scholarly journals The Alopecia Areata Phenotype Is Induced by the Water Avoidance Stress Test In cchcr1-Deficient Mice

Biomedicines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 840
Author(s):  
Qiaofeng Zhao ◽  
Satoshi Koyama ◽  
Nagisa Yoshihara ◽  
Atsushi Takagi ◽  
Etsuko Komiyama ◽  
...  
Keyword(s):  
Alopecia Areata ◽  
Gene Knockout ◽  
Stress Test ◽  
Coiled Coil ◽  
Risk Haplotype ◽  
Gene Expression Microarray ◽  
Wild Type ◽  
Gene Knockout Mice ◽  
Deficient Mice ◽  
Gene Expression Microarray Analysis

We recently discovered a nonsynonymous variant in the coiled-coil alpha-helical rod protein 1 (CCHCR1) gene within the alopecia areata (AA) risk haplotype. We also reported that the engineered mice with this risk allele exhibited. To investigate more about the involvement of the CCHCR1 gene in AA pathogenesis, we developed an AA model using C57BL/6N cchcr1 gene knockout mice. In this study, mice (6–8 weeks) were divided into two groups: cchcr1−/− mice and wild-type (WT) littermates. Both groups were subjected to a water avoidance stress (WAS) test. Eight weeks after the WAS test, 25% of cchcr1−/− mice exhibited non-inflammatory foci of alopecia on the dorsal skin. On the other hand, none of wild-type littermates cause hair loss. The foci resembled human AA in terms of gross morphology, trichoscopic findings and histological findings. Additionally, gene expression microarray analysis of cchcr1−/− mice revealed abnormalities of hair related genes compared to the control. Our results strongly suggest that CCHCR1 is associated with AA pathogenesis and that cchcr1−/− mice are a good model for investigating AA.

scholarly journals The alopecia areata phenotype is induced by the water avoidance stress test in cchcr1-deficient mice

2020 ◽  
Author(s):  
Qiao-Feng Zhao ◽  
Nagisa Yoshihara ◽  
Atsushi Takagi ◽  
Etsuko Komiyama ◽  
Akira Oka ◽  
...  
Keyword(s):  
Hair Loss ◽  
Alopecia Areata ◽  
Gene Knockout ◽  
Stress Test ◽  
Susceptibility Gene ◽  
Coiled Coil ◽  
Risk Haplotype ◽  
Gene Knockout Mice ◽  
Disease Susceptibility Gene ◽  
Deficient Mice

ABSTRACTBackgroundWe recently discovered a nonsynonymous variant in the coiled-coil alpha-helical rod protein 1 (CCHCR1) gene within the alopecia areata (AA) risk haplotype; mice engineered to carry the risk allele displayed a hair loss phenotype.ObjectiveTo further investigate the involvement of the CCHCR1 gene in AA pathogenesis.MethodsWe developed an AA model using C57BL/6N cchcr1 gene knockout mice. Mice (6-8 weeks) were divided into two groups: cchcr1-/- mice and wild-type (WT) mice. Both groups were subjected to a water avoidance stress (WAS) test.ResultsEight weeks after the WAS test, 25% of cchcr1 mice exhibited noninflammatory foci of alopecia on the dorsal skin. The foci resembled human AA in terms of gross morphology, trichoscopic findings and histological findings.ConclusionsOur results strongly suggest that CCHCR1 is associated with AA pathogenesis and that cchcr1-/- mice are a good model for investigating AA.Author summaryAlopecia areata is thought to affect 1-2% of the population. In severe alopecia areata, changes in appearance significantly reduce the patient’s quality of life, but there is no established treatment. Its pathogenic mechanism is thought to cause an autoimmune disease of the hair bulb of growing hair. Many genes and factors associated with AA onset form a background that easily causes such an immune reaction, and the causative genes related to alopecia areata are being elucidated worldwide. We were able to identify the CCHCR1 gene as one of the causative genes by genome analysis. In this study, we created CCHCR1-deficient mice using Cre/loxP technology and confirmed that 25% of CCHCR1-deficient mice that underwent the WAS test for psychological stimulation developed hair loss similar to that observed in human alopecia areata. This suggests that the CCHCR1 gene is a disease susceptibility gene for alopecia areata.

