normal human complement c3  (Quidel)

Journal: Infection and Immunity

Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus

doi: 10.1128/IAI.00212-20

Figure Lengend Snippet: Complement C3 interacts with the A. fumigatus conidial surface. (A) Flow cytometry showing the deposition of C3b on the conidial surfaces. Conidia were opsonized with pooled human serum and probed with monoclonal anti-C3b antibodies followed by FITC-conjugated secondary antibodies; opsonized conidia probed with FITC-conjugated secondary antibodies (Sec. Ab-FITC) served as the control. (B) Flow cytometry showing direct interaction of conidia with C3. Conidia opsonized with C3 were probed with anti-C3b antibodies and then with FITC-conjugated secondary antibodies; C3-opsonized conidia probed with FITC-conjugated secondary antibodies served as the control. (C) Confocal microscopy of the immunolabeled conidia showing C3b deposition on the conidial surface. C3-opsonized conidia were probed with monoclonal anti-C3b antibodies followed by FITC-conjugated secondary antibody and subjected to fluorescence microscopy; the control was the opsonized conidia probed only with FITC-conjugated secondary antibodies.

Article Snippet: Human complement C3 (purified from serum), lipopolysaccharide (LPS) from Escherichia coli , and polymyxin B-agarose were purchased from Sigma-Aldrich/Merck Millipore.

Techniques: Flow Cytometry, Confocal Microscopy, Immunolabeling, Fluorescence, Microscopy

Journal: Infection and Immunity

Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus

doi: 10.1128/IAI.00212-20

Figure Lengend Snippet: C3 interacts with A. fumigatus conidial surface in a concentration-dependent manner. (A) Flow cytometry. (B) ELISA showing concentration-dependent C3 binding to the conidial surface (C3 concentration, 1.95 to 250 ng/ml). (C) Unopsonized and serum-opsonized conidia were subjected to RodAp extraction using hydrofluoric acid (HF), and extracted protein was monitored by SDS-PAGE (12% gel with Coomassie brilliant blue staining). PM, protein markers; L2, unopsonized conidial HF extract; L3, serum-opsonized conidial HF extract. (D) Conidia were subjected to Western blotting (15% gel for protein separation) using polyclonal anti-RodAp antibodies (left: L1, recombinant RodAp; L2, serum-opsonized conidial HF extract; L3, unopsonized conidial HF extract), monoclonal anti-C3 antibodies (middle: serum-opsonized conidial HF extract), and monoclonal anti-C3b antibodies (right: L1, purified complement C3; L2, serum-opsonized conidial HF extract; L3, unopsonized conidial HF extract). Arrows indicate a common band recognized by both anti-C3b and anti-RodAp antibodies, suggesting a covalent interaction between RodAp and the complement protein C3.

Article Snippet: Human complement C3 (purified from serum), lipopolysaccharide (LPS) from Escherichia coli , and polymyxin B-agarose were purchased from Sigma-Aldrich/Merck Millipore.

Techniques: Concentration Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Binding Assay, SDS Page, Staining, Western Blot, Recombinant, Purification

Journal: Nanomedicine : nanotechnology, biology, and medicine

Article Title: Intratumoral Delivery of Antigen with Complement C3-bound Liposomes Reduces Tumor Growth in Mice

doi: 10.1016/j.nano.2018.10.009

Figure Lengend Snippet: Rhodamine-labeled OPSS-liposomes containing DQ-OVA incubated in human serum containing complement C3 (C3-liposomes) are taken up by (A) macrophages, (B) dendritic cells, and (C) B cells. Rhodamine-labeled control-liposomes (liposomes that contain DQ-OVA but without OPSS) and OPSS-liposomes lacking complement C3 are not taken up by cells. When compared with an equal amount of non-encapsulated DQ-OVA (Free DQ-OVA), C3-liposomes improve uptake and processing of DQ-OVA in macrophages and dendritic cells. B cells internalize C3-liposomes, but do not process DQ-OVA. Data are expressed as mean ± standard error (n=3). *p-value < 0.05, compared to Free DQ-OVA.

Article Snippet: Human serum with complement C3, and serum depleted of complement C3 were obtained from Quidel Corporation (Athens, OH, USA).

Techniques: Labeling, Incubation

Journal: Nanomedicine : nanotechnology, biology, and medicine

Article Title: Intratumoral Delivery of Antigen with Complement C3-bound Liposomes Reduces Tumor Growth in Mice

doi: 10.1016/j.nano.2018.10.009

Figure Lengend Snippet: Rhodamine-labeled OPSS-liposomes containing DQ-OVA incubated in human serum containing complement C3 (C3-liposomes) are taken up by (A) macrophages, (B) dendritic cells, and (C) B cells. Rhodamine-labeled control-liposomes (liposomes that contain DQ-OVA but without OPSS) and OPSS-liposomes lacking complement C3 are not taken up by cells. When compared with an equal amount of non-encapsulated DQ-OVA (Free DQ-OVA), C3-liposomes improve uptake and processing of DQ-OVA in macrophages and dendritic cells. B cells internalize C3-liposomes, but do not process DQ-OVA. Data are expressed as mean ± standard error (n=3). *p-value < 0.05, compared to Free DQ-OVA.

Article Snippet: Human serum with complement C3, and serum depleted of complement C3 were obtained from Quidel Corporation (Athens, OH, USA).

Techniques: Labeling, Liposomes, Incubation, Control