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( A ) The 5- to six-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either BC mRNA vaccine (0.05 μg; i.m.) or Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 adjuvanted with 50 μg poly I:C; i.n.). ( B – D ) At 1 month after boost, blood, BALF, and NLF were collected. Serum ( B ), BALF ( C ), and NLF ( D ) nAb titers against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 were determined by multiplex sVNT. ( E – H ) Spleen, lung, and NALT were harvested at 2 and 3 weeks after boost. ( E ) Percentage of ancestral SARS-CoV-2 RBD + swIg + B cells in spleen, lung, and NALT at 3 weeks after boost was determined by flow cytometry. Cross-reactivity of antigen-specific B cells toward XBB.1.5 and SARS-CoV-1 RBD is also shown. ( F – H ) Frequency of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( F ), lung cells ( G ), and pooled NALT cells ( H ) at 2 weeks after boost was determined by <t>FluoroSpot</t> upon restimulation with ancestral SARS-CoV-2, XBB.1.5, JN.1, RaTG13, Gx-P5L, SARS-CoV-1, and WIV-1 RBD peptides. Data in B – H are from 1 representative experiment performed twice with similar results; n = 4–6 per group/experiment. Symbols in B – E represent individual animals, and data shown are geometric means ( B and C ) and means ( D – H ) ± SD ( E – G ). Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( B , C , and E ) and Kruskal-Wallis test ( D and F – H ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in F and G represent statistical significance between corresponding groups.
Fluorospot Readers, supplied by Mabtech Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) The 5- to six-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either BC mRNA vaccine (0.05 μg; i.m.) or Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 adjuvanted with 50 μg poly I:C; i.n.). ( B – D ) At 1 month after boost, blood, BALF, and NLF were collected. Serum ( B ), BALF ( C ), and NLF ( D ) nAb titers against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 were determined by multiplex sVNT. ( E – H ) Spleen, lung, and NALT were harvested at 2 and 3 weeks after boost. ( E ) Percentage of ancestral SARS-CoV-2 RBD + swIg + B cells in spleen, lung, and NALT at 3 weeks after boost was determined by flow cytometry. Cross-reactivity of antigen-specific B cells toward XBB.1.5 and SARS-CoV-1 RBD is also shown. ( F – H ) Frequency of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( F ), lung cells ( G ), and pooled NALT cells ( H ) at 2 weeks after boost was determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, JN.1, RaTG13, Gx-P5L, SARS-CoV-1, and WIV-1 RBD peptides. Data in B – H are from 1 representative experiment performed twice with similar results; n = 4–6 per group/experiment. Symbols in B – E represent individual animals, and data shown are geometric means ( B and C ) and means ( D – H ) ± SD ( E – G ). Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( B , C , and E ) and Kruskal-Wallis test ( D and F – H ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in F and G represent statistical significance between corresponding groups.

Journal: The Journal of Clinical Investigation

Article Title: Intranasal DC-targeting vaccine booster elicits durable and cross-clade protective immunity against sarbecoviruses in mice

doi: 10.1172/JCI195784

Figure Lengend Snippet: ( A ) The 5- to six-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either BC mRNA vaccine (0.05 μg; i.m.) or Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 adjuvanted with 50 μg poly I:C; i.n.). ( B – D ) At 1 month after boost, blood, BALF, and NLF were collected. Serum ( B ), BALF ( C ), and NLF ( D ) nAb titers against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 were determined by multiplex sVNT. ( E – H ) Spleen, lung, and NALT were harvested at 2 and 3 weeks after boost. ( E ) Percentage of ancestral SARS-CoV-2 RBD + swIg + B cells in spleen, lung, and NALT at 3 weeks after boost was determined by flow cytometry. Cross-reactivity of antigen-specific B cells toward XBB.1.5 and SARS-CoV-1 RBD is also shown. ( F – H ) Frequency of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( F ), lung cells ( G ), and pooled NALT cells ( H ) at 2 weeks after boost was determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, JN.1, RaTG13, Gx-P5L, SARS-CoV-1, and WIV-1 RBD peptides. Data in B – H are from 1 representative experiment performed twice with similar results; n = 4–6 per group/experiment. Symbols in B – E represent individual animals, and data shown are geometric means ( B and C ) and means ( D – H ) ± SD ( E – G ). Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( B , C , and E ) and Kruskal-Wallis test ( D and F – H ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in F and G represent statistical significance between corresponding groups.

