cd8a knockout mice  (Jackson Laboratory)

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: CD8 + T‐cell deletion mitigated injury‐related neointimal hyperplasia in mice. (A)‐ Experimental design. Eightweekold male CD8a +/+ and CD8a −/− ‐mice were randomly subjected to a sham operation (Sham) or a carotid artery ligation+cuff (L + C) operation, followed by 14 days of either nonstress (NonStress) or chronic stress (Stress) prior to tissue collection and analysis. (B) Representative immunostaining and quantitative data show the infiltrated CD8a + macrophages ( red ) with DAPI nuclear counterstain ( blue ). Scale bar: 200 μm. (D, E) Representative hematoxylin and eosin (H&E) and quantitative data show the neointima/media area ratio, neointimal area, and intimal cell number ( n = 6 per group). Data are mean ± SEM ( n = 6/group). The significance of differences among the groups was analyzed by a one‐way ANOVA (C, D). NS, not significant.

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Ligation, Immunostaining

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: CD8 + T‐cell deficiency attenuated the injured vascular collagen deposition, cell proliferation, and macrophage infiltration. CD8a +/+ and CD8a −/− mice were subjected to a sham or L + C operation respectively and were sampled 14 days later. (A) Representative Masson's trichrome and PCNA immunostaining images of collagen deposition and PCNA + cells in the sham and L + C injured carotid arteries of mice of both genotypes. Scale bar: 200 μm. (B) The results of the quantitative analysis of collagen deposition, including the collagen fiber area ( left ), relative collagen content ( middle ), and number of PCNA + cells in the injured arteries of the four experimental groups. (C, D) Representative immunofluorescence images and quantitative data for the numbers of infiltrated CD68 + macrophages ( red ) in the carotid arteries of the four experimental groups. Scale bar: 200 μm. (D) The quantification of CD68 + cell numbers per section. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, D).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Immunostaining, Immunofluorescence

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: CD8 + T‐cell deficiency reduced the investigated molecular alterations in response to the L + C injury. (A, B) Representative western blot images and combined quantitative data for the levels of galectin3, AT1R, p‐Akt, pmTOR, and pP38MAPK in the carotid arteries of CD8a +/+ and CD8a −/− mice subjected to the sham or L + C double injury. (C) The quantitative polymerase chain reaction (qPCR) analysis revealed alterations in the levels of MCP1, ICAM1, VCAM1, MMP2, MMP9, cathepsin S, and cathepsin K in injured arteries of the mice in the four experimental groups. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, C).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Western Blot, Real-time Polymerase Chain Reaction

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: Chronic stress enhanced injury‐reduced CD8 + T‐cell infiltration and neointimal formation in the CD8a +/+ mice. The CD8a +/+ mice underwent a sham operation, L + C injury alone, or stress plus the L + C injury for 2 weeks and were then subjected to sampling for the histological analysis. (A, B) Representative H&E staining images and quantitative data showing the neointima/media area ratio ( left ), neointimal area ( middle ), and intimal cell counts ( right ). (C, D) Representative immunofluorescence images and quantitative data provide the numbers of infiltrated CD8a cells ( red ) in carotid artery cross‐sections of three experimental groups. Scale bar: 200 μm. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, D).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Sampling, Staining, Immunofluorescence

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: Stress accelerated IFN‐γ +/+ CD8 + T‐cell infiltration into the injured carotid artery tissues of CD8a +/+ mice. Double immunofluorescence used anti‐CD8a (red) and anti‐INF‐γ (green) antibodies was applied to identify the IFN‐γ +/+ CD8 + T cells in the three experimental groups. (A, B) Representative double immunofluorescence and quantitative data show the numbers of IFN‐γ +/+ CD8 + T cells in the uninjured and injured carotid artery tissues of the three experimental groups. Scale bar: 100 μm. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Immunofluorescence

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: CD8 + T‐cell deletion attenuated the stress/L + C–induced carotid artery neointimal hyperplasia. CD8a +/+ and CD8a −/− mice that had been subjected to chronic (14‐day) stress plus the L + C injury were subjected to sampling for the histological analyses. (A, B) Representative H&E, Masson's trichrome, and PCNA staining images and combined quantitative data show the neointima/media area ratio, neointimal area, intimal cell count, collagen fiber area, relative collagen content, and number of PCNA + cells in the intima of the two experimental groups. (C, D) Representative immunofluorescence images and quantitative data for the numbers of infiltrated CD68 ( red ) in carotid artery sections of the two experimental groups. Scale bar: 200 μm. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, D).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Sampling, Staining, Cell Counting, Immunofluorescence

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: CD8 + T‐cell deficiency suppressed the investigated molecular alterations in the carotid arteries in response to the stress plus L + C injury. (A, B) Representative western blot images and combined quantitative data for the levels of galectin3, AT1R, pAkt, pmTOR, and pP38MAPK proteins in the carotid arteries of both experimental groups. (C, D) The qPCR analyses revealed the levels of MCP1, VCAM1, ICAM1, MMP2, MMP9, cathepsin K, and cathepsin S genes. Data are mean ± SEM ( n = 6/group). Significance was assessed by unpaired Student's t ‐test (B, C).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Western Blot

