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cd11b fractions  (Miltenyi Biotec)


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    Miltenyi Biotec cd11b fractions
    F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. <t>CD11b</t> + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.
    Cd11b Fractions, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd11b+fraction/CD11b%2Fc+Antibody%2C+anti-rat%2C+REAfinity/pmc12741393-276-16-21
    Average 98 stars, based on 27 article reviews
    cd11b fractions - by Bioz Stars, 2026-10
    98/100 stars

    Images

    1) Product Images from "Lipid nanoparticle-encapsulated microRNA-192: An anti-inflammatory adjuvant that enhances vaccine efficacy in aged mice"

    Article Title: Lipid nanoparticle-encapsulated microRNA-192: An anti-inflammatory adjuvant that enhances vaccine efficacy in aged mice

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2025.102784

    F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. CD11b + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.
    Figure Legend Snippet: F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. CD11b + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Techniques Used: In Vivo, Quantitative RT-PCR, SDS Page, Injection, Enzyme-linked Immunosorbent Assay, Western Blot

    LNP-miR-192 improves vaccine efficacy in aged mice (A and B) K-LNP-empty, K-LNP-miR-192, F-LNP-NC, or F-LNP-miR-192 or S-LNP-miR-192 (20 μL of each) was mixed with SV (80 μL) and subcutaneously injected into aged BALB/c mice ( n = 6). A second dose of the same vaccine was administered 3 weeks later. Sera were collected 4 weeks after the first-dose vaccination, and serum HI titers for influenza type A and B were determined. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001: Log-transformed data were analyzed by Welch’s one-way ANOVA (A) and Welch’s t test (B). (C) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) mixed with 20 μL of the indicated LNPs. Four weeks after the first vaccination, sera were collected and HI titers were measured. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05: Kruskal-Wallis test with Dunn’s multiple comparisons test. (D) Aged BALB/c mice were subcutaneously vaccinated with SV (80 μL) mixed with F-LNP-miR-192 (20 μL). Two weeks later, draining lymph nodes were excised, and the number of GC B cells was measured via flow cytometry. Data are means ± SDs, with dots representing individual mice. ∗∗ p < 0.01; t test. (E and F) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) alone or SV formulated with 20 μL of F-LNP-miR-192 or F-LNP-NC, as indicated. Four weeks after the first vaccination, mice were challenged with influenza A virus and monitored daily for survival. Mice losing >25% of their initial body weight were euthanized and recorded as dead. (E) Survival curves for mice vaccinated with SV alone or SV combined with F-LNP-miR-192. A significant trend in survival across all four groups was detected via the log rank test for trends ( p < 0.05). (F) Survival comparison between the SV + F-LNP-miR-192 and SV + F-LNP-NC groups, with a significant difference in survival observed (log rank test, p < 0.05). (G) SV (80 μL) was mixed with one of the indicated LNPs (20 μL) and subcutaneously injected into aged mice. Sera were collected 24 and 48 h post-injection, and serum IL-6 concentrations were measured using ELISA. Data are means ± SDs (n = 3, nonsignificant [NS] > 0.05: one-way ANOVA). (H) Aged mice were subcutaneously vaccinated using the WVP vaccine (80 μL) of influenza A virus with PBS or F-LNP-miR-192 twice at a 2-week interval. One month after the first vaccination, sera were collected, and HI and neutralization titers (NT) were determined. Data are means ± SDs. (n = 5, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001: log-transformed data were analyzed by one-way ANOVA). (I, J) 20 μL of F-LNP was subcutaneously injected into aged mice. The skin tissues were excised at 2 days after infection (I) or at indicated days (J), and CD11b + cells were isolated using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.
    Figure Legend Snippet: LNP-miR-192 improves vaccine efficacy in aged mice (A and B) K-LNP-empty, K-LNP-miR-192, F-LNP-NC, or F-LNP-miR-192 or S-LNP-miR-192 (20 μL of each) was mixed with SV (80 μL) and subcutaneously injected into aged BALB/c mice ( n = 6). A second dose of the same vaccine was administered 3 weeks later. Sera were collected 4 weeks after the first-dose vaccination, and serum HI titers for influenza type A and B were determined. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001: Log-transformed data were analyzed by Welch’s one-way ANOVA (A) and Welch’s t test (B). (C) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) mixed with 20 μL of the indicated LNPs. Four weeks after the first vaccination, sera were collected and HI titers were measured. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05: Kruskal-Wallis test with Dunn’s multiple comparisons test. (D) Aged BALB/c mice were subcutaneously vaccinated with SV (80 μL) mixed with F-LNP-miR-192 (20 μL). Two weeks later, draining lymph nodes were excised, and the number of GC B cells was measured via flow cytometry. Data are means ± SDs, with dots representing individual mice. ∗∗ p < 0.01; t test. (E and F) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) alone or SV formulated with 20 μL of F-LNP-miR-192 or F-LNP-NC, as indicated. Four weeks after the first vaccination, mice were challenged with influenza A virus and monitored daily for survival. Mice losing >25% of their initial body weight were euthanized and recorded as dead. (E) Survival curves for mice vaccinated with SV alone or SV combined with F-LNP-miR-192. A significant trend in survival across all four groups was detected via the log rank test for trends ( p < 0.05). (F) Survival comparison between the SV + F-LNP-miR-192 and SV + F-LNP-NC groups, with a significant difference in survival observed (log rank test, p < 0.05). (G) SV (80 μL) was mixed with one of the indicated LNPs (20 μL) and subcutaneously injected into aged mice. Sera were collected 24 and 48 h post-injection, and serum IL-6 concentrations were measured using ELISA. Data are means ± SDs (n = 3, nonsignificant [NS] > 0.05: one-way ANOVA). (H) Aged mice were subcutaneously vaccinated using the WVP vaccine (80 μL) of influenza A virus with PBS or F-LNP-miR-192 twice at a 2-week interval. One month after the first vaccination, sera were collected, and HI and neutralization titers (NT) were determined. Data are means ± SDs. (n = 5, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001: log-transformed data were analyzed by one-way ANOVA). (I, J) 20 μL of F-LNP was subcutaneously injected into aged mice. The skin tissues were excised at 2 days after infection (I) or at indicated days (J), and CD11b + cells were isolated using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Techniques Used: Injection, Transformation Assay, Flow Cytometry, Virus, Comparison, Enzyme-linked Immunosorbent Assay, Neutralization, Infection, Isolation, SDS Page, Western Blot

