Visbiome Probiotics Differently Affect Gut-Associated Lymphoid Tissue Indolamine-2,3-Dioxygenase mRNA and Cerebrospinal Fluid Neopterin Levels in Antiretroviral-Treated HIV-1 Infected Patients
This is a Pilot Study involving 10 patients. The results showed that Visbiome probiotic when supplemented for 6 months in HIV-1-positive patients, is associated with a significant reduction of the CSF neopterin and GALT-associated IDO mRNA levels.
Patients were sampled for peripheral blood and colonoscopy before and after Visbiome probiotic supplementation. Colonic washing was carried out by polyethylene glycol-electrolyte solution (PEG) administration 24 hours before the examination.
The endoscopic procedure was performed with conscious sedation (midazolam 5 mg/iv) using large cup forceps (Radial Jaw 4, Boston Scientific, Natick, MA, USA). All HIV-1-positive patients underwent a total colonoscopy and retrograde ileoscopy for at least
10 cm of distal ileum with conventional or slim scope (model CF or PCF-160 AI, Olympus Medical Europe GmbH, Hamburg, Germany).
They obtained specimens (two biopsies from each site) from the terminal ileum, cecum, ascending, transverse, and descending colon. Lamina propria lymphocytes (LPLs) was stored as dried pellets for RNA extraction. Gut biopsies from each intestine site were pooled and processed.
Briefly, biopsies collected in RPMI 1640 were washed twice with EDTA wash media, resuspended, and incubated for 1 h at room temperature in EDTA solution 5 mM. Supernatant containing intraepithelial lymphocytes was removed, and biopsies
were digested by 1 h incubation at 37 ◦C with 1 mg/mL collagenase (Sigma-Aldrich, Milan, Italy) and 1.5 U DNAse I (Sigma-Aldrich), allowing the isolation of LPLs that were filtered through a 70 µm cell strainer.
CSF was collected by lumbar puncture, centrifuged, and cell-free supernatant samples were stored in aliquots at −80 ◦C.
These results using adjunctive therapies to reduce the levels of immune activation markers in CSF are unique in that they have so far been disappointing. For this reason they suggest following up by a larger confirmatory trial.
Carolina Scagnolari 1,2, Giuseppe Corano Scheri 3, Carla Selvaggi 2, Ivan Schietroma 3, Saeid Najafi Fard 3, Andrea Mastrangelo 3, Nseemi Giustini 3, Sara Serafino 3, Claudia Pinacchio 1, Paolo Pavone 3, Gianfranco Fanello 4, Giancarlo Ceccarelli 3, Vincenzo Vullo 3 and Gabriella d’Ettorre 3,*
Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Viale Regina Elena 291, 00161 Rome, Italy
Department of Molecular Medicine, Laboratory of Virology, Sapienza University of Rome, Viale di Porta Tiburtina 28, 00185 Rome, Italy
Department of Public Health and Infectious Diseases, Sapienza University of Rome, Viale del Policlinico 155, 00161 Rome, Italy
Department of Emergency Surgery, Emergency Endoscopic Unit, Policlinico Umberto I, Sapienza University of Rome, Viale del Policlinico 155, 00161 Rome, Italy