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.
Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, China.
Wei
W
Wang
Jing
J
Wang
Sheng-fu
SF
Dong
Chun-hong
CH
Liu
Paola
P
Italiani
Shu-hui
SH
Sun
Jing
J
Xu
Diana
D
Boraschi
Shi-ping
SP
Ma
Di
D
Qu
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