Supplemental data. Antibodies used for Flow Cytometry. BL = Biolegend (San Diego, CA), BD = BD Biosciences (San Jose, CA) PD-1 Panel
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1 Supplemental data Antibodies used for Flow Cytometry BL = Biolegend (San Diego, CA), BD = BD Biosciences (San Jose, CA) PD-1 Panel Brilliant Stain Buffer (BD Cat# ) CD3-BUV395 (BD Cat# ) CD4-BV65 (BL Cat# ) CD8-BV711 (BL Cat# 3143) CD45RO-BV421 (BL Cat# 34223) CD197-FITC (BL Cat# ) CD62L-PE-Cy7 (BL Cat# 34821) CD95-APC (BL Cat# 35611) CD279-PE (BL Cat# 32995) HLA*DR-APC-Cy7 (BL Cat# 37617) CD38-PerCP-Cy5.5 (BL Cat# ) NK Panel Brilliant Stain Buffer (BD Cat# ) CD3-BUV395 (BD Cat# ) CD45-FITC (BD Cat# ) CD16-PE-Cy7 (BL Cat# 3216) CD56-BV421 (BL Cat# ) NKp44-APC (BL Cat# 32519) NKp3-BV711 (BD Cat# ) NKp46-PE (BD Cat# )
2 Monocyte Panel Brilliant Stain Buffer (BD Cat# ) CD3-BUV395 (BD Cat# ) CD19-BV65 (BL Cat# 32237) CD14-BV711 (BL Cat# 31837) CD16- PE-Cy7 (BL Cat# 3216) CD163-PE (Trillium Cat# CD P) CD56-BV421 (BL Cat# ) CD312-FITC (SeroTech Cat# MCA233F) CD97-APC (ebiosciences Cat # ) CD26-APC-Cy7 (BL Cat# 32112) CD38- PerCP-Cy5.5 (BL Cat# ) HLA*DR-APC-Cy7 (BL Cat# 37617)
3 Supplemental Figure 1. Hamilton Anxiety and Depression and Pittsburgh Sleep Quality Index Scores in HIV positive patients receiving aprepitant The Wilcoxon signed-rank test was used to determine whether within subject median change from day is significantly different than. P values are indicated on the graphs. The Hamilton Anxiety Scale The Hamilton Anxiety Scale p=.8 p=.3 Day Day 28 Day The Hamilton Depression The HAM-D 17 p=.6 Day Day 28 Day Pittsburgh Sleep Quality Index Score Pittsburgh Sleep Quality Index Score 15 1 p=.3 5 Day Day 28 Day
4 Supplemental Table 1A. Adverse events by subject (Number of adverse events), n (% of total individuals with adverse event) Subject 1 Subject 3 Subject 4 Subject 6 Subject 7 Subject 11 Subject 12 Total Any Serious Adverse Events Yes () () () () () () () () No 3 (1) 9 (1) 1 (1) 4 (1) 1 (1) 3 (1) 1 (1) 22 (1) Total 3 (1) 9 (1) 1 (1) 4 (1) 1 (1) 3 (1) 1 (1) 22 (1) Severity Mild () 6 (66.7) 1 (1) 1 (25.) 1 (1) 2 (66.7) 1 (1) 12 (54.5) Moderate 3 (1) 3 (33.3) () 3 (75.) () 1 (33.3) () 1 (45.5) Severe () () () () () () () () Total 3 (1) 9 (1) 1 (1) 4 (1) 1 (1) 3 (1) 1 (1) 22 (1) Supplemental Table 1B. Adverse events details by subject, n (%) Subject 1 Subject 3 Subject 4 Subject 6 Total Relationship to Medication Not related () () () 1 (25.) 1 (5.9) Possible 3 (1) 9 (1) 1 (1) 2 (5.) 15 (88.2) Unlikely () () () 1 (25.) 1 (5.9) Action Taken None 3 (1) 9 (1) 1 (1) 4 (1) 17 (1) Outcome Improved () 2 (22.2) () () 2 (11.8) Recovered/resolved 3 (1) 6 (66.7) 1 (1) 3 (75.) 13 (76.5) Unchanged () 1 (11.1) () 1 (25.) 2 (11.8) Ongoing No 3 (1) 8 (88.9) 1 (1) 3 (75.) 15 (88.2) Yes () 1 (11.1) () 1 (25.) 2 (11.8)
5 Supplemental Figure 2. Hemoglobin, hematocrit and RBC count in HIV positive patients receiving aprepitant Wilcoxon signed-rank test was used to determine whether within subject median change from day is significantly different than. Significant p values (<.5) are indicated on the graphs. Hemoglobin (g/dl) Hemoglobin levels p=.1 Day Day 14 Day 28 Day Hematocrit (%) Hematocrit levels p=.3 Day Day 14 Day 28 Day RBC (x1 6 cells/µl) RBC p=.2 Day Day 14 Day 28 Day Supplemental Figure 3. Platelets in HIV positive patients receiving aprepitant Wilcoxon signed-rank test was used to determine whether within subject median change from day is significantly different than. P values are indicated on the graph. platelets (x1 3 /µl) platelets p=.6 p=.2 p=.3 Day Day 14 Day 28 Day
