hek bluetm il 6  (InvivoGen)

Journal: bioRxiv

Article Title: A modular mRNA–LNP vaccine platform enables integrated RNA, lipid and antigen design to protect against CCHFV

doi: 10.64898/2026.01.17.699915

Figure Lengend Snippet: Innate activation was compared across mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315 using NPmut mRNA as a common payload, and across matched empty LNP controls (iLNP; no mRNA) to isolate lipid-driven signaling. Reporter cells were exposed to a dose range of formulations (1 μg, 500 ng, 250 ng, 125 ng, and 62.5 ng; as indicated), and pathway activation was quantified as fold-change induction relative to baseline (dotted line). Bars denote mean responses with error bars indicating variability across replicates; selected pairwise comparisons are annotated with exact P values. a, NF-κB activation measured in THP1-Dual™ cells as induction of NF-κB–SEAP. Across doses, NPmut mRNA/LNPs and empty LNPs produced modest but consistent NF-κB induction, with broadly similar activation profiles across BP-104-, SM-102-, and ALC-0315–containing LNPs. b, IL-6 pathway signaling measured in HEKBlue–IL-6 reporter cells as fold-change induction of IL-6–dependent signaling. All three ionizable lipid formulations elicited comparable IL-6 activation across the dilution series, and empty LNP controls also induced IL-6 signaling, indicating that both lipid composition and mRNA cargo contribute to pathway engagement. c, Type I interferon signaling measured in IFN-α/β reporter HEK293 cells as fold-change induction of IFN-I activity. NPmut mRNA/LNPs triggered IFN-I responses across lipids, with ALC-0315 producing slightly higher induction at select concentrations (P values shown). Empty LNPs also elicited measurable IFN-I signaling, consistent with lipid-dependent innate stimulation independent of mRNA. Collectively, these data show that BP-104 engages canonical innate immune pathways (NF-κB, IL-6, and type I interferon) at levels comparable to SM-102 and ALC-0315 under controlled dosing conditions, supporting its suitability as an ionizable lipid for mRNA–LNP vaccine delivery.

Article Snippet: HEK-BlueTM IFN-α/β (InvivoGen, Cat# hkb-ifnabv2), HEK-BlueTM IL-6 (InvivoGen, Cat# hkb-hil6), and THP1-DualTM (InvivoGen, Cat# thpd-nfis) were employed to assess type I interferon activity and innate immune activation in response to NPmut mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315.

Techniques: Activation Assay, Produced, Activity Assay

Journal: bioRxiv

Article Title: A modular mRNA–LNP vaccine platform enables integrated RNA, lipid and antigen design to protect against CCHFV

doi: 10.64898/2026.01.17.699915

Figure Lengend Snippet: Primary murine dendritic cells (DCs) were loaded with NPmut mRNA formulated in lipid nanoparticles (LNPs) containing BP-104, SM-102, or ALC-0315, then co-cultured with naïve CD4⁺ and CD8⁺ T cells to quantify intracellular cytokines and soluble mediators in supernatants. a, Frequencies of cytokine-positive CD4⁺ and CD8⁺ T cells measured by intracellular cytokine staining, including IFN-γ and TNF-α (top), IL-2⁺ CD8⁺ cells (bottom left), and IL-17A⁺ CD4⁺ cells (bottom right). Bars show group means with individual biological replicates overlaid; statistical comparisons and corresponding P values are indicated. Across formulations, NPmut mRNA/LNP-loaded DCs elicited Th1-skewed responses (IFN-γ and TNF-α) with a measurable IL-17A⁺ CD4⁺ population, whereas negative control cultures remained low. ALC-0315 drove the strongest IFN-γ⁺ and TNF-α⁺ T cell responses across CD4⁺ and CD8⁺ compartments and increased IL-17A⁺ CD4⁺ frequencies, BP-104 produced intermediate activation, and SM-102 was comparatively attenuated. Lipid-dependent differences were also evident for IL-2⁺ CD8⁺ responses, indicating differential programming of proliferative/expansion-associated outputs. b, Secreted cytokine and chemokine profiles in DC–T cell co-culture supernatants (mean fluorescence intensity, MFI) measured for inflammatory and recruitment-associated mediators, including TNF-α, CXCL1, IFN-γ, IL-12, CCL5, IL-4, CXCL10, GM-CSF, IL-10, IL-6, IP-10, and MCP-1. Error bars indicate variability across replicates; selected pairwise comparisons are annotated with P values. Supernatant signatures mirrored intracellular T cell phenotypes: ALC-0315 induced the most pronounced inflammatory/recruitment program, BP-104 generated a coherent but moderated profile, similar to SM-102. Together, these data show that ionizable lipid identity imprints innate cytokine/chemokine cues that scale and shape downstream Th1/Th17 polarization under fixed mRNA payload and DC-loading conditions.

Article Snippet: HEK-BlueTM IFN-α/β (InvivoGen, Cat# hkb-ifnabv2), HEK-BlueTM IL-6 (InvivoGen, Cat# hkb-hil6), and THP1-DualTM (InvivoGen, Cat# thpd-nfis) were employed to assess type I interferon activity and innate immune activation in response to NPmut mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315.

Techniques: Cell Culture, Staining, Negative Control, Produced, Activation Assay, Co-Culture Assay, Fluorescence, Generated