Article Snippet: Spots were enumerated using the Mabtech IRIS ELISpot and FluoroSpot readers, and analysis was performed using Mabtech Apex software version 1.1.5.74.

Techniques: Multiplex Assay, Flow Cytometry, MANN-WHITNEY, Comparison

( A ) The 5- to 6-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 ) (i.n. or s.c.) or with an equivalent antigen dose of purified XBB.1.5 (1 μg) and SARS-CoV-1 (0.25 μg) rRBD, in combination with Clec9A-M2e (3.75 μg) (NTC). All the formulations were adjuvanted with 50 μg poly I:C. ( B – F ) Blood, BALF, and NLF were obtained at 1 month after boost. Serum ( B ), BALF ( C ), and NLF ( D ) nAb titers against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 were determined by multiplex sVNT. BALF ( E ) and NLF ( F ) anti-RBD IgA response against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 RBD was determined by ELISA. ( G – I ) Spleen, lung, and NALT were harvested at 2 weeks after boost. Frequency of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( G ), lung cells ( H ), and pooled NALT cells ( I ) was determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, RaTG13, and SARS-CoV-1 RBD peptides. Data in B – I are from 1 representative experiment performed twice with similar results; n = 4 per group/experiment. Symbols in B – F represent individual animals, and data shown are geometric means ( B and C ) and means ( D – I ) ± SD ( G and H ). Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( B and C ) and Kruskal-Wallis test ( D – H ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in G and H represent statistical significance between corresponding groups.

Journal: The Journal of Clinical Investigation

Article Title: Intranasal DC-targeting vaccine booster elicits durable and cross-clade protective immunity against sarbecoviruses in mice

doi: 10.1172/JCI195784

Figure Lengend Snippet: ( A ) The 5- to 6-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 ) (i.n. or s.c.) or with an equivalent antigen dose of purified XBB.1.5 (1 μg) and SARS-CoV-1 (0.25 μg) rRBD, in combination with Clec9A-M2e (3.75 μg) (NTC). All the formulations were adjuvanted with 50 μg poly I:C. ( B – F ) Blood, BALF, and NLF were obtained at 1 month after boost. Serum ( B ), BALF ( C ), and NLF ( D ) nAb titers against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 were determined by multiplex sVNT. BALF ( E ) and NLF ( F ) anti-RBD IgA response against ancestral SARS-CoV-2, XBB.1.5, and SARS-CoV-1 RBD was determined by ELISA. ( G – I ) Spleen, lung, and NALT were harvested at 2 weeks after boost. Frequency of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( G ), lung cells ( H ), and pooled NALT cells ( I ) was determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, RaTG13, and SARS-CoV-1 RBD peptides. Data in B – I are from 1 representative experiment performed twice with similar results; n = 4 per group/experiment. Symbols in B – F represent individual animals, and data shown are geometric means ( B and C ) and means ( D – I ) ± SD ( G and H ). Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( B and C ) and Kruskal-Wallis test ( D – H ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in G and H represent statistical significance between corresponding groups.

Article Snippet: Spots were enumerated using the Mabtech IRIS ELISpot and FluoroSpot readers, and analysis was performed using Mabtech Apex software version 1.1.5.74.

