Multiparametric analysis of peripheral blood in the normal pediatric population by flow cytometry

Wiener, Doris; Shah, Samveg; Malone, John I.; Lowell, Nancy E.; Lowitt, Saul; Dt, Rowlands

doi:10.1002/jcla.1860040305

Cited by 43 publications

(25 citation statements)

References 9 publications

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“…Then, T cell numbers continuously increase until age 30 years. After age 30 years, T cell numbers remain at a relatively stable level with a slow decline over time, as suggested by data (26,27). Our simulation results of T cell dynamics are within the normal range calculated from human data on cell numbers per mm 3 of blood, blood volume, and body weight (error bars in Fig.…”

Section: Predicting Total T Cell Numbers Total Sjtrec Levels and Sjsupporting

confidence: 85%

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Reevaluation of T Cell Receptor Excision Circles as a Measure of Human Recent Thymic Emigrants

Kirschner

2002

The Journal of Immunology

125

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The human thymus exports newly generated T cells to the periphery. As no markers have been identified for these recent thymic emigrants (RTE), it is presently impossible to measure human thymic output. T cell receptor excision circles (TREC) have been recently used to assess thymic output during both health and disease. Using a mathematical model, we quantify age-dependent changes both in the number of RTE generated per day and in TREC concentration during an 80-year lifespan. Through analyses, we demonstrate that RTE and peripheral T cell division have the same potential to affect TREC concentration at any age in healthy people. T cell death also influences TREC concentration, but to a lesser extent. During aging, our results indicate that thymic involution primarily induces an age-dependent decline in TREC concentrations within both CD4+ and CD8+ T cell populations. We further apply this model for studying TREC concentration during HIV-1 infection. Our analyses reveal that a decrease in thymic output is the major contributor to the decline in TREC concentration within CD4+ T cells, whereas both increased peripheral T cell division and decreased thymic output induce the decline in TREC concentration within CD8+ T cells. Therefore, we suggest that T cell turnover should be examined together with TREC concentration as a measure of RTE. If peripheral T cell division remains relatively unchanged, then TREC concentration indeed reflects thymic output.

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Section: Predicting Total T Cell Numbers Total Sjtrec Levels and Sjsupporting

confidence: 85%

“…5 T cells, we divide sjTREC/PBMCs with the ratios: total lymphocytes/PBMCs and T cells/total lymphocytes over time (26,33,34), and then multiply by 1.5 ϫ 10 5 T cells. sjTREC data from Douek et al and Zhang et al were both measured using blood samples.…”

Section: Shown Inmentioning

confidence: 99%

Reevaluation of T Cell Receptor Excision Circles as a Measure of Human Recent Thymic Emigrants

Kirschner

2002

The Journal of Immunology

125

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“…This finding was not confirmed in our study, however. In addition, published work from different laboratories showed that both CD4 ϩ and CD8 ϩ cell counts increase with age with an unchanged CD4/CD8 ratio (13,14).…”

Section: Discussionmentioning

confidence: 99%

Immunophenotyping of Peripheral Blood Lymphocytes in Saudi Men

Qouzi

Salamah

Rasheed

et al. 2002

Clin Vaccine Immunol

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Flow cytometry is a powerful tool in the diagnosis and management of congenital and acquired immunodeficiency syndromes. It is also used for the diagnosis of leukemia and lymphomas. Results of studies from different countries support the presence of variation in absolute and percent quantities of lymphocytes and their subsets (2,3,6,8,12). Several factors were found to affect these results, including age, gender, race, and environmental factors (2, 9, 11). For example, low CD4ϩ -T-cell counts were reported in Asians (1) and Chinese (7). In order to establish a reference range for the different lymphocyte populations, Shahabuddin (8) investigated a cohort of 150 male Saudi blood donors. He showed that there was a marked increase in the CD8 cell count, a decreased CD4/CD8 ratio, and a low natural killer (NK) cell count compared to 32 Caucasian males.In this report, we carried out the immunophenotyping of lymphocytes and their subsets in order to establish our own reference range in an adult Saudi population. MATERIALS AND METHODS Subjects.A total of 209 Saudi men were recruited for this study. These were random blood donors that were used as an internal control each time we conducted an immunophenotyping procedure.Blood collection. EDTA peripheral blood was collected and analyzed initially on Advia 120 (Bayer) hematological analyzer for total and differential blood counts. A blood smear was then prepared for a manual differential count. In brief, 100 l of whole blood was mixed and incubated, in the dark, with 20 l of each monoclonal antibody in separate tubes, at room temperature. Red blood cells were then lysed by adding 2 ml of lysing solution (Becton Dickinson), and the tubes were vortexed and incubated in the dark at room temperature for 10 min and finally centrifuged at 2,500 rpm for 5 min. The pellet was then washed once with 2 ml of phosphate-buffered saline (PBS), resuspended in 500 l of PBS, and finally analyzed with CellQuest software (Becton Dickinson). The FACScalibur was calibrated with Calibrite beads (Becton Dickinson) and AutoComp weekly.Statistical analysis. Data were entered and analyzed by using STATA v6.0 statistical software. The mean and standard deviation (SD) were calculated for each marker. The Student's t test was used to compare the means and SD of our results to those from other published sources.