scholarly journals Distinct Roles of CD28- and CD40 Ligand-Mediated Costimulation in the Development of Protective Immunity and Pathology during Chlamydia muridarum Urogenital Infection in Mice

2009 ◽  
Vol 77 (7) ◽  
pp. 3080-3089 ◽  
Author(s):  
Lili Chen ◽  
Wen Cheng ◽  
Pooja Shivshankar ◽  
Lei Lei ◽  
Xiaoyun Zhang ◽  
...  
Keyword(s):  
Primary Infection ◽  
Protective Immunity ◽  
Gene Knockout ◽  
Secondary Infection ◽  
Cd40 Ligand ◽  
Wild Type ◽  
Chlamydia Muridarum ◽  
Urogenital Infection ◽  
Costimulatory Signals ◽  
Deficient Mice

ABSTRACT Infection with Chlamydia muridarum in the mouse urogenital tract can induce both protective immunity and inflammatory pathologies, which has been used as a model for understanding the immune and pathogenic mechanisms of C. trachomatis infection. We compared the roles of CD28- and CD40 ligand (CD40L)-mediated costimulation in C. muridarum infection. Mice with CD28 or CD80/CD86 gene knockout (KO) displayed an infection course similar to that of wild-type mice during both primary and secondary infection, suggesting that CD28-mediated costimulation is not required for protection against C. muridarum infection. However, mice deficient in CD40L or CD40 displayed a prolonged infection course after primary or secondary infection, suggesting that CD40-CD40L costimulation plays an essential role in the development of anti-C. muridarum immunity. Interestingly, the CD28- or CD80/CD86-deficient mice displayed significantly lower levels of inflammatory pathologies in the upper genital tracts after primary infection, although the attenuation in inflammation was no longer significant during secondary infection. However, the CD40L or CD40 KO mice developed inflammatory pathologies as severe as those in wild-type mice following either primary or secondary infection despite the obvious deficits in adaptive immunity in these KO mice. The resistance of CD28 or CD80/CD86 KO mice to chlamydial infection correlated with production of gamma interferon, while the development of inflammatory pathologies in CD40L or CD40 KO mice correlated with the production of other proinflammatory cytokines in mouse urogenital tracts during the early stages of the infection. These observations together suggest that C. muridarum-induced protective immunity and inflammatory pathologies can be mediated by distinct costimulatory signals.

Effects of expression of p53 and Gadd45 on osmotic tolerance of renal inner medullary cells

2006 ◽  
Vol 291 (2) ◽  
pp. F341-F349 ◽  
Author(s):  
Qi Cai ◽  
Natalia I. Dmitrieva ◽  
Joan D. Ferraris ◽  
Luis F. Michea ◽  
Jesus M. Salvador ◽  
...  
Keyword(s):  
Gene Knockout ◽  
Collecting Duct ◽  
Wild Type ◽  
Growth And Survival ◽  
Osmotic Tolerance ◽  
Rate Of Increase ◽  
Gene Knockout Mice ◽  
Collecting Duct Cells ◽  
Urinary Concentrating Ability

