Journal: NPJ Vaccines

Article Title: Reduced dengue virus pre-membrane protein-HMGB1 interaction activates host cell transcription to attenuate infection

doi: 10.1038/s41541-026-01401-3

Figure Lengend Snippet: A Schematic diagram of the genetic differences between attenuated DENV-2 PDK53 and its wild-type DENV-2 16681 parent. Nucleotides and amino acids of wild-type 16681 and attenuated PDK53 represented in blue and red letters, respectively. Figure adapted from Guzman et al., 2010 . B , C Plaque phenotype of 16681, prM D29V mutant and PDK53 on BHK-21 cells. B Representative pictures of plaques. C Plaque sizes in mm 2 ; n ≥ 12. n represents the number of plaques. mean ± SEM. D DENV RNA levels relative to GAPDH mRNA levels in BHK-21 cells infected with either 16681, prM D29V mutant or PDK53 at MOI of 1 at 24 hpi; n = 3. n represents number of biological replicates. mean ± SD. E – H Plaque phenotype of 16681, prM D29V mutant and PDK53 on BHK-21 cells transfected with either siRNA targeting IRF3 (siIRF3) or scramble non-targeting siRNA as control (siControl). E Western blot analysis showing knockdown efficiency of IRF3. F Representative pictures of plaques. G Plaque sizes in mm 2 ; n ≥ 15. n represents number of plaques. mean ± SEM. H Plaque counts per well of 24-well plate; n ≥ 5. n represents number of wells of 24-well plate. mean ± SD. I – L DENV RNA levels relative to PPIA mRNA levels ( I ), and IFNβ ( J ), IFIT2 ( K ) and CXCL10 ( L ) mRNA levels relative to PPIA mRNA levels in primary moDCs infected with either 16681, prM D29V mutant or PDK53 at MOI of 1 at 48 hpi; n = 3. n represents number of biological replicates. mean ± SD. * p < 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001 in unpaired Student’s t test.

Article Snippet: The authors declare no competing financial interest but the following competing non-financial interests: E.E.O. has served as a dengue vaccine advisory board member for Takeda Pharmaceuticals, which has used DENV-2 PDK53 to develop TAK-003.

Techniques: Mutagenesis, Infection, Transfection, Control, Western Blot, Knockdown

Journal: NPJ Vaccines

Article Title: Reduced dengue virus pre-membrane protein-HMGB1 interaction activates host cell transcription to attenuate infection

doi: 10.1038/s41541-026-01401-3

Figure Lengend Snippet: A , B Plaque phenotype of 16681, PDK53 and prM V29D reverse mutant on BHK-21 cells. A Representative pictures of plaques. B Plaque sizes in mm 2 ; n ≥ 24. n represents number of plaques. mean ± SEM. C – F DENV RNA levels relative to PPIA mRNA levels ( C ), and IFNβ ( D ), IFIT2 ( E ) and CXCL10 ( F ) mRNA levels relative to PPIA mRNA levels in BHK-21 cells infected with either 16681, prM D29V mutant, PDK53 or prM V29D reverse mutant at MOI of 1 at 24 hpi; n = 3. n represents number of biological replicates. mean ± SD. * p < 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001 in unpaired Student’s t test.

Article Snippet: The authors declare no competing financial interest but the following competing non-financial interests: E.E.O. has served as a dengue vaccine advisory board member for Takeda Pharmaceuticals, which has used DENV-2 PDK53 to develop TAK-003.

