Establishment of the reference intervals of lymphocyte subsets for healthy Chinese Han adults and its influencing factors.

Cellular immune monitoring is becoming more critical in the clinic, but its application has not yet become sufficiently widespread. One reason may be the different reference intervals among clinical laboratories due to several factors. Percentage and number of lymphocyte subsets are standard indicators of cellular immune detection. The present study aimed to establish standardized reference intervals of lymphocyte subsets in the healthy Chinese Han adult population and examine such influencing factors as age, gender, region, and measurement instruments. A total of 496 healthy Chinese Han people aged 18-59 years from 3 China Mainland regions (north, east, and south) were enrolled. The sample of each center was simultaneously examined by three flow cytometers (FACSCantoTMII, FACSLyricTM, and FACSCaliburTM). A single-platform flow cytometry-based absolute count technique was used to quantify the percentage and number of each lymphocyte subset. The flow cytometry results were analyzed by variance analysis and Z test to determine the influence of age, gender, and instruments on lymphocyte subsets. Multi-center, age-specific, and gender-specific reference intervals of healthy Chinese Han adults' lymphocyte subsets were established. There was no statistical difference in the results from the three flow cytometers. Gender affected the results of CD4+ (%) and the absolute count of CD3-CD16+CD56+, where CD4+ (%) was higher in women, and the absolute count of CD3-CD16+CD56+ was higher in men. Age mainly affected the CD4+/CD8+ ratio, which was statistically higher in groups aged over 40 years; the percentage and number of CD3-CD19+ were more elevated in age groups below 30 years; however, the difference was not statistically significant. This study established the reference intervals of lymphocyte subsets for healthy Chinese Han adult populations under the standardized methods. This study was the first nationwide study in China to use a flow cytometry-based single-platform method to establish the reference intervals of lymphocyte subsets of the healthy Chinese Han adult population. Gender and age were shown to influence the results of lymphocyte subsets.

Xu K ，Miao L ，Chen W ，Wu H ，Gong Y ，Tu X ，Guo W ，Pan B ，Qu C ，Wu X ，Wang B ... -  《-》

Reference ranges of lymphocyte subsets in healthy adults and adolescents with special mention of T cell maturation subsets in adults of South Florida.

Analysis of peripheral blood lymphocyte subsets has become an essential tool in the evaluation of outcome of diagnostic and research related questions in immunological and pathological conditions. Periodic evaluation and establishment of normal lymphocyte reference ranges are required in clinical and research settings of various immunodeficiency disorders for evaluation of the significance of observations. It is also important that age and gender specific lymphocyte subset reference ranges should be locally established for meaningful comparison and accurate result interpretation as age plays a significant role in the development of immune system. We performed dual platform flow cytometry to determine reference ranges for lymphocyte subsets (CD3, CD4, CD8, CD19 [B cells] and CD16+CD56+ [Natural Killer - NK cells]) in 50 adolescents (age range: 12-18) and 100 adults (age range: 21-67) along with T cell maturation, activation and co-stimulatory molecules in healthy multiracial adult population of South Florida. The lymphocyte reference ranges percentages [absolute counts - Abs, cells/μl] unadjusted for gender differences for adolescents are: CD3: 49-83 [939-2959]; CD4: 27-53 [467-1563]; CD8: 16-40 [259-1262]; CD19+ B cells: 8-31 [169-1297] and CD16+CD56+ NK cells: 3-30 [59-1178] and for adults are: CD3: 65-88 [983-3572]; CD4: 26-62 [491-2000]; CD8: 14-44 [314-2,087]; CD19+ B cells: 2-27 [64-800] and CD16+CD56+ NK cells: 2-27 [27-693]. The ranges for CD4:CD8 ratio for adolescents and adults are 0.7-2.6 and 0.6-4.4, respectively. Gender based analysis of relative percentages of lymphocyte subsets showed no significant differences between adult and adolescent males and females. The mean CD4:CD8 ratio was significantly higher in adult females than males (P=0.04) and in adolescents this difference was not significant between genders. The mean CD3 and CD4 T cell percentages were higher and CD19 cell percentages were lower in adults compared to adolescents (P<0.0001). Absolute lymphocyte counts showed a positive correlation with the absolute counts of CD3+, CD4+, CD8+, CD19+, CD16+CD56+, CD45RO+ and CD45RA+ cells (all correlations with P<0.0001 except CD45RO [P=0.01] and CD45RA [P=0.03]). The reference values of peripheral blood lymphocyte subsets were analyzed in healthy adolescent and adult population of South Florida. This study indicates the need for periodic evaluation and establishment of lymphocyte reference ranges for patient population served based on gender and age since these could influence immune status and treatment outcome.

