Journal: Molecular immunology

Article Title: The Human Complement Receptor Type 2 (CR2)/CR1 Fusion Protein TT32, a Novel Targeted Inhibitor of the Classical and Alternative Pathway C3 Convertases, Prevents Arthritis in Active Immunization and Passive Transfer Mouse Models

doi: 10.1016/j.molimm.2018.09.013

Figure Lengend Snippet: Effect of TT32 on AP50 and CP50 activities. The AP50 and CP50 haemolytic complement activities were measured by using Rabbit blood cells (RBCs) and Sheep red blood cells lytic assays, respectively. The AH50 and CH50 were determined by analyzing the capacity of human and mouse serum incubated with various doses of TT32 or TT30 or SCR1–10 to lyse antibody coated RBCs or SBCs. The calculated percentage (%) of lysis of the control and serum samples from human and mouse were plotted against dilution factor. TT32 inhibits both human (A, B) and mouse (A) complement activation more efficiently than TT30 or CR1 1–10 using rabbit RBC alternative pathway (A, B) or sheep RBC classical pathway (C) lysis assay. HS, human serum; MS, mouse serum.

Article Snippet: Sensitization of sheep RBC (Bioreclamation, Westbury, NY) was achieved by incubation with rabbit anti-sheep hemolysin (Cedarlane Labs) in the presence of gelatin-veronal buffer (Boston Bioproducts) for 30 min at 37 ° C. Next, the optimal serum concentration to lyse erythrocytes via CP activation was determined by incubating sensitized SRBCs with serial dilutions of complement-preserved human serum and measuring hemoglobin release at 541 nm.

Techniques: Incubation, Lysis, Activation Assay