Anti-C1q antibody as a predictor of chronic HCV genotype 4 response to treatment in Egyptian children

Background: Hepatitis C virus [HCV] infection has been reported to be the most common blood born pathogen all over the world. The prevalence of HCV in children in developed countries ranges between 0.1 and 0.4%, and is generally lower than in adults. Combinedpegylated interferon and ribavirin is still the only standard of care treatment in spite of its side effects, high costs and low sustained virological response [SVR] rates. Hence, this provides a compelling reason for the identification of biomarker predictors of disease response to treatment

Objective: To evaluate anti-C1q antibody as a predictor of chronic HCV response to treatment with combined pegylated interferon alpha-2b and ribavirin in Egyptian children

Methodology: This study was conducted on forty-four chronic HCV-infected children [Male/Female: 30/14; aged 12.02 +/- 3.1 years] from the outpatient clinic, Pediatric Hepatology Department, National Liver Institute, Menoufiya University. They were given combined pegylated interferon alpha-2b [Peg-IFN-alpha-2b] and ribavirin [RBV] for 48 weeks and a quantitative polymerase chain reaction [PCR] for hepatitis C virus Ribonucleic acid was performed at 12, 24, 48 weeks during treatment and after another 24 weeks post-treatment. Anti-HCV antibody and Real-time PCR for HCV-RNA was performed [the detection limit was 15 IU/mL]. Anti-C1q antibodies were performed by enzyme-linked immunosorbent assay [ELISA]

Results: Serum levels of Anti-C1q antibodies were significantly higher [P = 0.001] in the Non-responders group [mean = 14.61 +/- 6.749ng/ml] compared to the SVR one [mean = 2.27 +/- 3.77ng/ml]. No statistically significant difference [P > 0.05] had been found between SVR and Non-responders regarding the age, ALT, viral load, or hepatic necroinflammatory activity and liver fibrosis. Anti-C1q at a cutoff value of 9.05 ng/ml, had sensitivity and specificity of 84.6% and 75% respectively and 92% positive predictive value. No significant correlation between the serum level of anti-C1q antibodies and the age, sex or HCV viral load, liver enzymes, and the degree of fibrosis and necroinflammatory activity was found [P> 0.05] for all parameters

Conclusion: Anti-C1q could be a good predictor for HCV treatment and should be included in pretreatment laboratory assessment for proper choice of chronic HCV children patients who will benefit from combination therapy

ESAT-6-ELISpot and interferon gamma in the diagnosis of pleural tuberculosis

Appropriate diagnostic methods for tuberculous pleural effusion are vital. The IFN-gamma tests using specific Mycobacterium Tuberculos is antigens in samples from the site of infection may be promising in diagnosis of tuberculosis. Objective we examined the ability of ELISpot test using circulating peripheral blood mononuclear cells [PBMC] and compartmentalized pleural fluid mononuclear cells [PFMC] for diagnosis of active TB infection in patients with tuberculous pleural effusion. Methods PBMC and PFMC-based ELISpot test for IFN-gamma test using specific M. tuberculosis antigen: Early Secretory Antigen Target-6 protein [ESAT-6] was used for diagnosis of active TB infection. Thirty-five patients with clinically suspected tuberculous pleural effusion were enrolled over a 12-month period. Results 11 patients out of 35 were positive by culture and PCR [31.4%]. Incubation of PBMC with ESAT-6 for 8 h showed sensitivity and specificity of 82% and 92%, respectively, for the PBMC-ELISpot as compared to PFMC-ELISpot that was 54% and 96% respectively. With 24 h incubation of ESAT-6 there was around 2.5 fold increase in the median number of spot forming cells [SFCs] in PFMC from 30 to 74, whereas there was minimal increase of median number of SFCs in PBMC from 55 to 60. Conclusion ESAT-6 - ELISpot using PBMC and PFMC is useful as a tool for diagnosis of TB effusion. PFMC needs longer period of incubation for processing of ESAT-6 than PBMC. Moreover, IFN-gamma in pleural effusion [PE] is another useful way for diagnosis of TB pleurisy which is sensitive, simple and cheap

Interferon gamma as a diagnostic marker for pleural tuberculosis

Background appropriate diagnostic methods for tuberculosis pleural effusion [TBPE] are vital. The IFN-gamma tests using specific M. Tuberculosis antigens in samples from the site of infection may be promising in diagnosis of tuberculosis

Objective we examined the ability of ELISpot test using circulating peripheral blood mononuclear cells [PBMC] and compartmentalized pleural fluid mononuclear cells [PFMC] for diagnosis of active TB infection in patients with TBPE

Methods PBMC and PFMC-based ELISpot test for IFN-gamma test using specific M. Tuberculosis antigen: Early Secretory Antigen Target-6 protein [ESA T-6] was used for diagnosis of active TB infection. Thirty-five patients with clinically suspected TBPE were enrolled over a 12-month period

Results eleven patients out of 35 were positive by culture and PCR [31.4%]. Incubation of PBMC with ESAT-6 for 8 hrs showed sensitivity and specficity of 82 % and 92 % respectively .for the PBMC-ELlSpot as compared to PFMC- ELlSpot that was 54% and 96 % respectively. With 24 hrs incubation of ESAT- 6 there was around 2.5 fold increase in the median number of spot forming cells [SFCs] in PFMC from 30 to 74, whereas there was minimal increase of median number of SFCs in PBMC from 55 to 60

Conclusion ESAT-6 - ELlSpot using PBMC and PFMC is useful as a tool for diagnosis of TB effusion. PFMC needs longer period of incubation for processing of ESAT-6 than PBMC. Moreover, IFN-gamma in pleural effusion [PE] is another useful way for diagnosis of TB pleurisy which is sensitive, simple and cheap

Pro-inflammatory CD14+ CD16++ monocytes in HCV persistent and spontaneously cleared infections

In human blood, two Monocytes [Mo] subpopulations have been distinguished: the classical CD 14+'DR' Mo and pro-inflammatory CD14'CD16-'DR' Mo. In HCV antibody positive with persistent HCY [positive HCV RNA], we found decreased numbers of the pro-inflammatory Mo in 20 patients with persistent HCV with an average of [34.1 f 22 cells/pl] in comparison to the numbers in the cleared patients [positive HCV Ab with negative HCV MA] [85 + 35 cells/d]. Intracellular TNF was measured by three-color immunofluorescence and flow cytometry after stimulation with Lipopolysaccharide [LPS]. In patients with persistent HCV infection, pro-inflammatory Mo showed one and half fold higher level of intracellular TNF staining. This indicates that the minor population of pro-inflammatory Mo plays pronounced roles in the HCV clearance or persistence