Interferon-α and inosine pranobex-mediated inhibition of reploication of human RNA viruses in vitro.

INTRODUCTION: Interferon- a (IFN-a), produced by immune cells, exhibits pleiotropic anti- viral activity. Inosine pranobex (PI), a synthetic derivative of a purine, shows direct anti- viral activity, and also acts indirectly, by activation of immune cells. The aim of this study was to evaluate an in vitro inhibition of Coxackievirus A16 (CAI6), enterovirus 71 (EV71) and human parainfluenza virus 4 (HPIV-4) replication by PI in combination with IFN-a. MATERIALS AND METHODS: In the present study we evaluated an in vitro effect of interferon-a and inosine pranobex on replication ofRNAviruses: CA-16, EV71, HPIV-4. Antiviral effects of IFN-a and IPwere assessed by phenotypic assays. The yield reduction assay (YRA), which evaluates the ability of the compounds to inhibit virus multiplication in cell cultures, was ap- plied. The Reed-Muench statistical method was used to determine the 50% end point (IC51). RESULTS: Our studies have shown that combination of IFN-a and inosine pranobex dis- play higher efficacy than treatment with either compound alone, and suggest syn- ergy that may increase therapeutic effectiveness. The reduction of the average viral ti- ters of EV71, CA-16 and HPIV-4 in A549 cell culture after applying 400 Ig/mL Ip and IFN-a (1000 IU/mL), in comparison to the viral titer in the control was reduced by 1,76 log,, TCID,,/ml, o 3,00 log, TCID50/ml , and 1,60 log,( TCID50/ml respec- tively. The antiviral activity of the tested compounds was also analyzed on the basis of IC., values. Application of 1000 IU/ml IFN-a, with PI after infection of A549 cells with mention above viruses reduced the IC,, by 3,5%, 41,3% and 29% respectively. CONCLUSIONS: Our study demonstrated that enhanced antiviral activity was observed when cells infected with RNA viruses were treated with combination of IFN-a and IP. The ef- fectiveness of IFN-a and IP under these conditions has not been previously reported. CA16 virus turned out to be the most sensitive to the action of used inhibitors.

The occurrence of herpes simplex viruses 1 and 2 in skin and mucosal lesions in patients with suspicion of genital herpes.

INTRODUCTION: Infection with herpes simplex viruses 1 and 2 (HSV 1 and 2 or Human herpesvirus HHV) are one of the most common infections in human. Real time PCR is a sensitive and specific method for diagnostics of HHV infections. The aim of the study was to investigate the occurrence of HHV 1 and HHV 2 DNA in patient with clinical symptoms suggesting HHV infection. METHODS: We used real time PCR to investigate swabs from genital and perianal lesions from 74 patients of the Department of Dermatology and Venereology Medical University Warsaw and of gynecological outpatient clinics in Warsaw 40 women and 34 men. RESULTS: The results were positive for HHV 2 in 25 cases (34%), for HHV 1 in 19 cases (26%) and for both viruses in 20 cases (27%). 10 samples were negative for both viruses. CONCLUSIONS: The results confirm that the main cause of symptomatic genital herpes is HHV 2, however the percentage of HHV 1 and specially of mixed HHV 1/HHV 2 infections was unexpectedly high.

[Comparison of three multiplex real-time PCR assays for the detection of herpes simplex viruses type 1 and type 2 DNA].

INTRODUCTION: Herpes simplex viruses type 1 and type 2 (HSV-1 and HSV-2) cause widespread infection worldwide with different course and intensity. Although the disease caused by this viruses mainly concern healthy children and adults, the HSV infections are much more dangerous for people with immunodeficiencies. The aim of this work was to compare the diagnostic value of two qPCR methods for detection HSV-1/2 DNA, based on TaqMan* and HybProbes chemistry with commercial HSV-1/2 Qual Kit. METHODS: DNA from 51 clinical samples was tested for presence of HSV-1/2 sequences on LightCycler 2.0 thermocycler, using two ,,in-house" developed multiplex real-time PCR assays and commercial test using SCORPIONSTM primers. RESULTS: The results showed high specificity and sensitivity of all molecular biology tests used. Statistically, there was no significant difference in the sensitivity of real-time PCR assays when using TaqMan* and HybProbes' chemistry and when using the commercial SCORPIONSTM based method (P>0.05). CONCLUSIONS: Obtained results show that all tested methods are highly specific and can possibly be used to simultaneously detect and differentiate HSV-1/2 DNA in clinical samples. The high detection rate and short duration of qPCR assayas has great importance for immunocompromised patients where quick application of effective and safe treatment is necessary. It is also important in the event of amorphous form of the infection and the occurrence of nonspecific and generalized symptoms.