Original Article

Alteration of the Inflammatory and Anti-Inflammatory Cytokine Profiles of Peripheral Blood Mononuclear Cell in Crohn’s Disease Patients after Following up


Background: Crohn's disease (CD) has a chronic course, which its recurrence varies widely among different patients. In this study we prospectively analyzed blood samples of 19 CD patients. Alteration in transcription of inflammatory and anti-inflammatory cytokines was analyzed compared with household members after three month follow up.

Methods: CD patients were diagnosed based on clinical symptoms, endoscopic and histopathologic characteristics. Nineteen CD patients and their households were evaluated from Jun 2019 to Feb 2021 at Tehran university hospitals. CD activity score, biological, clinical and demographic data of the patients were recorded at two time point intervals. Bacteriological tests were done using aerobic and anaerobic blood cultures. To investigate transcriptional alterations, peripheral blood mononuclear cells (PBMCs) were isolated using Ficol centrifugation method and relative quantitative real-time PCR was done to determine the expression level of IFN-γ, TNF-α, IL10, and FOXP3 cytokines.

Results: Our results showed a correlation between fecal calprotectin level (709.8 ± 554.6), C-reactive protein concentration (18.1 ± 15.9), and erythrocyte sedimentation rate (30.4 ± 17.9) with disease activity (Flare/remission). IL10 and Foxp3 anti-inflammatory gene’s expression were significantly (P = 0.003 for IL10 and P = 0.008 Foxp3) higher during the flare and remission in patients with active disease respectively. Bacteriological examination showed infection with Streptococcus spp. and Clostridium spp. in two CD patients during flares, which was correlated with upregulation and down-regulation of IL10, TNF-α, IFN-γ and FOXP3 proteins, respectively.

Conclusion: Occurrence of bacteremia, and higher amount of CAP, CRP and ESR are correlated with higher level of transcription for inflammatory cytokines, which could effectively reflect the disease activity. Raise in FoxP3 transcription proposed change in Treg sub-population in PBMC or its activity during the CD remission phase.


