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

Background: 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 infec...

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Autores principales: Saulle, Irma, Ibba, Salomè Valentina, Torretta, Enrica, Vittori, Cecilia, Fenizia, Claudio, Piancone, Federica, Minisci, Davide, Lori, Elisa Maria, Trabattoni, Daria, Gelfi, Cecilia, Clerici, Mario, Biasin, Mara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6646713/
https://www.ncbi.nlm.nih.gov/pubmed/31379846
http://dx.doi.org/10.3389/fimmu.2019.01648
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author Saulle, Irma
Ibba, Salomè Valentina
Torretta, Enrica
Vittori, Cecilia
Fenizia, Claudio
Piancone, Federica
Minisci, Davide
Lori, Elisa Maria
Trabattoni, Daria
Gelfi, Cecilia
Clerici, Mario
Biasin, Mara
author_facet Saulle, Irma
Ibba, Salomè Valentina
Torretta, Enrica
Vittori, Cecilia
Fenizia, Claudio
Piancone, Federica
Minisci, Davide
Lori, Elisa Maria
Trabattoni, Daria
Gelfi, Cecilia
Clerici, Mario
Biasin, Mara
author_sort Saulle, Irma
collection PubMed
description Background: 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. Methods: Human monocyte derived macrophages (MDMs) were differentiated from peripheral blood mononuclear cells (PBMCs) isolated from 6 HomoA healthy controls and stimulated with IFNγ and LPS. ERAP2-FL secretion was evaluated by mass spectrometry. PBMCs (14 HomoA and 16 HomoB) and CD8-depleted PBMCs (CD8(−)PBMCs) (4 HomoA and 4 HomoB) were in vitro HIV-infected in the absence/presence of recombinant human ERAP2-FL (rhERAP2) protein; p24 viral antigen quantification was used to assess viral replication. IFNγ and CD69 mRNA expression, as well as the percentage of perforin-producing CD8+ T Lymphocytes, were analyzed 3 and 7-days post in vitro 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. Results: ERAP2 can be secreted from human MDMs in response to IFNγ/LPS stimulation. Notably, the addition of rhERAP2 to PBMC and CD8(−)PBMC cultures resulted in the reduction of viral replication, though these differences were statistically significant only in PBMCs (p < 0.05 in both HomoA and HomoB). This protective effect was associated with an increase in IFNγ 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(−)DRII(+)) and Effector Memory/Terminally differentiated CD8(+) T cells ratio. Conclusions: 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.
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spelling pubmed-66467132019-08-02 Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection Saulle, Irma Ibba, Salomè Valentina Torretta, Enrica Vittori, Cecilia Fenizia, Claudio Piancone, Federica Minisci, Davide Lori, Elisa Maria Trabattoni, Daria Gelfi, Cecilia Clerici, Mario Biasin, Mara Front Immunol Immunology Background: 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. Methods: Human monocyte derived macrophages (MDMs) were differentiated from peripheral blood mononuclear cells (PBMCs) isolated from 6 HomoA healthy controls and stimulated with IFNγ and LPS. ERAP2-FL secretion was evaluated by mass spectrometry. PBMCs (14 HomoA and 16 HomoB) and CD8-depleted PBMCs (CD8(−)PBMCs) (4 HomoA and 4 HomoB) were in vitro HIV-infected in the absence/presence of recombinant human ERAP2-FL (rhERAP2) protein; p24 viral antigen quantification was used to assess viral replication. IFNγ and CD69 mRNA expression, as well as the percentage of perforin-producing CD8+ T Lymphocytes, were analyzed 3 and 7-days post in vitro 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. Results: ERAP2 can be secreted from human MDMs in response to IFNγ/LPS stimulation. Notably, the addition of rhERAP2 to PBMC and CD8(−)PBMC cultures resulted in the reduction of viral replication, though these differences were statistically significant only in PBMCs (p < 0.05 in both HomoA and HomoB). This protective effect was associated with an increase in IFNγ 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(−)DRII(+)) and Effector Memory/Terminally differentiated CD8(+) T cells ratio. Conclusions: 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. Frontiers Media S.A. 2019-07-16 /pmc/articles/PMC6646713/ /pubmed/31379846 http://dx.doi.org/10.3389/fimmu.2019.01648 Text en Copyright © 2019 Saulle, Ibba, Torretta, Vittori, Fenizia, Piancone, Minisci, Lori, Trabattoni, Gelfi, Clerici and Biasin. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Saulle, Irma
Ibba, Salomè Valentina
Torretta, Enrica
Vittori, Cecilia
Fenizia, Claudio
Piancone, Federica
Minisci, Davide
Lori, Elisa Maria
Trabattoni, Daria
Gelfi, Cecilia
Clerici, Mario
Biasin, Mara
Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title_full Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title_fullStr Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title_full_unstemmed Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title_short Endoplasmic Reticulum Associated Aminopeptidase 2 (ERAP2) Is Released in the Secretome of Activated MDMs and Reduces in vitro HIV-1 Infection
title_sort endoplasmic reticulum associated aminopeptidase 2 (erap2) is released in the secretome of activated mdms and reduces in vitro hiv-1 infection
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6646713/
https://www.ncbi.nlm.nih.gov/pubmed/31379846
http://dx.doi.org/10.3389/fimmu.2019.01648
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