Systemic T-cell activation in stable angina pectoris

Abstract

However, there have beenfew studies on the systemic cell-mediated immuneresponse in stable angina pectoris (SAP), and the dataobtained so far have not provided any strong supportfor continuous T-cell activation. This study examinesthe T-cell profi le, including subsets and activationstatus of circulating T cells, as well as serum levels ofthe soluble interleukin-2 receptor (sIL-2R) in SAP.•••We studied 38 men ( 60 years of age) who all hadangiographically verifi ed coronary artery disease with 2 coronary stenoses ( 50% reduction of lumendiameter). The diagnosis of SAP was defi ned as ef-fort-related angina of Canadian Cardiovascular Soci-ety functional class II and III. Patients with myocar-dial infarction or UAP within the 2 last months beforethe study were excluded. Other exclusion criteria werediabetes, immunologic disorders, neoplastic disease,evidence of acute or recent ( 2 months) infection,recent major trauma, surgery or coronary revascular-ization, and treatment with antibiotic, immunosup-pressive, or anti-infl ammatory agents. Forty-two pre-sumably healthy men of equivalent age served ascontrols. Informed consent was obtained from all sub-jects. The research protocol was approved by thelocally appointed ethics committee.Blood was obtained by vein puncture in the morn-ing after a 12-hour fast. T-lymphocyte subpopulationswere measured by 3 color fl ow cytometry usingFACScan (Becton Dickinson Immunocytometry Sys-tems, Jo¨nko¨ping, Sweden). The cells were stained bydifferent triples of monoclonal antibodies against thefollowing antigens: CD3 (all T cells), CD4 (T-helpercells), CD8 (T cytotoxic/suppressor cells), CD25 (IL-2R; an early marker of T-cell activation), andHLA-DR (a marker of late T-cell activation). Theantibodies were marked with 1 of 3 fl uorochromes:fl uorescein isothiocyanate, phycoerythrin, and peri-dinin chlorofyll protein. A sample of 20 l of eachmonoclonal reagent triple was added to 100 lofwhole blood in 12 75 test tubes, centrifuged gently,and incubated at 4°C for 30 minutes. Red blood cellswere lysed using 3 ml of FACS lysing solution during2 minutes incubation at room temperature in the dark.The cells were washed twice with phosphate-bufferedsaline with 0.1% sodium azide, resuspended in washbuffer, fi xed with 1% paraformaldehyde, and analyzedafter 2 to 3 hours. Data were analyzed using CELLQuest software (Becton-Dickinson, Stockholm, Swe-den). Instrument setups, adjustments, and viabilitycontrols were performed as previously described.