6 Supplemental Figure 4. Changes in total bilirubin, alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in HIV positive patients receiving aprepitant Results are presented as changes in marker levels between day and 14, day and 28 or day and 58 as indicated. Vertical bars are the widths of the 95% confidence intervals. P values by Wilcoxon signed-rank test are indicated on the plots. total bilirubin(mg/dl) p=.9 total bilirubin p=.1 p=.2 ALT (U/L) p=.9 ALT p=.8 p=.3 ALP (U/L) p=.2 ALP p=.3 p< day 14- day 28- day day 14- day 28- day day 14- day 28- day 58-
7 Supplemental Figure 5. Changes in CD4+ T-cell count in HIV positive patients receiving aprepitant Results are presented as changes in absolute CD4+ T-cell numbers between day and 14, day and 28 or day and 58 as indicated. Vertical bars are the widths of the 95% confidence intervals. P values by Wilcoxon signed-rank test are indicated on the plot. 4 CD4 p=.14 change in CD4 (cells per µl) p=.3 p=.6-6 day 14- day 28- day 58-
8 Supplemental Table 2. Viral loads by ultrasensitive HIV viral load assay in HIV positive patients receiving aprepitant HMMCgag (RNA copies/ml) PID Day Day 28 1 < <.75 < <.77 4 <.84 <.78 5 <.79 < < * 2.9 * * 11 <.93 < <.75 * HIV-1 DNA detected in controls without RT; values likely do not represent the true viral load.
9 Supplemental Figure 6. CD4+ Tcm and CD8+ Tcm in HIV positive patients receiving aprepitant Wilcoxon signed-rank test was used to determine whether within subject median change from day is significantly different than. P values are indicated on the graphs. CD4+ Tcm (%) CD4+ Tcm p=.2 p=.1 p=.8 Day Day 14 Day 28 Day CD8+ Tcm (%) CD8+ Tcm p=.7 p=.1 p=.1 Day Day 14 Day 28 Day
10 Supplemental Table 3. Complete list of proteins affected by aprepitant treatment. Likelihood ratio tests were performed using the ANOVA function of the car R package to calculate proteins that showed significant changes in their expression levels after treatment with aprepitant. The regression coefficient demonstrates both the direction and the magnitude of the change. False discovery rate (FDR) adjusted p-values that account for multiple comparisons are included in the table. Protein Name p-value regression coefficient FDR adjusted p-value Proteins with higher expression after aprepitant treatment Cathepsin.Z Complement.C Thioredoxin.domain.containing.protein Lysosomal.protective.protein Asialoglycoprotein.receptor Interleukin.1.receptor.like Leptin.receptor Vitamin.K.dependent.protein.C Vascular.endothelial.growth.factor.A Thyroid.Stimulating.Hormone Alpha.1.antitrypsin Cathepsin.F SPARC.like.protein CD29.antigen Protein.Z.dependent.protease.inhibitor Low.density.lipoprotein.receptor.related.protein.1.soluble Ectonucleoside.triphosphate.diphosphohydrolase Tenascin Hemojuvelin Dipeptidyl.peptidase Apolipoprotein.L Neutral.ceramidase Importin.subunit.alpha Carboxypeptidase.B Receptor.tyrosine.protein.kinase.erbB Arylsulfatase.B N.acylethanolamine.hydrolyzing.acid.amidase Complement.C5b.C6.complex Apolipoprotein.M Mannose.binding.protein.C Netrin Myeloperoxidase
11 Afamin Leucine.rich.repeats.and.immunoglobulin.like.domains.protein Coagulation.Factor.XI Ectonucleotide.pyrophosphatase.phosphodiesterase.family.member Trefoil.factor Carbohydrate.sulfotransferase Alpha.L.iduronidase X protein.sigma Coagulation.factor.VII Tyrosine.protein.kinase.receptor.TYRO Sex.hormone.binding.globulin Annexin.A Endoglin Cation.independent.mannose.6.phosphate.receptor Interleukin.1.receptor.type Antileukoproteinase Hepatocyte.growth.factor.receptor Apolipoprotein.E.isoform.E Tumor.necrosis.factor.receptor.superfamily.member X.ray.repair.cross.complementing.protein Fetuin.B Antithrombin.III Coagulation.Factor.V Serum.amyloid.P.component