Journal: Nature Communications

Article Title: Antibodies targeting the Crimean-Congo Hemorrhagic Fever Virus nucleoprotein protect via TRIM21

doi: 10.1038/s41467-024-53362-7

Figure Lengend Snippet: On D5 p.i., groups of mice ( N = 6) from Fig. were euthanized and evaluated for cellular immune responses to infection via ( a ) IFNу ELISpot shown as cumulative responses against peptides spanning the entire CCHFV NP (SFC: spot forming cells). For T-cell depletion study, WT C57BL6/J mice were ( b ) vaccinated with Sham or repNP RNA on day −28 relative to lethal CCHFV challenge. On days −5, −2, and +5 relative to CCHFV challenge, mice were treated with isotype or αCD4 and αCD8 antibody to deplete mice of T-cell populations. On D0, groups of mice were evaluated for immunological response to vaccine or treated with MAR1−5A3 antibody and infected with a lethal dose of 100 TCID 50 CCHFV strain UG3010. Mice ( N = 8) were ( c ) weighed daily and monitored for ( d ) survival until day 14 p.i., On D5 p.i., groups of mice ( N = 6) were evaluated for ( e ) depletion of CD4+ and CD8 + T-cell populations via Flow Cytometry. Antibody treatment achieved a 96.3% depletion of CD4 + T-cells and 98.5% depletion of CD8 + T-cells. In the second study, groups of WT C57BL6/J and CD8 −/− mice were vaccinated and infected as above. Mice ( N = 8) were ( f ) weighed daily and monitored for ( g ) survival until day 14 p.i. Dashed lines indicate limit of detection. Data shown as mean plus standard deviation. Significance was calculated using one-way ANOVA; ns P > 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Exact p -values: ( d ) both P = 0.0002, ( g ) both P = 0.0002.

Article Snippet: To measure CCHFV-specific T-cell responses, an IFNγ ELISpot kit (ImmunoSpot) was used with peptides spanning the entire CCHFV Hoti NP (GenScript).

Techniques: Infection, Enzyme-linked Immunospot, Flow Cytometry, Standard Deviation

Journal: NPJ Vaccines

Article Title: Replicon particle vaccination induces non-neutralizing anti-nucleoprotein antibody-mediated control of Crimean-Congo hemorrhagic fever virus

doi: 10.1038/s41541-024-00877-1

Figure Lengend Snippet: a Cytokine/chemokine responses in mice vaccinated with CCHF VRP, LASV VRP, or left unvaccinated (no VRP, given DMEM alone) 3 days prior to challenge and euthanized 3 or 6 days post infection (dpi) were analyzed using the ProcartaPlex Mouse Th1/Th2 Cytokine and Chemokine panel and 25 µL mouse plasma. b CCHFV-specific T-cell responses were evaluated using IFN-gamma ELISpot assay. Peptides covering the CCHFV IbAr10200 NP or Oman-98 GPC NSm-Gc domain (numbers represent aa positions) were used to stimulate splenocytes harvested from vaccinated animals. Data are reported as the number of spot-forming cells (SPC)/1.00 x 10 6 cells. c CCHFV-specific NP, Gn, Gc, and GP38 antibody responses (IgM and IgG) were evaluated via ELISA and are reported as endpoint dilution titers. Antibody titers, T-cell responses, and cytokine/chemokine levels were compared statistically between vaccine groups by timepoint (3 or 6 dpi) using multiple two-tailed t -tests (Mann-Whitney) and only statistically significant results are reported; * p < 0.5, ** p < 0.01. Individual animals are represented. Bars and error bars indicate mean ± SEM.

Article Snippet: Cell suspensions were frozen, thawed in RPMI, diluted to 2 × 10 6 cells/mL, and seeded in MabTech ELISpot plates containing peptides homologous to the VRP vaccine construct, covering the CCHFV IbAr10200 NP (AbClonal; 15-mers with 11 AA overlap) or Oman-98 GPC NSm-Gc domain (AbClonal; 15-mers with 4 AA overlap).

Techniques: Infection, Ifn? elispot, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

Journal: NPJ Vaccines

Article Title: Replicon particle vaccination induces non-neutralizing anti-nucleoprotein antibody-mediated control of Crimean-Congo hemorrhagic fever virus

doi: 10.1038/s41541-024-00877-1

Figure Lengend Snippet: a Cytokine/chemokine responses in mice vaccinated with CCHF VRP 28, 14, or 7 days prior to challenge or left unvaccinated (no VRP, given DMEM alone) and euthanized 3 or 6 days post infection (dpi) were analyzed using the ProcartaPlex Mouse Th1/Th2 Cytokine and Chemokine panel and 25 µL mouse plasma. b CCHFV-specific T-cell responses were evaluated using IFN-gamma ELISpot assay. Peptides covering the CCHFV IbAr10200 NP or Oman-98 GPC NSm-Gc domain (numbers represent aa positions) were used to stimulate splenocytes harvested from vaccinated animals. Data are reported as the number of spot-forming cells (SPC)/1 × 10 6 cells. c CCHFV-specific NP, Gn, Gc, and GP38 antibody responses (IgM and IgG) were evaluated via ELISA using mouse plasma and reported as endpoint dilution titers. Antibody titers, T-cell responses, and cytokine/chemokine levels were compared statistically between vaccine groups by timepoint (3 or 6 dpi) using multiple two-tailed t -tests (Mann-Whitney) and only statistically significant results are reported; * p < 0.5, ** p < 0.01. Individual animals are represented. Bars and error bars indicate mean ± SEM.

Article Snippet: Cell suspensions were frozen, thawed in RPMI, diluted to 2 × 10 6 cells/mL, and seeded in MabTech ELISpot plates containing peptides homologous to the VRP vaccine construct, covering the CCHFV IbAr10200 NP (AbClonal; 15-mers with 11 AA overlap) or Oman-98 GPC NSm-Gc domain (AbClonal; 15-mers with 4 AA overlap).

Techniques: Infection, Ifn? elispot, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY