The role of the BAFF/APRIL system in the T cell-independent specific response to blood stage Plasmodium falciparum hemozoin

Abstract

Background: The B cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are tumor necrosis factor family members that regulate B cell maturation, proliferation, survival and function. We have previously shown that blood-stage Plasmodium falciparum hemozoin (HZ) can act as a T-independent antigen (TI Ag) that induces the production of specific IgG to soluble crude P. falciparum Ag through the BAFF pathway. However, we have not yet clarified whether HZ need APRIL signaling in the TI response. Here, we aimed to clarify whether both BAFF and APRIL signaling pathways play roles in HZ induction of specific antibody production without T-cell help.

Methods: Normal monocytes alone or co-cultured with naïve B cells were stimulated by HZ (10 µM) in vitro. Naïve B cell cultures, with HZ alone or with exogenous recombinant BAFF (rBAFF) and recombinant APRIL (rAPRIL) plus recombinant IL-4 (rIL-4) for 6 and 10 days were used as controls to investigate activation of B cells. At various times, the levels of sBAFF, sAPRIL, and HZ-specific IgG in the culture supernatants were assessed by enzyme-linked immunosorbent assay. The BAFF and APRIL expression levels on the HZ-stimulated monocytes and their specific receptors on activated B cells, including the BAFF receptor (BAFF-R), the transmembrane activator and calcium-modulator and cyclophilin ligand interactor (TACI) and the B cell maturation antigen (BCMA), were determined by flow cytometry. mRNA expression levels for the receptors were validated using Real-Time quantitative PCR.

Results: HZ-activated monocytes released sBAFF and sAPRIL during the 72 h stimulation period. Increased mRNA encoding of their cognate receptors, BAFF-R, TACI, and BCMA, and increased HZ-specific IgG levels were also observed in HZ induction within the monocyte and B cell co-culture. The experiments under control conditions revealed that HZ alone could induce B cell culture to produce a small amount of the specific IgG compared with those in medium alone or rBAFF + rAPRIL + rIL-4.

Conclusion: Taken together, we suggest that in the TI response HZ stimulates monocyte and B cell co-culture to produce specific IgG through BAFF, APRIL and other independent complimentary signaling pathways.

Keywords: A proliferation-inducing ligand; B cell activating factor; B cell activating factor receptor; B cell maturation antigen; Hemozoin; Plasmodium falciparum; Transmembrane activator and calcium-modulator and cyclophilin ligand interactor.