    Related Articles

    FACS:

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids
    Article Snippet: .. Magnetic-activated cell sorting (MACS) was used to enrich for the CD11b + fraction (Miltenyi, 130049601) according to the manufacturer's protocol. .. The CD11b + cell fraction was collected in microglia culture medium [RPMI 1640 (Life Technologies, 21875034), 10% fetal bovine serum, 2 mM L- glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker) and 100 ng/ml IL-34 (Pepro Tech, 200-34)].

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids.
    Article Snippet: .. Neural organoids were generated using a modified version of a previously 5 published protocol.22 6 7 Single cell preparation and magnetic cell sorting (MACS) 8 Organoid dissociation was performed as described before.21 MACS was used to enrich for the 9 CD11b+ fraction (Miltenyi, 130049601) according to manufacturer’s protocol. .. The CD11b+ cell 10 fraction was collected in microglia culture medium (RPMI 1640 (Life technologies, 21875034), 11 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker, 12 Belgium) and 100 ng/ml IL-34 (Pepro Tech, 200-34)).

    Magnetic Cell Separation:

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids
    Article Snippet: .. Magnetic-activated cell sorting (MACS) was used to enrich for the CD11b + fraction (Miltenyi, 130049601) according to the manufacturer's protocol. .. The CD11b + cell fraction was collected in microglia culture medium [RPMI 1640 (Life Technologies, 21875034), 10% fetal bovine serum, 2 mM L- glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker) and 100 ng/ml IL-34 (Pepro Tech, 200-34)].

