Effect of Acid Suppression on Peripheral T-Lymphocyte Subsets and Immunohistochemical Esophageal Mucosal Changes in Patients With Gastroesophageal Reflux Disease.

BACKGROUND AND AIMS: Gastroesophageal reflux disease (GERD) is a common prevalent disease. We aimed to assess the dynamic changes in the peripheral T lymphocytes and lymphocytes infiltrating the esophageal mucosa after treatment with proton-pump inhibitor (PPI) in patients with GERD. PATIENTS AND METHODS: A total of 200 patients who presented with upper GIT symptoms were included in this prospective study. All patients were subjected to full history taking, clinical examination, and complete blood count. Upper endoscopy was performed to detect the grade of GERD, followed by 4 quadrant biopsies before and 1 month after acid suppressive drug therapy. Histopathologic and immunohistochemical examination were carried out for all biopsies. Flow cytometry analysis for the peripheral T lymphocytes and cytokine profile assay before therapy and after therapy were also carried out. RESULTS: In total, 200 patients comprising 132 male individuals (66%) and 68 female individuals (34%) with a mean age of 47.9±18.3 were included. The risk factors for development of GERD were smoking in 87 (43.5%), spicy food intake in 26 (13%), analgesics in 46 (23%), excessive tea and coffee in 35 (17.5%), and nondetected risk factors in 6 (3%). Endoscopic examination using Los Angeles grading system revealed that 102 patients (51%) were grade A, 57 patients (28.5%) were grade B, 38 patients (19%) were grade C, and 3 patients (1.5%) were grade D. No statistically significant differences could be detected in HGB levels and WBC, PLT, monocyte, granulocyte, and eosinophil counts before and after treatment with PPI. Histopathologic examination of esophageal biopsies showed significant posttreatment improvement in 132 cases (66%); however, 66 cases (33%) including the 2 cases (1%) of Barrett's esophagus showed nonsignificant pathologic improvement compared with the pretreatment picture. Immunohistochemical staining of esophageal biopsies with CD3 (T-cell marker) and CD20 (B-cell marker), before and 1 month after treatment, showed the presence of a very large number of infiltrating B cells in the esophageal mucosa (700±30/10 HPF) with large aggregations; in contrast, T-cell infiltration appeared less marked (570±23/10 HPF), and they formed smaller aggregates than those of B cells in pretreated patients, with P<0.01. However, 1 month after treatment with PPI, esophageal biopsies revealed a marked decrease in the number of both B (10±2/10 HPF) and T (290±12/HPF) cells in 66% of patients, with a P<0.01 in comparison with the pretherapy pattern. However, the remaining 33% of patients still showed a significantly high number of T cells (490±28/HPF), with a P <0.05 in comparison with the responder group that formed small aggregates with larger cell sizes, indicating their activation. Cytokine profiles before and after treatment revealed significant posttreatment reduction in their levels in the 132 cases with improvement in their clinical manifestations, and endoscopic and histopathologic findings, but there is no obvious change in the measured cytokine levels in 66 patients who simultaneously had no improvement in their endoscopic, histopathologic findings and mild improvement in their clinical manifestations. Moreover, significant posttreatment reduction of IL-8 and IL-1ß in the 98 (49%) patients with Los Angeles grading B, C, and D was observed. With regard to serum levels of IL-10 and IL-4, there were no statistically significant differences before and after treatment with PPI. Peripheral blood immunologic parameters revealed a statistically significant reduction of the total CD3 absolute count, T-helper lymphocyte (CD4/CD3) percentage, T-helper lymphocyte absolute count, and the percentage and absolute cytotoxic T-lymphocyte count (CD8/CD3) after treatment with PPI. Moreover, the same significant difference of peripheral blood lymphocytes was detected after exclusion of patients with Los Angeles grade A, which may be considered normal. CONCLUSIONS: Acid-induced T-cell-related cytokine production plays an important role in inflammation occurring in patients with GERD. Mucosal and peripheral inflammation reduces with PPI use.

Sofosbuvir-daclatasvir improves hepatitis C virus-induced mixed cryoglobulinemia: Upper Egypt experience.

