Assessment of faithful interleukin-3 production by novel bicistronic interleukin-3 reporter mice.

Interleukin-3 (IL-3) is an important hematopoietic growth factor and immunregulatory cytokine. Although activated T helper cells represent a main source of IL-3, other cell types have been reported to express this cytokine. However, precise identification and quantification of the cells that produce IL-3 in vivo have not been performed. Therefore, we used a CRISPR/Cas approach to engineer mice containing a bicistronic mRNA linking a readily identifiable reporter, enhanced green fluorescent protein (ZsGreen1), to IL-3 expression. To characterize these novel reporter mice, we first examined ZsGreen1 expression by CD4 T cells subsets primed and activated in vitro. We found that activated Th1 cells expressed ∼4-fold higher levels of ZsGreen1 as compared to Th0 and Th2 cells. Endogenous IL-3 expression remained intact although reporter Th1 cells secreted ∼33 % less IL-3 than similarly activated wild-type cells. To characterize the ability of reporter mice to accurately mark IL-3-producing cells in vivo, we infected mice with Nippostrongylus brasiliensis. Low but significant numbers of ZsGreen1+ CD4 T cells were detected in the mesenteric lymph nodes and lung following both primary and secondary infection. No difference in basophil and intestinal mast cell numbers were observed between infected reporter and wild-type mice indicating that reporter mice secreted IL-3 levels in vivo that results in IL-3-driven biological activities which are indistinguishable from those observed in corresponding wild-type mice. These IL-3 reporter mice will be a valuable resource to investigate IL-3-dependent immune responses in vivo.

A randomized controlled trial of a controlled breathing protocol on heart rate variability following myocardial infarction or coronary artery bypass graft surgery.

OBJECTIVES: To determine whether a controlled breathing programme increases heart rate variability following an acute myocardial infarction and/or coronary artery bypass graft surgery. RATIONALE: Heart rate variability is reduced following a myocardial infarction, and low heart rate variability is associated with a high mortality risk. By changing tidal volume and rate of breathing, individuals can alter beat-to-beat heart rate variability. It is hypothesized that heart rate increases with inspiration and decreases with exhalation, and that deep slow breathing enhances respiratory sinus arrhythmia, increasing heart rate variability. DESIGN: Randomized controlled trial. SETTING: Cardiac rehabilitation programme at a large academic medical centre in North Texas. SUBJECTS: From 2001 to 2005, 44 patients, age 46-65 years, who had a myocardial infarction and/or undergone coronary artery bypass graft surgery 1-8 weeks previously and were referred to the Cardiac Rehabilitation Program. INTERVENTION: Patients were randomized to either usual cardiac rehabilitation or cardiac rehabilitation with controlled breathing (6 breaths/min for 10 minutes twice daily during the eight-week treatment period). MAIN MEASURES: Weekly measurements of total power and standard deviation of the mean normal to normal RR interval (SDNN), and fortnightly measurements of respiratory sinus arrhythmia were taken using Biocom Technologies Heart Rhythm Scanner and Tracker software. RESULTS: No significant difference in change were seen between groups in SDNN (P = 0.3984), baseline respiratory sinus arrhythmia (P = 0.6556) or total power (P = 0.6184). CONCLUSION: Results suggest participation in the controlled breathing programme offered no additional benefit in increasing heart rate variability following myocardial infarction or coronary artery bypass graft surgery. However, 77% of study patients were on heart rate-lowering medications, which may have masked changes in heart rate variability.