Cytochrome P4502D6 (CYP2D6), target of liver kidney microsomal autoantibody type 1 (LKM1), characterizes autoimmune hepatitis type 2 (AIH2) but is also found in patients with chronic hepatitis C virus (HCV) infection. To provide a complete linear epitope B cell map of CYP2D6, we tested peptides spanning the entire sequence of CYP2D6. In addition to confirming previously described antigenic sites, we identified four new epitopes (193–212, 238–257, 268–287, and 478–497). CYP2D6193–212 is immunodominant and was the target of 12 of 13 (93%) patients with AIH2 and 5 of 10 (50%) HCV/LKM1-positive patients. Because LKM1 is present in both AIH2 and a viral infection, we tested whether Abs to CYP2D6193–212 arise through cross-reactive immunity between virus and self. We identified a hexameric sequence “RLLDLA” sharing 5 of 6 aa with “RLLDLS” of HCV2985–2990 and all 6 aa with CMV130–135. Of 17 CYP2D6193–212-reactive sera, 11 (7 AIH and 4 HCV) reacted by ELISA with the HCV homologue, 8 (5 AIH and 3 HCV) with the CMV homologue, and 8 (5 AIH and 3 HCV) showed double reactivity. Autoantibody binding to CYP2D6193–212 was inhibited by preincubation with HCV2977–2996 or CMV121–140. Recombinant HCV-nonstructural protein 5 and CMV-UL98 proteins also inhibited Ab binding to CYP2D6193–212. Affinity-purified CYP2D6193–212-specific Ab inhibited the metabolic activity of CYP2D6. The demonstrated similarity and cross-reactivity between CYP2D6193–212 and two unrelated viruses suggests that multiple exposure to viruses mimicking self may represent an important pathway to the development of autoimmunity.

Background and Aim: Lumpy skin disease (LSD), is a highly infectious viral disease of cattle, caused by LSD virus (LSDV) which belongs to the genus Capripoxvirus of family Poxviridae. In the summer of 2017, skin lesions suggestive of LSD were observed in cattle at several governorates in Egypt. This study aimed to detect LSDV in cattle specimens using rapid serological and molecular diagnostic assays. Materials and Methods: A total of 46 skin biopsies and uncoagulated blood samples were collected from cattle with LSD suggestive clinical signs, as well as 290 coagulated whole blood samples from cattle without skin lesion in different governorates in Egypt during the summer of 2017. Skin biopsies were used for virus isolation from the chorioallantoic membrane of 11-day-old specific pathogen-free embryonated chicken eggs (SPF-ECEs). LSDV was identified using conventional polymerase chain reaction (PCR), real-time PCR (RT-PCR), and fluorescent antibody technique (FAT) with specific hyperimmune serum against LSDV. Cattle sera were examined using indirect FAT (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA). Results: Skin nodules and sitfast lesions were significant clinical signs observed in all LSD suspect cattle. SPF-ECEs, from which positive isolations were made and it showed characteristic inflammatory and focal white pock lesions. The isolated viruses were identified as LSDV by FAT, conventional gel-based PCR, and RT-PCR. Among the skin biopsies and corresponding blood samples, LSDV-positive samples percentage were 39.13 and 36.95 by RT-PCR, followed 34.78 and 28.26 by conventional PCR and then 32.6 and 26.8 by FAT, respectively. The total positive percentage of LSDV antibody detected in cattle serum samples were 17.93 and 14.48 by indirect ELISA and IFAT. Conclusion: LSDV was detected and identified in skin biopsies and corresponding blood samples of naturally infected cattle, more LSDV-positive samples were detected by RT-PCR, followed by conventional PCR and then FAT. The indirect ELISA detected more antibody-positive samples than the IFAT from cattle serum samples. The RT-PCR assay is simple, sensitive, rapid, and reliable for the detection of LSDV in blood and skin nodule biopsies of suspected cattle.

Bovine herpesvirus 1 (BHV-1) is a highly contagious viral pathogen which causes infectious bovine rhinotracheitis in bovine worldwide. Currently, there is no antiviral prophylactic treatment available capable of the complete cure of the viral disease and facilitating recovery from latent infection in animals. The present study aimed to evaluate antiviral activities of Water Green Tea Extract (WGE) and Ethanol Propolis Extract (EPE) against BHV-1 virus comparing to commercial Acyclovir (ACV) in vitro in Madin-Darby Bovine Kidney (MDBK) cell line and in vivo in rabbits as a laboratory animal's model. The cytotoxicity assay was determined the safe dose of water green tea, and Ethanol propolis extracts and evaluated antiviral activity of each extract on infected MDBK with BHV-1. The fifteen rabbits were divided accidentally into five groups. Groups 1, 2 and 3 were inoculated with BHV-1 virus 10 7 TCID50/250 ul in nostrils and received propolis ethanol, water green tea extracts and ACV antiviral for 7 dpi respectively. Group 4 was inoculated with BHV-1 virus 107 TCID50/250 ul in nostrils without extracts or commercial drug. Group 5 was considered as control negative. Results of in-vitro study showed water green tea, and ethanol propolis extracts were potent inhibitor on BHV-1, which showed 80% protection against this virus and dropped in viral titer more than ACV. In vivo study of treated infected animals with WGE, EPE and ACV reduced clinical signs, elevated cytokines, and antibody production levels and failed re-isolated or detect DNA in blood or nasal samples swabs. Non treaded infected rabbits group developed respiratory clinical signs, humoral response and re-isolated BHV-1 and detected viral DNA of BHV-1 in blood, and nasal swabs from experimentally infected rabbits. In conclusion, propolis and green tea extracts were able to prevent virus replication and reduced CPE in MDBK cell cultures infected with BHV-1 and able to induce cytokines and antibodies levels production.

