Afiono Agung Prasetyo

Replication of chicken anemia virus (CAV) requires apoptin and is complemented by VP3 of human torque teno virus (TTV)

To test requirement for apoptin in the replication of chicken anemia virus (CAV), an apoptin-knockout clone, pCAV/Ap(-), was constructed. DNA replication was completely abolished in cells transfected with replicative form of CAV/Ap(-). A reverse mutant competent in apoptin production regained the full level of DNA replication. DNA replication and virus-like particle (VLP) production of CAV/Ap(-) was fully complemented by supplementation of the wild-type apoptin. The virus yield of a point mutant, CAV/ApT(108)I, was 1/40 that of the wild type, even though its DNA replication level was full. The infectious titer of CAV was fully complemented by supplementing apoptin. Progeny virus was free from reverse mutation for T(108)I. To localize the domain within apoptin molecule inevitable for CAV replication, apoptin-mutant expressing plasmids, pAp1, pAp2, pAp3, and pAp4, were constructed by deleting amino acids 10-36, 31-59, 59-88 and 80-112, respectively. While Ap1 and Ap2 were preferentially localized in nuclei, Ap3 and Ap4 were mainly present in cytoplasm. Although complementation capacity of Ap3 and Ap4 was 1/10 of the wild type, neither of them completely lost its activity. VP3 of TTV did fully complement the DNA replication and VLP of CAV/Ap(-). These data suggest that apoptin is inevitable not only for DNA replication but also VLP of CAV. The common feature of apoptin and TTV-VP3 presented another evidence for close relatedness of CAV and TTV.

Tracking the entry routes of hepatitis C virus as a surrogate of HIV in an HIV‐low prevalence country, the Philippines

From 2002 to 2007, 1,590 individuals were enrolled in an active surveillance program conducted in Metro Cebu, Philippines, where the anti‐HCV‐positive rate was significantly and constantly high among injecting drug users (83%, 793/960; 71–88%), especially among those living in downtown (89%, 683/770; 87–100%), despite the extremely low percentage of anti‐HIV‐positives (0.34%, 3/874). Sampling areas were then enlarged nationwide and the number of samples increased to 2,645 at the end of 2007. A total of 444 samples were positive for HCV RNA. Phylogenetic analysis based on NS5B and E1–E2 regions revealed that the most dominant HCV subtype was 1a, and followed by 2b, 2a, and 1b, and that the HCV strains had the largest variety in Metro Manila and its vicinity (Pu2009<u20090.01). Interestingly, subtype 1b was detected solely in Metro Manila, and four HCV strains collected in this area showed higher homology to specific foreign strains retrieved from the Genbank/EMBL/DDBJ database with bootstrap values of 68–95% comparing with other strains analyzed in this nationwide study. These data suggest that HCV strains may be introduced occasionally into the Philippines possibly through Metro Manila as a main entry point. Considering the fact that an HIV epidemic started primarily via contaminated needle sharing in Asia, the constantly high rate of HCV infections and the newly introduced foreign HCV strains in the absence of HIV epidemic warrant further investigation on HCV entry and spread for early detection of an HIV epidemic in the Philippines. J. Med. Virol. 81:1157–1162, 2009.

The -1082 A allele of the interleukin-10 single nucleotide polymorphism -1082 G/A is associated with hepatitis virus co-infection in Indonesian Javanese HIV patients

Objective: This study was conducted to determine the distribution of the single nucleotide polymorphism (SNP) at position -1082 of the proximal promoter region of the interleukin (IL)-10 gene and its association with hepatitis virus co-infection in Indonesian Javanese human immunodeficiency virus (HIV)-infected patients. Methods: Blood samples were collected from 156 Indonesian Javanese HIV-infected patients; tested with serological and molecular assays for hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatitis D virus (HDV); and subjected to SNP -1082 G/A polymorphism analysis. Results: Hepatitis B surface antigen (HBsAg) (+)/HBV DNA (+), HBsAg (-)/HBV DNA (+), anti-HCV antibodies (+)/HCV RNA (+), and anti-HCV antibodies (-)/HCV RNA (+) were detected from all of HIV-infected individuals at rates of 3.8% (6/156), 0.6% (1/156), 16.0% (25/156), and 11.5% (18/156), respectively. The frequencies of -1082 GG, -1082 GA, and -1082 AA were 14.7% (23/156), 60.3% (94/156), and 25% (39/156), respectively. HBV co-infection was found only in -1082 A allele carriers, and these patients also showed higher rates of HCV co-infection (odds ratio: 2.24, 95% confidence interval: 1.22–4.10, P=0.009).

