Complement fragments C3b and iC3b coupled to latex induce a respiratory burst in human neutrophils.
The complement fragments C3b and iC3b were purified from human serum by affinity chromatography with Sepharose-coupled monoclonal antibody against the C3d region of C3. The resulting preparations were more than 95% pure and contained less than 0.1% native IgG. Purified C3b and iC3b were coupled to latex beads (0.8 micron diameter) by means of F(ab')2 fragments of monoclonal antibodies against the beta chain or the C3d region of C3, thus orienting the C3b and the iC3b on the latex with the C3b- and iC3b-specific regions outwards. These particles were found to activate the respiratory burst of freshly isolated human neutrophils to 20-30% of the maximal capacity. Latex particles randomly coated with C3b or iC3b were about 3 times less stimulatory. C3b, iC3b and IgG coupled to latex in an oriented fashion were about equally effective in stimulating the respiratory burst. Neutrophils from a patient with a total deficiency of CR3 responded normally to C3b-coated latex but did not respond to iC3b-coated latex. A monoclonal antibody against the alpha chain of CR3 inhibited the activation by iC3b-coated latex and a polyclonal antibody against CR1 partially inhibited the activation by C3b-coated latex. We found an additive effect between IgG-coated latex and C3b-coated latex, regardless of the presence of IgG and C3b on the same particle or on different particles. Thus, binding of ligands to either CR1 or CR3 per se is sufficient to induce an activating signal to the NADPH oxidase in human neutrophils.
Hoogerwerf
M
M
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Weening
R S
RS
Hack
C E
CE
Roos
D
D
D015703:Antigens, CD / Q000276:immunology; D003179:Complement C3b / Q000276:immunology*; D003180:Complement C3b Inactivator Proteins; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007840:Latex; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D009895:Opsonin Proteins / Q000276:immunology; D010101:Oxygen Consumption; D011951:Receptors, Complement / Q000276:immunology; D017463:Receptors, Complement 3b
D015703:Antigens, CD; D003180:Complement C3b Inactivator Proteins; D007140:Immunoglobulin Fab Fragments; D007840:Latex; D009895:Opsonin Proteins; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D003179:Complement C3b