Effect of delayed cell processing and cryopreservation on immunophenotyping in multicenter population studies

Thyagarajan, Bharat; Barcelo, Hélène; Crimmins, Eileen M.; Weir, David R.; Minnerath, Sharon R.; Vivek, Sithara; Faul, Jessica D.

doi:10.1016/j.jim.2018.09.007

Cited by 33 publications

(21 citation statements)

References 18 publications

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“…We have attempted to minimize the effects of these limitations by imputing data for random errors and comparing cryopreserved data from this study with data obtained from a previous study on whole blood performed in MESA examination 4, 22 , 25 as well as literature on the use of cryopreserved cells. 38 An additional limitation relates to variable timing of blood draws for PBMC processing, which generally occurred in the mornings but not at the same time for each individual (eg, 8 am versus 11 am ). Given emerging data indicating intraindividual circadian variations in circulating immune cell subsets, 39 there is potential unmeasured confounding related to the timing of blood draws.…”

Section: Discussionmentioning

confidence: 99%

Nonclassical Monocytes (CD14dimCD16+) Are Associated With Carotid Intima-Media Thickness Progression for Men but Not Women

Feinstein

Doyle

Stein

et al. 2021

ATVB

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Objective: Few studies of population-based cohorts have investigated prospective associations of lymphoid and myeloid cell subsets in cardiovascular disease onset and progression. The purpose of this analysis was to determine associations of prespecified myeloid and lymphoid lineage cell subsets with common carotid artery intima-media thickness (IMT) progression. Approach and Results: We performed a prospective case-cohort study of 1195 participants from the Multi-Ethnic Study of Atherosclerosis who had peripheral blood mononuclear cells stored from the baseline examination. Key exposure variables were prespecified subsets of lymphoid and myeloid lineage immune cells, phenotyped by multicolor flow cytometry. The primary outcome was progression from baseline (Exam 1) to year 10 (Exam 5) in common carotid IMT. Higher proportions of nonclassical monocytes (CD14dimCD16++) were significantly associated with IMT progression over 10 years, but classical monocytes (CD14++CD16−), CD4+CD28− T cells, and T helper cells producing IL-17 (interleukin 17; T helper 17 cells) were not associated with significant changes in IMT over 10 years. There were significant interactions between monocyte subsets and sex with respect to IMT progression: in sex-stratified analyses, nonclassical monocytes were associated with significant IMT progression and classical monocytes were associated with significant IMT regression for men, whereas there were no significant associations of monocyte subsets with IMT change for women. Conclusions: Nonclassical monocytes were associated with progression of carotid IMT. There were significant sex differences in associations of monocyte subsets with IMT progression: for men, nonclassical monocytes were associated with IMT progression and classical monocytes were associated with regression, whereas these associations were null for women.

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Section: Discussionmentioning

confidence: 99%

Nonclassical Monocytes (CD14dimCD16+) Are Associated With Carotid Intima-Media Thickness Progression for Men but Not Women

Feinstein

Doyle

Stein

et al. 2021

ATVB

View full text Add to dashboard Cite

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“…The limitations include the observational nature of the data and technical issues with complex samples which resulted in some missing data. The use of cryopreserved PBMCs may result in different absolute levels of some of the subsets as compared with whole blood [ 25 ], although relative levels should be preserved [ 34 ]. Further, the immune cell distributions measured in this study from cryopreserved samples are similar to those previously measured in fresh whole blood obtained in MESA at Exam 4 (2005–2007) [ 8 , 9 ] among phenotypes measured in both studies.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Natural killer cells, gamma delta T cells and classical monocytes are associated with systolic blood pressure in the multi-ethnic study of atherosclerosis (MESA)

Delaney

Olson

Sitlani

et al. 2021

BMC Cardiovasc Disord

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Background Hypertension is a major source of cardiovascular morbidity and mortality. Recent evidence from mouse models, genetic, and cross-sectional human studies suggest increased proportions of selected immune cell subsets may be associated with levels of systolic blood pressure (SBP). Methods We assayed immune cells from cryopreserved samples collected at the baseline examination (2000–2002) from 1195 participants from the multi-ethnic study of atherosclerosis (MESA). We used linear mixed models, with adjustment for age, sex, race/ethnicity, smoking, exercise, body mass index, education, diabetes, and cytomegalovirus titers, to estimate the associations between 30 immune cell subsets (4 of which were a priori hypotheses) and repeated measures of SBP (baseline and up to four follow-up measures) over 10 years. The analysis provides estimates of the association with blood pressure level. Results The mean age of the MESA participants at baseline was 64 ± 10 years and 53% were male. A one standard deviation (1-SD) increment in the proportion of γδ T cells was associated with 2.40 mmHg [95% confidence interval (CI) 1.34–3.42] higher average systolic blood pressure; and for natural killer cells, a 1-SD increment was associated with 1.88 mmHg (95% CI 0.82–2.94) higher average level of systolic blood pressure. A 1-SD increment in classical monocytes (CD14++CD16−) was associated with 2.01 mmHG (95% CI 0.79–3.24) lower average systolic blood pressure. There were no associations of CD4+ T helper cell subsets with average systolic blood pressure. Conclusion These findings suggest that the innate immune system plays a role in levels of SBP whereas there were no associations with adaptive immune cells.

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“…Conflicting results might be derived from the choice of study material, as Colonna-Romano et al used separated PBMCs and our data are based on the analysis of fresh whole blood, which again illustrates the necessity of harmonization of flow cytometric methods in order to compare results from different studies in a meaningful way. Many authors have emphasized the critical part of methodical differences as the main source of variability in flow cytometric immunophenotyping [1,39,40].…”

Section: B Cellsmentioning

confidence: 99%

Comprehensive flow cytometric reference intervals of leukocyte subsets from six study centers across Europe

Oras

Quirant‐Sánchez

Popadić

et al. 2020

Clinical and Experimental Immunology

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A group of European FOCIS Centers of Excellence adapted panels of the Human Immunophenotyping Consortium (HIPC) for whole blood analysis. Using four core panels [T/regulatory T cell/B/natural killer (T/T reg /B/NK) and myeloid cells] the main leukocyte populations were analyzed in a clinical-diagnostic setting in a harmonized manner across different platforms. As a first step, the consortium presents here the absolute and relative frequencies of the leukocyte subpopulations in the peripheral blood of more than 300 healthy volunteers across six different European centers.

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Effect of delayed cell processing and cryopreservation on immunophenotyping in multicenter population studies

Cited by 33 publications

References 18 publications

Nonclassical Monocytes (CD14dimCD16+) Are Associated With Carotid Intima-Media Thickness Progression for Men but Not Women

Nonclassical Monocytes (CD14dimCD16+) Are Associated With Carotid Intima-Media Thickness Progression for Men but Not Women

Natural killer cells, gamma delta T cells and classical monocytes are associated with systolic blood pressure in the multi-ethnic study of atherosclerosis (MESA)

Comprehensive flow cytometric reference intervals of leukocyte subsets from six study centers across Europe

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