mouse anti mouse nk1 1 monoclonal antibody  (Bio X Cell)

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on standard diet (SD) or 60% high-fat diet (HFD) for a minimum of 12 weeks. (A-K) WT and HFD mice were inoculated by intranasal (IN) infection with 1000 tissue culture infectious dose-50 (TCID 50 ) of A/California/04/2009 (CA/09 H1N1) virus or PBS as a mock control. (A-E) NK cells were depleted with 300 μg of anti-NK1.1 PK136 antibody or an IgG2a isotype control days -2, -1, day of infection (d0), +1, and then every other day throughout the duration of the experiment. (A) Graphical summary and timeline of NK cell depletion studies made with Biorender (B) Probability of survival and median day survival time (n=9-10 per group ) and (C) Clinical scores post-infection. (D) Lungs were harvested at 3 days-post infection (dpi) and viral titers were determined via TCID 50 ( n=4-5 per group) (E) Representative gross lung images at 3 dpi with black arrows indicating areas of pulmonary hemorrhage. (F, I-K) At 1- or 3-dpi lungs were resected and processed for flow cytometry. Single-cell suspensions were stained for surface antibodies to characterize natural killer (NK) cells. (F) Number per 10 5 lung cells of NK cells (CD45 + , TCRβ - , NK1.1 + ) ( n=4-10 per group) (G) Lungs were processed for immunofluorescence at 3 dpi and stained for DAPI (blue), influenza NP protein (red) and NKp46 (NK cell marker, green) and (H) Number of NK cells per 250 2 μm was quantified. (I) Average expression of the NK cell markers CD27 and CD11b to assess maturation stage. (J) Frequency of CD11b + CD27 - lung NK cells and (K) Frequency of CD11b + CD27 + lung NK cells at baseline or at 1 or 3 dpi ( n=4-7 per group). Data are shown as mean ± SEM. Statistical significance was calculated using Mantel-Cox log-rank analysis (A) a two-way ANOVA (C), a One-way ANOVA with multiple comparison’s test (D, F, J, K) and a students t test with Welch’s correction (H) , * p < .05; ** p < .01; **** p < .0001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Infection, Virus, Control, Flow Cytometry, Single Cell, Staining, Immunofluorescence, Marker, Expressing

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on standard diet (SD) or 60% high-fat diet (HFD) for a minimum of 12 weeks. SD and HFD mice were inoculated by intranasal (IN) infection with 1000 tissue culture infectious dose-50 (TCID 50 ) of A/California/04/2009 (CA/09 H1N1) virus or PBS as a mock control. (A-C) NK cells were depleted with 300 μg of anti-NK1.1 PK136 antibody or an IgG2a isotype control days -2, -1, day of infection (d0) and then lungs or spleens were resected and processed for flow cytometry. (A) Example gating of lung NK cells (B) Frequency and number of NK cells (CD45 + Tcrβ - NK1.1 + DX5 + ) assessed by flow cytometry in the lung and (C) in the spleen. (D) NK cells were depleted with 300 μg of anti-NK1.1 PK136 antibody or an IgG2a isotype control days -2, -1, day of infection (d0), +1, and then every other day throughout the duration of the experiment and weight loss over time was assessed. Data are shown as mean ± SD (B, C) or SEM (D) . Statistical significance was calculated using a One-way ANOVA with multiple comparison’s test (B, C) and a Two-way ANOVA (D) , ** p < .01; *** p < .001; **** p < .0001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Infection, Virus, Control, Flow Cytometry

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on SD or 60% HFD for a minimum of 12 weeks. (A) Bone marrow was resected and processed for flow cytometry, including the total number of immune cells (CD45 + ), NK cells (CD45 + Tcrβ - NK1.1 + ) and intracellular staining for NK cell transcription factors including Tbet, EOMES, and GATA3, or (B) assessed in the spleen. (C, D) SD and HFD mice were inoculated by IN infection with 1000 TCID 50 of CA/09 H1N1 virus or PBS as a mock control and assessed for (C) frequency of lung NK cells. (D) Representative contour plot of CD11b and CD27 gating on lung NK cells. Data are shown as mean ± SEM (D) . Statistical significance was calculated using a student’s t test with Welch’s correction (A, B) or a One-way ANOVA with multiple comparison’s test (C) * p < .05; ** p < .01; *** p < .001.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Flow Cytometry, Staining, Infection, Virus, Control

