FIG 3.

Neutralization of Chandipura virus by complement proceeds via the classical pathway. (A) To identify the pathway mediating CHPV neutralization, 100 PFU of virus was incubated with either NHS, HI-NHS, 20 mM EDTA, 15 mM EGTA+2 mM MgCl₂, or NHS treated with EDTA or EGTA, and the remaining infectious titer was determined by plaque assays. Except the NHS-treated sample, none of the other treatments had any effect on the virus and, thus, were found to be statistically insignificant. (B) The total hemolytic activity of C4-depleted (C4 dep.) and -reconstituted serum (C4 dep.+C4) was tested against NHS using EA. (C) The significance of classical pathway in CHPV neutralization was tested by incubating CHPV (100 PFU) with either C4 dep. or C4 dep.+C4 with NHS serving as the control. The reduction in viral titer was determined by plaque assays. Statistical significance was calculated using values from six independent experiments, with ** denoting a P value of ≤0.005.