Respiratory Viruses and Torque Teno Virus in Adults with Acute Respiratory Infections

Objective: To define the molecular epidemiology of respiratory viral infections in adult patients. Methods: Nasal and throat swabs were collected from all adult patients with influenza-like illness (ILI), acute respiratory infection (ARI), or severe ARI (SARI) admitted to a tertiary hospital in Surakarta, Indonesia, between March 2010 and April 2011 and analyzed for 19 respiratory viruses and for torque teno virus (TTV) and human gyrovirus (HGyV). Results: Respiratory viruses were detected in 61.3% of the subjects, most of whom had ARI (90.8%, OR = 11.39), were hospitalized (96.9%, OR = 22.31), had asthma exacerbation (90.9%, OR = 8.67), and/or had pneumonia (80%, OR = 4.0). Human rhinovirus (HRV) A43 predominated. Influenza A H3N2, human metapneumovirus (HMPV) subtypes A1 and A2, the influenza B virus, human adenovirus B, and human coronavirus OC43 were also detected. All respiratory viruses were detected in the transition month between the rainy and dry seasons. No mixed respiratory virus infection was found. Coinfections of the influenza A H3N2 virus with TTV, HMPV with TTV, HRV with TTV, and human parainfluenza virus-3 with TTV were found in 4.7, 2.8, 19.8, and 0.9% of the samples, respectively. Conclusions: This study highlights the need to perform routine detection of respiratory viruses in adults hospitalized with ARI, asthma exacerbation, and/or pneumonia.

Molecular status of Human Immunodeficiency Virus, Hepatitis B virus, and Hepatitis C virus among injecting drug male commercial sex workers in Surakarta, Indonesia

Male commercial sex workers are one of the high-risk community for blood-borne viruses. However, there are no data concerning the molecular status of Human Immunodeficiency Virus (HIV), Hepatitis B Virus (HBV), and Hepatitis C Virus (HCV) circulated among male commercial sex workers with injecting drug habits in Surakarta, Indonesia. Blood samples obtained from injecting drug male commercial sex workers in Surakarta were examined for HIV antibodies, HBsAg, and HCV antibodies, respectively, by immunological assays. Blood samples were also subjected to viral nucleic acid extraction and molecular detection of HIV, HBV, and HCV by nested (RT) PCRs. The PCR products were purified from agarose gels, and the nucleotide sequences were retrieved and molecular analyzed. HIV, HBV, and HCV were detected in 29.4% (10/34), 17.6% (6/34), and 52.9% (18/34), respectively. HIV CRF01_AE and B were found to be circulating in the community. HBV genotype B3 was predominated, followed by C1. HCV genotype 1a was predominated, followed by 1c, 3a, 1b, and 4a. HIV, HBV, and HCV were found circulating in the male commercial sex workers with injecting drug habits in Surakarta, Indonesia.

Molecular analysis of Toxoplasma gondii Surface Antigen 1 (SAG1) gene cloned from Toxoplasma gondii DNA isolated from Javanese acute toxoplasmosis

Toxoplasma gondii Surface Antigen 1 (SAG1) is often used as a diagnostic tool due to its immunodominant-specific as antigen. However, data of the Toxoplasma gondii SAG1 protein from Indonesian isolate is limited. To study the protein, genomic DNA was isolated from a Javanese acute toxoplasmosis blood samples patient. A complete coding sequence of Toxoplasma gondii SAG1 was cloned and inserted into an Escherichia coli expression plasmid and sequenced. The sequencing results were subjected to bioinformatics analysis. The Toxoplasma gondii SAG1 complete coding sequences were successfully cloned. Physicochemical analysis revealed the 336 aa of SAG1 had 34.7 kDa of weight. The isoelectric point and aliphatic index were 8.4 and 78.4, respectively. The N-terminal methionine half-life in Escherichia coli was more than 10 hours. The antigenicity, secondary structure, and identification of the HLA binding motifs also had been discussed. The results of this study would contribute information about Toxoplasma gondii SAG1 and benefits for further works willing to develop diagnostic and therapeutic strategies against the parasite.

