1.Establishment of a Zika virus infection model in rats with type Ⅰ interferon receptor deficiency
Zeng CAI ; Qiaoyang XIAN ; Shan SU ; Zhang ZHANG ; Ziwen LONG ; Hongbin TANG
Chinese Journal of Microbiology and Immunology 2025;45(10):854-859
Objective:To establish a Zika virus-infected suckling SD rat model with type Ⅰ interferon receptor deficiency(SD-Ifnar1 -/-[cc])and provide a novel animal model for investigating Zika virus pathogenesis and developing therapeutic strategies. Methods:Seventeen-day-old male SD-Ifnar1 -/-[cc]rat pups were randomly divided into experimental and control groups( n=6). The experimental group received an intraperitoneal injection of Zika virus strain SZ-wiv01(5×10 4 PFU/rat,200 μl),while the control group received an equal volume of phosphate-buffered saline(PBS). Animals were euthanized via CO 2 asphyxiation on days 12 and 15 post-infection(dpi),and brain,spleen,liver,and testis tissues were harvested. Viral loads and cytokine expression levels were quantified using quantitative real-time PCR qRT-PCR),and histopathological evaluation was performed via HE staining. Results:qRT-PCR analysis revealed no detectable Zika virus RNA(Ct >35)in control tissues. In the experimental group,viral RNA(Ct <35)was detected in the brain,spleen,liver,and testis by day 12,with stable viral loads across tissues by day 15( P>0.05). Cytokine profiling demonstrated significant upregulation in the brain at day 12:IFN-β(5.58-fold, P<0.01),IL-6(7.11-fold, P<0.01),and CCL5(3.79-fold, P<0.01). By day 15,these levels remained elevated(IFN-β:3.07-fold;IL-6:4.04-fold;CCL5:5.22-fold;all P<0.01). In the liver,IFN-β mRNA decreased to 20% of the control level by day 15( P<0.05),while IL-6 declined to 24% and CCL5 mRNA dropped to 38% by day 12. No significant cytokine changes were observed in the spleen( P>0.05). Testicular tissues exhibited reduced IFN-β mRNA levels(43% of control at day 12,31% at day 15; P<0.05). Histopathological analysis revealed marked splenomegaly with disrupted splenic corpuscle architecture and lymphocyte depletion,significant inflammatory cell infiltration in hepatic portal areas,and testicular structural disorganization with inflammatory infiltration in Zika-infected rats. Conclusions:The SD-Ifnar1 -/-[cc]suckling rat model is successfully established and validated. This model recapitulates systemic Zika virus infection,tissue-specific pathology,and immune response dynamics,providing a robust platform for elucidating viral pathogenesis and advancing antiviral drug development.
2.Establishment of a Zika virus infection model in rats with type Ⅰ interferon receptor deficiency
Zeng CAI ; Qiaoyang XIAN ; Shan SU ; Zhang ZHANG ; Ziwen LONG ; Hongbin TANG
Chinese Journal of Microbiology and Immunology 2025;45(10):854-859
Objective:To establish a Zika virus-infected suckling SD rat model with type Ⅰ interferon receptor deficiency(SD-Ifnar1 -/-[cc])and provide a novel animal model for investigating Zika virus pathogenesis and developing therapeutic strategies. Methods:Seventeen-day-old male SD-Ifnar1 -/-[cc]rat pups were randomly divided into experimental and control groups( n=6). The experimental group received an intraperitoneal injection of Zika virus strain SZ-wiv01(5×10 4 PFU/rat,200 μl),while the control group received an equal volume of phosphate-buffered saline(PBS). Animals were euthanized via CO 2 asphyxiation on days 12 and 15 post-infection(dpi),and brain,spleen,liver,and testis tissues were harvested. Viral loads and cytokine expression levels were quantified using quantitative real-time PCR qRT-PCR),and histopathological evaluation was performed via HE staining. Results:qRT-PCR analysis revealed no detectable Zika virus RNA(Ct >35)in control tissues. In the experimental group,viral RNA(Ct <35)was detected in the brain,spleen,liver,and testis by day 12,with stable viral loads across tissues by day 15( P>0.05). Cytokine profiling demonstrated significant upregulation in the brain at day 12:IFN-β(5.58-fold, P<0.01),IL-6(7.11-fold, P<0.01),and CCL5(3.79-fold, P<0.01). By day 15,these levels remained elevated(IFN-β:3.07-fold;IL-6:4.04-fold;CCL5:5.22-fold;all P<0.01). In the liver,IFN-β mRNA decreased to 20% of the control level by day 15( P<0.05),while IL-6 declined to 24% and CCL5 mRNA dropped to 38% by day 12. No significant cytokine changes were observed in the spleen( P>0.05). Testicular tissues exhibited reduced IFN-β mRNA levels(43% of control at day 12,31% at day 15; P<0.05). Histopathological analysis revealed marked splenomegaly with disrupted splenic corpuscle architecture and lymphocyte depletion,significant inflammatory cell infiltration in hepatic portal areas,and testicular structural disorganization with inflammatory infiltration in Zika-infected rats. Conclusions:The SD-Ifnar1 -/-[cc]suckling rat model is successfully established and validated. This model recapitulates systemic Zika virus infection,tissue-specific pathology,and immune response dynamics,providing a robust platform for elucidating viral pathogenesis and advancing antiviral drug development.
3.Comparative analysis of specific IFN-γproduction induced by Mycobacterium tuberculosis in two dif-ferent nonhuman primates
Hongbin TANG ; Jing ZHANG ; Yan RAO ; Qiaoyang XIAN ; Ming GUO
Chinese Journal of Microbiology and Immunology 2015;35(7):486-490
Objective To analyze the differences of immune responses against Mycobacterium tu-berculosis antigens induced in two different nonhuman primates and to provide rationales for the selection of suitable animal models for vaccine efficacy evaluation.Methods Expression of functional surface markers including CD69 and HLA-DR, the activation markers on CD4+and CD8+T cells from in rhesus macaques and cynomolgus monkeys were measured by flow cytometry analysis.PBMCs were isolated from rhesus ma-caques and cynomolgus monkeys with Mycobacterium tuberculosis infection and stimulated with PPD and pep-tide pools ( ESAT-6/CFP-10) .Enzyme-linked imunospot ( ELISPOT) assay was performed to detect IFN-γproducing lymphocytes.Results The CD4+and CD8+T cells isolated from rhesus macaques without Myco-bacterium tuberculosis infection expressed higher levels of CD69 and HLA-DR than those from healthy cyno-molgus monkeys (P<0.01).The numbers of IFN-γspot forming cells/106 PBMCs in rhesus macaques with Mycobacterium tuberculosis infection for 10 and 11 months were respectively 3 and 3.5 times higher than that of cynomolgus monkeys upon after the stimulation of PBMCs with PPD.The levels of IFN-γproduction by the cells from rhesus macaque group were also higher than those from cynomolgus monkey group upon after the stimulation of PBMCs with ESAT-6 or CFP-10 peptide pools.Conclusion More IFN-γproducing cells were induced in rhesus macaques than that in cynimolgus monkeys after stimulation with Mycobacterium tu-berculosis antigens.Therefore, the rhesus macaques might be a better animal model for evaluating immune responses induced by Mycobacterium tuberculosis vaccines.

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