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Apoptosis in peripheral blood lymphocytes in intestinal tuberculosis and Crohn’s disease: Implications to diagnostic differentiation

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Indian Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Aim

Intestinal tuberculosis (ITb) and Crohn’s disease (CD) mimic each other often leading to misdiagnosis. We evaluated the difference between ITb and CD using the extent of apoptosis in peripheral blood lymphocytes.

Methods

CD4+ cells as a percentage of the lymphocytes and viable, dead, total apoptotic, early apoptotic, and late apoptotic CD4+ cells were assessed in the peripheral blood by flow cytometry in healthy controls and patients with confirmed active ITb and CD prior to initiating therapy. Early apoptotic and total apoptotic cells were further expressed as a proportion of the percentage of CD4+ cells.

Results

The percentages of CD4+ cells (6.5 [3.0, 8.7] vs. 13.40 [10.15, 13.40]; p < 0.001), total apoptotic cells (0.13 [0.0, 0.22] vs. 0.08 [0.0, 0.21]; p = 0.045), early apoptotic (1.24 [0.55, 2.54] vs. 0.71 [0.40, 1.30]; p = 0.037), and the proportion of the latter two parameters (17.18 [5.61, 57.33] vs. 4.84 [2.71, 9.83]; p-value 0.039) and (17.18 [7.4, 67.50] vs. 5.51 [3.10, 11.03]; p-value 0.036) were significantly different between patients with ITb and CD. The best sensitivity, specificity, and positive and negative predictive values for the diagnosis of ITb were seen with the CD4+ cell percentage (82.6%, 82.4%, 86.4%, 77.8%, respectively) and the proportion of early apoptotic cells (73.9%, 70.6%, 77.3%, 66.7%, respectively).

Conclusion

CD4+ cells as a percentage of peripheral blood lymphocytes and the proportion of early apoptotic CD4+ cells show promise to diagnostic differentiation between ITb and CD.

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References

  1. Pulimood AB, Amarapurkar DN, Ghoshal U, et al. Differentiation of Crohn’s disease from intestinal tuberculosis in India in 2010. World J Gastroenterol. 2011;17:433–43.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Ooi CJ, Makharia GK, Hilmi I, et al. Asia Pacific Consensus Statements on Crohn’s disease. Part 1: definition, diagnosis, and epidemiology: (Asia Pacific Crohn’s disease consensus--part 1). J Gastroenterol Hepatol. 2016;31:45–55.

    Article  PubMed  Google Scholar 

  3. Makharia GK, Srivastava S, Das P, et al. Clinical, endoscopic, and histological differentiations between Crohn’s disease and intestinal tuberculosis. Am J Gastroenterol. 2010;105:642–51.

    Article  PubMed  Google Scholar 

  4. Amarapurkar DN, Patel ND, Rane PS. Diagnosis of Crohn’s disease in India where tuberculosis is widely prevalent. World J Gastroenterol. 2008;14:741–6.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Molodecky NA, Shian SS, Rabi DM, et al. Increasing incidence and prevalence of the inflammatory bowel diseases with time, Based on Systematic Review. Gastroenterology. 2012;142:46–54.

    Article  PubMed  Google Scholar 

  6. Epstein D, Watermeyer G, Kirsch R. Diagnosis and management of CD in high-risk ITB: the clinical differences between CD and intestinal tuberculosis. Aliment Pharmacol Ther. 2007;25:1373–88.

    Article  CAS  PubMed  Google Scholar 

  7. Armand S, Vanhuls P, Delcroix G, Courcol R, Lemaître N. Comparison of the Xpert MTB/RIF test with an IS6110-TaqMan real-time PCR assay for direct detection of mycobacterium tuberculosis in respiratory and nonrespiratory specimens. J Clin Microbiol. 2011;49:1772–6.

  8. Pulimood AB, Peter S, Rook GW, Donoghue HD. In situ PCR for mycobacterium tuberculosis in endoscopic mucosal biopsy specimens of intestinal tuberculosis and Crohn disease. Am J Clin Pathol. 2008;129:846–51.

