Immunomodulatory activity of andrographolide on macrophage activation and specific antibody response.
OBJECTIVE
To investigate the immunomodulatory effects of andrographolide on both innate and adaptive immune responses.


METHODS
Andrographolide (10 microg/mL in vitro or 1 mg/kg in vivo) was used to modulate LPS-induced classical activated (M1) or IL-4-induced alternative activated (M2) macrophages in vitro and humor immune response to HBsAg in vivo. Cytokine gene expression profile (M1 vs M2) was measured by real-time PCR, IL-12/IL-10 level was detected by ELISA, and surface antigen expression was evaluated by flow cytometry, whereas phosphorylation level of ERK 1/2 and AKT was determined by Western blot. The level of anti-HBs antibodies in HBsAg immunized mice was detected by ELISA, and the number of HBsAg specific IL-4-producing splenocyte was enumerated by ELISPOT.


RESULTS
Andrographolide treatment in vitro attenuated either LPS or IL-4 induced macrophage activation, inhibited both M1 and M2 cytokines expression and decreased IL-12/IL-10 ratio (the ratio of M1/M2 polarization). Andrographolide down-regulated the expression of mannose receptor (CD206) in IL-4 induced macrophages and major histocompability complex/costimulatory molecules (MHC I, CD40, CD80, CD86) in LPS-induced macrophages. Correspondingly, anti-HBs antibody production and the number of IL-4-producing splenocytes were reduced by in vivo administration of andrographolide. Reduced phosphorylation levels of ERK1/2 and AKT were observed in macrophages treated with andrographolide.


CONCLUSIONS
Andrographolide can modulate the innate and adaptive immune responses by regulating macrophage phenotypic polarization and Ag-specific antibody production. MAPK and PI3K signaling pathways may participate in the mechanisms of andrographolide regulating macrophage activation and polarization.
Wang
Wei
W
Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, China.
Wang
Jing
J
Dong
Sheng-fu
SF
Liu
Chun-hong
CH
Italiani
Paola
P
Sun
Shu-hui
SH
Xu
Jing
J
Boraschi
Diana
D
Ma
Shi-ping
SP
Qu
Di
D
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects*; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004224:Diterpenes / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010766:Phosphorylation
D000276:Adjuvants, Immunologic; D017931:DNA Primers; D004224:Diterpenes; C030419:andrographolide