    Article Title: ATAXIN-2 intermediate-length polyglutamine expansions elicit ALS-associated metabolic and immune phenotypes.
    Article Snippet: .. CD11b+ fraction was isolated by MACS, as described above, and according to the manufacturer’s protocol (130049601, Miltenyi). ..

    Article Title: ATAXIN-2 intermediate-length polyglutamine expansions elicit ALS-associated metabolic and immune phenotypes
    Article Snippet: Motor function was tested every 4 weeks on an accelerating rotarod apparatus (47600 model, Ugo Basile, Varese, Italy) at four rounds per minute (rpm) to 40 rpm. .. CD11b+ fraction was isolated by MACS, as described above, and according to the manufacturer’s protocol (130049601, Miltenyi). ..

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids.
    Article Snippet: .. Neural organoids were generated using a modified version of a previously 5 published protocol.22 6 7 Single cell preparation and magnetic cell sorting (MACS) 8 Organoid dissociation was performed as described before.21 MACS was used to enrich for the 9 CD11b+ fraction (Miltenyi, 130049601) according to manufacturer’s protocol. .. The CD11b+ cell 10 fraction was collected in microglia culture medium (RPMI 1640 (Life technologies, 21875034), 11 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker, 12 Belgium) and 100 ng/ml IL-34 (Pepro Tech, 200-34)).

    Article Title: Regional desynchronization of microglial activity is associated with cognitive decline in Alzheimer’s disease
    Article Snippet: .. Astrocytes were isolated from the CD11b(-) fraction using ACSA2 MicroBeads (Miltenyi Biotec, 130–097-678) and a MACS separation system (Miltenyi Biotec). .. The prepared cell pellets were resuspended in 80 μL of AstroMACS separation buffer (Miltenyi Biotec, 130–117-336) per 10 7 total cells.

    Isolation:

    Article Title: ATAXIN-2 intermediate-length polyglutamine expansions elicit ALS-associated metabolic and immune phenotypes.
    Article Snippet: .. CD11b+ fraction was isolated by MACS, as described above, and according to the manufacturer’s protocol (130049601, Miltenyi). ..

    Article Title: ATAXIN-2 intermediate-length polyglutamine expansions elicit ALS-associated metabolic and immune phenotypes
    Article Snippet: Motor function was tested every 4 weeks on an accelerating rotarod apparatus (47600 model, Ugo Basile, Varese, Italy) at four rounds per minute (rpm) to 40 rpm. .. CD11b+ fraction was isolated by MACS, as described above, and according to the manufacturer’s protocol (130049601, Miltenyi). ..

    Article Title: Regional desynchronization of microglial activity is associated with cognitive decline in Alzheimer’s disease
    Article Snippet: .. Astrocytes were isolated from the CD11b(-) fraction using ACSA2 MicroBeads (Miltenyi Biotec, 130–097-678) and a MACS separation system (Miltenyi Biotec). .. The prepared cell pellets were resuspended in 80 μL of AstroMACS separation buffer (Miltenyi Biotec, 130–117-336) per 10 7 total cells.

    Incubation:

    Article Title: Neutrophil-derived catecholamines mediate negative stress effects on bone
    Article Snippet: MACS was performed using CD11b MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions. .. Briefly, cells were incubated with CD11b MicroBeads for 10 min at 4 °C and CD11b + cells were separated from the CD11b - fraction using a Quadro-MACS separator (Miltenyi Biotec) and moisturized LS columns (Miltenyi Biotec). .. Unfractured and fractured femora from mice euthanized 21 d post fracture were fixed in 4% formalin for 48 h and μCT scanning was performed using the Skyscan 1172 scanning tool (Bruker, Billerica, Massachusetts, USA) operating at 50 kV, 200 mA and a voxel resolution of 8 μm.

