CROI 2020 Abstract eBook

Abstract eBook

Poster Abstracts

alleles. CMV co-infection emerges as an important contributor to gut damage and microbial translocation and may contribute to non-AIDS comorbidities in EC. 233 FROM GUT TO BLOOD: REDISTRIBUTION OF ZONULIN IN HIV+ PATIENTS Max Augustin 1 , Carola Horn 1 , Meryem S. Ercanoglu 1 , Vincent Bondet 2 , Isabelle Suarez 1 , Seung-Hun Chon 1 , Dirk Nierhoff 1 , Elena Knops 1 , Eva Heger 1 , Gerd Fätkenheuer 1 , Jan Rybniker 1 , Darragh Duffy 2 , Michaela Müller-Trutwin 2 , Clara Lehmann 1 1 Cologne University Hospital, Cologne, Germany, 2 Institut Pasteur, Paris, France Background: Gastro-intestinal mucosal damage in HIV infection causes microbial translocation and immune activation, which in turn results in non- infectious comorbidities. Combined antiretroviral therapy (cART) restores this intestinal damage only partially. Various biomarkers of the epithelial barrier have been reported but some are not considered specific while others are impacted by cART. Zonulin is a modulator for epithelial tight junctions. Previous studies found an elevated level of circulating Zonulin in the blood during HIV infection, while others reported a decrease. We measured Zonulin in serum and intestinal tissue sections and compared it with inflammatory markers and the virus reservoir in the blood (PB), ileum (TI) and rectum (R) of HIV+ and controls. Methods: Biopsies and gut tissue sectionsfrom TI, R and PB were collected from 5 treatment naïve (HIV+NAÏVE) and 10 cART-treated (HIV+cART) HIV+ individuals and 11 controls (CTRL). Lamina propria mononuclear cells were isolated. Following flow cytometry and cell sorting of CD4+ T cells, total HIV-DNA was quantified in PB, TI and R. In serum circulating Zonulin was measured by ELISA (Immundiagnostik) and in gut sections by semi quantitative immunohistochemistry. Ultrasensitive digital ELISA (Simoa; Quanterix) was used to measure IFN-α in serum and tissue supernatants. Results: Median CD4+T cell count [cells/µl] in HIV+NAÏVEwas 70(30- 255) versus 426(293-787) in HIV+cART. Median time on ART[years]was 6(9-10). Circulating Zonulin levels [ng/ml]were highest in treatment-naïve HIV+when compared to cART-treated HIV+(p=0.04) or CTRL (p=0.0087; HIV+NAÏVE>HIV+cART>CTRL). Similarly, HIV+NAÏVE showed higher IFN-α and HIV-DNA levels in PB when compared to HIV+cART(IFN-α: p=0.04; HIV-DNA: p=0.04). In gut tissue sections however, Zonulin was higher in CTRL when compared to HIV patients. Circulating Zonulin in serumwas negatively correlated to plasma CD4 cell count (r= -0.54, p=0.04), CD4+T cell frequencies in TI (r= -0.58 p=0.04) and positively to IFN-α in TI (r=0.65, p=0.05). Conclusion: The data indicate that upon HIV infection, Zonulin levels decrease in gut, but increase in plasma. The latter were associated with loss of intestinal CD4+T cells and increased inflammation in the gut, suggesting that increased levels of systemic Zonulin correlate with intestinal damage. An increased understanding of the regulation of gut tight junctions during HIV infection may be crucial for the design of future therapies. 234 GUT BARRIER PROTECTANT INTESTINAL ALKALINE PHOSPHATASE IS REDUCED IN PEOPLE WITH HIV Charlie F. Saylor 1 , Jae H. Sim 1 , Lediya Cheru 1 , Yang Liu 1 , Paul Cavallaro 1 , Fatemeh Adiliaghdam 1 , Kiana Keller 1 , Shibani S. Mukerji 1 , Peter Carolan 1 , Kyle Background: Gastrointestinal (GI) microbial translocation in people with HIV (PWH) is associated with systemic immune activation and inflammation. The intestinal brush border enzyme intestinal alkaline phosphatase (IAP) is important for maintaining healthy GI barrier function. IAP promotes intestinal homeostasis by regulating the pH of the gut luminal surface via bicarbonate secretion and by detoxifying lipopolysaccharides (LPS). After IAP dephosphorylates the lipid moiety of LPS, the modified LPS is no longer active to induce proinflammatory responses through TLR4 and subsequent MyD88- dependent signaling pathways in the gut. IAP is reduced in human diseases in which intestinal dysbiosis has been implicated, such as inflammatory bowel disease and diabetes mellitus. Furthermore, exogenously administered IAP reversed intestinal inflammation and metabolic syndrome in animal models. We hypothesized that IAP would be lower in PWH given the known intestinal damage and dysbiosis in PWH. Methods: IAP activity was measured in fluid from the terminal ileum collected by colonoscopy in 30 participants with chronic HIV and 6 controls without HIV. For IAP activity, luminal fluid is mixed with phosphatase assay reagent containing p-nitrophenyl phosphate followed by determining optical density at 405nm. All participants did not have known GI disease, and participants with Staller 1 , James Richter 1 , Rich Hodin 1 , Janet Lo 1 1 Massachusetts General Hospital, Boston, MA, USA

