Peripheral Complement C4 Protein in Schizophrenia: Association with Gene Copy Number and Immune Cell Subtypes

The lack of disease-modifying treatments for schizophrenia necessitates the exploration of novel aspects of its pathophysiology, including innate immune mechanisms in the periphery. C4 protein activation, associated with the complement cascade of innate immunity, associates with symptoms and predicts outcomes. However, C4 protein activation does not coincide with expected changes to other proteins in the complement cascade, suggesting another source of C4 protein activation. Using a combination of fresh whole blood from ten anonymous donors and a large set of publicly available microarray data, we show, for the first time, that C4 protein is found and expressed primarily in neutrophils and monocytes. Then, we compared the correlation between C4 protein in neutrophils, classical monocytes, plasma and the number of C4A gene copies. We determined the number of C4A genes using digital droplet PCR, C4 protein in neutrophils (15 patients/21 controls) and plasma (30 patients/38 controls) using western blotting, and classical monocytes (30 patients/38 controls) using flow cytometry. We found a moderate positive correlation between the number of C4A gene copies and the amount of C4 protein only in neutrophils and only in the schizophrenia group (Spearman's rho, r = 0.64, p = 0.01, Cohen's d ∼1.67). Our results indicate a convergence of innate immunity mechanisms associated with schizophrenia. This novel mechanism of innate immunity in schizophrenia deserves further study to determine whether it could be a useful drug target.