Valiathan R ，Deeb K ，Diamante M ，Ashman M ，Sachdeva N ，Asthana D ... -  《-》

Age- and sex-related reference intervals of lymphocyte subsets in healthy ethnic Han Chinese children.

Immunophenotyping of blood lymphocytes has become an important tool in the diagnosis of immunologic and hematologic disorders such as immunodeficiencies, lymphoproliferative and autoimmune diseases. Lymphocyte subsets include total T-cells (CD3(+)), TH (T helper, CD3(+) CD4(+)), TC (cytotoxic T cells, CD3(+) CD8(+)), B-cells (CD3(-) CD19(+)), and NK-cells (CD3(-) CD16(+) CD56(+)). Specific lymphocyte subset reference intervals should be locally established for meaningful comparison and to obtain an accurate interpretation of the results. Reference intervals of lymphocyte subsets for Chinese children are scarce. We performed dual-platform flow cytometry to determine the reference intervals of the percentages and absolute counts of lymphocyte subsets, including total T-cells, TH cells, TC cells, B-cells, and NK-cells in 1,027 ethnic Han children aged 4 months to 7 years in Henan, China. The children were divided into seven age groups. The percentages and absolute counts differed significantly with age, with the percentages of TH cells and B cells and the CD4/CD8 ratio peaking during the first year, while the percentages of total T cells, TC cells, and NK cells were obviously increased with age; girls showed a trend toward having a higher percentage of TH cells and a higher CD4/CD8 ratio than boys. The absolute counts of lymphocyte subsets peaked during first year and then decreased steadily with age. The reference intervals of lymphocyte subsets among children from China differed from the reported values in Hong Kong, the United States, Cameroon, and Italy. The differences observed could be due to genetic and environmental factors, coupled with the methodology used. The reference intervals of lymphocyte subsets could be used as initial national reference ranges in guidelines for children aged 4 months to 7 years.

Jia L ，Li J ，Zhang Y ，Shi Y ，Yuan E ，Liu J ，Wang P ，Rong S ，Xing J ，Tian Y ，Li J ... -  《-》

Establishing Reference Values for Peripheral Blood Lymphocyte Subsets of Healthy Children in China Using a Single Platform.

Lymphocyte subsets significantly change during childhood; thus, age-matched reference values derived from healthy children are crucial. We established reference values for lymphocyte subsets, including T cells (CD3+), CD4 T cells (CD3 + CD4+), CD8 T cells (CD3 + CD8+), double negative T (DNT) cells (CD3 + CD4-CD8-), B cells (CD3-CD19+), NK cells (CD3-CD56+), and NKT-like cells (CD3 + CD56+) in the peripheral blood of 813 healthy children. We used the method of the international standard document (Clinical Laboratory Standard Institute C28-A3) to establish reference intervals with a single platform. First, we used the Skewness and Kurtosis test to analyze the normality of the data. The nonnormally distributed data was transformed into approximately normal distribution by the Box-Cox transformation. Second, we used the Tukey's method to eliminate outliers. Further, all the subjects were grouped into subgroups according to sex (male and female) and age (0-1 month, 2-12 months, 1-3 years, 4-6 years, and 7-18 years). We used the standard normal deviation test (Z-test) to evaluate whether age and sex were possible grouping factors. The analyses indicated age to be an important factor associated with changes in lymphocyte subsets. The absolute number of lymphocyte subsets and total number of lymphocytes, T cells, CD4 T cells, CD8 T cells, and B cells gradually increase from birth to 12 months and then gradually decrease with age. Furthermore, CD4 T cells and the ratio of CD4+/CD8+ gradually decrease with age. In contrast, CD8 T and DNT cells gradually increase with age. The percentage and number of NK and NKT-like cells gradually increase with age and remain stable between 1 and 18 years of age. In conclusion, the age-related reference intervals established in healthy children in this study can aid in monitoring and assessing the changes in immune levels in diseased conditions.

Zhang L ，Zhong H ，Wei B ，Fan J ，Huang J ，Li Y ，Liu W ... -  《-》

Characteristics of and reference ranges for peripheral blood lymphocytes and CD4+ T cell subsets in healthy adults in Shanxi Province, North China.

Xu D ，Wu Y ，Gao C ，Qin Y ，Zhao X ，Liang Z ，Wang Y ，Feng M ，Zhang C ，Liu G ，Luo J ... -  《-》