AM3, a natural glycoconjugate, induces the functional maturation of human dendritic cells.
OBJECTIVE
Dendritic cells (DCs) are dedicated antigen-presenting cells able to initiate specific immune responses and their maturation is critical for the induction of antigen-specific T-lymphocyte responses. Here, we have investigated the effects of Inmunoferon-active principle (AM3), the active agent of a commercial immunomodulatory drug, on human monocyte-derived DCs (MDDCs).


METHODS
MDDCs derived from healthy and hepatitis C virus (HCV)-infected patients were stimulated with AM3. We analysed the expression of cell surface proteins by flow cytometry, that of cytokine production by ELISA, and the expression of chemokines and chemokine receptors by RNase protection assays. T-lymphocyte proliferation was assessed in mixed lymphocyte reactions, protein expression by western blot and luciferase-based reporter methods, and Toll-like receptor (TLR)-blocking antibodies were employed to analyse TLR activity.


RESULTS
In MDDCs, AM3 induced or enhanced expression of CD54, CD83, CD86, HLA-DR, chemokines and chemokine receptors, interleukin (IL)-12p70 and IL-10. Furthermore, AM3 stimulated MDDCs to increase proliferation of allogenic T cells. AM3 triggered nuclear translocation of NF-kappaB and phosphorylation of p38 mitogen-activated protein kinase. AM3 promoted NF-kappaB activation in a TLR-4-dependent manner, and blocking TLR-4 activity attenuated the enhanced expression of CD80, CD83 and CD86 induced by AM3. AM3 enhanced the expression of maturation-associated markers in MDDCs from HCV-infected patients and increased the proliferation of T lymphocytes induced by these MDDCs.


CONCLUSIONS
These results underline the effects of AM3 in promoting maturation of MDDCs and suggest that AM3 might be useful in regulating immune responses in pathophysiological situations requiring DC maturation.
Martín-Vilchez
S
S
Liver Unit, Hospital Universitario de la Princesa, CIBER-EHD, Spain.
Molina-Jiménez
F
F
Alonso-Lebrero
J L
JL
Sanz-Cameno
P
P
Rodríguez-Muñoz
Y
Y
Benedicto
I
I
Roda-Navarro
P
P
Trapero
M
M
Aragoneses-Fenoll
L
L
González
S
S
Pivel
J P
JP
Corbí
A L
AL
López-Cabrera
M
M
Moreno-Otero
R
R
Majano
P L
PL
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000368:Aged; D015153:Blotting, Western; D002130:Calcium Phosphates / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D018925:Chemokines / Q000187:drug effects / Q000378:metabolism; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D006020:Glycopeptides / Q000494:pharmacology*; D006526:Hepatitis C / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D019707:Receptors, Chemokine / Q000187:drug effects / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000187:drug effects / Q000378:metabolism
D000276:Adjuvants, Immunologic; D002130:Calcium Phosphates; D018925:Chemokines; D006020:Glycopeptides; D019707:Receptors, Chemokine; D051197:Toll-Like Receptor 4; C038773:Immunoferon