Flow Cytometric Assessment of Neutrophil Oxidative Metabolism in Chronic Granulomatous Disease on Small Quantities of Whole Blood: Heterogeneity in Female Patients

A rapid and sensitive flow cytometric assay is presented for the quantitative estimation of the oxidative metabolic activity of individual polymorphonuclear leukocytes (PMN) on less than 100 td of whole blood. This procedure is a simplified version using whole blood of the method of Bass et al (J. Immunol. 130:1910, 1983) that estimated the metabolic burst activity of phorbol myristate acetate (PMA)-stimulated individual PMN as the intracellular generation of a fluorescence product by a flow cytometric assay. With this method, almost all the PMN from normal subjects responded to PMA as a single cell population generating bright intracellular fluorescence. PMN from a boy with chronic granulomatous disease (CGD), could not respond to PMA with any increase of their fluorescence intensity. His mother had two distinct PMN populations one functionally normal and the other defective, indicating a random lyonization in the carrier mother and the X-linked recessive mode of inheritance. In two female patients with CGD from unrelated families, their PMN responded to PMA, as a whole, with a minimal increase in the fluorescence intensity, but the metabolic defects in their PMN were not so complete as seen in a classical X-linked CGD boy. But, PMN from two female sibling patients from the other family responded to PMA as a single uniform cell population with a weak but definite fluorescence intensity. However, the genetic background of these female patients with CGD remains unclear, since PMN dysfunction could not be identified in their mothers with this method.This work was supported in part by a grant (No. 58440046) from the Ministry of Education of Japan

Differential expression of CD45RO (UCHL1) and its functional relevance in two subpopulations of circulating TCR-γ/δ+ lymphocytes

金沢大学大学院医学系研究科血管病態制御学We examined the developmental profile of TCR-γ/δ+ cells with respect to CD45RO expression. Although total TCR-γ/δ+ cells were negligible in the neonatal blood and increased with advancing age, most blood TCR-γ/δ+ cells markedly expressed CD45RO without a distinction of age, probably reflecting a different CD45RO expression of two subsets defined by BB3 and δTCS1 mAbs. The vast majority of BB3+ cells expressed CD45RO, whereas expression of CD45RO was virtually absent in the δTCS1+ population. Functional studies revealed that, while both TCR-γ/δ+ cell subsets showed CD3-mediated activation, only BB3+ (or TiγA+) cells, but not δTCS1+ cells, appeared to proliferate in response to PPD in PPD-reactive individuals. The results suggested that the CD45RO+ (BB3+ or TiγA+) subset among blood TCR-γ/δ+ cells may be mainly involved in the memory or primed component of the immune system responding to some foreign antigens

Why Are Outcomes Different for Registry Patients Enrolled Prospectively and Retrospectively? Insights from the Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF).

Background: Retrospective and prospective observational studies are designed to reflect real-world evidence on clinical practice, but can yield conflicting results. The GARFIELD-AF Registry includes both methods of enrolment and allows analysis of differences in patient characteristics and outcomes that may result. Methods and Results: Patients with atrial fibrillation (AF) and ≥1 risk factor for stroke at diagnosis of AF were recruited either retrospectively (n = 5069) or prospectively (n = 5501) from 19 countries and then followed prospectively. The retrospectively enrolled cohort comprised patients with established AF (for a least 6, and up to 24 months before enrolment), who were identified retrospectively (and baseline and partial follow-up data were collected from the emedical records) and then followed prospectively between 0-18 months (such that the total time of follow-up was 24 months; data collection Dec-2009 and Oct-2010). In the prospectively enrolled cohort, patients with newly diagnosed AF (≤6 weeks after diagnosis) were recruited between Mar-2010 and Oct-2011 and were followed for 24 months after enrolment. Differences between the cohorts were observed in clinical characteristics, including type of AF, stroke prevention strategies, and event rates. More patients in the retrospectively identified cohort received vitamin K antagonists (62.1% vs. 53.2%) and fewer received non-vitamin K oral anticoagulants (1.8% vs . 4.2%). All-cause mortality rates per 100 person-years during the prospective follow-up (starting the first study visit up to 1 year) were significantly lower in the retrospective than prospectively identified cohort (3.04 [95% CI 2.51 to 3.67] vs . 4.05 [95% CI 3.53 to 4.63]; p = 0.016). Conclusions: Interpretations of data from registries that aim to evaluate the characteristics and outcomes of patients with AF must take account of differences in registry design and the impact of recall bias and survivorship bias that is incurred with retrospective enrolment. Clinical Trial Registration: - URL: http://www.clinicaltrials.gov . Unique identifier for GARFIELD-AF (NCT01090362)

Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry.

BACKGROUND: The Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. METHODS AND RESULTS: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012-2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P < 0.001). Diabetes and coronary artery disease (CAD) were prevalent in 36.2% and 28.1% of patients as compared with global prevalence of 22.2% and 21.6%, respectively (P < 0.001 for both). Antiplatelet therapy was the most common antithrombotic treatment in India. With increasing stroke risk, however, patients were more likely to receive oral anticoagulant therapy [mainly vitamin K antagonist (VKA)], but average international normalized ratio (INR) was lower among Indian patients [median INR value 1.6 (interquartile range {IQR}: 1.3-2.3) versus 2.3 (IQR 1.8-2.8) (P < 0.001)]. Compared with other countries, patients from India had markedly higher rates of all-cause mortality [7.68 per 100 person-years (95% confidence interval 6.32-9.35) vs 4.34 (4.16-4.53), P < 0.0001], while rates of stroke/systemic embolism and major bleeding were lower after 1 year of follow-up. CONCLUSION: Compared to previously published registries from India, the GARFIELD-AF registry describes clinical profiles and outcomes in Indian patients with AF of a different etiology. The registry data show that compared to the rest of the world, Indian AF patients are younger in age and have more diabetes and CAD. Patients with a higher stroke risk are more likely to receive anticoagulation therapy with VKA but are underdosed compared with the global average in the GARFIELD-AF. CLINICAL TRIAL REGISTRATION-URL: http://www.clinicaltrials.gov. Unique identifier: NCT01090362