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. 2024 Jun;69(2):1253-1266.
doi: 10.1007/s11686-024-00851-w. Epub 2024 May 14.

MicroRNA mmu-miR-511-5p: A promising Diagnostic Biomarker in Experimental Toxoplasmosis Using Different Strains and Infective Doses in Mice with Different Immune States Before and After Treatment

Rasha Fadly Mady  1 , Mona Mohamed El-Temsahy  1 , Yasmine Amr Issa  2   3 , Aya Saied Zaghloul  1 , Safaa Ibrahim Khedr  4
Affiliations

MicroRNA mmu-miR-511-5p: A promising Diagnostic Biomarker in Experimental Toxoplasmosis Using Different Strains and Infective Doses in Mice with Different Immune States Before and After Treatment

Rasha Fadly Mady et al. Acta Parasitol. 2024 Jun.

Abstract

Purpose: Searching for a novel early diagnostic biomarker for toxoplasmosis, real-time-PCR was currently used to measure the serum mmu-miR-511-5p level in male Swiss-albino mice infected with either; ME49 or RH Toxoplasma gondii (T. gondii) strains.

Methods: Three mice groups were used; (GI) constituted the non-infected control group, while (GII) and (GIII) were experimentally infected with ME49 or RH strains, respectively. GII mice were orally infected using 10 or 20 ME49 cysts (ME-10 and ME-20), both were subdivided into; non-treated (ME-10-NT and ME-20-NT) and were further subdivided into; immunocompetent (ME-10-IC and ME-20-IC) [euthanized 3-days, 1, 2, 6 or 8-weeks post-infection (PI)], and immunosuppressed using two Endoxan® injections (ME-10-IS and ME-20-IS) [euthanized 6- or 8-weeks PI], and spiramycin-treated (ME-10-SP and ME-20-SP) that received daily spiramycin, for one-week before euthanasia. GIII mice individually received 2500 intraperitoneal RH strain tachyzoites, then, were subdivided into; non-treated (RH-NT) [euthanized 3 or 5-days PI], and spiramycin-treated (RH-SP) that were euthanized 5 or 10-days PI (refer to the graphical abstract).

Results: Revealed significant upregulation of mmu-miR-511-5p in GII, one-week PI, with gradually increased expression, reaching its maximum 8-weeks PI, especially in ME-20-NT group that received the higher infective dose. Immunosuppression increased the upregulation. Contrarily, treatment caused significant downregulation. GIII recorded significant upregulation 3-days PI, yet, treatment significantly decreased this expression.

Conclusion: Serum mmu-miR-511-5p is a sensitive biomarker for early diagnosis of ME49 and RH infection (as early as one-week and 3-days, respectively), and its expression varies according to T. gondii infective dose, duration of infection, spiramycin-treatment and host immune status.

Keywords: T. gondii; Immunosuppression; ME49 strain; RH strain; Spiramycin; mmu-miR-511-5p.

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Conflict of interest statement

The authors declare that they have no conflicts of interest of a financial or personal nature.

Figures

Fig. 1
Fig. 1
Experimental design used in the current study
Fig. 2
Fig. 2
a Amplification plots of mmu-miR-511-5p; b Amplification plots of housekeeping gene (U6)
Fig. 3
Fig. 3
The mean relative quantification (RQ) of the serum level of microRNA mmu-miR-511-5p in the experimental groups
See this image and copyright information in PMC

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