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CAM13K SOSIP immunization elicits neutralizing antibody responses in V2-apex bNAb precursor expressing knock-in mice. (A) CH01_RUA heavy chain (HC)-only knock-in mice  were immunized either with 20 μg CAM13K SOSIP trimer in 5 μg GLA-SE adjuvant ( n = 5) or with 5 μg GLA-SE adjuvant alone ( n = 3). Time points of immunization and bleeds are indicated. (B) Comparison of serum neutralization titers (reciprocal 50% inhibitory dilutions, ID 50 ) from trimer-immunized (red circles) and adjuvant-only (blue squares) knock-in mice (preimmunization and terminal bleeds are shown as open and filled symbols, respectively). Serum neutralizing activity was tested in the TZM-bl assay  against pseudoviruses carrying the immunogen-matched autologous CAM13K Env, three CH01_RUA sensitive HIV-1 Envs (C1080, CRF250, Q23), as well as N160 glycan knockout variants of CRF250 and Q23. MLV Env containing pseudovirus was used for control. Differences between immunized and adjuvant-only groups were assessed using a nonparametric Mann-Whitney test (Prism 9.4.0 GraphPad Software), with asterisks indicating P < 0.05.
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CAM13K SOSIP immunization elicits neutralizing antibody responses in V2-apex bNAb precursor expressing knock-in mice. (A) CH01_RUA heavy chain (HC)-only knock-in mice  were immunized either with 20 μg CAM13K SOSIP trimer in 5 μg GLA-SE adjuvant ( n = 5) or with 5 μg GLA-SE adjuvant alone ( n = 3). Time points of immunization and bleeds are indicated. (B) Comparison of serum neutralization titers (reciprocal 50% inhibitory dilutions, ID 50 ) from trimer-immunized (red circles) and adjuvant-only (blue squares) knock-in mice (preimmunization and terminal bleeds are shown as open and filled symbols, respectively). Serum neutralizing activity was tested in the TZM-bl assay  against pseudoviruses carrying the immunogen-matched autologous CAM13K Env, three CH01_RUA sensitive HIV-1 Envs (C1080, CRF250, Q23), as well as N160 glycan knockout variants of CRF250 and Q23. MLV Env containing pseudovirus was used for control. Differences between immunized and adjuvant-only groups were assessed using a nonparametric Mann-Whitney test (Prism 9.4.0 GraphPad Software), with asterisks indicating P < 0.05.
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CAM13K SOSIP immunization elicits neutralizing antibody responses in V2-apex bNAb precursor expressing knock-in mice. (A) CH01_RUA heavy chain (HC)-only knock-in mice  were immunized either with 20 μg CAM13K SOSIP trimer in 5 μg GLA-SE adjuvant ( n = 5) or with 5 μg GLA-SE adjuvant alone ( n = 3). Time points of immunization and bleeds are indicated. (B) Comparison of serum neutralization titers (reciprocal 50% inhibitory dilutions, ID 50 ) from trimer-immunized (red circles) and adjuvant-only (blue squares) knock-in mice (preimmunization and terminal bleeds are shown as open and filled symbols, respectively). Serum neutralizing activity was tested in the TZM-bl assay  against pseudoviruses carrying the immunogen-matched autologous CAM13K Env, three CH01_RUA sensitive HIV-1 Envs (C1080, CRF250, Q23), as well as N160 glycan knockout variants of CRF250 and Q23. MLV Env containing pseudovirus was used for control. Differences between immunized and adjuvant-only groups were assessed using a nonparametric Mann-Whitney test (Prism 9.4.0 GraphPad Software), with asterisks indicating P < 0.05.

Journal: mBio

Article Title: A Germline-Targeting Chimpanzee SIV Envelope Glycoprotein Elicits a New Class of V2-Apex Directed Cross-Neutralizing Antibodies

doi: 10.1128/mbio.03370-22

Figure Lengend Snippet: CAM13K SOSIP immunization elicits neutralizing antibody responses in V2-apex bNAb precursor expressing knock-in mice. (A) CH01_RUA heavy chain (HC)-only knock-in mice were immunized either with 20 μg CAM13K SOSIP trimer in 5 μg GLA-SE adjuvant ( n = 5) or with 5 μg GLA-SE adjuvant alone ( n = 3). Time points of immunization and bleeds are indicated. (B) Comparison of serum neutralization titers (reciprocal 50% inhibitory dilutions, ID 50 ) from trimer-immunized (red circles) and adjuvant-only (blue squares) knock-in mice (preimmunization and terminal bleeds are shown as open and filled symbols, respectively). Serum neutralizing activity was tested in the TZM-bl assay against pseudoviruses carrying the immunogen-matched autologous CAM13K Env, three CH01_RUA sensitive HIV-1 Envs (C1080, CRF250, Q23), as well as N160 glycan knockout variants of CRF250 and Q23. MLV Env containing pseudovirus was used for control. Differences between immunized and adjuvant-only groups were assessed using a nonparametric Mann-Whitney test (Prism 9.4.0 GraphPad Software), with asterisks indicating P < 0.05.

Article Snippet: Differences between immunized and adjuvant-only knock-in mouse groups were assessed using a nonparametric Mann-Whitney test using GraphPad Prism 9.4.0 software.

Techniques: Expressing, Knock-In, Neutralization, Activity Assay, Knock-Out, MANN-WHITNEY, Software