Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Surface binding of anti-PspA hkR36A MAbs with strains expressing family 1 PspA a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Binding Assay, Expressing

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Relative efficacy of anti-PspAhkR36A MAbs to protect mice against intravenous challenge. CBA/N mice were injected with purified anti-PspAhkR36A MAb M4F4, P1E11, or L5C8 (A and B), B3D12, B3H8, or L5F10 (C and D) or P2A4 or P2B5 (E and F) intraperitoneally at 5 mg/kg body weight (high dose). The corresponding isotype control MAb (IgG1 IC or IgG2a IC) was included in each set as the negative control. One hour later, mice were challenged with 107 CFU of BG8838 (A, C, and E) or 103 CFU of WU2 (B, D, and F), and mouse survival was recorded. The data for the group given anti-PspAhkR36A MAb were compared with those for the respective isotype control MAb using the log rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, statistically not significant.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Injection, Purification, Negative Control

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Anti-PspAhkR36A MAbs protect mice against pneumococcal infection even when given at a lower dose. CBA/N mice were injected intraperitoneally with 1.25 mg/kg body weight (low dose) of either anti-PspAhkR36A MAb M4F4, P1E11, or L5F10 (A and B) or P2A4 or P2B5 (C and D). The control group was given the respective isotype control MAb (IgG1 IC or IgG2a IC). Mice were challenged with BG8838 (A and C) or WU2 (B and D) 1 h later, and mouse survival was recorded. For other details, refer to the legend to Fig. 2.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Infection, Injection

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Flow cytometry-based analysis of complement C3 deposition on the surface of pneumococci in the presence of anti-PspA hkR36A MAbs a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Cytometry, Negative Control, Positive Control

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: In vivo protective efficacies of anti-PspA hkR36A MAbs correlate with the extent of complement deposition a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: In Vivo, Staining, Activity Assay

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: ELISA-based analysis for reactivity of anti-PspA hkR36A MAbs with recombinant PspAs representing the 6 PspA clades a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Surface binding of anti-PspA hkR36A MAbs with strains expressing family 1 PspA a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Binding Assay, Expressing

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Relative efficacy of anti-PspAhkR36A MAbs to protect mice against intravenous challenge. CBA/N mice were injected with purified anti-PspAhkR36A MAb M4F4, P1E11, or L5C8 (A and B), B3D12, B3H8, or L5F10 (C and D) or P2A4 or P2B5 (E and F) intraperitoneally at 5 mg/kg body weight (high dose). The corresponding isotype control MAb (IgG1 IC or IgG2a IC) was included in each set as the negative control. One hour later, mice were challenged with 107 CFU of BG8838 (A, C, and E) or 103 CFU of WU2 (B, D, and F), and mouse survival was recorded. The data for the group given anti-PspAhkR36A MAb were compared with those for the respective isotype control MAb using the log rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, statistically not significant.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Injection, Purification, Negative Control

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Anti-PspAhkR36A MAbs protect mice against pneumococcal infection even when given at a lower dose. CBA/N mice were injected intraperitoneally with 1.25 mg/kg body weight (low dose) of either anti-PspAhkR36A MAb M4F4, P1E11, or L5F10 (A and B) or P2A4 or P2B5 (C and D). The control group was given the respective isotype control MAb (IgG1 IC or IgG2a IC). Mice were challenged with BG8838 (A and C) or WU2 (B and D) 1 h later, and mouse survival was recorded. For other details, refer to the legend to Fig. 2.

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Infection, Injection

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: Flow cytometry-based analysis of complement C3 deposition on the surface of pneumococci in the presence of anti-PspA hkR36A MAbs a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: Cytometry, Negative Control, Positive Control

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Correlation between In Vitro Complement Deposition and Passive Mouse Protection of Anti-Pneumococcal Surface Protein A Monoclonal Antibodies

doi: 10.1128/CVI.00001-14

Figure Lengend Snippet: In vivo protective efficacies of anti-PspA hkR36A MAbs correlate with the extent of complement deposition a

