Out of the samples, 36 hBsAg positive and 9 randomly selected HBsAg negative were selected for HBV DNA extraction and PCR using established primers and protocols. The HBsAg prevalence was found to be 13.8% (51/369) with Oyo state having the highest (37.3%; N = 19/51) compared to other states. After amplification, HBs gene detection was 28.9%; (n = 13/45), HBc gene 33.3%; (n = 15/45), HB Pol gene 26.7%; (n = 12/45), and HBx gene 35.6%; (n = 16/45) and one OBI case (11.1%; n = 1/9). Considering the complications associated to HBV infections and the various changes being observed in the structure of the virus, more study is needed to enhance the diagnosis, manage- 230 ment, and control of hepatitis B.

To comply with regulatory guidelines and ensure that biopharmaceutical agents meet safety and efficacy, we developed an analytical method to detect anti-drug antibodies (ADAs) generated in response to HCW9218, a fusion molecule containing the human soluble TGF-β RII and IL-15/IL-15 Rα sushi domains. The method demonstrated a sensitivity of 34.6 ng/mL with a suitably high degree of specificity, selectivity, and precision. To assess the neutralizing capacity of ADAs, we developed cell-based assays specific to IL-15 and TGF-β RII. These assays effectively discriminated the neutralizing activity of sera spiked with neutralizing antibody (NAb). The determination of positive NAb was based on absorbances corresponding to a threshold value of 30% inhibition of IL-15 or TGF-β RII activity. Each assay was validated using human sera from ongoing clinical trials. ADAs were detected in most sera tested with titers less than 10,000. None of the sera inhibited IL-15 and TGF-β RII activity by more than 30%. Interestingly, circulating TGF-β present in sera mimicked the action of NAb by binding to soluble TGF-β RII resulting in higher baseline neutralization activity. These methods proved to be suitable for detection of ADAs and NAbs in HCW9218 clinical samples or analogs sharing similar receptor/cytokine subunits and/or downstream signaling pathways.

In chronic hepatitis C (CHC), the virus may induce autoimmune responses via autoantibodies production, including antinuclear antibodies (ANA). Former studies reported great ANA predisposition in CHC and these ANAs may be related to worse prognosis including hepatocellular carcinoma (HCC). We aimed to evaluate the association between ANA incidence and CHC-related HCC development and to evaluate these molecules effect on HCC severity including tumor size and advanced stages. Results revealed that ANA seropositivity was associated with disease severity. HCC patients (54%, OR = 9.7) were associated with ANA positivity more than liver cirrhosis (24.5%) and fibrosis (10.8%). ANA positivity was significantly high in patients with severe tumor features including macrovascular invasion (61.9%; OR = 8.1), large size (68.2%; OR = 2.4), Child C (83.3%; OR = 8.1), BCLC end stage (83.3%; OR = 8.6) and advanced CLIP stage (80.9%; OR = 7.9). ANA positivity were significantly (p < 0.05) correlated with some estimated liver fibrosis related biomarkers including EMA (r = 0.206), fibronectin (r = 0.273), cytokeratin-1 (r = 0.365) and collagen III (r = 0.324). In conclusion, our observation of increased ANA+ serum samples among CHC-related HCC might suggest the oncogenic role of ANA in such patients. Also, clinicians need to appreciate value of ANA testing among HCC patients as these molecules were associated with tumor severity and worse outcomes.

