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Poster presentations: Basic Science 2020

P081 The structure of the proteomic profile and gut microbiota in patients with primary immunodeficiency, common variable immunodeficiency and inflammatory bowel disease

A.A. Iakovlev, A. Volkov, O. Bashtovaya, I. Shcherbakova, T. Ghazudin, G. Leonova, G. Tarasova

Rostov State Medical University, Department of Gastroenterology, Rostov-on-Don, Russian Federation

Background

Infringement of intestinal microbiota in patients with primary immunodeficiency (PID), common variable immunodeficiency (CVID) and ulcerative colitis (UC) continues to be studied. The aim of the research is a comprehensive study of the structure of gut microbiota and mucosal protein profile (MPP) in patients with PID, CVID and UC in different phases of the disease course.

Methods

The research included 15 patients with PID, 22 CVID and 46 UC (28 with relapse and 18 with persistent remission of the disease), control group—20 healthy volunteers. Estimation of gut microbiota was performed by bacteriological seeding, hydrogen breath test (HBT) with lactulose. Content of short-chain fatty acids (SCFAs) and microbial lipid markers (MLM) in faeces and colon mucosa was determined by gas–liquid chromatographic and gas chromatography–mass spectrometry (GC–MS). MPP was based on isoelectric focusing techniques (SDS–PAGE, 2DGE). Mass spectrograms were obtained using MALDI-TOF-MS/MS (Bruker, USA).

Results

HBT showed a 4.5-, 8- and 11-fold increase in hydrogen production on a 150th-minute study. SCFA showed a 6-, 9- and 11-fold decrease in propionic and butyric acids, mainly in patients with PID and UC: 0.2 + 0.1, 0.14 + 0.03 and 0.04 + 0.02 mg/g, respectively. Microbiological analysis showed a decrease in titers of E. coli, bifido- and lactobacilli on average 4.6 + 0.8 Lg. The conditionally pathogenic microflora was represented by E. coli, lactose-negative strains (n = 51,105/g) and haemolytic strains (n = 34,104/g, Proteus spp. (n = 48,105/g), Staphylococcus spp. (n = 29,105–6/g), Candida (n = 51,104–6/g) and Clostridium spp. (n = 38,104/g) in stool culture. MLN GS-MS results showed a 5.5-, 3- and 6-fold increase in total bacterial load in patients with PID, CVID and UC, respectively, which was represented by resident anaerobic microflora: Streptococcus mutants, Clostridium difficile, Candida albicans. Results of MPP (detection rate of 75% or more) of the colon mucosa in patients with PID were detected: 1, 2, 4 okkludin, kininogen 1, interleukin-1B, interleukin 8, B2-glycoprotein, heat shock protein 27, in patients with CVID—translational elongation factor, apolipoprotein C-III. In patients with UC—NF-kB, alipoprotein C-III, TNF-α, interleukin-2 and 8 were presented.

Conclusion

In patients with PID, CVID and UC markers of colon excess growth and significant decrease in SCFA are recorded during the relapse period. MPP in patients with PID, CVID and UC was characterised by proteins characteristic of inflammation, apoptosis, proliferation and proteins reflecting the activity of the autoimmune process. Patients with UC in the remission have recovery trend of gut microbiota, production of SCFA, disappearance of specific components of altered MPP.