Paola Pileri

Decreased circulating T regulatory lymphocytes in obese patients undergoing bariatric surgery

Objective It has been previously demonstrated that T lymphocytes may be involved in the development of hypertension and microvascular remodeling, and that circulating T effector lymphocytes may be increased in hypertension. In particular, Th1 and Th 17 lymphocytes may contribute to the progression of hypertension and microvascular damage while T-regulatory (Treg) lymphocytes seem to be protective in this regard. However, no data is available about patients with severe obesity, in which pronounced microvascular alterations were observed. Design and methods We have investigated 32 severely obese patients undergoing bariatric surgery, as well as 24 normotensive lean subjects and 12 hypertensive lean subjects undergoing an elective surgical intervention. A peripheral blood sample was obtained before surgery for assessment of CD4+ T lymphocyte subpopulations. Lymphocyte phenotype was evaluated by flow cytometry in order to assess T-effector and Treg lymphocytes. Results A marked reduction of several Treg subpopulations was observed in obese patients compared with controls, together with an increased in CD4+ effector memory T-effector cells. Conclusion In severely obese patients, Treg lymphocytes are clearly reduced and CD4+ effector memory cells are increased. It may be hypothesized that they might contribute to the development of marked microvascular alterations previously observed in these patients.

[PP.09.17] REPRODUCIBILITY OF THE EVALUATION OF THE WALL TO LUMEN RATIO OF RETINAL ARTERIOLES WITH TWO DIFFERENT NON-INVASIVE APPROACHES: SCANNING LASER-DOPPLER FLOWMETRY AND ADAPTIVE OPTICS

Objective: The evaluation of the morphological characteristics of small resistance arteries in human beings in not easy. The gold standard is generally considered to be the evaluation of the media to lumen ratio of subcutaneous small vessels obtained by local biopsies and measured by wire or pressure micromyiography. However, non-invasive techniques for the evaluation of retinal arterioles were recently proposed, in particular two approaches seem to provide interesting information: scanning laser Doppler flowmetry and adaptive optics; both of them provide an estimation of the wall to lumen ratio (WLR) of retinal arterioles. The reproducibility of such measurements was previously stated to be acceptable (coefficient of variation <10% for SLDF, <4% for RTX-1), however, no direct comparison of the two techniques in the same population was previously performed. Design and method: Therefore, we evaluated 18 subjects and patients (10 normotensives, 8 hypertensives, 7/18 severely obese). In all of them an evaluation of the WLR of retinal arterioles was made by Scanning Laser Doppler Flowmetry (SLDF, Heidelberg Engineering, Heidelberg, Germany) and adaptive optics (RTX-1, Imagine Eyes, Orsay, France). The same operator evaluated the same acquired images in two different days (intra-observer variability), and two different operators evaluated the same images in the same day (inter-observer variability). Results: The results are reported in the Table (***p < 0.001). Variation coefficient of SLDF is much greater than that of AO. Figure. No caption available. Conclusions: It is clear how the reproducibility of the evaluation of the WLR with adaptive optics is far better, as compared with SLDF, since the variation coefficient are clearly lower. This may be important in terms of clinical evaluation of microvascular morphology in a clinical setting.

[OP.8C.03] COMPARISON BETWEEN THREE NON-INVASIVE TECHNIQUES OF EVALUATION MICROVASCULAR MORPHOLOGY VS. THE GOLD-STANDARD LOCALLY INVASIVE MICROMYOGRAPHY. PRELIMINARY DATA