Foot and Mouth Disease (FMD) is highly contagious disease affected cloven-hoofed animals which result in substantial economic losses. The present study was aimed to detect FMDV by different serological and molecular methods in cattle and buffaloes for providing an accurate and rapid diagnosis of FMD disease. 86 samples of tongue epithelium biopsies, fluid vesicles samples and saliva, as well as 86 coagulated and uncoagulated blood samples, were collected from 64 and 22 suspected cattle and buffaloes respectively in different governorates in Egypt, during August to December 2017. Serum samples were examined by 3ABC-ELISA for differentiating between infected and non-infected animals. While tissues biopsies and un-coagulated blood samples were examined by Sandwich ELISA, Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) as well as Real-Time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR). FMDV porotypes were identified by rRT-PCR in suspected cattle and buffaloes samples to FMDV serotype A, O and SAT2 and results showed that 54 samples positive for FMDV different serotypes while FMDV serotype differentiation in tissues biopsy of cattle were 18 (28.12%), 12 (18.75%), 3 (4.68 %) and 4 (6.25%). Also, the positive results of tissue samples from buffaloes examined by RT-PCR were 9 (40.09 %), 4 (6.25%), 2 (9.09 %) and 2 (9.09 %) for O, SAT2, serotype A and mixed serotypes respectively by different tests. The rRT-PCR provided an accurate and rapid laboratory diagnosis of FMDV as well as RT-PCR, and 3ABC-ELISA were given nearly the same results. Although the rRT-PCR generated results in less than 6 h and this is an important feature when definitive diagnostic results required in a short timescale during emergencies. Also, this study demonstrated the current situation of circulation FMDV type A, O, and SAT2 serotypes in cattle and buffaloes in Egypt.

Capri Pox Virus (Ca PV) is the causative agent of important diseases in sheep and goat with severe socioeconomic impact. Sheep Poxvirus (SPPV), Goat Poxvirus (GTPV) and Lumpy Skin Disease Virus (LSDV) are three members of the Capripox virus genus of Poxviridae family, which infect sheep, goats, and cattle, respectively. A rapid diagnostic assay for Ca PV by using conventional PCR RNA polymerase gene RP030 and real-time qPCR would be useful for disease surveillance, detection and differentiation of Ca PV in clinical and subclinical samples for management and treatments of outbreaks. The present study aimed to detect and identify Ca PV (SPPV and GTPV) in natural, infected scabs biopsy samples, which were collected from sheep and goats in different governorates in 2017 during outbreaks in Egypt using the conventional PCR RNA polymerase gene RP030 gene based and Real-Time qPCR fluorescent based. We collected eighty scabs from clinically affected animals (54 sheep and 26 goat) that were vaccinated in Chorio-Allantoic-Membranes (CAM) from 10-days-old embryonated-chicken eggs. The positive CAM showed pock lesions, which were observed with a thickening of the membrane after 2-3 passages post samples inoculation, and harvested positive CAMs, which were determined by Agar Gel Precipitation Test (AGPT) , Counter Immune Electrophoresis (CIE), and conventional PCR and real time qPCR were examined for the presences of Ca PVs. DNA extraction from clinical samples and positive CAM with pox lesions using DNA slandered references extraction kits compared to novel modification method (Microwave extraction). The PCR based RPO30 gene and the real-time qPCR showed 15 positive with percentage 27.77% in 54 sheep and 3 positive with percentage 12.5% in 26 goats. Although, AGPT and CIE gave lower result than molecular methods, they gave 11 and 13 positive samples from 54 sheep and in goats were 1 and 2 from 26 scab biopsy samples respectively, however they are useful for early confirmation of positive Ca PVs in low-income countries. PCR based RNA polymerase gene RP030 gene and real-time-PCR considered sensitive, rapid, and reliable methods for differentiating SPPV and GTPV from AGPT and CIE in CAM or in clinical samples without further isolation and propagation in embryonated-chicken eggs. The novel microwave method used to isolate high quality of DNA extracted from infected skin biopsy with SPPV and GPPV with no further purification steps required. It was done in 3 minutes only. The results of the current study confirmed that the suitability of the PCR-based RNA polymerase gene RP030 gene is suitable for differentiating between SPPV and GTPV; in one PCR run; without any post-processing steps.