Leukemia-related signaling pathways among HTLV-1 derived adult T-cell leukemia/lymphoma and asymptomatic carriers in comparison to normal group.

BACKGROUND AND OBJECTIVE: Human T-cell lymphotropic virus type 1 (HTLV-1) is associated with adult T-cell leukemia/lymphoma (ATLL), a fetal malignant infection. Recently, The HTLV-1 new Asymptomatic careers (ACs) have frequently been reported among blood donors. Reaching the profound concept of HTLV-1-associated molecular pathogenesis could result in finding novel therapeutic strategies. The current study aimed to determine leukemia-related signaling regulation in ATLL. MATERIALS AND METHOD: 30 participants were evaluated in three groups, including 10 ATLL patients, 10 ACs, and 10 normal controls. Blood samples were isolated without any chemotherapy history from ATLL patients. Also, blood samples were recovered from ACs and normal individuals. Lymphocyte isolation was done on collected blood samples. After this, RNA was extracted from the prepared samples and utilized for the cDNA synthesis. Tax and HBZ as viral genes and cellular genes, including MKP-1, EVI-1, JNK-1, FOXO-1, AKT-1, DEPTOR, MTOR, and JUN, were investigated using real-time PCR. RESULTS: The mean age of ATLL patients was 53.2 ± 7.32 years, and 9 (90%) were male. The EVI-1 and FOXO-1 expression levels were significantly associated with ATLL patients compared to internal control. However, the significant differences in expression of other genes in the remaining groups were not seen. CONCLUSION: Discovering viral and cellular signaling pathways that regulate HTLV-1 transformation is essential. A novel therapeutic strategy for ATLL regulating cellular signaling pathways in vivo could be considered. Therefore, clinical trials using activators and inhibitors of related cellular signaling pathways for cell therapy of ATLL are recommended. It is recommended more investigation on FOXO-1 and EVI-1 for targeting these genes to reveal the molecular pathogenesis of ATLL.

Prevalence and phylogenetic analysis of HTLV-1 in a segregated population in Iran.

Human T-lymphotropic virus type 1 (HTLV-1) infection is an important health issue that affects a variety of endemic areas. The Khorasan province, mainly its capital Mashhad in northeastern Iran, was reported to be as one of these endemic regions. Torbat-e Heydarieh, a large city Southwest border to Mashhad with a segregated population was investigated for the prevalence and associated risk factors of HTLV-1 infection in 400 randomly selected individuals. Blood samples were tested for the presence of HTLV-1 antibodies via the ELISA method and then were confirmed by an Immunoblot test. For the presence of HTLV-1 in lymphocytes of infected subjects, PCR was performed on LTR and TAX regions. DNA sequencing of LTR fragment was also carried out to determine the phylogenetic of HTLV-1, using the Maximum likelihood method. HTLV-1 sero-reactivity (sero-prevalence) among the study population was 2% (8/400), of which 1.25% had HTLV-1 provirus in lymphocytes (actual prevalence). HTLV-1 infection was significantly associated with the age, marital status, and history of blood transfusion (P < 0.05). However, there were no statistical differences between HTLV-1 infection, and gender, surgery, and hospitalization. In regression analysis, age showed the most significant correlation with the infection (P = 0.006, OR = 4.33). Based on our phylogenetic study, the HTLV-1 prevalent sequence type of Torbat-e Heydarieh belongs to the cosmopolitan subtype A. HTLV-1 prevalence in Torbat-e Heydarieh (1.25%) is low comparing to those of both Mashhad (2-3%) and Neishabour (3.5-5%) in the province of Khorasan. Thus, traveling mobility and population mixing such as marriage, bureaucratic affairs, occupation, and economic activities could be the usual routs of HTLV-1 new wave of spreading in this segregated city.

Detection of Specific Antibodies to HCV-ARF/CORE+1 Protein in Cirrhotic and Non-Cirrhotic Patients with Hepatitis C: A Possible Association with Progressive Fibrosis.

BACKGROUND: The role of different viral proteins in the progression of the disease to cirrhosis is not completely understood. The ARFP/F protein is a newly described protein synthesized from the +1 or -2 reading frames of the core protein gene, which its function remains unknown. The purpose of this study is to detect specific antibodies to HCV-ARF/Core+1 protein in cirrhotic and non-cirrhotic patients with HCV and investigate any possible association. METHODS: ARF/Core+1 recombinant proteins from HCV genotype 1a were expressed in Escherichia coli, and purified. Using an enzyme-linked immunosorbent assay, we assessed the prevalence of anti-ARF/Core+1 antibodies in 50 cirrhotic and 50 non-cirrhotic hepatitis C patients. RESULTS: All 50 cirrhotic patients were positive for anti-ARF/Core+1 antibody, while only 80% positive samples among non-cirrhotic patients were detected. The titer of anti-ARF/Core+1 antibody was also significantly higher in patients with cirrhosis than in non-cirrhotic patients. CONCLUSION: Compared to 80% positive samples among non-cirrhotic patients all 50 cirrhotic patients were positive for anti-ARF/Core+1 antibody and titer of anti-ARF/Core+1 antibody was significantly higher in patients with cirrhosis than in non-cirrhotic. These results suggest that ARF/Core+1 protein is associated with cirrhosis. A possible causative association between ARF/Core+1 and cirrhosis as well as the mechanism of this association needs to be further investigated.

A comparative approach between heterologous prime-boost vaccination strategy and DNA vaccinations for rabies.

BACKGROUND AND AIM: Rabies is a widespread neurological zoonotic disease causing significant mortality rates, especially in developing countries. Although a vaccine for rabies is available, its production and scheduling are costly in such countries. Advances in recombinant DNA technology have made it a good candidate for an affordable vaccine. Among the proteins of rabies virus, the Glycoprotein (RVG) has been the major target for new vaccine development which plays the principal role in providing complete protection against RV challenge. The aim of this study is to produce recombinant RVG which could be a DNA vaccine candidate and to evaluate the efficiency of this construct in a prime-boost vaccination regimen, compared to commercial vaccine. METHODS: Cloning to pcDNA3.1(+) and expression of rabies virus glycoprotein gene in BSR cell  line were performed followed by SDS-PAGE and Western blot analysis of the expressed glycoprotein. The resulting genetic construct was used as a DNA vaccine by injecting 80 µg of the plasmid to MNRI mice twice. Prime-Boost vaccination strategy was performed using 80 µg plasmid construct as prime dose and the second dose of an inactivated rabies virus vaccine. Production of rabies virus neutralizing antibody (RVNA) titers of the serum samples were determined by RFFIT. RESULTS: In comparisons between heterologous prime-boost vaccination strategy and DNA vaccinations, the potency of group D that received Prime-Boost vaccine with the second dose of pcDNA3.1(+)-Gp was enhanced significantly compared to the group C which had received pcDNA3.1(+)-Gp as first injection. CONCLUSION: In this study, RVGP expressing construct was used in a comparative approach between Prime-Boost vaccination strategy and DNA vaccination and compared with the standard method of rabies vaccination. It was concluded that this strategy could lead to induction of acceptable humoral immunity.