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on SD or 60% HFD for a minimum of 12 weeks and were intranasally inoculated with 1000 TCID 50 of CA/09 H1N1 virus or PBS (mock control) and lungs were resected and processed for flow cytometry at 1 or 3 dpi (n=4-11 per group). (A) Representative contour plots of EOMES expression in lung NK cells (CD45 + , TCRβ - NK1.1 + ) at 1 or 3 dpi (B) Frequency of intermediate EOMES expression in NK cells (C) Representative mean fluorescent intensity (MFI) plot normalized to mode of TBET expression in NK cells 1 dpi normalized to mode (D) Frequency of TBET expression in lung NK cells (E) Frequency of IFN-γ expression in NK cells (F) Representative contour plot of IFN-γ expression in NK cells in mock-infected controls or 3 dpi (G) Frequency of the early activation marker CD69 expression in NK cells (H) Representative MFI plot normalized to mode of Granzyme B expression and (I) MFI and frequency of Granzyme B expression in NK cells (J) IFNγ, Granzyme B, CD69, Tbet, and Ki67 expression displayed as spider plots. Data of NK cells from mock-infected WT mice in blue, mock-infected HFD mice in orange, WT 3 dpi in grey, and HFD 3 dpi in yellow. (K) Graphical summary of NK cell dysfunction in obesity during influenza infection, made with Biorender . Data are shown as mean ± SEM. Statistical significance was calculated using a one-way ANOVA with Tukey’s multiple comparison’s test ( B, D, E, G, & I) . * p < .05; ** p < .01; *** p < .001; **** p < .0001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Virus, Control, Flow Cytometry, Expressing, Infection, Activation Assay, Marker

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on SD or 60% HFD for a minimum of 12 weeks and were intranasally inoculated with 1000 TCID 50 of CA/09 H1N1 virus or PBS (mock control) and lungs were resected and processed for flow cytometry at 3 dpi to assess lung NK cells (CD45 + , TCRβ - NK1.1 + ). (A) Frequency of the marker of proliferation Ki-67 (n = 4-7/group) (B) Cell size as assessed by FSC-A (n = 8-9/group) (C) Frequency of c-MYC and (D) amino acid transporter CD98 expression (n=5-6/group) (E) Representative contour plots of GLUT1 expression on NK cells including fluorescence minus one control (FMO) and (F) Frequency of GLUT1 expression (G) Summary of mTOR signaling cascade and impact on NK cell function, made with Biorender . (H) Representative MFI plot normalized to mode of phosphorylated S6 kinase (pS6k) and (I) Frequency of pS6-kinase, (J) Representative MFI plot normalized to mode of phosphorylated mTOR and (K) Frequency of p-mTOR and (L) Representative MFI plot normalized to mode of phosphorylated AKT and (M) Frequency of p-AKT (n= 7-8/group). Data are shown as mean ± SEM. Statistical significance was calculated using a one-way ANOVA with Tukey’s multiple comparison’s test (A-D, F, I, K, M) . * p < .05; ** p < .01; *** p < .001; **** p < .0001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Virus, Control, Flow Cytometry, Marker, Expressing, Fluorescence, Cell Function Assay

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on SD or 60% HFD for a minimum of 12 weeks and were intranasally inoculated with 1000 TCID 50 of CA/09 H1N1 virus or PBS (mock control). At 3 dpi, lungs were resected and processed for various analyses. (A) Triglyceride levels in lung tissue (n= 3/group) or (B) Triglyceride species (881.7537 m/z +/- 10 ppm) visualized in lung tissue by MALDI spatial mass spectrometry normalized by TIC and relative intensity per pixel was quantified in (C) . (D) Normalized abundance of long chain fatty acids in lung tissue. (E & F) Lung tissue was processed for IF at 3 dpi (E) Representative tissue sections stained for DAPI (blue), influenza NP protein for active viral replication (green) or the lipid transporter CD36 (red) and (F) CD36 MFI was determined with multiple fields per lung for at least 2 mouse lungs per group (>= 112 fields analyzed). (G-I) Lungs were resected and processed for flow cytometry at 3 dpi (n=6/group) to assess CD36 expression on (G) total immune cells (CD45 + ) (H) Representative contour plots of CD36 expression on lung NK cells (CD45 + , TCRβ - NK1.1 + ) including an FMO control and (I) frequency of CD36 expression. (J-M) At 3 dpi with IAV or PBS (mock), lungs were resected and processed for single cell sorting to isolate lung NK cells by magnetic activated cell sorting for IF imaging of NK cells (NKp46+, green), DAPI (blue) and LipidTOX (red) (J & L) Representative images with (K & M) MFI quantitation of LipidTOX staining during mock infection or 3 dpi with IAV (n>+ 178 cells/group analyzed). (N, O) LipidTOX was also assessed by flow cytometry (n= 6-7/group) with (N) representative MFI plot normalized to mode with MFI expression and (O) Frequency of NK cells expressing LipidTOX. (P) Graphical summary of lipid driven mechanisms of NK cell dysfunction made with Biorender . Data are shown as mean ± SEM. Statistical significance was calculated using a students t test with Welch’s correction (A, K, M) , or a One-way ANOVA with Tukey’s multiple comparison’s test (B, F, G, I, N, O) . * p < .05; *** p < .001; **** p < .0001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Virus, Control, Mass Spectrometry, Staining, Flow Cytometry, Expressing, Single Cell, FACS, Imaging, Quantitation Assay, Infection

