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Peptron Inc peptide olp pool
A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation <t>of</t> <t>splenocytes.</t> Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV <t>OLP</t> peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.
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1) Product Images from "Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP"

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP

Journal: NPJ Vaccines

doi: 10.1038/s41541-026-01385-0

A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation of splenocytes. Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV OLP peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.
Figure Legend Snippet: A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation of splenocytes. Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV OLP peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.

Techniques Used: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison, Control, MANN-WHITNEY, In Vitro, Activation Assay, Staining

Sera were collected either before or after the challenge at 1, 3, and 7 dpi. Mock groups or pre-challenged mouse groups were used as controls for comparison. A Antibody responses were assessed by quantifying IgG1, IgG2b, IgG3, IgA, and IgM titers in BALB/c (left), C57BL/6 (middle), and IFNAR – / – (right) mice using immunoassays such as bead-based assay and P-ELISA. Data are means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with pre-challenge groups by Two-way ANOVA test with Holm–Sidak’s multiple comparisons. Comparison with the mock group was done using an unpaired t-test (Mann–Whitney test). B Splenocytes were collected from BALB/c, C57BL/6, and IFNAR – / – mice that were immunized with either mock or HSAmvac, before or after the challenge at 1, 3, and 7 dpi. Either mock groups or pre-challenged mouse groups were used as control comparisons. Cellular cytokine responses in CD4⁺ and CD8⁺ T cells to the SFTSV OLP peptide pool were evaluated by performing intracellular cytokine staining with activated splenocytes, followed by flow cytometry. Pre- and post-challenge levels of IFN-γ, IL-2, IL-4, and TNF-α were determined for CD4 ⁺ and CD8 ⁺ T cells. Data are presented as means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.001. Two-way ANOVA test with Holm–Sidak multiple comparison test, as com p ared to the corresponding control (Ctrl). Comparison with the low-dose group was done using an unpaired t-test (Mann–Whitney test).
Figure Legend Snippet: Sera were collected either before or after the challenge at 1, 3, and 7 dpi. Mock groups or pre-challenged mouse groups were used as controls for comparison. A Antibody responses were assessed by quantifying IgG1, IgG2b, IgG3, IgA, and IgM titers in BALB/c (left), C57BL/6 (middle), and IFNAR – / – (right) mice using immunoassays such as bead-based assay and P-ELISA. Data are means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with pre-challenge groups by Two-way ANOVA test with Holm–Sidak’s multiple comparisons. Comparison with the mock group was done using an unpaired t-test (Mann–Whitney test). B Splenocytes were collected from BALB/c, C57BL/6, and IFNAR – / – mice that were immunized with either mock or HSAmvac, before or after the challenge at 1, 3, and 7 dpi. Either mock groups or pre-challenged mouse groups were used as control comparisons. Cellular cytokine responses in CD4⁺ and CD8⁺ T cells to the SFTSV OLP peptide pool were evaluated by performing intracellular cytokine staining with activated splenocytes, followed by flow cytometry. Pre- and post-challenge levels of IFN-γ, IL-2, IL-4, and TNF-α were determined for CD4 ⁺ and CD8 ⁺ T cells. Data are presented as means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.001. Two-way ANOVA test with Holm–Sidak multiple comparison test, as com p ared to the corresponding control (Ctrl). Comparison with the low-dose group was done using an unpaired t-test (Mann–Whitney test).

Techniques Used: Comparison, Bead-based Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control, Staining, Flow Cytometry

Related Articles

In Vitro:

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP
Article Snippet: .. Splenocytes were restimulated in vitro with a commercially synthesized overlapping peptide (OLP) pool covering the full-length nucleoprotein (N) of SFTSV (Peptron, Inc., Korea) at a final concentration of 2 μg/mL per peptide. ..

Synthesized:

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP
Article Snippet: .. Splenocytes were restimulated in vitro with a commercially synthesized overlapping peptide (OLP) pool covering the full-length nucleoprotein (N) of SFTSV (Peptron, Inc., Korea) at a final concentration of 2 μg/mL per peptide. ..