1. Martinez-Fierro ML, Garza-Veloz I, Rocha-Pizaña MR, et al (2019). Serum cytokine, chemokine, and growth factor profiles and their modulation in inflammatory bowel disease. Medicine (Baltimore), 98 (38): e17208.
2. Kojima A, Nakano K, Wada K, et al (2012). Infection of specific strains of Streptococcus mutans, oral bacteria, confers a risk of ulcerative colitis. Sci Rep, 2:332.
3. Rolfe VE, Fortun PJ, Hawkey CJ, et al (2006). Probiotics for maintenance of remission in Crohn's disease. Cochrane Database Syst Rev, (4): Cd004826.
4. Reuter BK, Pizarro TT (2004). Commentary: the role of the IL-18 system and other members of the IL-1R/TLR superfamily in innate mucosal immunity and the pathogenesis of inflammatory bowel disease: friend or foe? Eur J Immunol, 34 (9): 2347-55.
5. Deretic V (2009). Links between autophagy, innate immunity, inflammation and Crohn’s disease. Dig Dis, 27 (3): 246-251.
6. Radford-Smith G, Jewell DP (1996). Cytokines and inflammatory bowel disease. Baillieres Clin Gastroenterol, 10 (1): 151-64.
7. Fais S, Capobianchi MR, Pallone F, et al (1991). Spontaneous release of interferon gamma by intestinal lamina propria lymphocytes in Crohn's disease. Kinetics of in vitro response to interferon gamma inducers. Gut, 32 (4): 403-407.
8. Alzahrani J, Hussain T, Simar D, et al (2019). Inflammatory and immunometabolic consequences of gut dysfunction in HIV: Parallels with IBD and implications for reservoir persistence and non-AIDS comorbidities. EBioMedicine, 46: 522-531.
9. Caër C, Wick MJ (2020). Human Intestinal Mononuclear Phagocytes in Health and Inflammatory Bowel Disease. Front Immunol, 11: 410.
10. Buttó LF, Schaubeck M, Haller D (2015). Mechanisms of Microbe-Host Interaction in Crohn's Disease: Dysbiosis vs. Pathobiont Selection. Front Immunol, 6: 555.
11. Witte AM, Veenendaal RA, Van Hogezand RA, et al (1998). Crohn's disease of the upper gastrointestinal tract: the value of endoscopic examination. Scand J Gastroenterol Suppl, 225: 100-5.
12. Best WR, Becktel JM, Singleton JW, et al (1976). Development of a Crohn's disease activity index. National Cooperative Crohn's Disease Study. Gastroenterology, 70 (3): 439-44.
13. Durko L, Stasikowska-Kanicka OA, Wagrowska-Danilewicz M, et al (2013). An analysis of the correlation of clinical, endoscopic and histological classifications in Crohn's disease. Prz Gastroenterol, 8 (6): 377-82.
14. Scott E (1951). A practical blood culture procedure. Am J Clin Pathol, 21 (3): 290-4.
15. Fuss IJ, Kanof ME, Smith PD, et al (2009). Isolation of whole mononuclear cells from peripheral blood and cord blood. Curr Protoc Immunol, 7:Unit7.
16. Schoepfer AM, Beglinger C, Straumann A, (2010) et al. Fecal calprotectin correlates more closely with the Simple Endoscopic Score for Crohn's disease (SES-CD) than CRP, blood leukocytes, and the CDAI. Am J Gastroenterol, 105 (1): 162-9.
17. Langhorst J, Elsenbruch S, Koelzer J, et al (2008). Noninvasive markers in the assessment of intestinal inflammation in inflammatory bowel diseases: performance of fecal lactoferrin, calprotectin, and PMN-elastase, CRP, and clinical indices. Am J Gastroenterol, 103 (1): 162-9.
18. Nell S, Suerbaum S, Josenhans C (2010). The impact of the microbiota on the pathogenesis of IBD: lessons from mouse infection models. Nat Rev Microbiol. 8 (8): 564-77.
19. Keighley M, Eastwood D, Ambrose N, et al (1982). Incidence and microbiology of abdominal and pelvic abscess in Crohn's disease. Gastroenterology, 83 (6): 1271-5.
20. Peach S, Lock M, Katz D, et al (1978). Mucosal-associated bacterial flora of the intestine in patients with Crohn's disease and in a control group. Gut, 19 (11): 1034-42.
21. Nakanishi Y, Sato T, Ohteki T (2015). Commensal Gram-positive bacteria initiates colitis by inducing monocyte/macrophage mobilization. Mucosal Immunology, 8 (1): 152-160.
22. Chamberlin W, Naser SA (2008). Blood cultures of 19 Crohn's disease patients. Am J Gastroenterol, 103 (3): 802-3.
23. Quera R, Espinoza R, Estay C, et al (2014). Bacteremia as an adverse event of fecal microbiota transplantation in a patient with Crohn's disease and recurrent Clostridium difficile infection. J Crohns Colitis, 8 (3): 252-3.
24. Daruwala C, Mercogliano G, Newman G, et al (2009). Bacteremia due to Clostridium difficile: case report and review of the literature. Clin Med Case Rep, 2: 5–9.
25. Ogawa K, Matsumoto T, Esaki M, et al (2012). Profiles of circulating cytokines in patients with Crohn's disease under maintenance therapy with infliximab. J Crohn's Colitis, 6 (5): 529-35.
26. Maeda M, Watanabe N, Neda H, et al (1992). Serum tumor necrosis factor activity in inflammatory bowel disease. Immunopharmacol Immunotoxicol, 14 (3): 451-61.
27. Sasaki T, Hiwatashi N, Yamazaki H, et al (1992). The role of interferony in the pathogenesis of Crohn’s disease. Gastroenterol Jpn, 27 (1): 29-36.
28. Nielsen O, Køppen T, Rüdiger N, et al (1996). Involvement of interleukin-4 and-10 in inflammatory bowel disease. Digestive Diseases and Sciences, 41 (9): 1786-1793.
29. Kucharzik T, Stoll R, Lügering N, et al (1995). Circulating antiinflammatory cytokine IL‐10 in patients with inflammatory bowel disease (IBD). Clin Exp Immunol, 100 (3): 452-6.
30. Marlow GJ, van Gent D, Ferguson LR (2013). Why interleukin-10 supplementation does not work in Crohn’s disease patients. World J Gastroenterol, 19 (25): 3931-3941.
31. Salas A (2009). Defective IL-10 production in severe phenotypes of Crohn’s. J Leukoc Biol, 85(5):896-903.
32. Lindsay J, Hodgson H (2001). The immunoregulatory cytokine interleukin‐10—a therapy for Crohn’s disease? Aliment Pharmacol Ther, 15 (11): 1709-16.
33. Negi S, Saini S, Tandel N, et al (2021). Translating Treg Therapy for Inflammatory Bowel Disease in Humanized Mice. Cells, 10 (8): 1847.
34. Wang Y, Liu XP, Zhao ZB, et al (2011). Expression of CD4+ forkhead box P3 (FOXP3)+ regulatory T cells in inflammatory bowel disease. J Dig Dis, 12 (4): 286-94.
IssueVol 51 No 7 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijph.v51i7.10099
Crohn’s disease Inflammation Cytokines Regulatory T cell Blood culture

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Ghasemi F, Basirat V, Izad M, Tavassolifar M, Yaseri M, Ebrahimi Daryani N, Alebouyeh M, Pourmand M. Alteration of the Inflammatory and Anti-Inflammatory Cytokine Profiles of Peripheral Blood Mononuclear Cell in Crohn’s Disease Patients after Following up. Iran J Public Health. 2022;51(7):1648-1657.