BACKGROUND AND AIMS: Hepatitis C virus (HCV) infection is associated with extrahepatic manifestations such as cryoglobulinemia and accounts for up to 90% of all cases of mixed cryoglobulinemia (MC). The present study aimed to evaluate the effect of sofosbuvir-daclatasvir therapy on symptomatic HCV-related MC and sustained virologic response (SVR) achievement. PATIENTS AND METHODS: This prospective cohort study was carried out on 120 patients with chronic HCV infection, clinically suspected to have MC, but only 63 of whom were positive for cryoglobulins. HCV-MC patients were treated with sofosbuvir 400 mg and daclatasvir 60 mg once daily for 3 months. The serum cryoglobulins levels, complement 3 (C3), complement 4 (C4) (using ELISA assay kits) and rheumatoid factor (RF) (using immunoturbidimetric assay kit), were measured in the included HCV infected patients (to confirm HCV-MC diagnosis), in addition to quantitave HCV-RNA assays, using real time PCR. All these measurements have been done before stating therapy and 12, 24 weeks post-therapy for assessments of immunological recovery, viral load and SVR. RESULTS: Significant increase in the serum cryoglobulin levels and RF with significant decrease in C3 and C4 serum levels were detected in only 63 out of 120 included HCV infected patients, upon whom the study has been completed. They showed significant decrease in their mean cryoglobulin levels from 41.47 µg/mL ±12.32 SD to 5.12 µg/mL ±3.59 SD then to 5.09 µg/mL ±3.02 SD, 12 to 24 weeks post-therapy respectively (p<0.001), with significant decline in RF concentrations and rise in C3 and C4 serum levels approaching the normal values. There were improvements in the presenting HCV-MC clinical manifestations in variable degrees, ranging from 5 (71.42%) in patients with glomerulonephritis to 62 (98.4%) in patients with purpura. Eighty-seven percent of the included patients showed complete response (clinical, virological and immunological recovery) and 13% showed partial response (virological and immunological recovery without clinical improvement of cryoglobulinemia associated manifestations). CONCLUSION: A combined therapy of sofosbuvir 400 mg and daclatasvir 60 mg once daily for 3 months was associated with a significant decrease in serum cryoglobulin levels and appears as a reasonable treatment option for HCV-associated MC.

Correlation between Serum Hyaluronic Acid with Steatosis, Non alcoholic Steato-hepatitis and Fibrosis in Patients with Chronic Hepatitis C Virus Infection.

Background: Hyaluronic acid (HA) is an attractive potential marker for noninvasive diagnosis of liver fibrosis instead of liver biopsy for both patients and physicians. Aim: To assess the role of HA for diagnosing the progression of steatosis to steato-hepatitis (SH) and fibrosis in patients with Chronic Hepatitis C virus (HCV) infection. Methods: 90 patients with chronic HCV infection, 77 (85.6%) males and 13 (14.4%) females, were included. Blood samples were collected for routine laboratory investigations, liver function tests and serum HA measurements. A liver biopsy was taken for histopathological examination. Results: Steatosis was found in 37 patients (41.1%), fibrosis in 29 patients (32.2%) and SH in 51 patients (56.7%). The mean serum HA for all patients was 86.4±48.2 ng/L. HA levels were significantly higher in patients with fibrosis (95.6±53 vs 54.5±3.5) and SH (88.7±52 vs 49.9±12) than those without (P value = 0.001and 0.001 respectively). HA levels were also significantly higher in patients with an advanced degree of fibrosis, SH and steatosis as compared to those with mild degrees (P value = 0.000, 0.001 and 0.01 respectively). Positive correlations were found between serum HA and the degree of fibrosis, SH and steatosis (P value =0.000 and r = + 0.758, 0.701and 0.727 respectively). The mean HA cut off value for the diagnosis of fibrosis and SH was taken to be 70 and 60 ng/L providing a diagnostic accuracy of 94.1% and 91.6% respectively. Conclusion: Serum HA level is a good noninvasive marker for the diagnosis of fibrosis and steato-hepatitis in patients with chronic HCV infection.

Role of portal haemodynamic parameters in prediction of oesophageal varices in cirrhotic patients.

BACKGROUND AND STUDY AIMS: Screening all cirrhotic patients by endoscopy for detection of varices is not cost-effective as the number of patients increases by time and half of them still would not have developed varices 10years after being diagnosed with cirrhosis. Therefore, this study aimed to evaluate hepatic haemodynamic Doppler parameters in predicting the presence of oesophageal varices (OVs) in cirrhotic patients for better selection of those actually needed for screening endoscopy. PATIENTS AND METHODS: Eighty-one patients with liver cirrhosis, 32 females and 49 males, with a mean age of 50.7±11.7years were recruited for the study. They included 61 patients with OVs and 20 patients without varices. The diagnosis of liver cirrhosis was based on clinical history, examination, and investigations. Liver function and kidney function tests and complete blood count (CBC) were performed for all patients. All patients underwent abdominal ultrasound (US), upper endoscopy, and hepatic Doppler US examination. RESULTS: The portal vein velocity (PVV) and liver vascular index (LVI) showed statistically significantly lower values in patients with OVs than those without OVs (p value=0.02 and 0.000, respectively). The congestion index (CI) of the portal vein, the portal hypertension index (PHI), and the splenoportal index (SPI) showed statistically significantly higher values in patients with OVs than those without OVs (p value=0.006, 0.001, and 0.001, respectively). CI and SPI were the best parameters that could predict the presence of OVs with high sensitivity, specificity, and diagnostic accuracy when cutoff values were set at >0.069 and 3.57, respectively (area under the curve=0.864 and 0.894, respectively). CONCLUSIONS: The CI of the portal vein and SPI are good predictors for the presence of OVs in cirrhotic patients, and could be used noninvasively to decrease the burden on the upper endoscopy unit by proper selection of those who are candidates for screening endoscopy.