Repulsive.guidance.molecule.A Low.affinity.immunoglobulin.gamma.Fc.region.receptor.II.b Proprotein.convertase.subtilisin.kexin.type Apolipoprotein.B Dynactin.subunit Vesicular.integral.membrane.protein.VIP Prolactin Platelet.activating.factor.acetylhydrolase Low.density.lipoprotein.receptor.related.protein.1B Kallikrein Hepatocyte.growth.factor.like.protein Ficolin Histone.lysine.N.methyltransferase.EHMT Acid.sphingomyelinase.like.phosphodiesterase.3a Polymeric.immunoglobulin.receptor Tumor.necrosis.factor.ligand.superfamily.member.13B Troponin.I.cardiac.muscle Tumor.necrosis.factor.ligand.superfamily.member gp41.c34.peptide.hiv SLAM.family.member Prothrombin Tyrosine.protein.kinase.JAK GDNF.family.receptor.alpha Heparan.sulfate.6.O.sulfotransferase
12 Bone.morphogenetic.protein Superoxide.dismutase.Mn.mitochondrial Vascular.endothelial.growth.factor.receptor Lactadherin Dipeptidyl.peptidase C.C.motif.chemokine Thrombin C.type.lectin.domain.family.4.member.K Inter.alpha.trypsin.inhibitor.heavy.chain.H Serum.albumin Plasma.protease.C1.inhibitor Osteopontin Leucine.rich.repeat.transmembrane.protein.FLRT Complement.factor.H.related.protein Interleukin.1.alpha Small.nuclear.ribonucleoprotein.F Interleukin.13.receptor.subunit.alpha Cadherin Lymphatic.vessel.endothelial.hyaluronic.acid.receptor Endoplasmic.reticulum.resident.protein Toll.like.receptor Apolipoprotein.E.isoform.E Interleukin.1.receptor.type Iduronate.2.sulfatase Complement.component.C Proteins with lower expression after aprepitant treatment RNA.binding.protein Lymphocyte.activation.gene.3.protein Fibroblast.growth.factor Thrombospondin Prolyl.endopeptidase.FAP Complement.component.C1q.receptor Proto.oncogene.tyrosine.protein.kinase.receptor.Ret Angiogenin Contactin Casein.kinase.II.subunit.alpha C.type.mannose.receptor Complement.C3d.fragment B.cell.receptor.CD Transforming.growth.factor.beta Dermatopontin Insulin.like.growth.factor.I Alpha.2.HS.glycoprotein Cadherin Mitochondrial.import.inner.membrane.translocase.subunit.TIM Bcl.2.like.protein Interleukin.2.receptor.subunit.alpha
13 Collectin Matrix.extracellular.phosphoglycoprotein Caspase Tyrosine.protein.kinase.Lck Ephrin.type.B.receptor A.disintegrin.and.metalloproteinase.with.thrombospondin.motifs C.C.motif.chemokine Dickkopf.related.protein Killer.cell.lectin.like.receptor.subfamily.F.member X3.hydroxyacyl.CoA.dehydrogenase.type Kin.of.IRRE.like.protein Brain.natriuretic.peptide Resistin Interleukin.17.receptor.A Fibroblast.growth.factor.receptor Macrophage.metalloelastase Protein.SET Fibroblast.growth.factor Complement.factor.B DnaJ.homolog.subfamily.B.member Collagen.alpha.1.XXIII..chain Cyclin.dependent.kinase.1.G2.mitotic.specific.cyclin.B1.complex Serine.threonine.protein.kinase.17B Cell.adhesion.molecule.related.down.regulated.by.oncogenes Transforming.growth.factor.beta SLIT.and.NTRK.like.protein Interleukin Serum.paraoxonase.arylesterase Hexokinase High.affinity.nerve.growth.factor.receptor Junctional.adhesion.molecule.B Integrin.alpha.IIb.beta.3.complex Growth.differentiation.factor Stem.cell.growth.factor.alpha Brevican.core.protein Complement.C3b.inactivated Platelet.derived.growth.factor.subunit.A Estrogen.receptor Angiopoietin Serine.threonine.protein.kinase.PAK Protein.tyrosine.kinase Dual.specificity.mitogen.activated.protein.kinase.kinase Reticulon.4.receptor Galectin Interleukin.23.receptor Fibroblast.growth.factor Interleukin Peptidyl.prolyl.cis.trans.isomerase.F.mitochondrial
14 Advanced.glycosylation.end.product.specific.receptor.soluble Matrix.metalloproteinase
x Lymphocyte count /µl CD8+ count/µl 800 Calculated
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