CROI 2017 Abstract e-Book

Abstract eBook

Poster and Themed Discussion Abstracts

Results: Treatment with Maraviroc resulted in a block in S phase of LX-2 cells with increased expression levels of cyclin D1 and p21 while the expression of p53 was reduced. Treatment with Maraviroc was also able to block the accumulation of fibrillar collagens and extracellular matrix proteins (ECM), as demonstrated by the decrease of specific markers as Collagen type I, α-SMA and TGF-β1. In addition we observed a down regulation of both metalloproteins (MMP-2, MMP-9), used for the degradation of the extracellular matrix and their inhibitors (TIMP-1, TIMP-2). Conclusion: The identification of a compound that may modulate the dynamic of liver fibrosis could be crucial in all chronic liver diseases. Maraviroc could play an important role because, in addition to its own anti-HIV activity, it could reduce the release of pro-inflammatory citokynes implicated in liver fibrogenesis, making this drug particularly recommended for antiretroviral regimens chosen to treat HIV/HCV coinfected patients. 586 CORRELATION OF BLOOD TRANSCRIPTOME WITH OUTCOME AFTER DAA TREATMENT FOR HCV INFECTION Cody Orr 1 , Paul Nietert 1 , Shyam Kottilil 2 , Gary Hardiman 1 , Eric G. Meissner 1 1 Med Univ of South Carolina, Charleston, SC, USA, 2 Univ of Maryland, Baltimore, MD, USA Background: Combination treatment with direct acting antivirals (DAA) results in a sustained virologic response (SVR) in most patients. For reasons that are poorly understood, some patients experience virologic relapse after treatment. The ability to predict relapse and identify biomarkers allowing shorter treatment would have practical implications in resource limited settings. We previously showed differences in hepatic expression of interferon-related genes that correlated with SVR in patients treated with sofosbuvir and ribavirin. Here, we hypothesized that whole blood gene expression would correlate with treatment outcome and suggest mechanisms of relapse. Methods: We analyzed cryopreserved paired whole blood samples collected in PAXgene tubes before and at the end of treatment (prior to relapse) from 40 chronically infected HCV patients (n=26 SVR, n=14 relapse) treated with 24 weeks of sofosbuvir and ribavirin in the NIH SPARE trial (NCT01441180). mRNA was extracted using the PAXgene Blood miRNA Kit (Qiagen) with quality determined by Agilent Bioanalyzer (median RIN 7.2, range 5.3-8.5). Expression of 579 unique immune-related transcripts was determined using the Nanostring Human Immunology v2 Panel. Data were analyzed with non-parametric approaches (SPSS software). NSolver Advanced Analysis Software 3.0 was used for immune cell type profiling of gene expression results. Results: Considering all 40 patients, expression of 251 of 579 genes changed significantly during treatment. Comparing SVR vs. relapse patients, differential expression was observed for 39 mRNAs pre-treatment, 27 mRNAs at the end-of-treatment, and 35 mRNAs over the course of treatment (post-pre). Intriguingly, genes involved in inhibition of host immunity had higher pre-treatment (CD244, CTLA4, SOCS1) and end-of-treatment (PD1, SOCS1) expression in relapsers. Immune cell type abundance based on transcriptional profiling suggested B-cell, CD4+ Th1-cell, and total, exhausted, and cytotoxic CD8+ T-cell frequencies decreased with treatment, while the neutrophil transcriptional profile increased, data that corresponded with changes in immune cell frequency determined by flow cytometry. Interestingly, there was a trend (p=0.08) towards higher transcriptional expression of an exhausted CD8+ T-lymphocyte profile at the end of treatment in relapsers. Conclusion: Whole blood gene expression profiles differ by outcome for DAA treatment of HCV infection and suggest a relationship of cellular exhaustion with relapse. 587 DO MAIT CELLS IMPACT FIBROSIS IN HCV AND HIV/HCV COINFECTED PATIENTS? Boris J. Beudeker 1 , Gertine van Oord 1 , Joop Arends 2 , Julian Schulze zur Wiesch 3 , Marieke van Heide - Mulder 1 , Robert de Knegt 1 , Annelies Verbon 1 , Andre Boonstra 1 , Mark Claassen 4 1 Erasmus Univ Med Cntr, Rotterdam, Netherlands, 2 Univ Med Cntr Utrecht, Utrecht, Netherlands, 3 Univ Med Cntr Hamburg-Eppendorf, Hamburg, Germany, 4 Rijnstate Hosp, Arnhem, Netherlands Background: Mucosal-associated invariant T (MAIT) cells are important innate T cells with anti-microbial and immunoregulatory activity that have been recently shown to be depleted in blood of patients with HIV and HCV mono-infections. In this study, we assessed the impact