    Article Title: Neutrophil-derived catecholamines mediate negative stress effects on bone.
    Article Snippet: MACS was performed using CD11b MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s instructions. .. Briefly, cells were incubated with CD11b MicroBeads for 10min at 4 °C and CD11b+ cells were separated from the CD11b- fraction using a Quadro-MACS separator (Miltenyi Biotec) and moisturized LS columns (Miltenyi Biotec). μCT analysis (Sets 1 and 4, 8–10) Unfractured and fractured femora from mice euthanized 21 d post fracture were fixed in 4% formalin for 48 h and μCT scanning was performed using the Skyscan 1172 scanning tool (Bruker, Billerica, Massachusetts, USA) operating at 50 kV, 200mA and a voxel resolution of 8 μm. .. Three-dimensional analysis was performed using CTAn and CTVol software (Bruker) according to the ASBMR guidelines44.

    Article Title: Nigrostriatal degeneration determines dynamics of glial inflammatory and phagocytic activity
    Article Snippet: CD11b + cells were separated in an autoMACS Pro Separator (Miltenyi Biotec). .. The CD11b − fraction was collected and incubated with FcR Blocking Reagent (1:10) and anti-ACSA2 MicroBeads (1:10, Miltenyi Biotec) in CB for 15 min at 4 °C and separated in an autoMACS to isolate astrocytes. .. CD11b + and ACSA2 + cells were pelleted and resuspended in the lysis buffer from the Dynabeads mRNA Direct Kit (Ambion, Foster City, CA, USA).

    Blocking Assay:

    Article Title: Nigrostriatal degeneration determines dynamics of glial inflammatory and phagocytic activity
    Article Snippet: CD11b + cells were separated in an autoMACS Pro Separator (Miltenyi Biotec). .. The CD11b − fraction was collected and incubated with FcR Blocking Reagent (1:10) and anti-ACSA2 MicroBeads (1:10, Miltenyi Biotec) in CB for 15 min at 4 °C and separated in an autoMACS to isolate astrocytes. .. CD11b + and ACSA2 + cells were pelleted and resuspended in the lysis buffer from the Dynabeads mRNA Direct Kit (Ambion, Foster City, CA, USA).

    Generated:

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids.
    Article Snippet: .. Neural organoids were generated using a modified version of a previously 5 published protocol.22 6 7 Single cell preparation and magnetic cell sorting (MACS) 8 Organoid dissociation was performed as described before.21 MACS was used to enrich for the 9 CD11b+ fraction (Miltenyi, 130049601) according to manufacturer’s protocol. .. The CD11b+ cell 10 fraction was collected in microglia culture medium (RPMI 1640 (Life technologies, 21875034), 11 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker, 12 Belgium) and 100 ng/ml IL-34 (Pepro Tech, 200-34)).

    Modification:

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids.
    Article Snippet: .. Neural organoids were generated using a modified version of a previously 5 published protocol.22 6 7 Single cell preparation and magnetic cell sorting (MACS) 8 Organoid dissociation was performed as described before.21 MACS was used to enrich for the 9 CD11b+ fraction (Miltenyi, 130049601) according to manufacturer’s protocol. .. The CD11b+ cell 10 fraction was collected in microglia culture medium (RPMI 1640 (Life technologies, 21875034), 11 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker, 12 Belgium) and 100 ng/ml IL-34 (Pepro Tech, 200-34)).

    Single Cell:

    Article Title: PU.1 restores microglial dysfunction caused by C9ORF72 repeat expansions in neural organoids.
    Article Snippet: .. Neural organoids were generated using a modified version of a previously 5 published protocol.22 6 7 Single cell preparation and magnetic cell sorting (MACS) 8 Organoid dissociation was performed as described before.21 MACS was used to enrich for the 9 CD11b+ fraction (Miltenyi, 130049601) according to manufacturer’s protocol. .. The CD11b+ cell 10 fraction was collected in microglia culture medium (RPMI 1640 (Life technologies, 21875034), 11 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin (BioWhittaker, 12 Belgium) and 100 ng/ml IL-34 (Pepro Tech, 200-34)).