Concentration Assay:

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP
Article Snippet: .. Splenocytes were restimulated in vitro with a commercially synthesized overlapping peptide (OLP) pool covering the full-length nucleoprotein (N) of SFTSV (Peptron, Inc., Korea) at a final concentration of 2 μg/mL per peptide. ..



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A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation <t>of</t> <t>splenocytes.</t> Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV <t>OLP</t> peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.
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A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation <t>of</t> <t>splenocytes.</t> Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV <t>OLP</t> peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.
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A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation <t>of</t> <t>splenocytes.</t> Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV <t>OLP</t> peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.
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A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation of splenocytes. Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV OLP peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.

Journal: NPJ Vaccines

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP

doi: 10.1038/s41541-026-01385-0

Figure Lengend Snippet: A . Immunoglobulin level analysis. Total levels of IgG1, IgG2b, IgG3, IgA, and IgM in BALB/c (top), C57BL/6 (middle), and IFNAR – / – (bottom) mice were examined using flow cytometry or P-ELISA before and after receiving either 1 or 10 µg of HSAmvac on days 0, 14, 28, and 42. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, ** p < 0.001, and **** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test was performed when comparing the low-dose (1 µg) group. B Neutralizing antibody titers (FRNT₅₀) were measured on days 0, 14, and 28 in BALB/c, C57BL/6, and IFNAR⁻/⁻ mice vaccinated with either 1 µg or 10 µg of HSAmvac. Data are presented as means ± SD for three mice. Statistical analysis was performed using a two-way or one-way ANOVA with Holm–Sidak multiple comparison test, comparing results to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison to the group that received 1 µg of HSAmvac. C BALB/c, C57BL/6, and IFNAR – / – mice received a prime–booster immunization with HSAmvac at 2-week intervals. The SFTSV antigen-specific cytokine response was assessed following in vitro activation of splenocytes. Cells were restimulated with an overlapping peptide pool (2 μg/mL per peptide) spanning the SFTSV nucleoprotein (N) using the SFTSV OLP peptide pool. Cells were harvested from mice pre- and post-SFTSV challenge for intracellular cytokine staining. The percentage of cells producing cytokines, including IFN-γ, IL-2, IL-4, and TNF-α, was determined by flow cytometry. Data are presented as means ± SD for three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and ** p < 0.0001. A two-way or one-way ANOVA with Holm–Sidak multiple comparison test was used, as compared to the corresponding control (Ctrl; day 0). An unpaired t-test (Mann–Whitney test) was used for comparison with the group that received 1 µg of HSAmvac.

Article Snippet: Splenocytes were restimulated in vitro with a commercially synthesized overlapping peptide (OLP) pool covering the full-length nucleoprotein (N) of SFTSV (Peptron, Inc., Korea) at a final concentration of 2 μg/mL per peptide.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison, Control, MANN-WHITNEY, In Vitro, Activation Assay, Staining

Sera were collected either before or after the challenge at 1, 3, and 7 dpi. Mock groups or pre-challenged mouse groups were used as controls for comparison. A Antibody responses were assessed by quantifying IgG1, IgG2b, IgG3, IgA, and IgM titers in BALB/c (left), C57BL/6 (middle), and IFNAR – / – (right) mice using immunoassays such as bead-based assay and P-ELISA. Data are means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with pre-challenge groups by Two-way ANOVA test with Holm–Sidak’s multiple comparisons. Comparison with the mock group was done using an unpaired t-test (Mann–Whitney test). B Splenocytes were collected from BALB/c, C57BL/6, and IFNAR – / – mice that were immunized with either mock or HSAmvac, before or after the challenge at 1, 3, and 7 dpi. Either mock groups or pre-challenged mouse groups were used as control comparisons. Cellular cytokine responses in CD4⁺ and CD8⁺ T cells to the SFTSV OLP peptide pool were evaluated by performing intracellular cytokine staining with activated splenocytes, followed by flow cytometry. Pre- and post-challenge levels of IFN-γ, IL-2, IL-4, and TNF-α were determined for CD4 ⁺ and CD8 ⁺ T cells. Data are presented as means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.001. Two-way ANOVA test with Holm–Sidak multiple comparison test, as com p ared to the corresponding control (Ctrl). Comparison with the low-dose group was done using an unpaired t-test (Mann–Whitney test).

