Skip to main content

Encephalitozoon intestinalis Infection Impacts the Expression of Apoptosis-Related Genes in U937 Macrophage Cells

Abstract

Purpose

Encephalitozoon intestinalis affects many physiological processes of host cells to survive, proliferate, and spread to different regions within the body. In this study, the effects of the parasite on host cell apoptosis and proliferation were investigated.

Methods

To determine the impact of the parasite on the host cell apoptosis, changes in the expression profile of genes were investigated with the qPCR array using the Human Apoptosis Panel in infected and non-infected macrophage cells. Also, the rate of apoptosis in the cells was determined by Giemsa staining method. Cell proliferation was determined by measuring the DNA concentration in infected and non-infected cells.

Results

The thirty-six of apoptosis-related genes were down-regulated, while 20 of apoptosis-related genes were up-regulated in infected cells compared to uninfected cells. However, there were no significant changes detected in 32 analyzed genes between infected and control groups. E. intestinalis was determined to decrease cell proliferation in U937 macrophage cells. Unexpectedly, Giemsa staining showed an increase in the rate of apoptosis in infected cells.

Conclusion

Regulated genes after infection are involved in many different biological pathways and various components of the cell. This suggests that the parasite uses highly sophisticated ways to maintain the viability of the cell.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Anane S, Attouchi H (2010) Microsporidiosis: epidemiology, clinical data and therapy. Gastroenterol Clin Biol 34(8–9):450–464. https://doi.org/10.1016/j.gcb.2010.07.003

    CAS  Article  PubMed  Google Scholar 

  2. Cetinkaya Ü, Hamamcı B, Kaynar L, Kuk S, Şahin İ, Yazar S (2015) Investigation of the presence of Encephalitozoon intestinalis and Enterocytozoonbieneusi in bone marrow transplant patients by IFA-MAbs method. Mikrobiyol Bul 49(3):432–438. https://doi.org/10.5578/mb.9809

    Article  PubMed  Google Scholar 

  3. Hamamcı B, Çetinkaya Ü, Berk V, Kaynar L, Kuk S, Yazar S (2015) Prevalence of Encephalitozoon intestinalis and Enterocytozoonbieneusi in cancer patients under chemotherapy. Mikrobiyol Bul 49(1):105–113. https://doi.org/10.5578/mb.8787

    Article  PubMed  Google Scholar 

  4. Oğuz Kaya İ, Doğruman Al F, Mumcuoğlu İ (2018) Investigation of Microsporidia prevalence with calcofluor white and uvitex 2B chemiluminescence staining methods and molecular analysis of species in diarrheal patients. Mikrobiyoloji Bül 52:401–412. https://doi.org/10.5578/mb.67363

    Article  Google Scholar 

  5. Curry A (2005) Microsporidiosis. In: Cox FEG, Wakelin D, Gillespie SH, Despommier DD (eds) Topley and Wilson’s Microbiology and Microbial Infections; Parasitology, 10th edn. ASM press, Washington, D.C., pp 529–555

    Google Scholar 

  6. Franzen C (2005) How do microsporidia invade cells? Folia Parasitol (Praha) 52(1–2):36–40. https://doi.org/10.14411/fp.2005.005

    Article  Google Scholar 

  7. Fasshauer V, Gross U, Bohne W (2005) The parasitophorous vacuole membrane of Encephalitozoon cuniculi lacks host cell membrane proteins immediately after invasion. Eukaryot Cell 4(1):221–224. https://doi.org/10.1128/EC.4.1.221-224.2005

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. del Aguila C, Izquierdo F, Granja AG, Hurtado C, Fenoy S, Fresno M, Revilla Y (2006) Encephalitozoon microsporidia modulates p53-mediated apoptosis in infected cells. Int J Parasitol 36:869–876. https://doi.org/10.1016/j.ijpara.2006.04.002

    CAS  Article  PubMed  Google Scholar 

  9. Scanlon M, Shaw AP, Zhou CJ, Visvesvara GS, Leitch GJ (2000) Infection by microsporidia disrupts the host cell cycle. J Eukaryot Microbiol 47:525–531. https://doi.org/10.1111/j.1550-7408.2000.tb00085.x

    CAS  Article  PubMed  Google Scholar 

  10. Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516. https://doi.org/10.1080/01926230701320337

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. Reed JC (2000) Mechanisms of apoptosis. Am J Pathol 157(5):1415–1430. https://doi.org/10.1016/S0002-9440(10)64779-7

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Cetinkaya U, Charyyeva A, Gurbuz E (2018) Evaluation of the reproductive potential of Encephalitozoon intestinalis in four different cell line. Mikrobiyol Bul 52(4):390–400. https://doi.org/10.5578/mb.67383

