Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction

<b>Background:</b> Human Immunodeficiency Virus (HIV) infections remain a source of cardiopulmonary complications among people receiving antiretroviral therapy. Still to this day, pulmonary hypertension (PH) severely affects the prognosis in this patient population. The persistent expres...

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Main Authors: Amanda K. Garcia, Noelia C. Lujea, Javaria Baig, Eli Heath, Minh T. Nguyen, Mario Rodriguez, Preston Campbell, Isabel Castro Piedras, Edu Suarez Martinez, Sharilyn Almodovar
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Infectious Disease Reports
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Online Access:https://www.mdpi.com/2036-7449/17/3/65
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author Amanda K. Garcia
Noelia C. Lujea
Javaria Baig
Eli Heath
Minh T. Nguyen
Mario Rodriguez
Preston Campbell
Isabel Castro Piedras
Edu Suarez Martinez
Sharilyn Almodovar
author_facet Amanda K. Garcia
Noelia C. Lujea
Javaria Baig
Eli Heath
Minh T. Nguyen
Mario Rodriguez
Preston Campbell
Isabel Castro Piedras
Edu Suarez Martinez
Sharilyn Almodovar
author_sort Amanda K. Garcia
collection DOAJ
description <b>Background:</b> Human Immunodeficiency Virus (HIV) infections remain a source of cardiopulmonary complications among people receiving antiretroviral therapy. Still to this day, pulmonary hypertension (PH) severely affects the prognosis in this patient population. The persistent expression of HIV proteins, even during viral suppression, has been implicated in vascular dysfunction; however, little is known about the specific effects of these proteins on the pulmonary vasculature. This study investigates the impact of Nef variants derived from HIV-positive pulmonary hypertensive and normotensive donors on pulmonary vascular cells in vitro. <b>Methods:</b> We utilized well-characterized Nef molecular constructs to examine their effects on cell adhesion molecule gene expression (<i>ICAM1</i>, <i>VCAM1</i>, and <i>SELE</i>), pro-apoptotic gene expression (<i>BAX</i>, <i>BAK</i>), and vasoconstrictive endothelin-1 (<i>EDN1</i>) gene expression in endothelial nitric oxide synthase (eNOS) nitric oxide and the production and secretion of pro-inflammatory cytokines over 24, 48, and 72 h post-transfections with Nef variants. <b>Results:</b> HIV Nef variants SF2, NA7, and PH-associated Fr17 and 3236 induced a significant increase in adhesion molecule gene expression of <i>ICAM1</i>, <i>VCAM1</i>, and <i>SELE</i>. Pulmonary normotensive Nef 1138 decreased <i>ICAM1</i> gene expression, but had increased <i>VCAM1</i>. PH Nef ItVR showed a consistent decrease in <i>ICAM1</i> and no changes in <i>SELE</i> and <i>VCAM1</i> expression. Further gene expression analyses of pro-apoptotic genes <i>BAX</i> and <i>BAK</i> demonstrated that Nef NA7, SF2, normotensive Nef 1138, and PH Nef Fr8, Fr9, Fr17, and 3236 variants significantly increased gene expression for apoptosis. Normotensive Nef 1138, as well as PH Nef Fr9 and ItVR, all displayed a statistically significant decrease in <i>BAX</i> expression. The expression of <i>EDN1</i> had a statistically significant increase in samples treated with Nef NA7, SF2, normotensive Nef 2044 and PH Nef 3236, Fr17, and Fr8. Notably, PH-associated Nef variants sustained pro-inflammatory cytokine production, including IL-2, IL-4, and TNFα, while anti-inflammatory cytokine levels remained insufficient. Furthermore, eNOS was transiently upregulated by all Nef variants except for normotensive Nef 2044. <b>Conclusions:</b> The distinct effects of Nef variants on pulmonary vascular cell biology highlight the complex interplay between Nef, host factors, and vascular pathogenesis according to the variants.
