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 • 2007
Source Paper
Tomasz J. Guzik, Nyssa E. Hoch, Kathryn A. Brown, Louise A. McCann, Ayaz Rahman et al.
The Journal of Experimental Medicine • 2007
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: Induction of hypertension using DOCA-salt treatment and assessment of hypertension-related outcomes 40 days after induction
This is a DOCA-Salt Hypertension Model protocol using mouse as the model organism. The procedure involves 8 procedural steps, 3 equipment items, 5 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 cell transfer (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. Maintain all mice in sterile environment and regularly screen for infections.
Note: All mice on C57BL/6 background. Protocols approved by institutional Animal Care and Use Committee at Emory University.
“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 cell transfer before beginning angiotensin II infusion and blood pressure monitoring.
Note: This step only applies 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) intraperitoneally 3 days before angiotensin II infusion begins.
Note: This step is only for experiments testing TNF-alpha or IFN-gamma inhibition
“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. Measure blood pressure both invasively and noninvasively throughout infusion period.
Note: Blood pressure measured by both invasive and noninvasive methods
“490 ng/min/kg angiotensin II was infused and blood pressure was measured both invasively and noninvasively”
Continue administering etanercept or neutralizing IFN-gamma antibody intraperitoneally every 3 days during the angiotensin II infusion period.
Note: This step is only for experiments testing TNF-alpha or IFN-gamma inhibition
“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.”
Create DOCA-salt hypertension in mice using previously described methods.
Note: Specific protocol details referenced as previously described
“DOCA-salt hypertension was created as previously described”
Allow 40 days to elapse after DOCA-salt hypertension induction before performing experimental studies.
Note: This is the critical timepoint for this specific experiment
“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.
Induction of hypertension using DOCA-salt treatment and assessment of hypertension-related outcomes 40 days after induction
Objective
Induction of hypertension using DOCA-salt treatment and assessment of hypertension-related outcomes 40 days after induction
Subjects
From papermouse • C57BL/6, RAG-1 -/-, AT1a -/-, p47 phox -/- • unknown • Not specified • 30g (referenced for antibody dosing)
Cohort notes
From paperC57BL/6 background for all strains.
Animal acquisition and housing (Not specified)
Adoptive cell transfer (if applicable) (Not specified)
Wait period after adoptive transfer (3 weeks)
Pre-treatment with etanercept or anti-IFN-gamma antibody (if applicable) (Single injection 3 days before infusion)
Blood pressure (invasive measurement)
From paperNot specified in methods section
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Blood pressure (noninvasive measurement)
From paperNot specified in methods section
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Hypertension development and maintenance
From paperNot specified in methods section
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Blood pressure (invasive measurement)
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 (noninvasive measurement)
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 development and maintenance
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 methods section
Scoring or quantification
Quantify the primary readouts for this experiment: Blood pressure (invasive measurement); Blood pressure (noninvasive measurement); Hypertension development and maintenance.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Blood pressure (invasive measurement), Blood pressure (noninvasive measurement), Hypertension development and maintenance.
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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Not specified • Not specified • Not specified • Not specified
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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
Not specified • Not specified • Not specified • Not specified
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Source access
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Steps
8
Evidence Quotes
16
Protocol Items
8
Linked Products
1
Canonical Sync
Pending
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Steps
8
Evidence
16
Specific Products
1/1
Canonical Sync
Pending
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