Objective To investigate the therapeutic effects of interleukin 23p19(IL-23p19) monoclonal antibody in the MRL/lpr lupus-like mouse model. Methods A total of 36 female MRL/lpr mice aged 8 weeks were randomly divided into 6 groups: PBS group (blank control), IgG group (isotype IgG), dexamethasone (DEX) group (positive control), and three IL-23p19 monoclonal antibody treatment groups with different dose gradients: low dose (LD, 1 mg/kg), medium dose (MD, 3 mg/kg), and high dose (HD, 10 mg/kg). Drug intervention began at 12 weeks of age via tail vein injection. Urine protein levels were measured using urine protein test strips; serum anti-dsDNA antibody levels were detected by ELISA; serum creatinine and blood urea nitrogen levels were measured using an automatic biochemical analyzer; renal histopathological changes were analyzed by H&E and PAS staining; immunofluorescence was used to assess IgG and C3 immune complex deposition in kidney tissues; flow cytometry was employed to examine the expression of T helper 1(Th1), Th2, Th17, T follicular helper (Tfh), and regulatory T cells(Treg) cell subsets in the spleen; and RT-qPCR was used to detect the expression of related transcription factors in the spleen. Results IL-23p19 monoclonal antibody reduced urine protein levels, alleviated splenomegaly, improved renal function, and decreased anti-dsDNA antibody levels in MRL/lpr mice. It also mitigated glomerulonephritis and reduced renal immune complex deposition. Furthermore, IL-23p19 monoclonal antibody significantly suppressed the proportion of Th1 and Th17 cells while upregulating Treg cell proportion in the spleen. Additionally, it downregulated T-bet and retinoic acid receptor-related orphan receptor γt (RORγt) mRNA levels and upregulated forkhead box P3(FOXP3) mRNA levels in the spleen. Conclusions IL-23p19 monoclonal antibody demonstrates significant therapeutic effects in MRL/lpr mice, likely through modulation of the Th17/Treg cell balance.
Animals ; Female ; Mice, Inbred MRL lpr ; T-Lymphocytes, Regulatory/drug effects* ; Th17 Cells/drug effects* ; Antibodies, Monoclonal/therapeutic use* ; Interleukin-23 Subunit p19/immunology* ; Mice ; Lupus Nephritis/drug therapy* ; Kidney/drug effects* ; Antibodies, Antinuclear/blood*

Objective:To investigate impact of PD-L2 gene knockout on splenic immune phenotype of C57BL/6 mice.Methods:Body weight of female and male wild-type(WT)and PD-L2 gene knockout(KO)mice aged 3～12 weeks were continuously measured.Tissue structure differences in heart,liver,spleen,lung,kidney,thymus,ovary,uterus and testis of four groups of mice were observed by HE staining.Urease UV rate method was employed to measure urea level in mouse serum,and creatinine concentra-tion in serum was determined by creatine oxidase method.Flow cytometry was used to assess proportions of immune cells and cytokine differences in spleens of mice.Results:PD-L2 gene knockout had no apparent effect on body weight or organ structure of mice.Serum urea and creatinine levels were higher in female KO mice compared to WT mice,but no significant differences were observed among males.Female KO mice exhibited reduced proportions of CD4+T cells,CD8+T cells,Treg,NK cells and dendritic cells(DC)in spleen compared to WT mice.Conversely,proportions of Tfh,B cells,plasma cells,na?ve B cells and B2 cells were elevated,proin-flammatory cytokines such as IL-17,IL-6 and IL-22 were increased,while anti-inflammatory cytokines like IL-10 and IL-4 were decreased.Differences among male mice were either not significant or absent.Conclusion:PD-L2 gene knockout primarily affects splenic immune phenotype of female mice,leading to increased inflammatory responses and weakened immune suppression after knockout.