The response of renal inner medullary (IM) collecting duct cells (mIMCD3) to high NaCl involves increased expression of Gadd45 and p53, both of which have important effects on growth and survival of the cells. However, mIMCD3 cells, being immortalized by SV40, proliferate rapidly, which is known to sensitize cells to high NaCl, whereas IM cells in situ proliferate very slowly and survive much higher levels of NaCl. In the present studies, we have examined the importance of Gadd45 and p53 for survival of normal IM cells in their usual high-NaCl environment by using more slowly proliferating second-passage mouse inner medullary epithelial (p2mIME) cells and comparing cells from wild-type and gene knockout mice. Acutely elevating NaCl (and/or urea) reduces Gadd45a, but increases Gadd45b and Gadd45g mRNA, depending on the mix of NaCl and urea and the rate of increase of osmolality. Nevertheless, p2mIME cells from Gadd45b−/−, Gadd45g−/−, and Gadd45bg−/− mice survive elevation of NaCl (or urea) essentially the same as do wild-type cells. p53−/− Cells do not tolerate as high a concentration of NaCl (or urea) as p53+/+ cells, but urinary concentrating ability of p53−/− mice is normal, as is the histology of inner medullas from p53−/− and Gadd45abg−/− mice. Thus although Gadd45 and p53 may play roles in osmotically stressed mIMCD3 cells, we do not find that their expression makes an important difference, either for Gadd45 in slower proliferating p2mIME cells or for Gadd45 or p53 in normal inner medullary epithelial cells in situ.

scholarly journals Alopecia areata susceptibility variant identified by MHC risk haplotype sequencing reproduces symptomatic patched hair loss in mice

2018 ◽  
Author(s):  
Akira Oka ◽  
Atsushi Takagi ◽  
Etsuko Komiyama ◽  
Shuhei Mano ◽  
Kazuyoshi Hosomichi ◽  
...  
Keyword(s):  
Hair Loss ◽  
Alopecia Areata ◽  
Complex Disease ◽  
Susceptibility Gene ◽  
Coiled Coil ◽  
Alternative Mechanism ◽  
Risk Haplotype ◽  
Associated Proteins ◽  
Allele Specific ◽  
Synonymous Variant

AbstractBackgroundAlopecia areata (AA) is a highly heritable multifactorial and complex disease. However, no convincing susceptibility gene has yet been pinpointed in the major histocompatibility complex (MHC), a region in the human genome known to be associated with AA as compared to other regions.ResultsBy sequencing MHC risk haplotypes, we identified a variant (rs142986308, p.Arg587Trp) in the coiled-coil alpha-helical rod protein 1 (CCHCR1) gene as the only non-synonymous variant in the AA risk haplotype. Using CRISPR/Cas9 for allele-specific genome editing, we then phenocopied AA symptomatic patched hair loss in mice engineered to carry the Cchcr1 risk allele. Skin biopsies of these alopecic mice showed strong up-regulation of hair-related genes, including hair keratin and keratin-associated proteins (KRTAPs). Using transcriptomics findings, we further identified CCHCR1 as a novel component of hair shafts and cuticles in areas where the engineered alopecic mice displayed fragile and impaired hair.ConclusionsThese results suggest an alternative mechanism for the aetiology of AA based on aberrant keratinization, in addition to generally well-known autoimmune events.

scholarly journals Caspase-3/NLRP3 signaling in the mesenchymal stromal niche regulates myeloid-biased hematopoiesis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jing Zhang ◽  
Yaozhen Chen ◽  
Dandan Yin ◽  
Fan Feng ◽  
Qunxing An ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Knockout Mice ◽  
Caspase 3 ◽  
Ex Vivo ◽  
Gene Knockout ◽  
Hematopoietic Stem ◽  
Nlrp3 Gene ◽  
Gene Knockout Mice ◽  
Deficient Mice