Techniques: Mutagenesis, Infection

Journal: NPJ Vaccines

Article Title: Reduced dengue virus pre-membrane protein-HMGB1 interaction activates host cell transcription to attenuate infection

doi: 10.1038/s41541-026-01401-3

Figure Lengend Snippet: Whole genome microarray was performed on primary moDCs infected with either 16681, prM D29V mutant, NS1 G53D mutant or PDK53 at MOI of 1 at 48 hpi; n = 3. n represents number of biological replicates. A Table (left) and proportional Venn diagram (right), showing the number of genes differentially expressed during attenuated DENV-2s infection against wild-type 16681 infection. Differentially expressed genes (DEGs) were defined as genes with fold change more than 2 and adjusted p-value less than 0.05. Proportional Venn diagram created according to the diagram made by using DeepVenn ( https://www.deepvenn.com/ ) . B – E Pathway enrichment analysis on the DEGs shared among three attenuated DENV-2s ( B , C ), and between prM D29V mutant and PDK53 only ( D , E ). B , D Pie charts of the percentages of the up- and down-regulated DEGs. C , E Pathway enrichment analysis conducted by using the EnrichR tool against the Gene Ontology Biological Process (GOBP) database , . Significantly enriched pathways were defined as pathways with adjusted p -value less than 0.05 and comprising at least 5 genes. The left Y-axis and X-axis indicate enriched pathways and the number of DEGs enriched in each pathway, respectively. Significantly enriched pathways are further classified into biological functions and annotated on the right Y-axis.

Article Snippet: The authors declare no competing financial interest but the following competing non-financial interests: E.E.O. has served as a dengue vaccine advisory board member for Takeda Pharmaceuticals, which has used DENV-2 PDK53 to develop TAK-003.

Techniques: Microarray, Infection, Mutagenesis

Journal: NPJ Vaccines

Article Title: Reduced dengue virus pre-membrane protein-HMGB1 interaction activates host cell transcription to attenuate infection

doi: 10.1038/s41541-026-01401-3

Figure Lengend Snippet: A – E A549 cells transfected with either siRNA targeting HMGB1 (siHMGB1) or scramble non-targeting siRNA as control (siControl) were infected with either 16681, prM D29V mutant or PDK53 at MOI of 1 at 24 hpi; n = 3. n represents number of biological replicates. A Western blot analysis showing knockdown efficiency of HMGB1. B – D NanoString nCounter analysis was performed on infected siRNA-treated A549 cells using the nCounter Human Immunology panel. Volcano plots of differential immune gene expressions in siHMGB1-treated A549 cells versus siControl-treated A549 cells during 16681 infection ( B ), prM D29V mutant infection ( C ), and PDK53 infection ( D ). Each point represents the average value of each immune gene from three biological replicates. The expression difference is considered significant for log2 fold change more than 1.3 (outer vertical dashed black lines) and p-value less than 0.05 (above horizontal solid red line). Points are colored according to their average expressions. Blue represents significantly downregulated genes. Red represents significantly upregulated genes. Gray represents not significantly differentially expressed genes. E DENV RNA levels relative to GAPDH mRNA levels in siRNA-treated A549 cells following infection. mean ± SD. F , G Plaque phenotype of DENV-3 wild type and DENV-3 prM S29V mutant on BHK-21 cells. F Representative pictures of plaques. G Plaque sizes in mm2; n ≥ 18. n represents number of plaques. mean ± SEM. H – K DENV RNA levels relative to GAPDH mRNA levels ( H ), and IFNβ ( I ), IFIT2 ( J ) and CXCL10 ( K ) mRNA levels relative to GAPDH mRNA levels in Huh-7 cells infected with either DENV-3 wild type or DENV-3 prM S29V mutant at MOI of 1 at 48 hpi; n = 3. n represents number of biological replicates. mean ± SD. L DENV RNA levels relative to GAPDH mRNA levels in primary moDCs infected with either TAK-003 DENV-3 or its prM S29V mutant at MOI of 1 over the course of 48 hours after infection; n = 3. n represents number of biological replicates. mean ± SD. * p < 0.05; ** p ≤ 0.01; *** p ≤ 0.001 in unpaired Student’s t test.

Article Snippet: The authors declare no competing financial interest but the following competing non-financial interests: E.E.O. has served as a dengue vaccine advisory board member for Takeda Pharmaceuticals, which has used DENV-2 PDK53 to develop TAK-003.

Techniques: Transfection, Control, Infection, Mutagenesis, Western Blot, Knockdown, Expressing