Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: ( a ) Phylogenetic tree of NA amino acid sequence depicting color-coded AGs. Tex12 and Per09 belong to AG1 (dark yellow); Sin16 and Kan17 to AG2 (green); Hel823 to AG3 (Blue), and Ind11 to AG4 (purple). ( b-f ) The relative body weight, survival and LD50 of DBA/2J mice inoculated with ( b ) HxNA/Perth/16/2009nib-64, ( c ) HxNA/Texas/50/2012, ( d ) HxNA/Helsinki/823/2013, ( e ) HxNA/Indiana/08/2011, and ( f ) HxNA/Kansas/14/2017. The LD50 values were calculated using the Reed–Muench method.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

Techniques: Sequencing, Endpoint Dilution Assay

Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: ( a ) Relative body weight of BALB/c mice inoculated with serial dilution of HxNPer09. ( b ) Relative body weight of DBA/2J mice inoculated with a serial dilution of HxNSin16.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

Techniques: Serial Dilution

Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: DBA/2J mice were prime-boosted in a two-week interval regimen with 0.1 µg of AF03-adjuvanted Perth09, Tex12, Sing16, Hel823, or Ind11 tetNA. ( a ) ELISA was performed with sera obtained two weeks after the boost using Perth09 tetNA that was captured in nickel-coated plates. ( b ) NAI was determined using HxN2 HxNA/Perth/16/2009 and HxN2s with homologous NA used in the immunization ( c ). Two weeks after the boost mice were challenged with 5LD50 of the reassortant HxNPer09. The graphs show the percentage of initial body weight and mortality ( d ). Statistical comparison of the ELISA endpoint dilution titers was performed using Mann-Whitney test. Differences between NAI titers were analyzed using Welch’s t test. Data points in a-c are from individual mice except for the PBS group, which represent pools of 5 mice each, and horizontal solid lines represent means. Dashed lines represent the limit of detection. Data points in d represent averages of individual mice and error bars SEM. Statistical analysis of relative body weight changes was assessed by an approximate F-test and survival using Mantel-Cox rank. Data in d and e are pooled from 2 independent experiments except for NA Tx12, NA Sin16, and NA Hel823, which are from 1 experiment. Dashed lines represent the limit of detection. ** P <0.01,*** P <0.001, **** P <0.0001 compared with the PBS immunized group.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

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

Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: DBA/2J mice (n = 5 per group per experiment) were prime-boosted in a two week interval with 0.1 µg of AF03-adjuvanted tetNA or PBS as indicated. ELISA and NAI assays were performed with sera isolated two weeks after the boost immunization. ( a ) ELISA was performed using Tex12 tetNA that was captured in nickel coated plates. ( b ) NAI of immune sera determined against HxNTex12 and ( c ) HxNPer09. ( d ) Two weeks after the boost, mice were challenged with 5 LD50 of HxNTex12. The graphs show the percentage of initial body weight and mortality. ( e ) ELISA was performed against Sin16 tetNA. NAI was determined against ( f ) HxNKan17 ( g ) and HxNPer09. ( h ) Relative body weight and mortality of mice challenged with 5LD50 of the HxNKan17 virus. ( i ) ELISA was performed against Hel823 tetNA. NAI determined against ( j ) HxNHel823 and ( k ) HxNPer09. ( l ) Relative body weight and mortality of mice challenged with 5LD50 of HxNHel823 virus. ( m ) ELISA against Ind11 tetNA. NAI determined against ( n ) HxNInd11 or ( o ) HxNPer09. Relative body weight and mortality of mice challenged with ( p ) 5 or ( q ) 2 LD50 of HxNInd11. Statistical comparison of the ELISA endpoint dilution titers was performed using Mann-Whitney test. Differences between NAI titers were analyzed using Welch’s t test. Statistical analysis of relative body weight changes was assessed by an approximate F -test and statistical analysis of survival using Mantel-Cox rank. Dashed lines represent the limit of detection. All experiments were repeated once and the data pooled for analysis. * P <0.05, ** P <0.01,*** P <0.001, **** P <0.0001 compared with the PBS immunized group, or as indicated.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Virus, Comparison, MANN-WHITNEY

Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: Serum used for passive transfer was obtained from DBA/2J mice that had been immunized in a prime-boost regimen with 1 µg of AF03-Adjuvanted tetNA or PBS (naïve serum). One day after serum transfer, mice were challenged with 4 different HxN2 viruses. ( a ) Relative body weight and survival of mice challenged with 2 LD50 of HxNPer09. ( b ) ELISA performed using Per09 tetNA that was captured in nickel coated plates. ( c ) NAI determined against the HxNPer09 challenge virus. ( d ) Relative body weight and survival of mice challenged with 2 LD50 of HxNInd11. ( e ) ELISA performed using Sin16 tetNA that was captured in nickel coated plates. ( f ) NAI determined using HxNSin16 virus, a reassortant with NA that belongs to the same AG as the NA of A/Kansas/14/2017. ( g ) Relative body weight and survival of mice challenged with 2 LD50 of HxNHel823. ( h ) ELISA performed using Hel823 tetNA that was captured in nickel coated plates. ( i ) NAI determined against HxNKan17. ( j ) Relative body weight and survival of mice challenged with 2 LD50 of HxNInd11. ( k ) ELISA performed using Ind11 tetNA that was captured in nickel coated plates. (k) NAI determined against HxNInd11. The graphs in panels a, d, g, and j show the percentage of initial body weight and mortality. Statistical analysis of relative body weight changes was analyzed as repeated measurements using method of residual maximum likelihood (REML) and survival using Mantel-Cox rank. Challenge experiments were performed twice and data from the 2 independent experiments were pooled for analysis. Dashed lines represent the limit of detection. ** P <0.01; **** P <0.0001 compared with the PBS immunized group.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

Techniques: Enzyme-linked Immunosorbent Assay, Virus

Journal: bioRxiv

Article Title: The breadth of protection mediated by anti-N2 neuraminidase antibody responses relies on cross-reactive and not cross-inhibiting antibodies

doi: 10.64898/2026.02.02.703223

Figure Lengend Snippet: Pathogenicity of A/Singapore/1/1957 in DBA/2J mice. ( a ) Relative weight and survival of mice inoculated with a 3-fold serial dilution of parental Sin57. ( b ) Relative weight and survival of mice inoculated with a 3-fold serial dilution of mouse-adapted A/Singapore/1/1957.

Article Snippet: Female DBA/2J mice, aged 7-8 weeks, were purchased from Janvier (France).

Techniques: Serial Dilution