Abstract Background The fate of hematopoietic stem cells (HSCs) is determined by a complex regulatory network that includes both intrinsic and extrinsic signals. In the past decades, many intrinsic key molecules of HSCs have been shown to control hematopoiesis homeostasis. Non-hematopoietic niche cells also contribute to the self-renewal, quiescence, and differentiation of HSCs. Mesenchymal stromal cells (MSCs) have been identified as important components of the niche. However, the regulatory role of MSCs in hematopoiesis has not been fully understood. Methods Caspase-3 and NLRP3 gene knockout mice were generated respectively, and hematopoietic development was evaluated in the peripheral circulation and bone marrow by flow cytometry, colony formation assay, and bone marrow transplantation. Bone-associated MSCs (BA-MSCs) were then isolated from gene knockout mice, and the effect of Caspase-3/NLRP3 deficient BA-MSCs on hematopoiesis regulation was explored in vivo and ex vivo. Results We report that Caspase-3 deficient mice exhibit increased myelopoiesis and an aberrant HSC pool. Ablation of Caspase-3 in BA-MSCs regulates myeloid lineage expansion by altering the expression of hematopoietic retention cytokines, including SCF and CXCL12. Interestingly, NLRP3 gene knockout mice share phenotypic similarities with Caspase-3 deficient mice. Additionally, we found that NLRP3 may play a role in myeloid development by affecting the cell cycle and apoptosis of hematopoietic progenitors. Conclusions Our data demonstrate that the Caspase-3/NLRP3 signaling functions as an important regulator in physiological hematopoiesis, which provides new insights regarding niche signals that influence hematopoiesis regulation in the bone marrow.

scholarly journals Specificity of aminergic status in skin and tumor of C57BL/6-PlautmI.IBug-This Plau6FDhu/GFDhu mice with B16/F10 melanoma

2020 ◽  
Vol 66 (6) ◽  
pp. 707-711
Author(s):  
Oleg Kit ◽  
Elena Frantsiyants ◽  
Irina Kaplieva ◽  
Ekaterina Surikova ◽  
Irina Neskubina ◽  
...  
Keyword(s):  
Biogenic Amines ◽  
Knockout Mice ◽  
Gene Knockout ◽  
Local Stress ◽  
Plasminogen Activation ◽  
Intact Skin ◽  
Gene Knockout Mice ◽  
Hydroxyindoleacetic Acid ◽  
Solitary Metastases ◽  
Deficient Mice

Background. Systems of plasminogen activation and biogenic amines are involved in carcinogenesis, but their relationship has not been established. Aim – study of the quantitative specificity of biogenic amines in the skin and melanoma in urokinase-gene knockout mice. Materials and methods.  Levels of catecholamines, histamine, serotonin and 5-hydroxyindoleacetic acid (HIAA) were determined by ELISA in the skin and В16/F10 melanoma of urokinase (uPA) -gene knockout mice of both genders (n=24); С57ВL/6 mice (n=64) were controls (C). Results. Differential characteristics of melanoma development in uPA-deficient mice included: an earlier onset of the primary tumor and its slow growth, more pronounced in females, in combination with hemorrhages in the lungs of males and solitary metastases in the lungs of females. This was facilitated by higher levels of norepinephrine in the skin of males/females – by 4.8/4.9 times, histamine – by 3.6/1.7 (p<0.05) times and serotonin – by 3.4/8.3 times. Dopamine accumulated in melanoma in all uPA-gene knockout mice: in females –1.6 times (p<0.05), in males – 2.1 times higher than in intact skin, with a 2.5 times reduction of norepinephrine in females. Levels of histamine decreased, but exceeded controls: in females – by 1.8 times (p<0.05), in males –by 3.5 times. Levels of serotonin in uPA-deficient females were as high, while in males they were 3.4 times lower than in intact skin. Conclusions. The specificity of the aminergic system in the skin of uPA-gene knockout mice demonstrated the inhibition of local stress and contributed to the reduction of malignant potential of melanoma by increasing immune properties of the skin.

scholarly journals Experimental Pseudomonas aeruginosa Keratitis in Interleukin-10 Gene Knockout Mice

2003 ◽  
Vol 71 (3) ◽  
pp. 1328-1336 ◽  
Author(s):  
Nerida Cole ◽  
Mark Krockenberger ◽  
Fiona Stapleton ◽  
Shamila Khan ◽  
Emma Hume ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gene Knockout ◽  
Bacterial Load ◽  
Interleukin 10 ◽  
Wild Type ◽  
Cytokine Levels ◽  
Infiltrating Cells ◽  
Gene Knockout Mice ◽  
Corneal Angiogenesis ◽  
Kinetics Of