Journal: NPJ Vaccines

Article Title: Development of a Prophylactic mRNA Vaccine for Severe Fever with Thrombocytopenia Syndrome Using HSA based LNP

doi: 10.1038/s41541-026-01385-0

Figure Lengend Snippet: Sera were collected either before or after the challenge at 1, 3, and 7 dpi. Mock groups or pre-challenged mouse groups were used as controls for comparison. A Antibody responses were assessed by quantifying IgG1, IgG2b, IgG3, IgA, and IgM titers in BALB/c (left), C57BL/6 (middle), and IFNAR – / – (right) mice using immunoassays such as bead-based assay and P-ELISA. Data are means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with pre-challenge groups by Two-way ANOVA test with Holm–Sidak’s multiple comparisons. Comparison with the mock group was done using an unpaired t-test (Mann–Whitney test). B Splenocytes were collected from BALB/c, C57BL/6, and IFNAR – / – mice that were immunized with either mock or HSAmvac, before or after the challenge at 1, 3, and 7 dpi. Either mock groups or pre-challenged mouse groups were used as control comparisons. Cellular cytokine responses in CD4⁺ and CD8⁺ T cells to the SFTSV OLP peptide pool were evaluated by performing intracellular cytokine staining with activated splenocytes, followed by flow cytometry. Pre- and post-challenge levels of IFN-γ, IL-2, IL-4, and TNF-α were determined for CD4 ⁺ and CD8 ⁺ T cells. Data are presented as means ± SD of three mice. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.001. Two-way ANOVA test with Holm–Sidak multiple comparison test, as com p ared to the corresponding control (Ctrl). Comparison with the low-dose group was done using an unpaired t-test (Mann–Whitney test).

Article Snippet: Splenocytes were restimulated in vitro with a commercially synthesized overlapping peptide (OLP) pool covering the full-length nucleoprotein (N) of SFTSV (Peptron, Inc., Korea) at a final concentration of 2 μg/mL per peptide.

Techniques: Comparison, Bead-based Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control, Staining, Flow Cytometry

( A ) Correlations between the peak S1 IgG Ab titer and T cell responses to spike protein (left) or N protein (right) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 75). ( B ) Top panel; Cumulative magnitude of the T cell response to spike and N proteins, mapped epitope peptide (MEP and MEP2) panels and overlapping peptide (OLP) panels (top panel) ordered by increasing magnitude of nAb response (bottom panel) or ( C ) Magnitude of nAb response (top panel) ordered by increasing cumulative magnitude of T cell response to spike and N proteins, MEP/MEP2 panels and OLP panels (bottom panel) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 70). HCW with no nAb (IC50 titer less than 50) are indicated by black arrows. + and * denote individuals with no T cell response to any protein or peptide pool. ( D ) Proportion of HCW with a nAb titer (IC50) or T cell response to spike and N proteins within given ranges stratified by symptom group; ≥1 COVID-19 case definition symptoms (n = 49 or 48), non-case definition symptoms (n = 19) or asymptomatic (n = 8) ( A ) Spearman’s rank correlation. HCW, health care workers; M, Membrane; ORF, open reading frame; N, nucleocapsid; nAb, neutralizing antibody; S1, spike subunit 1; SFC, spot forming cells per 10 6 PBMC.

Journal: Science Immunology

Article Title: Discordant neutralizing antibody and T cell responses in asymptomatic and mild SARS-CoV-2 infection

