Identification of Host Cellular Protein as hnRNP K

<div><p>(A) Specific protein spots were cored out and de-stained, following which the gel plug was digested with trypsin. Sequence query of peptide fragments was carried out by Proteomic Research Services, using LC/MS/MS analysis. Results of the 21 sequenced peptides are illustrated.</p> <p>(B) Results of peptide sequencing of the 56-kDa protein by LC/MS/MS showed high homology scores to hnRNP K in sequence alignments.</p></div

Evidence for the Involvement of a Host Cellular Protein in Enh II Activity and HBV Replication

<div><p>(A) Electrophoretic mobility shift assays were performed using HepG2 nuclear extracts with four different probes. Probe 1, lanes 1–4; probe 2 (1752A), lanes 5–8; probe 3, lanes 9–12; probe 4 (1752G), lanes 13–16. Each set of probes contains increasing concentrations (0.0 μg, 0.05 μg, 0.10 μg, and 0.15 μg) of non-specific competitor DNA [poly-(dI)-poly-(dC)], respectively.</p> <p>(B) 40 μg of nuclear protein extracts obtained from HepG2 cells was allowed to bind onto 5 mg Dynabeads M-280 streptavidin-biotin-oligonucleotides in the presence of 2:1 (w/w) ratio of non-specific competitor DNA poly (dI–dC). 1-D isoelectric focusing was followed by 2-D vertical separation on SDS-PAGE (10%). The estimated molecular weight of the specific protein spots detected by silver staining (arrow) is indicated.</p></div

hnRNP K siRNAs Down-Regulate HBV Viral Replication

<div><p>(A) HepG2 cells were co-transfected with 1752A full-length replicative HBV clone either with or without hnRNP K siRNA (2 μg). Non-silencing (Non-T) and lamin A/C (Lamin) siRNAs were used as controls. hnRNP K expression was measured by quantitative real-time RT-PCR.</p> <p>(B) HBV viral load was quantitated by real-time PCR in cells transfected as described in (A).</p> <p>(C) Lamin A/C expression was measured from real-time RT-PCR. Ratios were normalized to 100% for the non-transfected cells. The results represent two independent samples; standard deviations are shown.</p> <p>Black columns represent either non-transfected cells or cells transfected with non-silencing siRNA. White columns represent cells co-transfected with HBV and lamin A/C siRNA. Grey columns represent cells co-transfected with HBV and hnRNP K siRNAs (A, Dharmacon; B, Qiagen; C, Proligo).</p></div

Distinct Segregation between High and Low Viremic HBV Individuals Is Correlated to Changes at Nucleotide Position 1752

<p>Comparison of DNA sequences from nucleotide 1720–1769 with the HBV DNA concentration levels of the participants are illustrated. Sera was collected from 60 participants; DNA was isolated from sera and amplified with two rounds of PCR. Results of the sequences were aligned and compared.</p

hnRNP K Is Involved in Modulating Viral Replication

<p>HepG2 cells were co-transfected with full-length replicative HBV clones (indicated by “+”) 1752A, 1752ΔG, 1752ΔT, and 1752ΔC (see <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0020163#s2" target="_blank">Methods</a> and <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0020163#sg001" target="_blank">Figure S1</a>) with increasing dosages (50, 250, and 1,250 ng/μl) of hnRNP K variant 2 (v2) or variant 3 (v3) as indicated. pcDNA 3.1 serves as a control. Transfections were performed in duplicate; standard deviations are shown.</p

Defining the structural basis for human leukocyte antigen reactivity in clinical transplantation

The current state-of-the-art technology employed to assess anti-human leukocyte antigen antibodies (Anti-HLA Ab) for donor-recipient matching and patient risk stratification in renal transplantation is the single antigen bead (SAB) assay. However, there are limitations to the SAB assay as it is not quantitative and due to variations in techniques and reagents, there is no standardization across laboratories. In this study, a structurally-defined human monoclonal alloantibody was employed to provide a mechanistic explanation for how fundamental alloantibody biology influences the readout from the SAB assay. Performance of the clinical SAB assay was evaluated by altering Anti-HLA Ab concentration, subclass, and detection reagents. Tests were conducted in parallel by two internationally accredited laboratories using standardized protocols and reagents. We show that alloantibody concentration, subclass, laboratory-specific detection devices, subclass-specific detection reagents all contribute to a significant degree of variation in the readout. We report a significant prozone effect affecting HLA alleles that are bound strongly by the test alloantibody as opposed to those bound weakly and this phenomenon is independent of complement. These data highlight the importance for establishing international standards for SAB assay calibration and have significant implications for our understanding of discordance in previous studies that have analyzed its clinical relevance.Published versio

Rapid production of clinical-grade SARS-CoV-2 specific T cells.

10.1002/acg2.101Adv Cell Gene Ther34e101

Dendritic cell therapy with CD137L-DC-EBV-VAX in locally recurrent or metastatic nasopharyngeal carcinoma (NPC).

Annual Meeting of the American-Society-of-Clinical-Oncology (ASCO)381