Source Paper
Role of the T cell in the genesis of angiotensin II–induced hypertension and vascular dysfunction
Tomasz J. Guzik, Nyssa E. Hoch, Kathryn A. Brown, Louise A. McCann, Ayaz Rahman et al.
The Journal of Experimental Medicine •
Source Paper
Tomasz J. Guzik, Nyssa E. Hoch, Kathryn A. Brown, Louise A. McCann, Ayaz Rahman et al.
The Journal of Experimental Medicine •
Hypertension promotes atherosclerosis and is a major source of morbidity and mortality. We show that mice lacking T and B cells (RAG-1−/− mice) have blunted hypertension and do not develop abnormalities of vascular function during angiotensin II infusion or desoxycorticosterone acetate (DOCA)–salt. Adoptive transfer of T, but not B, cells restored these abnormalities. Angiotensin II is known to stimulate reactive oxygen species production via the nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase in several cells, including some immune cells. Accordingly, adoptive transfer of T cells lacking the angiotensin type I receptor or a functional NADPH oxidase resulted in blunted angiotensin II–dependent hypertension and decreased aortic superoxide production. Angiotensin II increased T cell markers of activation and tissue homing in wild-type, but not NADPH oxidase–deficient, mice. Angiotensin II markedly increased T cells in the perivascular adipose tissue (periadventitial fat) and, to a lesser extent the adventitia. These cells expressed high levels of CC chemokine receptor 5 and were commonly double negative (CD3+CD4−CD8−). This infiltration was associated with an increase in intercellular adhesion molecule-1 and RANTES in the aorta. Hypertension also increased T lymphocyte production of tumor necrosis factor (TNF) α, and treatment with the TNFα antagonist etanercept prevented the hypertension and increase in vascular superoxide caused by angiotensin II. These studies identify a previously undefined role for T cells in the genesis of hypertension and support a role of inflammation in the basis of this prevalent disease. T cells might represent a novel therapeutic target for the treatment of high blood pressure.
Objective: To induce hypertension through continuous angiotensin II infusion and measure blood pressure changes using both invasive and noninvasive methods
This is a Angiotensin II Infusion and Blood Pressure Measurement protocol using mouse as the model organism. The procedure involves 8 procedural steps, 3 equipment items, 4 materials. Extracted from a 2007 paper published in The Journal of Experimental Medicine.
Model and subjects
mouse • C57BL/6, RAG-1 -/-, AT1a -/-, p47 phox -/- • unknown • Not specified • 30g (referenced for antibody dosing)
Study window
~5.7 week study window
Core workflow
Animal acquisition and housing • Adoptive transfer procedure (if applicable) • Wait period after adoptive transfer
Primary readouts
Key equipment and reagents
Verified items
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Obtain C57BL/6, RAG-1 -/-, and AT1a -/- mice from Jackson ImmunoResearch Laboratories and p47 phox -/- mice from Taconic. House animals in sterile environment and regularly screen for infections.
Note: All mice maintained on C57BL/6 background. Institutional Animal Care and Use Committee approval required.
“C57BL/6, RAG-1 -/-, and AT1a -/- mice were obtained from Jackson ImmunoResearch Laboratories. The p47 phox -/- mice and their appropriate controls were obtained from Taconic. Animals were maintained in a sterile environment and were regularly screened for infections.”
Anesthetize mice with xylazine/ketamine and inject cells via tail vein for adoptive transfer experiments.
Note: This step is only for experiments involving adoptive transfer
“For adoptive transfer, mice were anesthetized with xylazine/ketamine and cells were injected via tail vein.”
Allow 3 weeks to elapse after adoptive transfer before beginning angiotensin II infusion and blood pressure monitoring.
Note: This step applies only to adoptive transfer experiments
“Angiotensin II infusion and blood pressure monitoring was begun 3 wk after adoptive transfer.”
Administer etanercept (8 mg/kg) or neutralizing IFN-gamma antibody (0.5 mg per injection per 30g mouse, clone R4-6A2) intraperitoneally 3 days before angiotensin II infusion begins.
Note: This step is only for experiments using these treatments. Etanercept from AmGen, antibody from eBioscience.
“8 mg/kg etanercept (AmGen) or a neutralizing IFN-gamma antibody (eBioscience; clone R4-6A2; 0.5 mg per injection per 30 g mouse) was administered i.p. 3 d before and every 3 d during angiotensin II infusion.”
Start continuous infusion of angiotensin II at 490 ng/min/kg body weight.
Note: Infusion method and duration not specified in text
“490 ng/min/kg angiotensin II was infused and blood pressure was measured both invasively and noninvasively”
Simultaneously measure blood pressure using both invasive and noninvasive methods during angiotensin II infusion.
Note: Specific measurement intervals and duration not provided in text
“blood pressure was measured both invasively and noninvasively, as previously described”
Administer etanercept or IFN-gamma antibody injections every 3 days during the angiotensin II infusion period.
Note: Only for experiments using etanercept or antibody treatments
“8 mg/kg etanercept (AmGen) or a neutralizing IFN-gamma antibody (eBioscience; clone R4-6A2; 0.5 mg per injection per 30 g mouse) was administered i.p. 3 d before and every 3 d during angiotensin II infusion.”
For some experiments, create DOCA-salt hypertension as an alternative to angiotensin II infusion using previously described methods.
Note: This is an alternative experimental approach, not combined with angiotensin II infusion
“In some mice, DOCA-salt hypertension was created as previously described, and studies were performed after 40 d from the induction of hypertension”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
To induce hypertension through continuous angiotensin II infusion and measure blood pressure changes using both invasive and noninvasive methods
Objective
To induce hypertension through continuous angiotensin II infusion and measure blood pressure changes using both invasive and noninvasive methods
Subjects
From papermouse • C57BL/6, RAG-1 -/-, AT1a -/-, p47 phox -/- • unknown • Not specified • 30g (referenced for antibody dosing)
Cohort notes
From paperAll mice on C57BL/6 background.
Animal acquisition and housing (Not specified)
Adoptive transfer procedure (if applicable) (Not specified)
Wait period after adoptive transfer (3 weeks)
Pre-treatment with etanercept or IFN-gamma antibody (if applicable) (Single injection 3 days before infusion)
Blood pressure measurements (invasive)
From paperNot specified in text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Blood pressure measurements (noninvasive)
From paperNot specified in text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Hypertension induction and progression
From paperNot specified in text
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Blood pressure measurements (invasive)
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Blood pressure measurements (noninvasive)
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Hypertension induction and progression
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Not specified in text
Scoring or quantification
Quantify the primary readouts for this experiment: Blood pressure measurements (invasive); Blood pressure measurements (noninvasive); Hypertension induction and progression.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Blood pressure measurements (invasive), Blood pressure measurements (noninvasive), Hypertension induction and progression.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Tomasz J. Guzik et al. (2007). Role of the T cell in the genesis of angiotensin II–induced hypertension and vascular dysfunction. The Journal of Experimental Medicine
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AmGen • Not specified • Not specified • Not specified
eBioscience • Clone R4-6A2 • Not specified • Not specified
Not specified • Not specified • Not specified • Not specified
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Source access
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Steps
8
Evidence Quotes
15
Protocol Items
7
Linked Products
1
Canonical Sync
Pending
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Steps
8
Evidence
15
Specific Products
1/1
Canonical Sync
Pending
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