Reference values for T, B and NK human lymphocyte subpopulations in adults

The data presented in this paper are reference ranges for frequencies of thirty-eight subpopulations of T, B and NK lymphocytes, established from a cohort of 253 healthy blood donors aged from 19 to 67. When relevant, the influence of age or sex was taken into account to calculate these reference values. This article is related to the research article entitled “Influence of age, sex and HCMV-serostatus on blood lymphocyte subpopulations in healthy adults” (Apoil et al., 2017) [1]. Immunophenotyping data obtained from each individual is made publicly available for extended analyses.


a b s t r a c t
The data presented in this paper are reference ranges for frequencies of thirty-eight subpopulations of T, B and NK lymphocytes, established from a cohort of 253 healthy blood donors aged from 19 to 67. When relevant, the influence of age or sex was taken into account to calculate these reference values. This article is related to the research article entitled "Influence of age, sex and HCMVserostatus on blood lymphocyte subpopulations in healthy adults" (Apoil et al., 2017) [1]. Immunophenotyping data obtained from each individual is made publicly available for extended analyses.  Reference ranges were adjusted for age or sex when these parameters impact the values.

Data
Data in the following Tables present the reference values for 38 distinct human T, B or NK lymphocyte subpopulations. When sex or age has a significant impact on these subpopulations, separate reference values are given for male, female, younger (19-44) or older (45-67) individuals. Data are expressed either as absolute numbers of cells in G/L (a), or as the percentage of cells relative to total CD3 þ (b), CD3 þ CD4 þ (c), CD3 þ CD8 þ (d) T lymphocytes, Tregs (e), total B (f) or NK cells (g); DN: CD4-CD8-double-negative T cells; DP: CD4 þ CD8 þ double-positive T cells.

Cohort assembly
Healthy blood donors aged 19-67 (median age 44 y.o.) were recruited between 2011 and 2013 in Toulouse (Etablissement Français du Sang Pyrénées-Méditerranée, Southwest of France). All subjects were negative in serological tests for blood-transmissible infections (HIV, hepatitis B and C, HTLV, syphilis) and were exempt from any pathology or treatment which could interfere with leukocyte parameters (history of cancer or of autoimmune disease, active or recent systemic infection, immunosuppressive or immunomodulating therapy, severe allergy, or a vaccine administered less than 3 months ago). For each enrolled blood donor, a single 7 mL EDTA tube of peripheral whole blood was collected between 8 a.m. and 11 a.m. Reference values were calculated from a cohort of 253 individuals, adjusted for sex ratio and frequency of HCMV-seropositivity, which was made from a larger cohort of 283 blood donors (complete immunophenotyping data for these 283 individuals is presented in Appendix A: Supplementary material). Details about the composition of this cohort are available online in Supplementary material from [1].

Immunophenotyping and cytometry data analysis
Immunophenotyping was performed by multicolour flow cytometry: samples were labeled with 4 distinct antibody panels and absolute counts of T, B and NK cells were determined by using BD Trucount s tubes. Cytometric data was acquired on a BD CANTO II s flow cytometer (BD Biosciences, Le Pont De Claix, France) and was analyzed with BD Diva and FlowJo s software (LLC, Ashland, OR). Antibody panels and gating strategies are detailed in Ref. [1]. Mean values and reference ranges were calculated according to CLSI guidelines [2] (including suspected outliers in the calculation) by using the Reference Value Advisor software [3], from the entire population sample, or from subgroups of more than 120 individuals (males/females and younger/older). To evaluate the influence of age and sex, Mann-Whitney non-parametric tests were carried-out after removal of outliers, and the corrected ranges are indicated when po0.05.

Funding sources
This work was supported by Paul Sabatier University (Toulouse, France), Région Midi-Pyrénées and by the French Ministry of Higher Education and Research.