doi: 10.1126/sciimmunol.abf3698

Figure Lengend Snippet: ( A ) Correlations between the peak S1 IgG Ab titer and T cell responses to spike protein (left) or N protein (right) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 75). ( B ) Top panel; Cumulative magnitude of the T cell response to spike and N proteins, mapped epitope peptide (MEP and MEP2) panels and overlapping peptide (OLP) panels (top panel) ordered by increasing magnitude of nAb response (bottom panel) or ( C ) Magnitude of nAb response (top panel) ordered by increasing cumulative magnitude of T cell response to spike and N proteins, MEP/MEP2 panels and OLP panels (bottom panel) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 70). HCW with no nAb (IC50 titer less than 50) are indicated by black arrows. + and * denote individuals with no T cell response to any protein or peptide pool. ( D ) Proportion of HCW with a nAb titer (IC50) or T cell response to spike and N proteins within given ranges stratified by symptom group; ≥1 COVID-19 case definition symptoms (n = 49 or 48), non-case definition symptoms (n = 19) or asymptomatic (n = 8) ( A ) Spearman’s rank correlation. HCW, health care workers; M, Membrane; ORF, open reading frame; N, nucleocapsid; nAb, neutralizing antibody; S1, spike subunit 1; SFC, spot forming cells per 10 6 PBMC.

Article Snippet: For experiments involving T cell stimulation with OLP peptide pools and spike MEP2 pool ELISpot plates (Merck-Millipore, MSIP4510) were coated with human anti-IFNγ Ab (1-D1K, Mabtech; 10μg/ml) in PBS overnight at 4°C.

Techniques: Infection

( A-D ) Top panels; Magnitude of the T cell response to spike protein (n = 75) ( A ) Cumulative magnitude of T cell responses to spike protein and spike mapped epitope peptide (MEP and MEP2) pools (n = 70) ( B ) N protein and N, M and ORF3a/7a MEP pools (n = 75) ( C ) or N1, N2, M and ORF3a overlapping peptide (OLP) pools (n = 70) ( D ) ordered by increasing cumulative magnitude of T cell responses in HCW with laboratory-confirmed SARS-CoV-2 infection. Bottom panels; nAb titers (IC50) in HCW with laboratory-confirmed SARS-CoV-2 infection, ordered by corresponding top panel. ( E ) The number of reactive SARS-CoV-2 proteins or peptide pools (top panel) and nAb titer (IC50; bottom panel) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 70). Top panel ordered by cumulative magnitude; bottom panel ordered by top panel. HCW with no nAb (IC50 titer less than 50) are indicated by black arrows. + and * denote two individuals with no T cell response to any protein or peptide pool. HCW, healthcare workers; M, Membrane; N, nucleocapsid; nAb, neutralizing antibody; ORF, open reading frame; SFC, spot forming cells per 10 6 PBMC.

Journal: Science Immunology

Article Title: Discordant neutralizing antibody and T cell responses in asymptomatic and mild SARS-CoV-2 infection

doi: 10.1126/sciimmunol.abf3698

Figure Lengend Snippet: ( A-D ) Top panels; Magnitude of the T cell response to spike protein (n = 75) ( A ) Cumulative magnitude of T cell responses to spike protein and spike mapped epitope peptide (MEP and MEP2) pools (n = 70) ( B ) N protein and N, M and ORF3a/7a MEP pools (n = 75) ( C ) or N1, N2, M and ORF3a overlapping peptide (OLP) pools (n = 70) ( D ) ordered by increasing cumulative magnitude of T cell responses in HCW with laboratory-confirmed SARS-CoV-2 infection. Bottom panels; nAb titers (IC50) in HCW with laboratory-confirmed SARS-CoV-2 infection, ordered by corresponding top panel. ( E ) The number of reactive SARS-CoV-2 proteins or peptide pools (top panel) and nAb titer (IC50; bottom panel) in HCW with laboratory-confirmed SARS-CoV-2 infection (n = 70). Top panel ordered by cumulative magnitude; bottom panel ordered by top panel. HCW with no nAb (IC50 titer less than 50) are indicated by black arrows. + and * denote two individuals with no T cell response to any protein or peptide pool. HCW, healthcare workers; M, Membrane; N, nucleocapsid; nAb, neutralizing antibody; ORF, open reading frame; SFC, spot forming cells per 10 6 PBMC.

Article Snippet: For experiments involving T cell stimulation with OLP peptide pools and spike MEP2 pool ELISpot plates (Merck-Millipore, MSIP4510) were coated with human anti-IFNγ Ab (1-D1K, Mabtech; 10μg/ml) in PBS overnight at 4°C.

Techniques: Infection