Objective:To investigate whether kisspeptin can influence the maternal-fetal interface regulatory T cells(Treg),thereby participating in the pathogenesis of recurrent spontaneous abortion(RSA).Methods:Normal pregnancy(NP)and RSA mice models were established,where NP mice received a tail vein injection of PBS(NP-PBS group),and RSA mice received a tail vein in-jection of PBS(RSA-PBS group)and active fragment of kisspeptin KP10(RSA-KP10 group),observing embryo absorption rates.Im-munohistochemistry was employed to assess expressions of kisspeptin and Foxp3 in mice uterine tissues.Peripheral blood Treg cells were isolated and expanded through magnetic bead separation.Intervention with KP10 and KP234(kisspeptin receptor antagonist)was administered,and flow cytometry was used to detect levels of IL-10 and TGF-β1 secretion by Treg cells,as well as differences in proliferation and apoptosis.RNA-Seq transcriptomic sequencing was conducted on uterine tissues from RSA-PBS group and RSA-KP10 group of mice.Differentially expressed genes(DEGs)were subjected to GO,KEGG and GSEA enrichment analyses.Results:Embryo absorption rate in RSA mice was higher than that in NP mice,the embryo absorption rate was decreased after tail vein injec-tion of KP10.Expressions of kisspeptin and Foxp3 in uterus of RSA mice was lower than that in NP mice,while increased after injec-tion of KP10.Kisspeptin could modulate the secretion of IL-10 and TGF-β1 by Treg cells,influencing their proliferation without affect-ing apoptosis.Enrichment analysis results showed that DEGs were mainly enriched in reproductive structure development,IL-17,and TGF-β signaling pathways.Conclusion:Kisspeptin can influence both the quantity and function of Treg cells,offering a new theoreti-cal foundation for investigating the pathogenesis and treatment of RSA.

Objective:To investigate whether kisspeptin can influence the maternal-fetal interface regulatory T cells(Treg),thereby participating in the pathogenesis of recurrent spontaneous abortion(RSA).Methods:Normal pregnancy(NP)and RSA mice models were established,where NP mice received a tail vein injection of PBS(NP-PBS group),and RSA mice received a tail vein in-jection of PBS(RSA-PBS group)and active fragment of kisspeptin KP10(RSA-KP10 group),observing embryo absorption rates.Im-munohistochemistry was employed to assess expressions of kisspeptin and Foxp3 in mice uterine tissues.Peripheral blood Treg cells were isolated and expanded through magnetic bead separation.Intervention with KP10 and KP234(kisspeptin receptor antagonist)was administered,and flow cytometry was used to detect levels of IL-10 and TGF-β1 secretion by Treg cells,as well as differences in proliferation and apoptosis.RNA-Seq transcriptomic sequencing was conducted on uterine tissues from RSA-PBS group and RSA-KP10 group of mice.Differentially expressed genes(DEGs)were subjected to GO,KEGG and GSEA enrichment analyses.Results:Embryo absorption rate in RSA mice was higher than that in NP mice,the embryo absorption rate was decreased after tail vein injec-tion of KP10.Expressions of kisspeptin and Foxp3 in uterus of RSA mice was lower than that in NP mice,while increased after injec-tion of KP10.Kisspeptin could modulate the secretion of IL-10 and TGF-β1 by Treg cells,influencing their proliferation without affect-ing apoptosis.Enrichment analysis results showed that DEGs were mainly enriched in reproductive structure development,IL-17,and TGF-β signaling pathways.Conclusion:Kisspeptin can influence both the quantity and function of Treg cells,offering a new theoreti-cal foundation for investigating the pathogenesis and treatment of RSA.

Objective:To investigate impact of PD-L2 gene knockout on splenic immune phenotype of C57BL/6 mice.Methods:Body weight of female and male wild-type(WT)and PD-L2 gene knockout(KO)mice aged 3～12 weeks were continuously measured.Tissue structure differences in heart,liver,spleen,lung,kidney,thymus,ovary,uterus and testis of four groups of mice were observed by HE staining.Urease UV rate method was employed to measure urea level in mouse serum,and creatinine concentra-tion in serum was determined by creatine oxidase method.Flow cytometry was used to assess proportions of immune cells and cytokine differences in spleens of mice.Results:PD-L2 gene knockout had no apparent effect on body weight or organ structure of mice.Serum urea and creatinine levels were higher in female KO mice compared to WT mice,but no significant differences were observed among males.Female KO mice exhibited reduced proportions of CD4+T cells,CD8+T cells,Treg,NK cells and dendritic cells(DC)in spleen compared to WT mice.Conversely,proportions of Tfh,B cells,plasma cells,na?ve B cells and B2 cells were elevated,proin-flammatory cytokines such as IL-17,IL-6 and IL-22 were increased,while anti-inflammatory cytokines like IL-10 and IL-4 were decreased.Differences among male mice were either not significant or absent.Conclusion:PD-L2 gene knockout primarily affects splenic immune phenotype of female mice,leading to increased inflammatory responses and weakened immune suppression after knockout.