Torque teno virus infection in male commercial sex workers in Surakarta Indonesia

The molecular epidemiology data of torque teno virus (TTV) in Indonesia is very rare. This study evaluated the prevalence of TTV in male commercial sex workers, as one of the high risk community for blood borne viruses pathogens in Surakarta, Indonesia. All blood samples collected from male commercial sex workers in Surakarta in 2009-2013 were tested by nested polymerase chain reaction (PCR). The amplified PCR products were molecularly cloned and subjected to sequence analysis. TTV DNA was detected in 80.9% (72/89) samples. Furthermore, the molecular characterization revealed that the most prevalent was genogroup 3, followed by genogroup 2 and l, respectively. TTV was detected in male commercial sex workers in Surakarta with high infection rate. Further investigation about TTV circulation in Indonesian population is needed in order to provide additional information about the genetic variability and TTV epidemiology in Indonesia, especially in the high risk communities.

HLA-DQBl*0402 alleles polymorphisms detected in Javanese HIV patients with positive anti-Toxoplasma gondii IgM

The human leukocyte antigen (HLA)-DQB1 gene polymorphisms may associated with the infection risk of Toxoplasma gondii in HIV patients. The HLA-DQB1*0402 in HIV-1-positive patients could be considered risk factors for developing neurological opportunistic infections, mainly Toxoplasma encephalitis. However, the HLA-DQB1*0402 gene polymorphisms status in the Javanese HIV patients is unknown. This study evaluated the prevalence of HLA-DQB*0402 alleles polymorphisms in Javanese HIV patients with positive anti-Toxoplasma gondii IgM status. Since 2009 our research group performing a molecular epidemiology of blood borne viruses in Central Java Indonesia, by collecting the epidemiological and clinical data from the high risk communities. All blood samples were screened for blood borne pathogens by serological and molecular assays including for HIV and Toxoplasma gondii. The genomic DNA was isolated from the whole blood samples. Genetic polymorphisms of HLA-DQB1*0402 alleles were detected with polymerase chain re...

Molecular status of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus among transgender commercial sex workers in Surakarta, Indonesia

Sexual contact and other risk behavior among transgender working as commercial sex workers are important factors for sexual and blood-borne virus (BBV) infections. However, there no data concerning the molecular status of human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) circulated among transgender working as commercial sex workers. Blood samples obtained from transgender working as commercial sex workers in Surakarta were examined for HIV antibodies, HBsAg and HCV antibodies, respectively, by immunological assays. All blood samples were also subjected for viral nucleic acid extraction and molecular detection of HIV, HBV and HCV by nested RT-PCR. The PCR products were purified from agarose gels, and the nucleotide sequences were retrieved and molecular analyzed. HIV, HBV and HCV was detected in 26.9% (7/26), 19.2% (5/26) and 46.2% (12/26), respectively. HIV CRF01_AE and B were found to be circulating in the community. HBV genotype B3 predominated, followed by C1. HCV...

Sequence conservation, HLA-E-Restricted peptide, and best-defined CTL/CD8+ epitopes in gag P24 (capsid) of HIV-1 subtype B

Human immunodeficiency virus type 1 (HIV-1) remains a cause of global health problem. Continuous studies of HIV-1 genetic and immunological profiles are important to find strategies against the virus. This study aimed to conduct analysis of sequence conservation, HLA-E-restricted peptide, and best-defined CTL/CD8+ epitopes in p24 (capsid) of HIV-1 subtype B worldwide. The p24-coding sequences from 3,557 HIV subtype B isolates were aligned using MUSCLE and analysed. Some highly conserved regions (sequence conservation ≥95%) were observed. Two considerably long series of sequences with conservation of 100% was observed at base 349-356 and 550-557 of p24 (HXB2 numbering). The consensus from all aligned isolates was precisely the same as consensus B in the Los Alamos HIV Database. The HLA-E-restricted peptide in amino acid (aa) 14-22 of HIV-1 p24 (AISPRTLNA) was found in 55.9% (1,987/3,557) of HIV-1 subtype B worldwide. Forty-four best-defined CTL/CD8+ epitopes were observed, in which VKNWMTETL epitope (aa 18...