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    F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. <t>CD11b</t> + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.
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    F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. CD11b + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Lipid nanoparticle-encapsulated microRNA-192: An anti-inflammatory adjuvant that enhances vaccine efficacy in aged mice

    doi: 10.1016/j.omtn.2025.102784

    Figure Lengend Snippet: F-LNP-miR-192 downregulates STAT1 and STAT3 in vivo (A, B) RAW264.7 cells were treated with mock, 5 μL of F-LNP-NC (100 pmol), or 5 μL of F-LNP-miR-192 (100 pmol) for 24 h, followed by stimulation with 200 ng/mL of LPS for 12 h. Total RNA was extracted, and STAT1 mRNA levels were measured via RT-qPCR, normalized to GAPDH levels (A). Cell lysates prepared from stimulated cells were subjected to SDS-PAGE, and proteins were detected with the indicated antibodies (B). (C) GSEA was performed for the cytokine and cytokine receptor pathway, as well as the cellular senescence pathway, with normalized enrichment scores (NES) and each p value was calculated. (D) SV (80 μL) was mixed with 20 μL of F-LNP-NC, F-LNP-miR-192, or PBS and then subcutaneously injected into young BALB/c mice. Sera were collected 24 h after injection, and serum IL-6 levels were measured using ELISA. (E) F-LNP-miR-192 (20 μL) was subcutaneously injected into young mice, and then the skin tissues were excised on day 0, 1, 2, or 3 post-injection. CD11b + and CD11b − cells were sorted using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Article Snippet: Cells were washed with magnetic-activating cell sorting (MACS) buffer and separated into with CD11b + and CD11b − fractions using MicroBeads (Miltenyi) and MS column.

    Techniques: In Vivo, Quantitative RT-PCR, SDS Page, Injection, Enzyme-linked Immunosorbent Assay, Western Blot

    LNP-miR-192 improves vaccine efficacy in aged mice (A and B) K-LNP-empty, K-LNP-miR-192, F-LNP-NC, or F-LNP-miR-192 or S-LNP-miR-192 (20 μL of each) was mixed with SV (80 μL) and subcutaneously injected into aged BALB/c mice ( n = 6). A second dose of the same vaccine was administered 3 weeks later. Sera were collected 4 weeks after the first-dose vaccination, and serum HI titers for influenza type A and B were determined. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001: Log-transformed data were analyzed by Welch’s one-way ANOVA (A) and Welch’s t test (B). (C) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) mixed with 20 μL of the indicated LNPs. Four weeks after the first vaccination, sera were collected and HI titers were measured. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05: Kruskal-Wallis test with Dunn’s multiple comparisons test. (D) Aged BALB/c mice were subcutaneously vaccinated with SV (80 μL) mixed with F-LNP-miR-192 (20 μL). Two weeks later, draining lymph nodes were excised, and the number of GC B cells was measured via flow cytometry. Data are means ± SDs, with dots representing individual mice. ∗∗ p < 0.01; t test. (E and F) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) alone or SV formulated with 20 μL of F-LNP-miR-192 or F-LNP-NC, as indicated. Four weeks after the first vaccination, mice were challenged with influenza A virus and monitored daily for survival. Mice losing >25% of their initial body weight were euthanized and recorded as dead. (E) Survival curves for mice vaccinated with SV alone or SV combined with F-LNP-miR-192. A significant trend in survival across all four groups was detected via the log rank test for trends ( p < 0.05). (F) Survival comparison between the SV + F-LNP-miR-192 and SV + F-LNP-NC groups, with a significant difference in survival observed (log rank test, p < 0.05). (G) SV (80 μL) was mixed with one of the indicated LNPs (20 μL) and subcutaneously injected into aged mice. Sera were collected 24 and 48 h post-injection, and serum IL-6 concentrations were measured using ELISA. Data are means ± SDs (n = 3, nonsignificant [NS] > 0.05: one-way ANOVA). (H) Aged mice were subcutaneously vaccinated using the WVP vaccine (80 μL) of influenza A virus with PBS or F-LNP-miR-192 twice at a 2-week interval. One month after the first vaccination, sera were collected, and HI and neutralization titers (NT) were determined. Data are means ± SDs. (n = 5, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001: log-transformed data were analyzed by one-way ANOVA). (I, J) 20 μL of F-LNP was subcutaneously injected into aged mice. The skin tissues were excised at 2 days after infection (I) or at indicated days (J), and CD11b + cells were isolated using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Lipid nanoparticle-encapsulated microRNA-192: An anti-inflammatory adjuvant that enhances vaccine efficacy in aged mice