Hypothyroidism encompasses both autoimmune forms, notably Hashimoto's thyroiditis (HT), and nonimmune hypothyroidism (NIHT), each driven by distinct molecular mechanisms. Despite overlapping clinical features, their specific molecular differences remain underexplored. This study aimed to compare the expression of inflammatory biomarkers (Calprotectin, Cyclophilin A, Endocan, Procalcitonin) and microRNAs (miR-223, miR-711) among HT, NIHT, and healthy individuals, evaluating their diagnostic relevance. A total of 120 participants were enrolled: 40 with HT, 40 with NIHT, and 40 healthy controls. Serum inflammatory markers were quantified via ELISA, and miRNA levels assessed using qRT-PCR normalized to U6 RNA. Statistical analyses included ANOVA, correlation, and ROC curve evaluation. Significant group differences were found in inflammatory marker levels (p < 0.001), with NIHT patients showing higher concentrations than HT and controls. miR-223 was notably upregulated in both HT and NIHT groups (p = 0.006), whereas miR-711 showed a non-significant downregulation. ROC analysis revealed miR-223 had the highest diagnostic value in distinguishing HT (AUC = 0.84) and NIHT (AUC = 0.85) from controls, outperforming other markers. Cyclophilin A also demonstrated strong discriminatory capability. These findings suggest that combined profiling of inflammatory markers and miRNAs - especially miR-223 and Cyclophilin A holds promise for improved diagnosis and understanding of hypothyroid subtypes.

Participation of TRPM8 in hepatocellular carcinoma (HCC) development was not precisely declared. CD47 mediates immune escape and macrophage phagocytosis of tumors. CDK4 mediates oncogenesis. Synergistic implications of TRPM8, CD47, and CDK4 in HCC were not declared. This research aims to demonstrate the expressions of TRPM8, CD47and CDK4 in HCC and to declare correlations and significance. Paraffin blocks from 101 hCC and 82 adjacent non-tumorous liver were immunostained using TRPM8, CD47 and CDK4 antibodies. HCC showed highly significant increased TRPM8, CD47, and CDK4 expressions than control liver tissue (p < 0.001) for all. TRPM8, CD47, and CDK4 were significantly associated with poor prognostic criteria as high tumor grade, advanced stage, microvascular invasion, and necrosis. There was a significant association between cirrhotic and non-cirrhotic adjacent liver regarding positive TRPM8 (p < 0.02) and high CDK4 expressions (p < 0.045). There were significant direct relationships between each immunohistochemical antibody and the other two. Prolonged overall survival was significantly associated with low CDK4 (p = 0.019). In conclusion, TRPM8, CD47, and CDK4 may regulate synergistic functions in HCC oncogenesis and accomplish unfavorable prognostic significance. TRPM8 and CDK4 might share in development of HCC from cirrhosis. TRPM8, CD47, and CDK4 could be therapeutic targets in HCC.

Introduction: Gastric cancer (GC) is a leading cause of cancer-related mortality worldwide. Dysregulation of molecular pathways, including β-Catenin-mediated Wnt signaling, epithelial-to-mesenchymal transition (EMT), and E-Cadherin-modulated cell adhesion, plays critical roles in gastric carcinogenesis. This study assesses the expression patterns of β-Catenin and E-Cadherin in GC and explores their prognostic significance.

Methods: This retrospective, multi-center study analyzed GC cases diagnosed between 2009 and 2019 at the pathology departments of Security Forces and Rabta Hospitals. Tissue microarray (TMA) paraffin blocks from 48 GC cases were immunohistochemically stained using antibodies for β-Catenin (Leica, 17C2) and E-Cadherin (Leica, 36B5). β-Catenin expression was scored as membranous, cytoplasmic, or nuclear, with overexpression defined as ≥ 50% positive cells. E-Cadherin staining was categorized from absent (score 0) to marked membranous staining (score 3), with scores 0-2 considered aberrant. Statistical analysis was performed using SPSS version 23.

Results: Of the 48 cases, β-Catenin overexpression was observed in 50% of cases, significantly associated with tumor differentiation (p = 0.033), age > 60 years (p = 0.042), and male sex (p = 0.028). Aberrant E-Cadherin expression was found in 65% of cases, linked to poorly cohesive and diffuse subtypes (p = 0.053), poor differentiation (p = 0.042), and recurrence (p = 0.043), with a trend toward reduced survival (p = 0.056).

Conclusion: β-Catenin overexpression and aberrant E-Cadherin expression are frequent in GC, reflecting their roles in tumor progression via Wnt signaling and EMT. These findings highlight their potential as prognostic biomarkers and therapeutic targets, particularly for Wnt pathway-directed therapies in personalized GC management.