    Article  CAS  PubMed  Google Scholar 

  9. Ting J, Baoying F, Yu Z, Xujun H. The diagnostic value of polymerase chain reaction for mycobacterium tuberculosis to distinguish intestinal tuberculosis from Crohn's disease: a meta-analysis. Saudi J Gastroenterol. 2017;23:3–10.

    Article  Google Scholar 

  10. Sellin JH, Shah RR. the promise and pitfalls of serologic testing in inflammatory bowel disease. Gastroenterol Clin N Am. 2012;41:463–82.

  11. Pulimood AB, Ramakrishna BS, Kurian G, et al. Endoscopic mucosal biopsies are useful in distinguishing granulomatous colitis due to CD from tuberculosis. Gut. 1999;45:537–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Pulimood AB, Peter S, Ramakrishna B, et al. Segmental colonoscopic biopsies in the differentiation of ileocolic tuberculosis from CD. J Gastroenterol Hepatol. 2005;20:688–96.

    Article  PubMed  Google Scholar 

  13. Kumar S, Bopanna S, Kedia S, et al. Evaluation of Xpert MTB/RIF assay performance in the diagnosis of abdominal tuberculosis. Intest Res. 2017;15:187–94.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Bu P, Keshavarzian A, Stone DD, et al. Apoptosis: one of the mechanisms that maintains unresponsiveness of the intestinal mucosal immune system. J Immunol. 2001;66:6399–403.

    Article  Google Scholar 

  15. Sturm A, Leite AZ, Danese S, et al. Divergent cell cycle kinetics underlie the distinct functional capacity of mucosal T cells in Crohn's disease and ulcerative colitis. Gut. 2004;53:1624–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Tiede I, Fritz G, Strand S, et al. CD28-dependent Rac1 activation is the molecular target of azathioprine in primary human CD4+T lymphocytes. J Clin Invest. 2003;111:1133–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Doering J, Begue B, Lentze MJ, et al. Induction of T lymphocyte apoptosis by sulphasalazine in patients with Crohn's disease. Gut. 2004;53:1632–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Sabatino D, Ciccocioppo R, Cinque B, et al. GR. Defective mucosal T cell death is sustainably reverted by infliximab in a caspase dependent pathway in Crohn’s disease. Gut. 2004;53:70–7.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Hirsch CS, Toossi Z, Vanham G, et al. Apoptosis and T cell hyporesponsiveness in pulmonary tuberculosis. J Infect Dis. 1999;179:945–53.

    Article  CAS  PubMed  Google Scholar 

  20. Hirsch CS, Toossi Z, Johnson JL, et al. Augmentation of apoptosis and interferon-γ production at sites of active mycobacterium tuberculosis Infection in Human Tuberculosis. J Infect Dis. 2001;183:779–88.

    Article  CAS  PubMed  Google Scholar 

  21. Fayyazi A, Eichmeyer B, Soruri A, et al. Apoptosis of macrophages and T cells in tuberculosis associated caseous necrosis. J Pathol. 2000;191:417–25.

    Article  CAS  PubMed  Google Scholar 

  22. Ina K, Itoh J, Fukushima K, et al. Resistance of Crohn’s disease T cells to multiple apoptotic signals is associated with a Bcl-2/Bax mucosal imbalance. J Immunol. 1999;163:1081–90.

    CAS  PubMed  Google Scholar 

  23. Mustafa T, Bjune TG, Jonsson R,  Pando RH, Nilsen R. Increased expression of Fas ligand in human tuberculosis and leprosy lesions: a potential novel mechanism of immune evasion in mycobacterial infection. Scand J Immunol. 2001;54:630–9.

    Article  CAS  PubMed  Google Scholar 

  24. Davoudi S, Rasoolinegad M, Younesian M, et al. CD4+ cell counts in patients with different clinical manifestations of tuberculosis. Braz J Infect Dis. 2008;12:483–6.