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: WT mice were placed on SD or 60% HFD for a minimum of 12 weeks and were intranasally inoculated with 1000 TCID 50 of CA/09 H1N1 virus or PBS (mock control). At 3 dpi, lungs were resected and processed for flow cytometry. (A) Representative MFI plot and MFI of CD36 expression on NK cells (B) LipidTOX staining for total immune cells, (CD45 + ), T cells (CD45 + Tcrβ + NK1.1 - ), other immune cells (CD45 + Tcrβ - NK1.1 - ), and NK T cells (CD45 + Tcrβ + NK1.1 + ) (C) IFN-γ expression by T cells (CD45 + Tcrβ + NK1.1 - ), other immune cells (CD45 + Tcrβ - NK1.1 - ), and NK T cells (CD45 + Tcrβ + NK1.1 + ) and (D) CD36 expression. (E, F) At 3 dpi with PBS (mock) (E) or IAV (F) , spleens were resected and processed for single cell sorting to isolate lung NK cells by magnetic activated cell sorting for IF imaging of NK cells (NKp46+, green), DAPI (blue) and LipidTOX (red) with representative images and MFI quantitation of LipidTOX staining. Data are shown as mean ± SEM. Statistical significance was calculated using a One-way ANOVA with Tukey’s multiple comparison’s (A-D) or a student’s t test (E, F) . * p < .05; ** p < .01; *** p < .001; ns = not significant.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Virus, Control, Flow Cytometry, Expressing, Staining, Single Cell, FACS, Imaging, Quantitation Assay

Journal: bioRxiv

Article Title: Obesity-Driven Lung Lipidome Remodeling Suppresses NK Cell Activation and Antiviral Immunity to Influenza Infection

doi: 10.64898/2026.03.06.710186

Figure Lengend Snippet: (A) Timeline of diet and infection made with Biorender (B) Weights over time at start of diet-switch (HFD->SD) for all groups measured weekly (C) Total cholesterol (D) high-density lipoprotein (HDL) and (E) low-density lipoprotein (LDL) in blood measured weekly post-diet switch (n=11-15/group). (F) Representative images of gross anatomy livers, (G) liver weights (n=7-10/group) and (H) representative images of gross anatomy lungs with fatty deposits (black arrows) at 4 weeks post-diet switch. (I) Mouse weight at week 4 post-diet switch including SD and HFD controls at time of IAV infection. (J-T) Mice were intranasally inoculated with 1000 TCID 50 of CA/09 H1N1 virus and assessed (n=16-26/group) for (J) survival (K) clinical scores and (L) weight loss, with the diet-switch group weight loss adjusted for weight loss only without infection average weekly weight loss (M) Summary of NK cell characterization by flow cytometry made in Biorender (N-T) At 3dpi, lungs were resected and processed for flow cytometry to assess lung NK cells (CD45 + , TCRβ - NK1.1 + ) (n= 6-7/group) including frequency of (N) Granzyme B (O) IFN-γ (P) phosphorylated mTOR (Q) phosphorylated AKT (R) phosphorylated S6 kinase and (S) MFI of intracellular LipidTOX staining. (T) Graphical summary of results from short term weight loss experiment and impact on NK cell function made with Biorender . Data are shown as mean ± SEM. Statistical significance was calculated a One-way ANOVA with Tukey’s multiple comparison’s test (G, I, N-S) , a Mantel-Cox log-rank analysis (J) and a two-way ANOVA (K & L). * p < .05; ** p < .01; *** p < .001; **** p < .0001.

Article Snippet: For in vivo NK cell depletion, mice were administered 300 μg anti-NK1.1 monoclonal antibody (PK136, BioXCell) or the corresponding InVivoPlus mouse IgG2a isotype control (BioXCell) by intraperitoneal injection -2, -1, day of infection, +1, then every other day until the conclusion of the experiment.

Techniques: Infection, Virus, Cell Characterization, Flow Cytometry, Staining, Cell Function Assay