ABSTRACT Pseudomonas aeruginosa keratitis is one of the most destructive diseases of the cornea. The host response to this infection is critical to the outcome. The cytokine interleukin-10 (IL-10) is thought to play an important role in modulating excessive inflammation and antimicrobial defenses. We have found that in IL-10−/− mice there is a significant decrease in bacterial load in corneas at 7 days postchallenge with P. aeruginosa. This decrease was accompanied by a reduction in neutrophil numbers in the cornea and changes in cytokine levels compared to those of wild-type mice. A characteristic increase in neovascularization in the cornea was found in the IL-10−/− mice. This increased angiogenesis correlated with an increased expression of KC, whereas the kinetics of macrophage inflammatory peptide 2 expression correlated with neutrophil numbers. This finding suggests that KC may play a role in corneal angiogenesis. The source of IL-10 in mouse corneas was identified as a subpopulation of infiltrating cells and keratocytes. This study demonstrates that IL-10 plays an important role in regulating the balance of inflammatory mediators during P. aeruginosa infection of the cornea.

scholarly journals Comparison of Ocular Pathologies in Vitamin A–Deficient Mice and RPE65 Gene Knockout Mice

2011 ◽  
Vol 52 (8) ◽  
pp. 5507 ◽  
Author(s):  
Yang Hu ◽  
Ying Chen ◽  
Gennadiy Moiseyev ◽  
Yusuke Takahashi ◽  
Robert Mott ◽  
...  
Keyword(s):  
Vitamin A ◽  
Knockout Mice ◽  
Gene Knockout ◽  
Gene Knockout Mice ◽  
Deficient Mice

scholarly journals Critical Role of Cytotoxic T Lymphocytes in Immune Clearance of Rickettsial Infection

2001 ◽  
Vol 69 (3) ◽  
pp. 1841-1846 ◽  
Author(s):  
David H. Walker ◽  
Juan P. Olano ◽  
Hui-Min Feng
Keyword(s):  
T Lymphocytes ◽  
Knockout Mice ◽  
Gene Knockout ◽  
Wild Type ◽  
Spleen Cells ◽  
Rickettsial Infection ◽  
Cd8 T Lymphocytes ◽  
Gene Knockout Mice ◽  
Ifn Γ ◽  
Ctl Activity

ABSTRACT Cytotoxic T-lymphocyte (CTL) activity developed against the major infected target cells of rickettsial infections, endothelial cells and macrophages. Spleen cells from mice immune to Rickettsia conorii exerted specific major histocompatibility complex (MHC) class I-matched CTL activity against R. conorii-infected SVEC-10 endothelial cells, with peak activity on day 10. Similarly, spleen cells from Rickettsia australis-immune mice exerted specific CTL activity against an R. australis-infected macrophage-like cell line. Gamma interferon (IFN-γ) gene knockout mice were more than 100-fold more susceptible to R. australis infection than wild-type C57BL/6 mice. MHC class I gene knockout mice were the most susceptible, more than 50,000-fold more susceptible to a lethal outcome of R. australis infection than wild-type C57BL/6 mice. These results indicate that CTL activity was more critical to recovery from rickettsial infection than were the effects of IFN-γ. The observation that perforin gene knockout mice were more than 100-fold more susceptible than wild-type C57BL/6 mice indicates that perforin-mediated activity accounts for a large component, but not all, of the CTL-mediated antirickettsial effect. CTL activity was expressed by immune CD8 T lymphocytes. Adoptive transfer of immune CD8 T lymphocytes from IFN-γ gene knockout mice intoR. australis-infected IFN-γ gene knockout mice dramatically reduced the infectious rickettsial content in the organs, confirming that CD8 T lymphocytes provide immunity against rickettsiae besides that provided by the secretion of IFN-γ. CTLs appear to be crucial to recovery from rickettsial infection.

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