Expression of Immune Checkpoint Regulators, Cytotoxic T-Lymphocyte Antigen-4, and Programmed Death-Ligand 1 in Epstein-Barr Virus-associated Nasopharyngeal Carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is the most common cancer arising from the nasopharynx with a poor prognosis. Targeting immune checkpoint is one of the new promising lines in cancer treatment. Cytotoxic T-lymphocyte antigen-4 (CTLA-4) and programmed death-ligand 1 (PD-L1) are immune checkpoints that regulate T-cell immune function. AIM: This work aimed to assess the immunohistochemical expression of PD-L1 and CTLA-4 in NPC and their ability to predict survival and response therapy and to check their validity as immunotherapy targets. Twenty-six cases of NPC were studied by immunohistochemistry for PD-L1 and CTLA-4 and by nested polymerase chain reaction followed by DNA sequencing for the presence of EBNA-1 gene of Epstein-Barr virus (EBV). All investigated cases were diagnosed and treated in the Zagazig University Hospital in the period from August 2015 to July 2018. EBNA-1 gene was identified in 84.6% of the cases. Whereas the expression of PD-L1 was noted in 46.2% of all cases studied, 54.6% of EBV-associated NPCs were found to express PD-L1. There was a significant association between PD-L1 expression and the advanced stage of the tumor (P<0.001). CTLA-4 expression was observed in 88.4% of all NPC cases as cytoplasmic staining in both tumor cells and tumor-infiltrating lymphocytes. CTLA-4 expression in lymphocytes was associated with the presence of EBV. A significant association was detected between CTLA-4 and tumor-infiltrating lymphocyte expression on one side and the stage of the tumor on the other. High expression of CTLA-4 was significantly associated with disease progression and worse overall survival. CONCLUSION: PD-L1 and CTLA-4 are adverse prognostic markers in NPC. The authors propose that targeted therapy against PD-L1 and CTLA-4 will be a hopeful therapy for cases of NPC with resistance to concurrent chemoradiation treatment in Egypt, especially EBV-associated cases.

Evaluation of Procalcitonin, C-Reactive Protein, and Interleukin-6 as Early Markers for Diagnosis of Neonatal Sepsis.

BACKGROUND: Neonatal sepsis diagnosis is a challenge because of its nonspecific presentation together with low sensitivity of the time-consuming bacterial cultures. So, many sepsis markers, like C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6), are emerging to improve its diagnosis. AIM: This study was done to investigate the role of CRP, PCT, and IL-6 in promoting the early diagnosis of neonatal sepsis in an attempt to decrease morbidity and mortality. METHODS: This cross-sectional study was conducted on 50 neonates suspected with sepsis enrolled from the neonatal intensive care unit (NICU) of Zagazig University Hospitals, Egypt. Blood cultures for these neonates were done before starting antibiotics. Also, bacterial DNA was revealed from the blood by broad-range 16S rDNA polymerase chain reaction (PCR). Measurements of CRP using the immunoturbidimetry method, PCT using fluorescence immunoassay quantitative method, and IL-6 using commercially available ELISA kit were done to all enrolled neonates. RESULTS: Forty-one neonates with proved sepsis were found to be positive in blood culture and/or PCR for bacterial 16S rDNA. The most common isolated organisms were Klebsiella (61.3%), followed by E. coli (9.7%) and CONS (9.7%). We detected much significant higher levels of PCT, CRP, and IL-6 in the proved sepsis group than the suspected neonatal sepsis cases (p ≤ 0.001, 0.001, and 0.004, respectively). Serum PCT levels showed the highest sensitivity, specificity, PPV, NPV, and accuracy of 97.6%, 89%, 97%, 88.9%, and 96% than other studied sepsis markers. CONCLUSION: PCT has satisfactory characteristics as a good marker than IL-6 and CRP for the diagnosis of neonatal sepsis.

Assessment of The Role of Serum Neutrophil Gelatinase-Associated Lipocalin in Egyptian Childhood Bronchial Asthma: One Centre Study.

Neutrophil gelatinase-associated lipocalin (NGAL) is emerging as a potential biomarker in many medical conditions including asthma. The aim of this study was to assess the role of serum NGAL in Egyptian childhood bronchial asthma. The study included 156 patients and 39 apparently healthy control children. Full clinical examination, pulmonary function tests; CBC, CRP, IgE, liver function tests, and renal function tests, and serum NGAL level were performed. The difference between the studied groups was statistically significant regarding IgE, eosinophils and NGAL (P= 0.001 for each). In addition, the difference between the subgroup with severe persistent asthma and the subgroup with mild intermittent asthma was significant (P=0.001). ROC curve analysis showed that at a cut-off value of 0.884 the sensitivity and specificity of differentiating severe bronchial asthma patients from controls was 82 % and 76 %, respectively. In conclusion, NGAL may represent a potential marker of bronchial asthma in children with severe disease.

Prevalence of occult hepatitis C virus infection in patients who achieved sustained virologic response to direct-acting antiviral agents.

