Assay of classical and alternative pathway activities of murine complement using antibody-sensitized rabbit...

Methods for measurement of classical complement pathway activity (CH50) and alternative complement pathway activity (ACH50) in mouse serum using rabbit erythrocytes sensitized with guinea pig anti-rabbit erythrocyte antibody have been established. The assays measured CH50 values in mouse sera that could hardly be determined by the conventional method using antibody-sensitized sheep red blood cells. Mouse serum ACH50 values determined by the method were also 5-7 times higher than those obtained in conventional assays with rabbit erythrocytes. Both the CH50 and ACH50 values varied with the strain among the 25 different strains of mice studied. BALB/c (nu/nu, male), LT/SuJ and Jcl-ICR27 strains exhibited higher CH50 values, and NIH (nu/+), ICR (nu/nu), NOD (male) and AKR strains showed lower values. The ACH50 was higher in C3H/HeN (male), C57BL/6J (male), Jcl-ICR27 and BALB/c (nu/nu, male) mice, and lower in ICR (nu/nu), NOD (female) and AKR mice. Sera from 16 out of the 25 mouse strains showed ACH50 values comparable to or higher than those in man. As for CH50, however, even the highest value seen in BALB/c (nu/nu, male) mice corresponded to about three-fifths of an average value in man. It is concluded that the complement system of mice, especially the alternative pathway of complement activation, functions as actively as that in man. It was also found that male mice have higher CH50 and ACH50 values than female mice. The differences in these parameters between males and females were only slight at the age of 4 weeks and became conspicuous after 6 weeks at which time both the CH50 and ACH50 virtually reached their respective peak levels of activity.