DOI: 10.5061/dryad.rj61p
title: Data from: Peripheral TNFa elevations in abstinent alcoholics are associated with Hepatitis C infection

description: Substantial evidence supports the view that inflammatory processes
 contribute to brain alterations in HIV infection. Mechanisms recently
 proposed to underlie neuropathology in Alcohol Use Disorder (AUD) include
 elevations in peripheral cytokines that sensitize the brain to the
 damaging effects of alcohol. This study included 4 groups: healthy
 controls, individuals with AUD (abstinent from alcohol at examination),
 those infected with HIV, and those comorbid for HIV and AUD. The aim was
 to determine whether inflammatory cytokines are elevated in AUD as they
 are in HIV infection. Cytokines showing group differences included
 interferon gammainduced protein 10 (IP-10) and tumor necrosis factor α
 (TNFα). Follow-up t-tests revealed that TNFα and IP-10 were higher in AUD
 than controls but only in AUD patients who were seropositive for Hepatitis
 C virus (HCV). Specificity of TNFα and IP10 elevations to HCV infection
 status was provided by correlations between cytokine levels and HCV viral
 load and indices of liver integrity including albumin/globulin ratio,
 fibrosis scores, and AST/platelet count ratio. Because TNFα levels were
 mediated by HCV infection, this study provides no evidence for elevations
 in peripheral cytokines in "uncomplicated", abstinent
 alcoholics, independent of liver disease or HCV infection. Nonetheless,
 these results corroborate evidence for elevations in IP-10 and TNFα in HIV
 and for IP-10 levels in HIV+HCV coinfection.
description: Zahr cytokines in
 pg-per-mlThese are the raw, pg/mL
 data from the shortly to be published PlosOne paper: Peripheral TNFα
 elevations in Alcohol Use Disorders are associated with Hepatitis C
 Infection, Natalie M. Zahr, Priya Asok, Edith V. Sullivan, & Adolf
 Pfefferbaum. Based on the recommendations of the Human Immune Monitoring
 Center (HIMC), http://iti.stanford.edu/himc.html, we used mean florescence
 intensity (MFI) in the manuscript. However, here we release the pg/mL data
 as that seems to be more useful for general use. Whole blood samples
 (n=223), collected in lavender EDTA tubes between March 2013 and October
 2016 were centrifuged (500 rcf at room temperature for 10min). Plasma was
 transferred to 1.5mL conical tubes, centrifuged at 13,000 rcf at room
 temperature for another 10min, and the resulting supernatant was
 transferred to 1.5mL conical tubes for storage at −80° C until analysis by
 the HIMC. The HIMC, which continually benchmarks processes to minimize
 technical variability (Maecker et al., 2005), performed immunological
 assays. Human 41-plex kits (HCYTOMAG-60K, 7 kits, each able to run 42
 samples) were purchased from EMD Millipore and used according to the
 manufacturer’s recommendations with modifications as described. Briefly,
 samples were mixed with antibody-linked magnetic beads on a 96-well plate
 and incubated overnight incubation at 4°C with shaking. Cold and Room
 temperature incubation steps were performed on an orbital shaker at
 500-600 rpm. Plates were washed twice with wash buffer in a Biotek ELx405
 washer. Following one hour incubation at room temperature with
 biotinylated detection antibody, streptavidin fluorochrome (i.e.,
 streptavidin-PE) was added for 30 minutes with shaking. Plates were washed
 as above and PBS added to wells for reading in the Luminex 200 Instrument
 with a lower bound of 50-100 beads per sample per cytokine. Each sample
 was measured in duplicate. Custom assay control beads by Radix
 Biosolutions were added to all wells.Zahr_cytokines in
 pg-per-ml.csv