The reappearance of HCV infection months or years after sustained virologic response (SVR) may be due to the persistence of HCV in tissue cells in spite of being undetected in serum. This situation is known as occult hepatitis C infection (OCI). We aimed to assess the prevalence of OCI in Egyptian patients with chronic hepatitis C (CHC) who achieved SVR after treatment with direct-acting antiviral agents (DAA). We carried out a cross-sectional study at the Advanced Center for Liver Diseases of Zagazig University Hospitals and Al-Ahrar Viral Hepatitis Treatment Center, Sharkia Governorate, Egypt. One hundred and fifty adult patients with CHC, who achieved SVR 12-24 weeks after end of treatment with sofosbuvir/daclatasvir ± ribavirin (139 patients, 92.67%), sofosbuvir/ledipasvir ± ribavirin (eight patients, 5.33%), sofosbuvir/simeprevir (two patients, 1.33%), and ombitasvir/ paritaprevir/ritonavir + ribavirin (one patient, 0.67%), according to the Egyptian National Committee for Control of Viral Hepatitis, were included in the study. We tested these patients for HCV RNA in peripheral blood mononuclear cells (PBMCs) immediately after confirmation of SVR12-24 weeks. Statistical analysis was performed by means of the Shapiro-Wilk test, Mann-Whitney U test, Chi-square test, and Fisher's exact test. Seventeen patients (11.33%) were positive for PBMNCs HCV RNA. The prevalence of OCI was highest in patients treated with simeprevir/sofosbuvir (2/2 patients). There is a substantially high prevalence of OCI after treatment with DAAs. We recommend dual testing for HCV RNA in both serum and PBMCs at the end of treatment of HCV infection with DAAs and during validation of the SVR following the initial response.

Evaluation of Soluble HLA-G Serum Level as Diagnostic Biomarker in Allergic Rhinitis Patients and its Association with Specific IgE Levels.

About 30 % of people are affected by allergic rhinitis (AR). Allergic rhinitis is an inflammatory condition defined by a disturbance of immunoregulation generating an amplified T helper "Th-2" response. An inflammatory mechanism mediated by HLA-G is thought to be involved through initiation of Th2 cytokine profile releasing "interleukin IL-3, IL-4, and IL-10". We evaluated levels of soluble HLA-G "sHLA-G" and studied its association with "allergen specific IgE" in AR patients. Twenty-five AR patients and twenty healthy subjects were enrolled. sHLA-G levels (U/ml) were measured by an immunoenzymatic method, while specific IgE levels (IU/ml) of respiratory allergens were analyzed using immunoblotting assay. The results revealed that AR group has significantly higher serum sHLA-G levels than normal subjects, P < 0.001. There was a highly significant and positive correlation between sHLA-G levels and specific IgE levels of Alternaria alternate and Dermatophagoides pteronyssinus (P < 0.01). Besides, there was a significant (P < 0.05) positive correlation between sHLA-G levels and specific IgE levels of brich and mixed grasses. We concluded that serum sHLA-G level is significantly increased in AR patients and that serum sHLA-G level could be a diagnostic biomarker in AR patients for clinical severity assessment.

T Regulatory Cells Response to Allergen Specific Immunotherapy in Patients with Allergic Airway Diseases: A Prospective Study.

Forkhead box P3 (FoxP3) T regulatory (Treg) cells modulate the immune system by blocking other types of T-cells. They maintain tolerance to self-antigens and help in inducing tolerance to foreign antigens. A deregulation of FoxP3 Tregs seems to play an important role in allergic disorders. The aim of this work was to study the response of FoxP3 Treg cells and their FoxP3 expression in patients, attending the Allergy Unit and the Chest Outpatient Clinic, Faculty of Medicine, Zagazig University, with allergic airway diseases, before and 1 year after receiving subcutaneous allergen specific immunotherapy (SIT). This prospective study was carried out on 25 patients with allergic airway diseases, confirmed by positive skin test, and that showed clinical improvement one year after SIT. All cases were subjected to total immunoglobulin E quantitation by ELISA. FoxP3 Treg cells frequency and FoxP3 relative fluorescence intensity, as an indicator of Tregs function, were assessed by flowcytometry. The results were compared before and after SIT. Twenty five age and sex matched apparently healthy volunteers were enrolled as controls. Our findings demonstrated that in comparison to the control group, the count of FoxP3 T regulatory cells was higher; however, the function was lower among the enrolled patients (P= 0.007 and P< 0.001, respectively). When FoxP3 Tregs were compared in the patients before and one year after SIT, it was found that both the count and FoxP3 expression showed statistically significant increase (P< 0.001). An inverse correlation was found between FoxP3 Tregs count and FoxP3 expression. It is concluded that patients with allergic airway diseases have increased number of FoxP3 Treg cells but with defective function. SIT plays a role in increasing the number of FoxP3 Tregs and improving their suppressive function, which leads to control of airway inflammation and thus clinical improvement.