    doi: 10.1016/j.omtn.2025.102784

    Figure Lengend Snippet: LNP-miR-192 improves vaccine efficacy in aged mice (A and B) K-LNP-empty, K-LNP-miR-192, F-LNP-NC, or F-LNP-miR-192 or S-LNP-miR-192 (20 μL of each) was mixed with SV (80 μL) and subcutaneously injected into aged BALB/c mice ( n = 6). A second dose of the same vaccine was administered 3 weeks later. Sera were collected 4 weeks after the first-dose vaccination, and serum HI titers for influenza type A and B were determined. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001: Log-transformed data were analyzed by Welch’s one-way ANOVA (A) and Welch’s t test (B). (C) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) mixed with 20 μL of the indicated LNPs. Four weeks after the first vaccination, sera were collected and HI titers were measured. Data are means ± SEMs, with dots representing individual mice. ∗ p < 0.05: Kruskal-Wallis test with Dunn’s multiple comparisons test. (D) Aged BALB/c mice were subcutaneously vaccinated with SV (80 μL) mixed with F-LNP-miR-192 (20 μL). Two weeks later, draining lymph nodes were excised, and the number of GC B cells was measured via flow cytometry. Data are means ± SDs, with dots representing individual mice. ∗∗ p < 0.01; t test. (E and F) Aged BALB/c mice were subcutaneously vaccinated once or twice with SV (80 μL) alone or SV formulated with 20 μL of F-LNP-miR-192 or F-LNP-NC, as indicated. Four weeks after the first vaccination, mice were challenged with influenza A virus and monitored daily for survival. Mice losing >25% of their initial body weight were euthanized and recorded as dead. (E) Survival curves for mice vaccinated with SV alone or SV combined with F-LNP-miR-192. A significant trend in survival across all four groups was detected via the log rank test for trends ( p < 0.05). (F) Survival comparison between the SV + F-LNP-miR-192 and SV + F-LNP-NC groups, with a significant difference in survival observed (log rank test, p < 0.05). (G) SV (80 μL) was mixed with one of the indicated LNPs (20 μL) and subcutaneously injected into aged mice. Sera were collected 24 and 48 h post-injection, and serum IL-6 concentrations were measured using ELISA. Data are means ± SDs (n = 3, nonsignificant [NS] > 0.05: one-way ANOVA). (H) Aged mice were subcutaneously vaccinated using the WVP vaccine (80 μL) of influenza A virus with PBS or F-LNP-miR-192 twice at a 2-week interval. One month after the first vaccination, sera were collected, and HI and neutralization titers (NT) were determined. Data are means ± SDs. (n = 5, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001: log-transformed data were analyzed by one-way ANOVA). (I, J) 20 μL of F-LNP was subcutaneously injected into aged mice. The skin tissues were excised at 2 days after infection (I) or at indicated days (J), and CD11b + cells were isolated using MACS beads. Total protein content in each sample was adjusted to 30 μg, subjected to SDS-PAGE, and proteins were detected via western blotting using indicated antibodies.

    Article Snippet: Cells were washed with magnetic-activating cell sorting (MACS) buffer and separated into with CD11b + and CD11b − fractions using MicroBeads (Miltenyi) and MS column.

    Techniques: Injection, Transformation Assay, Flow Cytometry, Virus, Comparison, Enzyme-linked Immunosorbent Assay, Neutralization, Infection, Isolation, SDS Page, Western Blot