Objective: The evaluation of the morphological characteristics of small resistance arteries in human beings in not easy. The gold standard is generally considered to be the evaluation of the media to lumen ratio (M/L) of subcutaneous small vessels obtained by local biopsies and evaluated by wire or pressure micromyiography. However, non-invasive techniques for the evaluation of retinal arterioles were recently proposed, in particular two approaches seem to provide interesting information: scanning laser Doppler flowmetry (SLDF) and adaptive optics (AO); both of them provide an estimation of the wall to lumen ratio (WLR) of retinal arterioles. A non-invasive measurement of basal and total capillary density may be obtained by videomicroscopy/capillaroscopy. No direct comparison of the three non-invasive techniques in the same population was previously performed, in particular AO was never validated against micromyography. Design and method: In the present study we enrolled 12 normotensive subjects and 8 hypertensive patients undergoing an election surgical intervention; 11/20 were severely obese). All patients underwent a biopsy of subcutaneous fat during surgery. Subcutaneous small resistance artery structure was assessed by wire myography and the M/L was calculated. WLR of retinal arterioles was obtained by Scanning Laser Doppler Flowmetry and AO (SLDF, Heidelberg Engineering, Heidelberg, Germany and RTX-1, Imagine Eyes, Orsay, France). Functional (basal) and structural (total) microvascular density were evaluated by capillaroscopy (Videocap 3, DS Medica, Milan, Italy) before and after venous congestion. Results: The results are summarized in the Table (slope of the relation: p < 0.01 RTX-1 vs. SLDF). Figure. No caption available. Conclusions: Our data suggest that AO has a substantial advantage over SLDF in terms of evaluation of microvascular morphology, since it is more closely correlated with the M/L of subcutaneous small arteries, considered a gold-standard approach but limited in its clinical application by the local invasiveness of the procedure.

RELATIONSHIP BETWEEN MICROVASCULAR T REGULATORY LYMPHOCYTES AND CIRCULATING LYMPHOCYTES

Objective: Both innate and adaptive immune systems may contribute to the pathogenesis of cardiovascular disease and vascular remodeling through inflammation and oxidative stress. Particularly, the balance between Th1 effector lymphocytes (producing interferon-&ggr;), Th 17 (producing IL-17) and T regulatory (Treg) lymphocytes, which elicit an anti-inflammatory activity, may be crucial for blood pressure elevation and organ damage development, at least in experimental models. Tregs have been previously demonstrated to inversely correlate with subcutaneous small resistance artery media to lumen ratio (M/L) and retinal arteriole wall to lumen ratio (W/L) (unpublished data). Design and method: Therefore, we evaluated the possible relationship between Treg detected in small resistance arteries and circulating levels of Treg. We enrolled 11 normotensive subjects and 4 hypertensive patients undergoing an election surgical intervention (usually removal of adrenal gland for a non-producing adenoma). No sign of local or systemic inflammation was present in any subjects or patients. All patients underwent a biopsy of subcutaneous fat during surgery and small resistance arteries were isolated. We extracted genomic DNA from small resistance arteries and analyzed methylation status of the FoxP3 gene promoter involved in Treg lymphocytes activation. Unmethylated FoxP3 has been demonstrated to be specific for Treg lymphocytes. A peripheral blood sample was obtained before surgery for assessment of T lymphocyte subpopulations. Lymphocyte phenotype was evaluated by flow cytometry after 5 days in vitro activation in order to assess Th17 lymphocytes. Results: A significant positive correlation was detected between Treg in small resistance arteries and circulating levels of Treg (R = 0.42, p∼0.05) whereas an inverse correlation was observed between Th17 lymphocytes and Treg in small resistance arteries (R = -0.46, p < 0.05). Conclusions: Our data suggest that Treg lymphocytes detected in subcutaneous small resistance artery wall are related to circulating Treg which were previously observed to inversely correlate with subcutaneous small resistance artery media to lumen ratio and retinal arteriole wall to lumen ratio. This suggest that Treg may be protective against microvascular damage confirming an involvement of adaptive immune system on microvascular remodeling.

Aging and structural alterations of subcutaneous small resistance arteries in hypertensive patients

To cite this article: Carolina De Ciuceis, Claudia Rossini, Claudia Agabiti Rosei, Enzo Porteri, Alice Gavazzi, Stefano Caletti, Paola Pileri, Maria Antonietta Coschignano, Enrico Agabiti Rosei, Damiano Rizzoni* (2015) P2.1: AGING AND STRUCTURAL ALTERATIONS OF SUBCUTANEOUS SMALL RESISTANCE ARTERIES IN HYPERTENSIVE PATIENTS, Artery Research 12:C, 6–6, DOI: https://doi.org/10.1016/j.artres.2015.10.209