Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection

<p><strong>Background:</strong> Haplotype-specific alternative splicing of the endoplasmic reticulum (ER) aminopeptidase type 2 (ERAP2) gene results in either full-length (FL, haplotype A) or alternatively spliced (AS, haplotype B) mRNA. HapA/HapA homozygous (HomoA) subjects show a reduced susceptibility to HIV-1 infection, probably secondary to the modulation of the antigen processing/presenting machinery. ERAP1 was recently shown to be secreted from the plasma membrane in response to activation; we investigated whether ERAP2 can be released as well and if the secreted form of this enzyme retains its antiviral function.</p> <p><strong>Methods:</strong> Human monocyte derived macrophages (MDMs) were differentiated from peripheral blood mononuclear cells (PBMCs) isolated from 6 HomoA healthy controls and stimulated with IFN&gamma; and LPS. ERAP2-FL secretion was evaluated by mass spectrometry. PBMCs (14 HomoA and 16 HomoB) and CD8-depleted PBMCs (CD8^&minus;PBMCs) (4 HomoA and 4 HomoB) were <em>in vitro</em> HIV-infected in the absence/presence of recombinant human ERAP2-FL (rhERAP2) protein; p24 viral antigen quantification was used to assess viral replication. IFN&gamma; and CD69 mRNA expression, as well as the percentage of perforin-producing CD8+ T Lymphocytes, were analyzed 3 and 7-days post <em>in vitro</em> HIV-1-infection, respectively. The effect of rhERAP2 addition in cell cultures on T cell apoptosis, proliferation, activation, and maturation was evaluated as well on 24 h-stimulated PBMCs.</p> <p><strong>Results:</strong> ERAP2 can be secreted from human MDMs in response to IFN&gamma;/LPS stimulation. Notably, the addition of rhERAP2 to PBMC and CD8^&minus;PBMC cultures resulted in the reduction of viral replication, though these differences were statistically significant only in PBMCs (<em>p</em> &lt; 0.05 in both HomoA and HomoB). This protective effect was associated with an increase in IFN&gamma; and CD69 mRNA expression and in the percentage of perforin-expressing CD107⁺CD8⁺ cells. RhERAP2 addition also resulted in an increase in CD8⁺ activated lymphocyte (CD25⁺HLA^&minus;DRII⁺) and Effector Memory/Terminally differentiated CD8⁺ T cells ratio.</p> <p><strong>Conclusions:</strong> This is the first report providing evidence for the release of ERAP2 in the secretome of immunocompetent cells. Data herein also indicate that exogenous ERAP2-FL exerts its protective function against HIV-1 infection, even in HomoB subjects who do not genetically produce it. Presumably, this defensive extracellular feature is only partially dependent on immune system modulation.</p>