Article Snippet: None of the MAbs bound PspAs representing families 2 (clades 3, 4, and 5) and 3 (clade 6), suggesting that the anti-PspA hkR36A MAbs were family 1 specific. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Hybridoma Fold change for pneumococcal strain (PspA family/clade): L82016 (1/1) b R36A (1/2) b TIGR4 (2/3) JCP#56 (2/4) ATCC 6303 (2/5) BG9300 (3/6) B3D12 16.5 17.2 0.8 0.8 0.8 1.2 B3H8 1.8 2.1 1.0 0.9 1.0 0.8 L5C8 17.1 16.7 0.8 0.8 0.8 0.8 L5F10 19.5 26.9 0.8 0.7 0.9 1.2 M4F4 30.0 32.5 1.1 1.2 1.6 1.2 P1E11 29.2 31.5 1.4 1.4 1.7 1.6 D1A5 10.6 11.9 0.9 0.8 0.7 0.9 K1B12 15.9 16.5 0.7 0.7 0.6 0.9 M6B2 13.5 14.5 1.0 1.4 1.3 1.9 P2F9 9.8 13.4 0.9 1.0 0.9 1.0 P2A4 21.6 19.9 1.0 1.5 0.9 1.0 P2B5 22.0 24.0 1.0 1.2 1.2 1.2 J4C1 28.2 29.3 1.0 0.8 0.7 0.8 P2C2 16.2 17.0 0.6 0.6 0.6 0.6 A1D9 2.0 2.7 0.6 0.7 0.7 0.6 C4B4 18.1 18.1 0.8 0.6 0.6 0.7 F4B6 24.7 26.3 1.0 0.9 0.8 1.1 D3H6 4.0 5.7 0.7 0.7 0.7 1.1 Open in a separate window a The culture supernatants from the 18 anti-PspA hkR36A MAb-secreting hybridomas were tested for reactivity with recombinant PspAs representing the 6 clades of PspA (extracellular domain with or without the proline-rich region) by an ELISA.

Techniques: In Vivo, Staining, Activity Assay

Journal: EMBO Molecular Medicine

Article Title: Mannose receptor‐derived peptides neutralize pore‐forming toxins and reduce inflammation and development of pneumococcal disease

doi: 10.15252/emmm.202012695

Figure Lengend Snippet: A ELISA showing the dose‐dependent binding of plate‐bound MRC‐1 peptides P2, P3, and the control peptides CP1 and CP2 to PLY (0–0.5 μM). BSA was used as negative control to show the binding specificity. Data are mean ± s.e.m. of two independent experiments, each containing three replicates per condition. B Hemolysis assay ( n = 4) of 1 μg/ml purified PLY in the presence of increasing concentrations of MRC‐1 peptides, P2, scrambled P2, P3, and control peptide CP2 (1–1000 μM). Data represent mean ± s.e.m. * P < 0.05 by one‐way ANOVA with Dunnett's post hoc test for multiple comparisons. Exact P values are shown in Appendix Table S4 . C LDH cytotoxicity assay in human THP‐1 macrophages stimulated with purified PLY, LLO, or SLO (0.5 μg/ml) in the presence or absence of 100 μM peptides P2, scrambled P2, P3, or control peptide CP2 for 18 h. Cholesterol (100 μM) was used as positive control to inhibit hemolysis. Data are mean ± s.e.m. from 4 independent experiments. **** P < 0.0001 by two‐way ANOVA with Bonferroni post hoc test for multiple comparisons. n.s. denotes not significant. Exact P values are shown in Appendix Table S4 . D Binding of FITC‐labeled peptides P2 and CP2 to wild‐type pneumococci, TIGR4 (T4), and its isogenic PLY mutant (T4Δ ply ) was visualized by fluorescence microscopy. Scale bars, 10 μm. In magnified images, scale bars, 1 μm. Images are representative of three independent experiments. E The hemolytic activity of wild‐type pneumococci, TIGR4 (T4) and the PLY mutant, T4Δ ply in the presence of 100 μM peptide P2 and CP2. Data are the mean ± s.e.m. of three independent experiments. *** P < 0.001 by one‐way ANOVA with Bonferroni post hoc test for multiple comparisons. n.s. denotes not significant. Exact P values are shown in Appendix Table S4 . Source data are available online for this figure.

