htlv type i p19  (ZeptoMetrix corporation)

Journal: Journal of Medical Virology

Article Title: Persistent Human T‐Lymphotropic Virus Type 1 (HTLV‐1) Infection in the Placenta of Pregnant Women

doi: 10.1002/jmv.70585

Figure Lengend Snippet: Syncytin‐1 may mediate HTLV‐1 infection of placental trophoblasts. (A) Expression of syncytin‐1 receptors ASCT1 and ASCT2 on MT‐2 cells was measured by flow cytometry. (B) A schematic diagram of the coculture protocol for HTLV‐1 transfer to trophoblasts. BeWo cells or PCTs were cocultured with MT‐2 cells (10:1 ratio of MT‐2 and target cells) for 48 h, followed by washing and further incubation. (C) Representative phase‐contrast images (scale bar = 200 μm) show reduced confluency in BeWo cells cocultured with MT‐2 cells. (D) Protein levels of HTLV‐1 p19, syncytin‐1, and β‐actin in BeWo cells and PCTs under basal conditions or after forskolin induction (20–50 μM for BeWo; 50–100 μM for PCTs), with or without HRB1‐mediated syncytin‐1 cell fusion blockade. The relative intensity of p19 expression is shown below with data normalized to the BeWo or PCT cocultured with MT‐2 cells without forskolin treatment. Syncytin‐1 expression was normalized to the corresponding group without forskolin treatment.

Article Snippet: Protein expression was assessed using the following antibodies: anti‐HTLV type I p19 clone 45/6.11.1.3 (1:2000, ZeptoMetrix), syncytin‐1 polyclonal antibody (1:3000, Bioss), and β‐actin antibody (1:2500, Cell Signaling).

Techniques: Infection, Expressing, Flow Cytometry, Incubation

Journal: Nature Communications

Article Title: High-resolution analysis of the human T-cell leukemia virus capsid protein reveals insights into immature particle morphology

doi: 10.1038/s41467-025-67129-1

Figure Lengend Snippet: a Representative HTLV-1 immature particles. Shown are two representative HTLV-1 immature particles from the cryo-EM dataset, where the red circles represent a simplified model of the particle picking scheme, highlighting all regions of the Gag lattice used in the reconstruction. Scale bar, 50 nm. b Single particle reconstruction. Shown is the top view of the single particle lattice reconstruction for HTLV-1 Gag. The map was locally sharpened using deepEMhancer . c Side view of the immature Gag lattice. The CA-NTD and CA-CTD are colored as blue and gold, respectively. d Local resolution estimates. Shown is a cross-section through the central CA hexamer. The density is colored by the local resolution estimate. e Annotated side views of the modeled HTLV-1 CA regions. Shown is a modeled region of a CA monomer with annotated helices. The CA-NTD and CA-CTD are colored in blue and gold, respectively. f Helix model fitting into the cryo-EM density. Shown is helix 4, highlighting the agreement of the fitted model into the density.

Article Snippet: Blots were incubated with 1:2000 anti-HTLV-1 p24 antibody (6G9, Santa Cruz) or anti-HIV-1 p24 antibody (24-4, Santa Cruz), washed 5x for 3 min. each in TBST, then incubated with 1:2000 anti-mouse IgG StarBright TM Blue 700 (Bio-Rad, CA).

Techniques: Cryo-EM Sample Prep, Single Particle

Journal: Nature Communications

Article Title: High-resolution analysis of the human T-cell leukemia virus capsid protein reveals insights into immature particle morphology

doi: 10.1038/s41467-025-67129-1

Figure Lengend Snippet: a Top view of the central hexamer model fitted into a lower resolution and unsharpened density. The red boxes highlight the small density present at the center of each hexamer. b Molecular dynamics simulation of HTLV-1 CA coordinating IP6. Six copies of R13 and K18 in the CA-NTD can coordinate IP6. The central pore density from ( a ) is shown for the placement of IP6 in the simulation. c , d In vitro HTLV-1 CA assembly assay. Shown are SDS-PAGE analysis of purified HTLV-1 CA after the addition of IP6 or different polyanion at varying concentrations to WT CA or predicted IP6 binding mutants. The gels are representative from three-independent experiments e Negative stain TEM. Shown are representative negative stain TEM micrographs from one replicate of the resuspended pellet from the corresponding polyanion type, polyanion concentration, and HTLV-1 CA type from ( c ) to ( d ). Scale bar, 200 nm. f Immunoblot analysis of immature particle production. Cell culture supernatants from cells producing immature HTLV-1 or HIV-1 particles were analyzed by immunoblot, shown is one representative blot from three-independent experiments. Particle production was compared between WT 293T and IP6-depleted cells. g Quantification of the immunoblot analysis from ( f ) was done from three-independent replicates, and the significance was determined using an unpaired two-sided t -test by comparing particle production from the IP6-depleted cells to that from WT cells for each virus. *, P < 0.001. The error bars represent the standard error of the mean (SEM). The exact p-values are: HIV-1 IPPK KO = 0.0005 and HIV-1 IPMK KO < 0.0001. h Immature particle morphology. Cryo-EM micrographs of immature HTLV-1 particles purified from IPPK KO cells that were co-transfected with the expression plasmid encoding the MINPP1 phosphatase. Scale bar, 100 nm. Micrographs are representative images from one purification.

Article Snippet: Blots were incubated with 1:2000 anti-HTLV-1 p24 antibody (6G9, Santa Cruz) or anti-HIV-1 p24 antibody (24-4, Santa Cruz), washed 5x for 3 min. each in TBST, then incubated with 1:2000 anti-mouse IgG StarBright TM Blue 700 (Bio-Rad, CA).

Techniques: In Vitro, SDS Page, Purification, Binding Assay, Staining, Concentration Assay, Western Blot, Cell Culture, Virus, Cryo-EM Sample Prep, Transfection, Expressing, Plasmid Preparation