Techniques: Purification, Multiplex Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Comparison

( A ) The 5- to 6-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either BC mRNA vaccine (0.05 μg; i.m.) or Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 adjuvanted with 50 μg poly I:C; i.n.). ( B – E ) Spleen, BM from femur/tibia, and lung were harvested at 6 months after boost. ( B and C ) Percentages of AIM + Tfh ( B ) and RBD + GC ( C ) B cells in spleen and lung were determined by flow cytometry. ( D and E ) Frequency of BM RBD-specific IgG + LLPCs and non-LLPCs (normalized to total IgG) ( D ) and pooled lung RBD-specific IgG + and IgA + LLPCs and non-LLPCs (normalized to total IgG and IgA) ( E ) was determined by B cell ELISpot. ( F – J ) Spleen, lung, and NALT were harvested at 4 months after boost. ( F – H ) Frequencies of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( F ), lung cells ( G ), and pooled NALT cells ( H ) were determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, JN.1, RaTG13, Gx-P5L, SARS-CoV-1, and WIV-1 RBD peptides. ( I and J ) Percentages of lung ( I ) and NALT ( J ) CD4 + and CD8 + T RM cells were determined by flow cytometry. Data in B – J are from 1 representative experiment performed twice with similar results; n = 4–5 per group/experiment. Symbols in B – E , I , and J represent individual animals, and data shown in B – J are means. Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( D ) and Kruskal-Wallis test ( B , C , F , G , I , and J ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in F and G represent statistical significance between corresponding groups.

Journal: The Journal of Clinical Investigation

Article Title: Intranasal DC-targeting vaccine booster elicits durable and cross-clade protective immunity against sarbecoviruses in mice

doi: 10.1172/JCI195784

Figure Lengend Snippet: ( A ) The 5- to 6-week-old BALB/c mice were immunized with original Comirnaty mRNA vaccine as described in Figure 1. At 3 months after the second immunization dose, mice were boosted with either BC mRNA vaccine (0.05 μg; i.m.) or Clec9A OMNI (4 μg Clec9A XBB + 1 μg Clec9A CoV1 adjuvanted with 50 μg poly I:C; i.n.). ( B – E ) Spleen, BM from femur/tibia, and lung were harvested at 6 months after boost. ( B and C ) Percentages of AIM + Tfh ( B ) and RBD + GC ( C ) B cells in spleen and lung were determined by flow cytometry. ( D and E ) Frequency of BM RBD-specific IgG + LLPCs and non-LLPCs (normalized to total IgG) ( D ) and pooled lung RBD-specific IgG + and IgA + LLPCs and non-LLPCs (normalized to total IgG and IgA) ( E ) was determined by B cell ELISpot. ( F – J ) Spleen, lung, and NALT were harvested at 4 months after boost. ( F – H ) Frequencies of IFN-γ–, IL-2–, and/or TNF-α–secreting splenocytes ( F ), lung cells ( G ), and pooled NALT cells ( H ) were determined by FluoroSpot upon restimulation with ancestral SARS-CoV-2, XBB.1.5, JN.1, RaTG13, Gx-P5L, SARS-CoV-1, and WIV-1 RBD peptides. ( I and J ) Percentages of lung ( I ) and NALT ( J ) CD4 + and CD8 + T RM cells were determined by flow cytometry. Data in B – J are from 1 representative experiment performed twice with similar results; n = 4–5 per group/experiment. Symbols in B – E , I , and J represent individual animals, and data shown in B – J are means. Statistical analysis: nonparametric 2-tailed Mann-Whitney test ( D ) and Kruskal-Wallis test ( B , C , F , G , I , and J ) with Dunn’s multiple-comparison test. * P < 0.05, ** P < 0.01. Asterisk colors in F and G represent statistical significance between corresponding groups.

Article Snippet: Spots were enumerated using the Mabtech IRIS ELISpot and FluoroSpot readers, and analysis was performed using Mabtech Apex software version 1.1.5.74.

Techniques: Flow Cytometry, Enzyme-linked Immunospot, MANN-WHITNEY, Comparison