Extracellular allograft inflammatory factor-1 (AIF-1) potentiates Th1 cell differentiation and inhibits Treg response in human peripheral blood mononuclear cells from normal subjects

We identified the presence of AIF-1 (allograft inflammatory factor-1) in human peripheral blood mononuclear cells (PBMCs) from normal subjects by immunocytological methods. After isolation of different types of mononuclear cells by FACS (Fluorescence-activated cell sorting) with > 95% purity, we studied the transcript levels of AIF-1 using qPCR. We observed the following order of AIF-1 mRNA expression in mononuclear cells: T-lymphocytes (>) Monocytes (>) B-lymphocytes (>) NK. After T cell expansion of isolated PBMCs using anti-CD3-CD28 magnetic beads (Dynabeads), AIF-1 increased intracellularly in the presence of brefeldin A; this finding, along with an increase in the medium in the absence of the drug, suggests that AIF-1 is processed in the Golgi apparatus and may be secreted extracellularly. In another set of experiments, interleukin-12 and anti-interleukin-4 were added to PBMCs during T cell expansion to promote Th1 polarization and to inhibit Th2 differentiation. In this case, the presence of 6 nM of rhAlF-1 (recombinant human AIF-1) increased the mRNA expression of interferon-gamma and interleukin-2. In the same set of experiments, the incubation of PBMCs with rhAIF-1 (6 nM) promoted the decrease of mRNA expression of IL-10 and TGF-beta, along with the decrease of CD25 and Foxp3 proteins. Furthermore, extracellular rhAIF-1 (6 nM) increased the survival of naive and effector T cells during Th1 polarization by inhibition of apoptosis, without causing changes in cell cycle rate and in retinoblastoma-cyclin-dependent kinase (Rb-CDK) activation. Taken together, rhAIF-1 treatment of PBMCs potentiates Th1 response and inhibits functionally suppressive CD25 + Foxp3 + Treg, which suggests an important immunomodulatory role in governing T cell response.