A set of plasmatic microRNA related to innate immune response highly predicts the onset of Immune Reconstitution Inflammatory Syndrome (IRIS) in tuberculosis co-infected HIV individuals (ANRS-12358 study)

Polidy PEAN^1*Ratana MENG¹Eliott Benichou^1,2Srey Pichsovannary³Bunnet DIM⁴Laurence Borand^4,5Olivier Marcy⁶Didier Laureillard⁷François-Xavier Blanc⁸Tineke Cantaert¹Yoann Madec⁹Laurence Weiss¹⁰Daniel Scott-Algara¹¹

• ¹Immunology Unit, Pasteur Institute of Cambodia, Phnom Penh, Phnom Penh Municipality, Cambodia
• ²Université Paris Saclay, Université de Versailles Saint-Quentin-en-Yvelines, Versailles, Île-de-France, France
• ³Sihanouk Hospital Center of HOPE, Phnom Penh, Phnom Penh Municipality, Cambodia
• ⁴Clinical Research Team, Epidemiology and Public Health Unit, Institut Pasteur du Cambodge, Phnom Penh, Cambodia
• ⁵Center for Tuberculosis Research, Johns Hopkins University, Baltimore, Maryland, United States
• ⁶Research Institute for Sustainable Development (IRD) EMR 271, National Institute for Health and Medical Research (INSERM) UMR 1219, University of Bordeaux, 33000 Bordeaux, France
• ⁷Infectious and Tropical Diseases Department, University Hospital, 30900 Nimes, France
• ⁸Nantes Université, CHU Nantes, Service de Pneumologie, l'institut du thorax, Nantes, France
• ⁹Epidemiology of Emerging Diseases, Institut Pasteur, Université de Paris, 75000 Paris, France
• ¹⁰Université Paris Cité, Immunology, 75000 Paris, France
• ¹¹International Affairs Departement, Institut Pasteur, 75000 Paris, France

Background: After initiation of combination antiretroviral treatment (cART), HIV-1/tuberculosis coinfected patients are at high risk of developing tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS). MicroRNAs, small molecules of approximately 22 nucleotides, which regulate post-transcriptional gene expression and their profile has been proposed as a biomarker for many diseases. We tested whether the microRNA profile could be a predictive biomarker for TB-IRIS.Methods: Twenty-six selected microRNAs involved in the regulation of the innate immune response were investigated. Free plasmatic and microRNA-derived exosomes were measured by flow cytometry. The plasma from 74 HIV-1+TB+ individuals (35 IRIS and 39 non-IRIS) at the time of the diagnosis and before any treatment (baseline) of CAMELIA trial (ANRS1295-CIPRA KH001-DAIDS-ES ID10425); 15 HIV+TB-and 23 HIV-TB+, both naïve of any treatment; and 20 HIV-TB-individuals as controls were analysed.Results: At baseline, both IRIS and non-IRIS HIV+/TB+ individuals had similar demographic and clinical characteristics, including sex, age, body mass index, very low CD4+ cell counts (27 cells/mm 3 ), and plasma HIV RNA load levels (5.76 log copies/ml). Twenty out of 26 plasmatic-microRNAs tested were no different between IRIS and controls. Twelve of the 26 tested microRNAs showed statistically significant differences between IRIS and non-IRIS patients (p-values ranging from p <0.05 to p <0.0001). Among these, five could discriminate between IRIS and non-IRIS individuals using ROC curve analysis (AUC scores ranging from 0.74 to 0.92). The combination of two (hsa-mir-29c-3p and has-mir-146a-5p) or three microRNAs (hsa-mir-29c-3p, hsa-mir-29a-3p, and hsa-mir-146a-5p) identified IRIS with 100% sensitivity and high specificity (95% and 97%, respectively).The combination of at least two or three plasmatic microRNAs known to regulate inflammation and/or cytokine responses could be used as biomarkers to discriminate IRIS from non-IRIS in HIV-TB co-infected individuals at the time of diagnosis and prior to any treatment.