  25. Mahon RN, Rojas RE, Fulton SA, Franko JL, Harding CV, Boom WH. Mycobacterium tuberculosis Cell Wall glycolipids directly inhibit CD4+ T-cell activation by interfering with proximal T-cell-receptor signaling. Infect Immun. 2009;77:4574–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Tiwari V, Kedia S, Garg SK, et al. CD4 + CD25 + FOXP3 + T cell frequency in the peripheral blood is a biomarker that distinguishes intestinal tuberculosis from Crohn’s disease. PLoS One. 2018;13:e0193433.

  27. Balcha TT, Skogmar S, Sturegård E, et al. A clinical scoring algorithm for determination of the risk of tuberculosis in HIV-infected adults: a cohort study performed at Ethiopian health centers. Open Forum Infect Dis. 2014;1:ofu095.

  28. Senju M, Hulstaert F, Lowder J, Jewell DP. Flow cytometric analysis of peripheral blood lymphocytes in ulcerative colitis and Crohn's disease. Gut. 1991;32:779–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Maini MK, Gilson RJC, Chavda N, et al. Reference ranges and sources of variability of CD4 counts in HIV-seronegative women and men. Genitourin Med. 1996;72:27–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Dayama A, Pandit D, Mudaliar S, et al. A pilot study on CD4 & CD8 cell counts in healthy HIV seronegative pregnant women. Indian J Med Res. 2003;117:198–200.

    CAS  PubMed  Google Scholar 

  31. Shete A, Thakar M, Abraham PR, Paranjape R. A review on peripheral blood CD4+ T lymphocyte counts in healthy adult Indians. Indian J Med Res. 2010;132:667–75.

  32. Boirivant M, Marini M, Di Felice G, et al. Lamina propria T cells in Crohn’s disease and other gastrointestinal inflammation show defective CD2 pathway-induced apoptosis. Gastroenterology. 1999;116:557–65.

    Article  CAS  PubMed  Google Scholar 

  33. Molodecky NA, Gilaad GK. Environmental risk factors for inflammatory bowel disease. Gastroenterol Hepatol. 2010;6:339–46.

    Google Scholar 

  34. Boirivant M, Pica R, DeMaria R,  Testi R, Pallone F, Strober W. Stimulated human lamina propria T cells manifest enhanced Fas-mediated apoptosis. J Clin Investig. 1996;98:2616–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Wang F, Hou H, Xu L, et al. Mycobacterium tuberculosis-specific TNF-α is a potential biomarker for the rapid diagnosis of active tuberculosis disease in Chinese population. PLoS One. 2013;8:e79431.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Singh UB, Pandey P, Mehta G, et al. Genotypic, phenotypic and clinical validation of GeneXpert in extra-pulmonary and pulmonary tuberculosis in India. PLoS One. 2016;11:e0149258.

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Acknowledgments

We thank our colleagues Shiran Shetty and Girisha Balaraju, from the Department of Gastroenterology and Hepatology, Kasturba Medical College, Manipal, Anurag Shetty, Department of Gastroenterology, Kasturba Medical College, Mangalore, for the help in identifying patients for the study. We also thank Ashma Dorothy Monteiro, Department of Statistics, Manipal University, for the help with statistical analysis.

Funding

This work was supported by grants from the Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Government of India.

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, 576 104, India

    Suprabha Suresh Nayak, Mamatha Vishwanatha Shetty & Cannanore Ganesh Pai

  2. School of Life Sciences, Kasturba Medical College, Manipal Academy of Higher Education, Manipal 576 104, India

    Kanive Parashiva Guruprasad & Kapaettu Satyamoorthy

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  1. Suprabha Suresh Nayak
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  2. Mamatha Vishwanatha Shetty
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Correspondence to Cannanore Ganesh Pai.

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SSN, MVS, CGP, KPG, and KS declare that they have no conflict of interest.

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Nayak, S.S., Shetty, M.V., Pai, C.G. et al. Apoptosis in peripheral blood lymphocytes in intestinal tuberculosis and Crohn’s disease: Implications to diagnostic differentiation. Indian J Gastroenterol 39, 338–345 (2020). https://doi.org/10.1007/s12664-019-01011-z

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