PURPOSE: Basophils expressing FcɛRIα, the alpha chain of the total IgE high affinity receptor, appear to play a role in antigen (Ag) sensitization, although dendritic cells (DCs) are the principal Ag-presenting cells (APCs). To investigate whether these two types of APCs are involved differentially in Ag-sensitization when distinct types of adjuvants are utilized. METHODS: To investigate whether basophils and DCs serve as APCs in vitro, whole splenocytes, FcɛRIα(+) basophil-depleted splenocytes, and CD11c(+) DC-depleted splenocytes from naïve DO11.10 mice were stimulated in vitro with ovalbumin (OVA) or OVA peptide 323-339 to evaluate Ag-induced proliferation. To investigate whether basophils function as APCs in vivo, BALB/c mice were actively immunized with ragweed (RW) emulsified in alum or Complete Freund's Adjuvant (CFA) followed by an RW challenge in eye drops. An anti-FcɛRIα antibody (Ab) or a control Ab was injected intraperitoneally during the sensitization phase. Twenty-four hours after the RW challenge, conjunctivas and spleens were harvested for histological analysis to evaluate conjunctival eosinophilia and cytokine production, respectively. RESULTS: Depletion of basophils or DCs from naïve DO11.10 splenocytes significantly suppressed proliferative responses to either OVA or OVA peptide. Treatment with the anti-FcɛRIα Ab suppressed the conjunctival eosinophilia when alum, but not CFA, was utilized as the adjuvant. Similarly, the anti-FcɛRIα Ab inhibited cytokine production by splenocytes when alum was used as the adjuvant. CONCLUSION: Adjuvants determine which APCs are utilized in Ag-sensitization.
PURPOSE: Basophils expressing FcɛRIα, the alpha chain of the total IgE high affinity receptor, appear to play a role in antigen (Ag) sensitization, although dendritic cells (DCs) are the principal Ag-presenting cells (APCs). To investigate whether these two types of APCs are involved differentially in Ag-sensitization when distinct types of adjuvants are utilized. METHODS: To investigate whether basophils and DCs serve as APCs in vitro, whole splenocytes, FcɛRIα(+) basophil-depleted splenocytes, and CD11c(+) DC-depleted splenocytes from naïve DO11.10 mice were stimulated in vitro with ovalbumin (OVA) or OVA peptide 323-339 to evaluate Ag-induced proliferation. To investigate whether basophils function as APCs in vivo, BALB/c mice were actively immunized with ragweed (RW) emulsified in alum or Complete Freund's Adjuvant (CFA) followed by an RW challenge in eye drops. An anti-FcɛRIα antibody (Ab) or a control Ab was injected intraperitoneally during the sensitization phase. Twenty-four hours after the RW challenge, conjunctivas and spleens were harvested for histological analysis to evaluate conjunctival eosinophilia and cytokine production, respectively. RESULTS: Depletion of basophils or DCs from naïve DO11.10 splenocytes significantly suppressed proliferative responses to either OVA or OVA peptide. Treatment with the anti-FcɛRIα Ab suppressed the conjunctival eosinophilia when alum, but not CFA, was utilized as the adjuvant. Similarly, the anti-FcɛRIα Ab inhibited cytokine production by splenocytes when alum was used as the adjuvant. CONCLUSION: Adjuvants determine which APCs are utilized in Ag-sensitization.