Article Snippet: The encapsulated S. pneumoniae serotype 4 strain TIGR4 (T4; ATCC BAA‐334) and the type 2 strain D39 were used in this study, as well as the isogenic capsule and pneumolysin deletion mutants, T4R (Fernebro et al , ), T4Δ ply , and T4RΔ ply (Littmann et al , ), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Control, Negative Control, Hemolysis Assay, Purification, LDH Cytotoxicity Assay, Positive Control, Labeling, Mutagenesis, Fluorescence, Microscopy, Activity Assay

Journal: EMBO Molecular Medicine

Article Title: Mannose receptor‐derived peptides neutralize pore‐forming toxins and reduce inflammation and development of pneumococcal disease

doi: 10.15252/emmm.202012695

Figure Lengend Snippet: A IL‐8 released by human THP‐1 macrophages stimulated with purified PLY, LLO, or SLO (0.5 μg/ml) in the presence or absence of 100 μM peptides P2, P3, or control peptide CP2 for 18 h. Cholesterol (100 μM) was used as positive control to inhibit hemolysis. Data are mean ± s.e.m. from three independent experiments. **** P < 0.0001 by two‐way ANOVA with Bonferroni post hoc test for multiple comparisons. n.s. denotes not significant. Exact P values are shown in Appendix Table S4 . B Schematic showing the cellular architecture of the 3D lung epithelial model. C 3D volume images of the GFP‐lung epithelial models at 1 and 3 h post‐stimulation with 1 μg/ml PLY in the presence or absence of 100 μM peptide P2 or the control peptide CP2. Images are representative of two independent experiments with n = 3 models/condition. D Invasion of wild‐type pneumococci T4 (TIGR4) or its isogenic PLY mutant T4Δ ply into the lung epithelial models ( n = 3/condition) in the presence or absence of 100 μM peptide P2 or the control peptide CP2 at 2 h post‐infection was measured using CFU viability assay following gentamicin killing of extracellular bacteria. Anti‐PLY was used as control to test the effect of blocking PLY. Data are mean ± s.e.m. of n = 3 models/condition from two independent experiments. % bacterial entry = (bacteria uptaken/input) × 100. ** P < 0.01 by one‐way ANOVA with Dunnett's post hoc test for multiple comparisons. n.s. denotes not significant. Exact P values are shown in Appendix Table S4 . E Human DCs were infected with type 4 and type 2 pneumococci, T4 and D39, respectively, at MOI of 10 in the presence or absence of 100 μM peptides, P2 or CP2, and intracellular bacteria were counted at 3 h post‐infection following gentamicin killing of extracellular bacteria. Cytochalasin D (0.5 mM) was used as negative control to inhibit phagocytosis. Anti‐PLY was used as control to test the effect of blocking PLY. Data are mean ± s.e.m. of three independent experiments. **** P < 0.0001 by two‐way ANOVA with Bonferroni post hoc test for multiple comparisons. n.s. denotes not significant. Exact P values are shown in Appendix Table S4 . F DCs were infected with the unencapsulated pneumococcal strain T4R in the presence of 100 μM peptides, P2 or CP2 at MOI of 10 for 2 h. Immunofluorescence microscopy images show that in DCs treated with peptide P2 (but not the control peptide CP2), intracellular T4R (green) do not co‐localize with MRC‐1 (red), but with the autophagy protein LC3B (cyan). Images are representative of three independent experiments. Scale bars, 10 μm. In magnified images, scale bars, 5 μm. Arrows indicate regions of co‐localization of intracellular T4R with MRC‐1 and LC3B. G Quantification of percentage of intracellular S. pneumoniae ( n = 50) in infected DCs that co‐localize with MRC‐1 and LC3B. Data are mean ± s.e.m. from two independent experiments. **** P < 0.0001 by two‐way ANOVA with Bonferroni post hoc test for multiple comparisons. n.s. denotes not significant. P values are shown in Appendix Table S4 . Source data are available online for this figure.

Article Snippet: The encapsulated S. pneumoniae serotype 4 strain TIGR4 (T4; ATCC BAA‐334) and the type 2 strain D39 were used in this study, as well as the isogenic capsule and pneumolysin deletion mutants, T4R (Fernebro et al , ), T4Δ ply , and T4RΔ ply (Littmann et al , ), respectively.

Techniques: Purification, Control, Positive Control, Mutagenesis, Infection, Viability Assay, Bacteria, Blocking Assay, Negative Control, Immunofluorescence, Microscopy