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spelling doaj-art-b4ffadfdd04c442eaa94fed0b8bb34c52025-08-20T03:27:07ZengMDPI AGInfectious Disease Reports2036-74492025-06-011736510.3390/idr17030065Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell DysfunctionAmanda K. Garcia0Noelia C. Lujea1Javaria Baig2Eli Heath3Minh T. Nguyen4Mario Rodriguez5Preston Campbell6Isabel Castro Piedras7Edu Suarez Martinez8Sharilyn Almodovar9Department of Immunology & Molecular Microbiology, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USADepartment of Immunology & Molecular Microbiology, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USADepartment of Immunology & Molecular Microbiology, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USADepartment of Chemical Engineering, Edward E. Whitacre Jr. College of Engineering, Texas Tech University, Lubbock, TX 79430, USACollege of Arts and Sciences, Texas Tech University, Lubbock, TX 79430, USACenter for Biotechnology & Genomics, Texas Tech University, Lubbock, TX 79430, USACollege of Arts and Sciences, Texas Tech University, Lubbock, TX 79430, USACenter for Biotechnology & Genomics, Texas Tech University, Lubbock, TX 79430, USADepartment of Biology, University of Puerto Rico at Ponce, Ponce, PR 00732, USADepartment of Immunology & Molecular Microbiology, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA<b>Background:</b> Human Immunodeficiency Virus (HIV) infections remain a source of cardiopulmonary complications among people receiving antiretroviral therapy. Still to this day, pulmonary hypertension (PH) severely affects the prognosis in this patient population. The persistent expression of HIV proteins, even during viral suppression, has been implicated in vascular dysfunction; however, little is known about the specific effects of these proteins on the pulmonary vasculature. This study investigates the impact of Nef variants derived from HIV-positive pulmonary hypertensive and normotensive donors on pulmonary vascular cells in vitro. <b>Methods:</b> We utilized well-characterized Nef molecular constructs to examine their effects on cell adhesion molecule gene expression (<i>ICAM1</i>, <i>VCAM1</i>, and <i>SELE</i>), pro-apoptotic gene expression (<i>BAX</i>, <i>BAK</i>), and vasoconstrictive endothelin-1 (<i>EDN1</i>) gene expression in endothelial nitric oxide synthase (eNOS) nitric oxide and the production and secretion of pro-inflammatory cytokines over 24, 48, and 72 h post-transfections with Nef variants. <b>Results:</b> HIV Nef variants SF2, NA7, and PH-associated Fr17 and 3236 induced a significant increase in adhesion molecule gene expression of <i>ICAM1</i>, <i>VCAM1</i>, and <i>SELE</i>. Pulmonary normotensive Nef 1138 decreased <i>ICAM1</i> gene expression, but had increased <i>VCAM1</i>. PH Nef ItVR showed a consistent decrease in <i>ICAM1</i> and no changes in <i>SELE</i> and <i>VCAM1</i> expression. Further gene expression analyses of pro-apoptotic genes <i>BAX</i> and <i>BAK</i> demonstrated that Nef NA7, SF2, normotensive Nef 1138, and PH Nef Fr8, Fr9, Fr17, and 3236 variants significantly increased gene expression for apoptosis. Normotensive Nef 1138, as well as PH Nef Fr9 and ItVR, all displayed a statistically significant decrease in <i>BAX</i> expression. The expression of <i>EDN1</i> had a statistically significant increase in samples treated with Nef NA7, SF2, normotensive Nef 2044 and PH Nef 3236, Fr17, and Fr8. Notably, PH-associated Nef variants sustained pro-inflammatory cytokine production, including IL-2, IL-4, and TNFα, while anti-inflammatory cytokine levels remained insufficient. Furthermore, eNOS was transiently upregulated by all Nef variants except for normotensive Nef 2044. <b>Conclusions:</b> The distinct effects of Nef variants on pulmonary vascular cell biology highlight the complex interplay between Nef, host factors, and vascular pathogenesis according to the variants.https://www.mdpi.com/2036-7449/17/3/65HIV proteinsendothelial cell dysfunctionpulmonary vascular remodelingHIV Nef<i>ICAM1</i><i>VCAM1</i>
spellingShingle Amanda K. Garcia
Noelia C. Lujea
Javaria Baig
Eli Heath
Minh T. Nguyen
Mario Rodriguez
Preston Campbell
Isabel Castro Piedras
Edu Suarez Martinez
Sharilyn Almodovar
Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
Infectious Disease Reports
HIV proteins
endothelial cell dysfunction
pulmonary vascular remodeling
HIV Nef
<i>ICAM1</i>
<i>VCAM1</i>
title Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
title_full Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
title_fullStr Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
title_full_unstemmed Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
title_short Differential Effects of Human Immunodeficiency Virus Nef Variants on Pulmonary Vascular Endothelial Cell Dysfunction
title_sort differential effects of human immunodeficiency virus nef variants on pulmonary vascular endothelial cell dysfunction
topic HIV proteins
endothelial cell dysfunction
pulmonary vascular remodeling
HIV Nef
<i>ICAM1</i>
<i>VCAM1</i>
url https://www.mdpi.com/2036-7449/17/3/65
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