Length of sedimentation reaction in undiluted blood (erythrocyte sedimentation rate): Variations with sex and age and reference limits

Although the length of sedimentation reaction in blood (LSRB) (commonly, but improperly called erythrocyte sedimentation rate, ESR) has long been used in clinical laboratories because it is simple and low-cost, its sensitivity and specificity are unsatisfactory. Usually, the values are reported using the Westergren method with sodium citrate-anticoagulated specimens. We used a new procedure, the TEST1(TM), which measures the length of sedimentation reaction in undiluted K(3)EDTA anticoagulated blood samples following ICSH (International Committee for Standardization in Haematology) recommendations. Samples obtained from 840 reference individuals (430 females and 410 males, mean age 44 and 46.5 years respectively, range 1 to 90 years) were utilised to estimate the reference limits. The subjects, classified by sex, were subdivided into four statistically different age groups to determine the reference limits (2.5th and 97.5th percentiles). Sex difference was statistically significant in two age groups, from 14 to 50 (p<0.0001) and from 50 to 70 years (p<0.01). We did not observe significant sex difference within the age bracket from 1 to 14 years and from 70 to 90 years. In both sexes LSRB values increased with age, in significant correlation with fibrinogen concentration (p<0.0001). and became significantly higher in subjects older than 70 compared to all the younger subjects (p<0.01 in females and p<0.02 in males). Thus, we defined adequate reference ranges in elderly

Biological variability of lymphocyte subsets of human adults' blood.

Abstract BACKGROUND: Alterations in lymphocyte subpopulations are present in several immune diseases, and clinicians and researchers recognise the importance of investigating the distribution and changes in lymphocyte subsets over relatively long periods of time in order to evaluate the effectiveness of treatment and follow the course of disease. Yet further insight is required on the biological variability (BV) of lymphocyte subsets, which is crucial to the correct interpretation of longitudinal changes and provides essential information for setting desirable quality specifications and defining the usefulness of reference values. METHODS: Four-colour-flow cytometry was used to investigate the BV of lymphocyte populations (LP) in the peripheral blood of 20 healthy adults recruited from our laboratory staff and followed for three months. The total lymphocyte count was measured, and the relative frequencies determined for T-cells (CD3+), T-helper cells (CD3+CD4+), cytolytic T-cells (CD3+CD8+), B-cells (CD3-CD19+), NK-cells (CD3-CD16+/56+), non-MHC restricted cytolytic T-cells (CD3+CD56+) and activated T-cells (CD3+HLA-DR+). RESULTS AND CONCLUSIONS: Data on the components of BV were applied to set quality specifications for allowable precision, bias and total error. Analytical performances were established, and they were more than desirable for all the markers considered in our study. By comparing within-subject and between-subjects BV, we were able to define the uselessness of reference ranges in the evaluation of changes in CD serial results. Data on within-subject BV and analytical precision were thus used to determine the reference change values, in order to identify the significance of changes in serial results. The findings made in the present study provide further evidence of the relevance of BV in the evaluation of immunological markers of LP

The TEST 1 automated system: a new method for measuring the erythrocyte sedimentation rate.

We evaluated performance of the TEST 1 (SIRE Analytical Systems, Udine, Italy), a fully automated analyzer for the measurement of the erythrocyte sedimentation rate (ESR). Intra-assay reproducibility was satisfactory for a wide range of ESR values, whereas there was a significant decrease in ESR data when the samples were stored at 4 degrees C for up to 24 hours. We compared TEST 1 with the Westergren ESR method approved by the International Council for Standardization in Haematology (ICSH) and with the Diesse Ves-Matic analyzer Linear regression analysis comparing the TEST 1 and the ICSH reference method yielded satisfactory correlations for K3 EDTA- and sodium citrate-anticoagulated samples. Bland-Altman analysis showed no evidence of a systematic bias between the TEST 1 and the reference method. A close correlation was found between the TEST 1 system and the Diesse Ves-Matic analyzer despite a significant positive systematic bias. Reference values for men and women were analyzed according to nonparametric statistics. The TEST 1 was easy to use, had a satisfactory operative practicability required minimal maintenance, and reduced contact with potential biohazards. This system enables the determination of ESR with any common standard-sized tube; the use of samples anticoagulated with K3 EDTA can widely reduce the workload in clinical laboratories

Lymphocytes subsets reference values in childhood.

Immunophenotyping of blood lymphocyte subsets and activation markers is a basic tool in the diagnostic process of primary immunodeficiency diseases, its use becoming more and more widespread as the knowledge about these illnesses increases. However, the availability of reliable reference values, which need to be age-matched for the pediatric population, is a pre-requisite for the reliable interpretation of immunophenotyping data. Aim of this study is to analyze the lymphocyte subsets and activation markers distribution in children aged 0-18 years referring to the University Hospital of Padova and to create age-matched reference values expressed by percentiles, thus providing a valuable guideline for the interpretation of the immunophenotype

Flow cytometry CD4+CD26 12CD38+ lymphocyte subset in the microenvironment of Hodgkin lymphoma-affected lymph nodes

Hodgkin lymphoma (HL) is traditionally diagnosed by the presence of neoplastic Hodgkin and Reed-Sternberg (HRS) cells found in minority within a typical inflammatory microenvironment. It is now recognized that the majority of these T CD4 cells are T regulatory (Treg) and play an important immunosuppressive role and contribute to tumour persistence. Flow cytometric immunophenotyping of lymphocytes was performed on lymph node samples over a 12-year period (2000-2012) to identify the Hodgkin-specific subset and potential biomarkers related to Treg cells. CD3, CD19 and T CD4+CD26-CD38+ subsets were measured in the lymphocytic infiltrate of 108 consecutive lymph node samples concurrently diagnosed histologically as HL and in 43 cases of benign reactive lymphoid hyperplasia (BRLH). HL, compared to BRLH, shows statistically significant differences within the reactive microenvironmental population: decreased CD19+ cells (23 % vs 39 %; p\u2009<\u20090.001), increased CD3+ (74 % vs 58 %; p\u2009<\u20090.001) and CD4+CD26-CD38+ cells (38 % vs 11.5 %; p\u2009<\u20090.001). By using the co-expressed markers CD38 and CD26 for logistic analysis, the obtained receiver operating characteristic (ROC) curves confirm that the CD4+CD26-CD38+ subset is strongly expressed in HL (ROC AUC\u2009=\u20090,8639). Flow cytometric detection of CD4+CD26-CD38+ cells seems able to identify the cellular microenvironmental pattern in HL and to distinguish it from BRLH. Although there is extensive experience in flow cytometric analysis of non-HL, it is not routinely applied in cases of HL and our findings suggest that it may be useful in quickly and easily characterizing its cellular para-neoplastic inflammatory background