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: IFN‐γ signaling promoted CD8 + T‐cell recruitment and neointimal formation in mice subjected to the 2‐week chronic stress conditions. IFN‐γ +/+ and IFN‐γ −/− mice that had received double injuries underwent the chronic stress and were then subjected to sampling for immunoblotting. (A, B) Representative western blot images and quantitative data show the levels of galectin‐3 and AT1R proteins in the carotid arteries of the mice in both experimental groups. IFN‐γ +/+ mice that had received double injuries underwent the stress+vehicle intervention or stress+IFN‐γ neutralizing antibody (Anti‐ IFN‐γ) intervention, respectively, for 2 weeks and were then sampled for the histological analysis. (C, D) Representative H&E staining and CD8a immunofluorescence images and quantitative data present the neointima/media area ratio, neointimal area, intimal cell counts, and CD8 + T cell numbers in the injured arteries of IFN‐γ +/+ mice. CD8a −/− mice that had received double injuries underwent stress plus an adoptive transfer with CD8 + T cells isolated from stressed IFN‐γ +/+ or stressed IFN‐γ −/− donor mice, respectively for 2 weeks and were then subjected to sampling for the histological analysis. (E, F) Representative H&E and CD8a immunofluorescence and quantitative data show the neointima/media area ratio, neointimal area, and intimal cell numbers in the carotid arteries of both experimental groups. Data are mean ± SEM ( n = 6/group). Significance was assessed by unpaired Student's t ‐test (B, D, F).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Sampling, Western Blot, Staining, Immunofluorescence, Adoptive Transfer Assay, Isolation

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: IFN‐γ depletion and deletion reduced CD8a + T‐cell infiltration into the injured carotid arteries of the stressed mice. (A, B) Representative immunofluorescence images and quantitative data for the numbers of infiltrated CD8a ( red ) in injured carotid artery sections of the vehicle and IFN‐γ depletion groups. (A, B) Representative immunofluorescence images and quantitative data for the numbers of infiltrated CD8a ( red ) in injured carotid artery sections of IFN‐γ +/+ CD8 + T‐cell and IFN‐γ −/− CD8 + T‐cell mice. Scale bar: 200 μm. Data are mean ± SEM ( n = 6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, D).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Immunofluorescence

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: The effects of mouse S‐serum on VSMCs' intracellular signaling and their function. Mouse VSMCs were treated with 5% Non‐serum/CD8a +/+ , 5% Non‐serum/CD8a −/− , 5% S‐serum/CD8a +/+ , or 5% S‐serum/CD8a −/− for 12 h and then subjected to a western blotting assay. (A, B) Representative western blot images and combined quantitative data show the levels of p‐Akt and p‐mTOR in the four experimental groups ( n = 4 per group). Here, 5% Non‐serum/CD8a +/+ , 5% Non‐serum/CD8a −/− , 5% S‐serum/CD8a +/+ , and 5% S‐serum/CD8a −/− were respectively added to the out‐wells of transwells. Mouse VSMCs were seeded onto the inner chambers with serum‐free DMEM and then cultured (for 6 h for the migration assay or overnight for the invasion assay). (C, D) Representative microscopy and quantitative data show the numbers of migrated and invaded VSMCs. Scale bar: 75 μm. Data are mean ± SEM ( n = 4–6 for each group). Significance was assessed by a one‐way ANOVA followed by Tukey's post hoc tests (B, D).

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Western Blot, Cell Culture, Migration, Invasion Assay, Microscopy

Journal: The FASEB Journal

Article Title: CD8 + T‐Cell Deletion Suppressed the Development of Injury‐Induced Experimental Neointimal Hyperplasia in Mice With or Without Chronic Stress

doi: 10.1096/fj.202502380R

Figure Lengend Snippet: Graphical illustration of the mechanism of vascular remodeling under chronic stress. CD8+ T‐cell functions as an important mediator of injury‐related experimental neointimal hyperplasia via the modulation of inflammation, proteolysis and proliferation that is mediated by the IFN‐γ/AT1R‐galactin‐3 and mTOR/Akt‐p38MAPK axes. MCP‐1, monocyte chemoattractant protein‐1; ICAM‐1, intracellular adhesion molecular‐1; VCAM‐1, vascular cellular adhesion molecular‐1; MMP‐2, matrix metalloproteinase‐2, MMP‐9, matrix metalloproteinase‐9, Cat S, cathepsin S; Cat K, cathepsin K; AT1R, angiotensin type 1 receptor; p‐Akt, protein kinase‐B; p‐mTOR, phospho‐mammalian target of rapamycin; p‐p38, phospho‐p38 mitogen activation protein kinase.

Article Snippet: Male CD8a knockout mice (CD8a −/−19 ) and IFN‐γ knockout mice (IFN‐γ −/−19 ) (both on C57BL/6 background) were obtained from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; protocol no. 2021‐W5‐2174).