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“…With respect to T-cells, however, although some studies have reported changes in T cell levels, especially a decrease in the CD8 cells with increasing age [2,10,11,[32][33][34], other (older) have reported increase in CD4+ and /or CD8+ [24,35,36]. The impact of age on lymphocyte subsets is not well established, with contradictory results from various studies.…”

Section: A U T H O R P R O O F Future Science Groupmentioning

confidence: 99%

Reference Ranges of Lymphocyte Subsets in Healthy Qatari Adults

Althani¹,

Hamdi²,

Al-Marwani³

et al. 2014

Biomark. Med.

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Aim: Determine reference ranges of lymphocyte subsets in healthy Qatari adults. Patients & Methods: Blood samples (n = 150) were investigated using four colors flow cytometery. Results: The mean percentage and absolute values of the lymphocyte subsets were: CD3: 73.6 ± 6.1%; 2,072 ± 644 cells/μl; CD4: 41.1 ± 7.9%; 1,167 ± 396 cells/ μl; CD8: 33.8 ± 7.1%; 967 ± 364 cells/μl; B cells (CD19+): 11.3 ± 3.9%; 318 ± 144 cells/μl and natural killer (NK) cells (CD16/56+): 16.5 ± 6.3%, 440 ± 401 cells/μl, respectively. The CD4/CD8 ratio was 1.3 ± 0.5. The ratio, CD3+ and CD4+ percentage and CD4+ cell counts were significantly higher in females; CD8+ and NK cell count and CD8+ percentage were significantly higher in males. The impact of age was less certain. Conclusion:The reference values of lymphocyte subsets appear to be specific to this population, demonstrating the importance of establishing local reference ranges. Keywords: B cells • CD3 • CD4 • CD8 • flow cytometry • lymphocyte subsets • natural killer cells • reference ranges, IntroductionLymphocyte immunophenotyping by flow cytometry is an important tool in differential diagnosis of congenital and acquired immune deficiencies, lymphoproliferative disorders and acute leukemias [1]. In addition, it is used for monitoring treatment response and disease progression with anti-retroviral and immunosuppressive therapy. Various studies suggest that lymphocyte subsets reference ranges are influenced by age, sex, ethnicity, and environment [2][3][4][5]. The variations in the values of the lymphocyte subsets emphasize the importance of establishing local reference ranges to allow for meaningful comparisons and accurate interpretation of immunophenotyping data across different cultures and ethnic groups. Lifestyle factors, e.g. smoking, have also been shown to influence lymphocyte subset values [6,7].Indeed, reference range for peripheral blood lymphocyte subsets have been established in many studies across different countries, revealing variations in the normal range for lymphocyte subsets according to age, sex, race, and environmental factors [3,4,[8][9][10][11][12][13][14][15][16]. Establishment of an internal normal range of lymphocyte subsets is a important for each population considering the effect of environmental factors, age, ethnical diversity on the immune system. Hence, this study aims to determine the laboratory reference ranges for lymphocyte subsets in healthy Qatari adults using flow cytometry.This article reports the local reference range for peripheral blood lymphocyte subsets, both percentages and absolute counts, in a representative sample of 150 healthy adult Qatari population.The results have been stratified by age, sex, and smoking status. The population selected is largely homogenous (100% Qatari Arabs) so effect of race was not investigated. The results are briefly compared with other population data from published studies. Materials & methods SubjectsA total of 150 healthy Qatari volunteers (18-55 yr old; all Qatari Arabs) were recruited for the stud...

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Multiparametric analysis of peripheral blood in the normal pediatric population by flow cytometry

Cited by 43 publications

References 9 publications

Reevaluation of T Cell Receptor Excision Circles as a Measure of Human Recent Thymic Emigrants

Reevaluation of T Cell Receptor Excision Circles as a Measure of Human Recent Thymic Emigrants

Immunophenotyping of Peripheral Blood Lymphocytes in Saudi Men

Reference Ranges of Lymphocyte Subsets in Healthy Qatari Adults

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