influenza a h1n1 nucleoprotein antibody  (Bio-Techne corporation)

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: A homogeneous polysaccharide from Houttuynia cordata (HCPM) and crude polysaccharides from Houttuynia cordata (HCP) mitigate pneumonia severity in virus–bacterial coinfection mice. (A, B) C57BL/6 mice were infected intranasally with 0.2 LD 50 influenza A virus (H1N1) and then infected 10 7 CFU methicillin-resistant Staphylococcus aureus (MRSA) three days later (11 mice in each group). C57BL/6 mice were treated with the indicated drugs once daily for seven days via gavage, beginning 2 h after viral infection. Survival rate (A) and body weight (B) were continuously monitored for 14 days. (C–N) C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA 3 days later. Mice were treated with indicated drugs once daily for 5 days (6 mice in each group). (C) Lung index (the ratio of lung weight to body weight). (D) Histopathological injury and inflammatory infiltration of representative lung sections in each group. Scale bar = 50 μm. (E) Western blotting analysis of edema clearance-related proteins (Na,K-ATPase α 1) in lung homogenates ( n = 3). (F) The expression of viral nucleoprotein (NP) ( n = 3). (G) Bacterial loads in mice lungs ( n = 6). The concentration of the cytokines including TNF- α (H), IL-6 (I), IFN- α (J), IFN- β (K), IFN- γ (L), and MPO (M) in lungs were determined using ELISA ( n = 5). Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Virus, Infection, Western Blot, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: HCPM and HCP alleviate intestinal injury in viral–bacterial coinfection mice. C57BL/6 mice were infected intranasally with 0.2LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Mice were treated with indicated drugs once daily for five days (6 mice in each group). (A) Morphological observation of intestinal epithelial integrity. Histopathological examination of small intestines through H&E staining ( n = 4). Scale bar = 200 μm. (B) Mucin (blue) was detected by alcian blue & nuclear fast red staining kit ( n = 4). Scale bar: 50 μm. (C–E) Immunofluorescent staining of TJ proteins including ZO-1 (C), Occludin (D), and Claudin-1 (E) in the small intestine. Images are representative of three individual experiments. Scale bar = 20 μm. (F–H) Mean fluorescence intensity of ZO-1 (F), Occludin (G), and Claudin-1 (H) ( n = 3). (I–K) The concentration of IL-6 (I), IFN- γ (J), and MPO (K) in small intestine homogenates were determined using ELISA ( n = 5). Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Staining, Fluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: HCPM and HCP inhibit the overactivation of intestinal complement in viral–bacterial coinfection mice. C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Mice were treated with indicated drugs from Day 0 to Day 4 following H1N1 infection ( n = 6). The small intestines were obtained from different groups on Day 4 and Day 5. (A) Representative images of immunohistochemical staining for C3a in small intestines ( n = 4). (B, C) Quantitative analysis of the C3a-stained immunohistochemical images by ImageJ ( n = 4). (D) Representative images of immunohistochemical staining for C5a in small intestines ( n = 4). (E, F) Quantitative analysis of the C5a-stained immunohistochemical images by ImageJ ( n = 4). Scale bar = 50 μm. Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗∗ P < 0.01 and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Immunohistochemical staining, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: HCPM and HCP restrain the overactivation of intestinal NLR family pyrin domain-containing 3 (NLRP3) inflammasome in viral–bacterial coinfection mice. C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Mice were treated with indicated drugs for five days. The small intestines were obtained from different groups on Day 4 and Day 5. (A, H) The protein expression of NLRP3, cleaved-Caspase-1, Caspase-1, and ASC in small intestines was detected by Western blotting ( n = 3). Densitometric analysis of NLRP3 (B, I), and cleaved-Caspase-1 (C, J) protein was performed. The contents of IL-18 and IL-1 β in small intestinal tissue on Day 4 (D, E) and Day 5 (K, L) were detected by ELISA ( n = 6). (F, M) Representative images of NLRP3 immunostaining in small intestine ( n = 4). Scale bar = 50 μm. (G, N) Representative images of cleaved-Caspase-1 immunostaining in the small intestine ( n = 4). Scale bar = 50 μm. Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗∗ P < 0.01 and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunostaining

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: The recovery effect of HCPM on Treg/Th17 cell imbalance may be related to its effect on NLRP3 inflammasome signaling in viral–bacterial coinfection mice. C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Coinfection mice were treated with MCC950 (5 mg/kg, intraperitoneal injection) or HCPM (80 mg/kg, intragastric administration) from Day 0 to Day 4 following H1N1 infection. The intestinal lamina propria and lung were obtained from different groups on Day 4 and Day 5. The frequency of Th17 (CD4 + IL-17A + ) cells and Treg (CD4 + Foxp3 + ) cells in intestinal lamina propria and lung were assayed using flow cytometry. (A, B) The changes of Th17 cells in intestinal lamina propria on Day 4 and Day 5 ( n = 4). (C, D) The changes of Treg cells in intestinal lamina propria on Day 4 and Day 5 ( n = 4). (E) The ratio of Treg/Th17 cells in intestinal lamina propria on Day 4 and Day 5 ( n = 4). (F, G) The changes of Th17 cells in lungs on Day 4 and Day 5 ( n = 4). (H, I) The changes of Treg cells in lungs on Day 4 and Day 5 ( n = 4). (J) The ratio of Treg/Th17 cells in lungs on Day 4 and Day 5 ( n = 4). Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Injection, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: HCPM and HCP regulate the balance of Treg and Th17 cells in the lung and intestinal mucosa during viral–bacterial coinfection. C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Mice were treated with indicated drugs for five days. (A–D) The frequency of Th17 (CD4 + IL-17A + ) cells and Treg (CD4 + Foxp3 + ) cells in the lungs and small intestines were assayed using flow cytometry ( n = 4). (E) The ratio of Treg/Th17 cells in the lungs ( n = 4). (F) The ratio of Treg/Th17 cells in small intestines ( n = 4). Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: An anti-complement homogeneous polysaccharide from Houttuynia cordata ameliorates acute pneumonia with H1N1 and MRSA coinfection through rectifying Treg/Th17 imbalance in the gut–lung axis and NLRP3 inflammasome activation

doi: 10.1016/j.apsb.2025.04.008

Figure Lengend Snippet: HCPM and HCP restore the frequency of Treg and Th17 cells in Peyer's patches (PPs)–mesenteric lymph nodes (MLNs)–lung axis in viral–bacterial coinfection mice. C57BL/6 mice were infected intranasally with 0.2 LD 50 H1N1 and then infected with 10 7 CFU MRSA three days later. Mice were treated with indicated drugs for five days. The PPs, MLNs, and lungs were obtained from different groups on Day 4 and Day 5. (A–D) The frequency of Th17 (CD4 + IL-17A + ) cells and Treg (CD4 + Foxp3 + ) cells in PPs, MLNs, and lungs were assayed using flow cytometry ( n = 4). (E) The dynamic changes of Th17 cells in PPs, MLNs, and lungs ( n = 4). (F) The dynamic changes of Treg cells in PPs, MLNs, and lungs ( n = 4). Data are represented as mean ± SD and analyzed by one-way ANOVA followed by Dunnett's multiple comparisons test. P values < 0.05 were considered statistically significant, ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. coinfection; not significant (ns).

Article Snippet: H1N1 NP (#NB100-56570) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Infection, Flow Cytometry