Techniques: Activation Assay

Journal: Journal of Hypertension

Article Title: CD8 + T-cell deficiency protects mice from abdominal aortic aneurysm formation in response to calcium chloride 2

doi: 10.1097/HJH.0000000000003823

Figure Lengend Snippet: CD8 + T-cell deletion mitigated the CaCl 2 -induced AAA formation and the investigated protein levels in aortic tissues. A,B: Representative photographs and data of the maximal external aortic diameter in CD8 +/+ and CD8 −/− mice treated with NaCl or CaCl 2 , respectively (n = 8/group). C–F: Representative western blotting images and quantitative data for the levels of NLRP3, caspase-1, gp91 phox , collagen I, and collagen III proteins in four experimental groups (n = 4/group). G,H: The ELISA results showing the serum levels of IFN-γ and IL-1β in the four groups (n = 6/group).

Article Snippet: Male CD8 −/− (Cd8a knockout [KO]) and IFN-γ −/− (IFN-γ knockout [KO]) mice were purchased from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; Protocol no. 2021-W5–2174) and were bred for the preparation of male CD8 −/− mice and male IFN-γ −/− of the same age.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Hypertension

Article Title: CD8 + T-cell deficiency protects mice from abdominal aortic aneurysm formation in response to calcium chloride 2

doi: 10.1097/HJH.0000000000003823

Figure Lengend Snippet: CD8 + T-cell deficiency alleviated the histopathological changes and recruitment of inflammatory cells. A: The histopathological analysis (H&E, EVG, and Masson trichrome staining) of abdominal aortas of CD8 +/+ and CD8 −/− mice treated with NaCl or CaCl 2 , respectively. Scale bar: upper layer 100 μm, lower layer 50 μm. B: Quantification of the collagen deposition area of the abdominal aortas (n = 5/group). C: Quantification of the elastin degradation score of the abdominal aortas (n = 5/group). D-F: Representative images of IFN-γ + /CD8a + T cell and MMP-2 + / and MMP-9 + /CD68 + macrophage in the adventitia of CD8 +/+ and CD8 −/− AAA mice (n = 5/group). Scale bar: 50 μm. L, Lumen.

Article Snippet: Male CD8 −/− (Cd8a knockout [KO]) and IFN-γ −/− (IFN-γ knockout [KO]) mice were purchased from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; Protocol no. 2021-W5–2174) and were bred for the preparation of male CD8 −/− mice and male IFN-γ −/− of the same age.

Techniques: Staining

Journal: Journal of Hypertension

Article Title: CD8 + T-cell deficiency protects mice from abdominal aortic aneurysm formation in response to calcium chloride 2

doi: 10.1097/HJH.0000000000003823

Figure Lengend Snippet: IFN-γ loading accelerated the CaCl 2 -induced AAA formation in the CD8 −/− mice. A,B: Representative photographs and quantitative data of the maximal external aortic diameter in the PBS-injected (Vehicle) or IFN-γ-injected CD8 −/− mice after CaCl 2 aneurysm induction (n = 8/group). C,D: Representative immunoblotting images and quantitative data for the levels of NLRP3, caspase-1, gp91 phox , collagen I, and collagen III proteins in the two groups (n = 3/group). E: The ELISA results showing the serum levels of IL-1β in both groups (n = 6/group).

Article Snippet: Male CD8 −/− (Cd8a knockout [KO]) and IFN-γ −/− (IFN-γ knockout [KO]) mice were purchased from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; Protocol no. 2021-W5–2174) and were bred for the preparation of male CD8 −/− mice and male IFN-γ −/− of the same age.

Techniques: Injection, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Hypertension

Article Title: CD8 + T-cell deficiency protects mice from abdominal aortic aneurysm formation in response to calcium chloride 2

doi: 10.1097/HJH.0000000000003823

Figure Lengend Snippet: IFN-γ accelerated the histopathological changes and the recruitment of inflammatory cells in the CaCl 2 -induced AAAs in the CD8 −/− mice. A: The histopathological analysis (H&E, EVG, and Masson trichrome staining) of abdominal aortas of PBS-injected (Vehicle) and IFN-γ-injected CD8 −/− mice after CaCl 2 aneurysm induction. Scale bar: upper layer 100 μm, lower layer 50 μm. B: Quantification of the collagen deposition area of the abdominal aortas (n = 5/group). C: Quantification of the elastin degradation score of the abdominal aortas (n = 6/group). D-F: Representative images of IFN-γ + /CD8a + T cell and MMP-2 + / and MMP-9 + /CD68 + macrophage in the adventitia of CD8 −/− AAA mice treated with PBS and IFN-γ (n = 5/group). Scale bar: 50 μm. L, lumen.

Article Snippet: Male CD8 −/− (Cd8a knockout [KO]) and IFN-γ −/− (IFN-γ knockout [KO]) mice were purchased from Shanghai Biomodel Organism Science & Technology Development Co. (Shanghai, China; Protocol no. 2021-W5–2174) and were bred for the preparation of male CD8 −/− mice and male IFN-γ −/− of the same age.

Techniques: Staining, Injection