HIV were treated with stable ART > 6 months and had suppressed HIV RNA. IL-1β was measured in the terminal ileum fluid by ELISA. IAP and IL-1β were both normalized to total protein measured in the intestinal fluid. Results: In PWH, IAP activity was significantly lower compared to controls (6.25±3.69 [mean±SD] vs 10.98±2.10, p=0.0009). Proinflammatory IL-1β trended to be higher in the intestinal fluid of PWH compared to controls (33.16±73.55 vs. 5.97±8.96 pg/mg protein, p=0.099). BMI (27.3±4.3 vs 26.9±4.2 kg/m 2 , p=0.83), and HbA1c (5.5±0.3 vs 5.5±0.4 %, p=0.86) were similar between the groups. Peripheral CD4+ cell count was 729±234 cells/µL in PWH. Conclusion: We demonstrated significantly lower IAP in the terminal ileum of PWH compared to uninfected controls, which has not been reported previously. This novel finding of reduced IAP in PWH may provide additional insight into the pathogenesis of intestinal barrier dysfunction and its associated comorbidities in PWH. Future studies are needed to further elucidate the role of IAP in HIV- associated GI dysfunction and the potential use of exogenous IAP to reduce LPS-mediated inflammation in PWH.

Poster Abstracts

235 IMPACT OF INTRAVENOUS HEROIN AND HIV ON GUT INTEGRITY AND IMMUNE ACTIVATION Corrilynn O. Hileman 1 , Emily Bowman 2 , Janelle Gabriel 2 , Aaren Kettelhut 2 , Julia C. Kosco 3 , Danielle Labbato 3 , Theresa O. Rodgers 3 , Cheryl A. Smith 1 , Nicholas Funderburg 2 , Grace A. McComsey 3 1 MetroHealth Medical Center, Cleveland, OH, USA, 2 The Ohio State University, Columbus, OH, USA, 3 University Hospitals Cleveland Medical Center, Cleveland, OH, USA Background: Altered gut integrity and translocation of microbial products appear to be central in HIV-related immune activation. Opioid use may promote similar changes in gut permeability potentially augmenting immune activation in HIV-infected opioid users. Injection as a route administration may also heighten inflammation. Excess immune activation may increase risk of co- morbid metabolic conditions and contribute to the increased risk of mortality in people with HIV who inject opioids. Methods: HIV+ and HIV- heroin users and HIV+ and HIV- never heroin users were prospectively enrolled. Never users were matched to HIV+ heroin users by sex, age and CD4+ count (HIV+ only). Soluble markers of systemic inflammation, monocyte activation, gut integrity and microbial translocation were quantified by ELISA. ANOVA and multivariable linear regression were used to compare markers between groups and to test for effect modification by HIV status. Results: 100 enrolled (19 HIV+ Heroin+; 38 HIV- Heroin+; 19 HIV+ Heroin-; 24 HIV- Heroin-). Groups were similar except HIV+ Heroin+ had lower trunk fat (p<0.01) and lower current (p=0.02), but similar nadir CD4+ counts. Heroin+ groups were more likely to be Hispanic (p<0.01), have active hepatitis c (p<0.01) and be current smokers (p<0.01). Overall, median age was 42 years and 75%were men. For HIV+ groups, median known duration of HIV was 13 years and all but 3 had HIV-1 RNA <200 copies/ml. For Heroin+ groups, 98% were current smokers; 49% also used cocaine and 11% used methamphetamine. Active heroin use was associated with higher soluble tumor necrosis factor alpha receptors-I and –II (sTNF-RI and –II), high sensitivity C-reactive protein (hsCRP), D-dimer, soluble CD14 (trend only), soluble CD163, LPS binding protein (LBP) and beta-D-glucan independent of HIV status, age, sex, race, trunk fat, hepatitis c and smoking. HIV was only associated with sTNF-RII (trend), sCD163 and zonulin in adjusted models. HIV status tended to modify the effect between heroin use and hsCRP and LBP (p<0.10 for interactions); however, in stratified models, HIV- Heroin+ had larger difference in both markers compared to Heroin-. While some

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CROI 2020