Breast is a major global health issue and the most common cancer in women. Identifying vascular invasion is challenging due to the need to distinguish true invasion from artifacts. This study explored lymphatic embolism in invasive breast carcinoma using the monoclonal antibody D2-40 as a prognostic indicator. A total of 100 patients with invasive breast carcinoma from 2009 to 2011 were included in the study. Tissue microarray technique (TMA) was used on patient tissue, constructing three paraffin blocks from each participant's histological data. Immunohistochemistry with D2-40 and CD34 antibodies was performed to identify lymphatic and blood emboli, respectively, and results were compared with previous findings. A prior report using hematoxylin-eosin staining found fewer patients with lymphatic emboli (34) compared to our study (56) using D2-40. Lymphatic emboli correlated with axillary metastases, with an odds ratio (OR) of 3.50, a 95% confidence interval (CI) of 1.92-5.08, and a p-value of 0.001, whereas hematoxylin-eosin alone showed OR = 1.42, 95% CI = 0.40-3.47, and p-value = 0.23. TMA with D2-40 staining detected more lymphatic emboli than hematoxylin-eosin staining alone. Higher embolic expression rates are linked to increased tumor aggressiveness, worse prognosis and shorter overall survival.

Introduction: Epidermal Growth Factor Receptor (EGFR) expression is not well-studied in Human Epidermal Growth Factor Receptor 2 (HER2) positive breast cancer. We aim to study the prevalence of EGFR immunohistochemical expression in HER2-positive breast carcinomas and to correlate this expression with different anatomo-clinical parameters.

Methods: It was a retrospective study involving cases of HER2-positive breast carcinoma collected at the Immuno-Histo-Cytology Department of Salah Azaïez Institute of Tunis between 2018 and 2020. An immunohistochemical study using the anti-human EGFR monoclonal antibody was performed. Cases with an overall score ≥1+ were considered positive.

Results: Fifty patients were included. EGFR expression in HER2-positive breast carcinomas was more likely to occur in patients under the age of 50 (p = 0.063). It was significantly associated with the absence of lymphovascular invasion (p = 0.047). In multivariate analysis, young age, absence of lympho-vascular invasion, and high Ki67 proliferation index (>60%) were independently associated with positive EGFR expression (p = 0.047, p = 0.040, and p = 0.050, respectively).

Conclusion: Through this first Tunisian study, our data revealed that the immunohistochemical expression of EGFR is associated with young age, absence of lymphovascular invasion, and a high mitotic index (Ki67), which may suggest a potential predictive value for chemotherapy response.

The identification of biomarkers for pulmonary exacerbations in cystic fibrosis (CF) is inevitable. We aimed to evaluate the serum levels of substance p (SP), neuropeptide Y (NPY), and Pituitary Adenylate Cyclase Activating Polypeptide (PACAP), at time of pulmonary exacerbation and after treatment with antibiotics. Twenty cystic fibrosis patients with mean age of 8.83 years (±4.37) were enrolled. Serum samples were taken at the time of admission and two weeks post-antibiotic therapy. Serum levels of target markers were determined using ELISA. Serum SP, NPY, and PACAP levels were significantly higher at exacerbation (229.22 ± 73.86 pg/ml, 1869.89 ± 787.14 pg/ml, and 32,261.51 ± 22283.78 fg/ml, respectively) than post-antibiotic therapy (206.29 ± 77.83 pg/ml, 1412.95 ± 647.09 pg/ml, and 17,359.39 ± 10105.39 fg/ml, respectively; p < 0.05). Positive correlations were observed between serum levels of SP and PACAP (r = 0.515, p = 0.020) and NPY (r = 0.779, p < 0.001), and between NPY and PACAP (r = 0.513, p = 0.021). A negative correlation was found between NPY and BMI z-score (r = -0.503, p = 0.024). As a conclusion, serum SP, NPY and PACAP levels are potential biomarkers for CF pulmonary exacerbations and response to antibiotic therapy.