    Article  PubMed  Google Scholar 

  13. Ovalle-Bracho C, Franco-Muñoz C, Londoño-Barbosa D, Restrepo-Montoya D, Clavijo-Ramírez C (2015) Changes in macrophage gene expression associated with Leishmania (Viannia) braziliensis infection. PLoS ONE 10(6):e0128934. https://doi.org/10.1371/journal.pone.0128934

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Sokolova YY, Bowers LC, Alvarez X, Didier ES (2019) Encephalitozoon cuniculi and Vittaforma corneae (Phylum Microsporidia) inhibit staurosporine-induced apoptosis in human THP-1 macrophages in vitro. Parasitology 146(5):569–579. https://doi.org/10.1017/S0031182018001968

    CAS  Article  PubMed  Google Scholar 

  15. Hofland LJ, van Koetsveld PM, Lamberts SW (1990) Percoll density gradient centrifugation of rat pituitary tumor cells: a study of functional heterogeneity within and between tumors with respect to growth rates, prolactin production and responsiveness to the somatostatin analog SMS 201–995. Eur J Cancer 26(1):37–44. https://doi.org/10.1016/0277-5379(90)90254-q

    CAS  Article  PubMed  Google Scholar 

  16. Heussler VT, Kuenzi P, Rottenberg S (2001) Inhibition of apoptosis by intracellular protozoan parasites. Int J Parasitol 31:1166–1176. https://doi.org/10.1016/s0020-7519(01)00271-5

    CAS  Article  PubMed  Google Scholar 

  17. Higes M, Juarranz Á, Dias-Almeida J, Lucena S, Botías C, Meana A, García-Palencia P, Martín-Hernández R (2013) Apoptosis in the pathogenesis of Nosema ceranae (Microsporidia: Nosematidae) in honey bees (Apismellifera). Environ Microbiol Rep 5(4):530–536. https://doi.org/10.1111/1758-2229.12059

    Article  PubMed  Google Scholar 

  18. Knodler LA, Finlay BB (2001) Salmonella and apoptosis: to live or let die? Microbes infect 3:1321–1326. https://doi.org/10.1016/s1286-4579(01)01493-9

    CAS  Article  PubMed  Google Scholar 

  19. Johnstone RW, Ruefli AA, Lowe SW (2002) Apoptosis: a link between cancer genetics and chemotherapy. Cell 108:153–164. https://doi.org/10.1016/s0092-8674(02)00625-6

    CAS  Article  PubMed  Google Scholar 

  20. Cianciulli A, Porro C, Calvello R, Trotta T, Panaro MA (2018) Resistance to apoptosis in Leishmania infantum-infected human macrophages: a critical role for anti-apoptotic Bcl-2 protein and cellular IAP1/2. Clin Exp Med 18(2):251–261. https://doi.org/10.1007/s10238-017-0482-1

    CAS  Article  PubMed  Google Scholar 

  21. Johnstone RW, Frew AJ, Smyth MJ (2008) The TRAIL apoptotic pathway in cancer onset, progression and therapy. Nat Rev Cancer 8(10):782–798. https://doi.org/10.1038/nrc2465

    CAS  Article  PubMed  Google Scholar 

  22. Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A (1996) Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271:12687–12690. https://doi.org/10.1074/jbc.271.22.12687

    CAS  Article  PubMed  Google Scholar 

  23. Fulda S, Meyer E, Debatin KM (2002) Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene 21(15):2283–2294. https://doi.org/10.1038/sj.onc.1205258

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgements

This study was financially supported by the Erciyes University Scientific Research Projects Unit, Kayseri, Turkey (No. TCD-2016-7042).

Author information

Authors and Affiliations

Authors

Contributions

UC, AC and ME designed the study and made the critical revision of the article. UC and MS performed the in vitro experiment. Collecting test data, drafting the article and getting a final approval of the version to be published were done by UC, ME and AC, as well as data analysis and interpretation was done by AC. In addition, UC and ME were responsible for supervision, and project administration.

Corresponding author

Correspondence to Ülfet Çetinkaya.

Ethics declarations

Conflict of Interest

The authors report no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Çetinkaya, Ü., Caner, A., Charyyeva, A. et al. Encephalitozoon intestinalis Infection Impacts the Expression of Apoptosis-Related Genes in U937 Macrophage Cells. Acta Parasit. 66, 397–405 (2021). https://doi.org/10.1007/s11686-020-00288-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11686-020-00288-x

Keywords

  • Parasites
  • Microsporidia
  • Apoptosis
  • Gene
  • Human macrophage
  • Cell proliferation