of HIV, HCV and HCV/HIV co-infection on circulating and intrahepatic MAIT cells and correlations with liver fibrosis. Methods: In this cross-sectional study, 9 healthy subjects, 9 HIV, 20 HCV and 22 HCV/HIV co-infected patients were included. Blood and liver fine needle aspirate biopsies (FNAB) were studied using flowcytometry for CD3+CD161+Vα7.2+MAIT cell frequency, phenotype and function in HCV mono-infected and HCV/HIV co-infected patients without or with mild fibrosis (Metavir-score F0-F1) or severe fibrosis to cirrhosis (Metavir-score F3-F4). Results: Circulating MAIT cells were decreased in blood of HCV, HIV and HCV/HIV patients without or with only mild liver fibrosis. In HCV/HIV co-infected individuals with F3-F4, the frequency of circulating MAIT cells was even further depleted, whereas their function was comparable to HCV/HIV co-infected patients with low or absent fibrosis. In contrast, in HCV mono-infected patients, MAIT cell frequencies were not related to fibrosis severity; however, MAIT cell function was impaired in mono-infected patients with more fibrosis. More advanced liver fibrosis in HCV or HCV/HIV-infected patients was not reflected by increased accumulation of MAIT cells in the affected liver. Conclusion: Severe liver fibrosis is associated with dysfunctional MAIT cells in blood of HCV mono-infected patients, and lower MAIT frequencies in blood of HCV/HIV co-infected patients, without evidence for accumulation in the liver. 588 PATHOGENESIS OF HIV-INDUCED LIVER DISEASE IN DUAL IMMUNE AND LIVER HUMANIZED MICE Raghubendra S. Dagur , Weimin Wang, Yan Cheng, Weizhe Li, Edward Makarov, Natalia Osna, Santhi Gorantla, Larisa Y. Poluektova Univ of Nebraska, Omaha, NE, USA Background: HIV-1 infection has a negative impact on liver function, and liver diseases have become a leading cause of mortality in infected patients. Mechanisms of liver damage remain unclear. In vitro cell culture experiments usually have limitation; moreover, collection of clinical data depends on multiple boundaries: time of infection, co- infections with hepatitis viruses, drugs of abuse, and treatment of patients. To overcome such limitations, we utilized chimeric mice dually reconstituted with human blood and liver. Methods: TK-NOG (CIEA, Japan) males were simultaneously transplanted with human CD34+ hematopoietic stem cells (HSCs) and hepatocytes (Hep) at 6-8 weeks of age, intrasplenically. Levels of human liver and immune reconstitution were monitored by human albumin (Alb) in plasma (ELISA) and the presence of human immune cells in blood, spleen and liver tissues (FACS). At 5 months post-transplantation, mice were infected with HIV-1 and euthanized at 5 weeks post infection. Liver pathomorphology was assessed by staining for CK-18, apoptotic Hep (M30), αSMA (fibrosis), GFAP (stellate cells activation), human HLA-DR, CD4, CD8 infiltration, and HIV-1 p24 antigen. Infection was confirmed by detection of virus in plasma and HIV-gag mRNA expression in primary hepatocytes. Changes in liver infiltrating immune cells milieu were evaluated by comparison of the expression of TLR receptors, chemokines and their receptors, and acute and chronic inflammatory responses by RT-PCR array. To further dissect the mechanisms of Hep damage at cellular and functional levels, primary Hep cultures were exposed to HIV-1. Results: HIV-infected mice showed a decline of human Alb concentration (42%,p < 0.05) and significant reduction in CK18+ Hep (50%,p< 0.05), compared to uninfected. The decrease in human albumin level was correlated with CD4+ cells in liver (R²=0.72,p < 0.05) and increase in HIV-1 viral load in plasma (R²=0.77, p < 0.05). In vitro and in vivo, HIV-1 caused apoptosis of Hep. In infected-liver tissues, up-regulation of TLR7(5.9 fold) > 9(4.1)>1(2.0)>3(1.9)> 4(1.5) and increased expression of chemokines involved in the formation of lymphoid aggregates (CCL7(3.3 fold), CCL23(2.8) and CCL21(2.6)), attraction of lymphocytes (CXCL10(15.9)) and neutrophils (CXCR1(3.6fold) and CXCR2(3.2)), as well as an elevation of IL23A (2.3), compared to uninfected were evident. Conclusion: Dual reconstituted TK-NOG mice are a feasible platform to study in vivo HIV-mediated human liver damage and therapeutics development. 589 PERIPHERAL ENDOTHELIAL FUNCTION IS SUBOPTIMAL IN HCV-UNTREATED, HIV-SUPPRESSED ADULTS Kara W. Chew 1 , Eric Daar 2 , Jordan E. Lake 3 , Chrisandra L. Shufelt 4 , Zhaoping Li 1 , Diana Liao 1 , Wilbert Jordan 5 , C. Noel Bairey Merz 4 , Judith S. Currier 1 , Debika Bhattacharya 1

Poster and Themed Discussion Abstracts

CROI 2017 247