Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma

Rachael P. Jackman, Garth H Utter, John W. Heitman, Dale F. Hirschkorn, Jacqueline P. Law, Nelly Gefter, Michael P. Busch, Philip J. Norris

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Measurement of peripheral blood cytokines and other immunomodulatory proteins is a useful and popular tool for assessing human immune responses to a wide range of assaults. A common challenge in this work is obtaining fresh, high-quality samples and limiting the time between blood collection and the separation of plasma or serum from cells. In this study we sought to determine the effect of sample age at the time of processing on the measured levels of 41 soluble immune mediators. Two cohorts were examined: healthy lab donors and trauma patients, who have significant immune perturbation. Whole-blood samples were aliquoted, and plasma was isolated, at days 0, 1, 2, and 3 after collection. Multiplexing techniques were used to measure protein concentrations, and general estimating equations were used to determine if there was a significant change over time. Over the 3-day period examined, only 15 of the 41 proteins showed no significant change in either cohort. Among the remaining proteins both increases and decreases were observed, with changes ranging from 2.4% per day to 325% per day. Proteins with significant changes in one cohort did not always show significant changes in the other group. These results support the need to separate plasma or serum from whole blood as quickly as possible and/or to standardize the length of time to processing within a given study of peripheral blood protein concentrations. When this is not possible, care should be taken to account for differences due to sample age.

Original languageEnglish (US)
Pages (from-to)318-326
Number of pages9
JournalClinical and Vaccine Immunology
Volume18
Issue number2
DOIs
StatePublished - Feb 2011

Fingerprint

Plasma (human)
Blood
Cytokines
Proteins
Plasmas
Processing
Serum
Multiplexing
Blood Proteins
Tissue Donors
Wounds and Injuries

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Immunology
  • Immunology and Allergy
  • Microbiology (medical)

Cite this

Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma. / Jackman, Rachael P.; Utter, Garth H; Heitman, John W.; Hirschkorn, Dale F.; Law, Jacqueline P.; Gefter, Nelly; Busch, Michael P.; Norris, Philip J.

In: Clinical and Vaccine Immunology, Vol. 18, No. 2, 02.2011, p. 318-326.

Research output: Contribution to journalArticle

Jackman, RP, Utter, GH, Heitman, JW, Hirschkorn, DF, Law, JP, Gefter, N, Busch, MP & Norris, PJ 2011, 'Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma', Clinical and Vaccine Immunology, vol. 18, no. 2, pp. 318-326. https://doi.org/10.1128/CVI.00465-10
Jackman, Rachael P. ; Utter, Garth H ; Heitman, John W. ; Hirschkorn, Dale F. ; Law, Jacqueline P. ; Gefter, Nelly ; Busch, Michael P. ; Norris, Philip J. / Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma. In: Clinical and Vaccine Immunology. 2011 ; Vol. 18, No. 2. pp. 318-326.
@article{9c5defa19745461886bb67b4ede9a25b,
title = "Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma",
abstract = "Measurement of peripheral blood cytokines and other immunomodulatory proteins is a useful and popular tool for assessing human immune responses to a wide range of assaults. A common challenge in this work is obtaining fresh, high-quality samples and limiting the time between blood collection and the separation of plasma or serum from cells. In this study we sought to determine the effect of sample age at the time of processing on the measured levels of 41 soluble immune mediators. Two cohorts were examined: healthy lab donors and trauma patients, who have significant immune perturbation. Whole-blood samples were aliquoted, and plasma was isolated, at days 0, 1, 2, and 3 after collection. Multiplexing techniques were used to measure protein concentrations, and general estimating equations were used to determine if there was a significant change over time. Over the 3-day period examined, only 15 of the 41 proteins showed no significant change in either cohort. Among the remaining proteins both increases and decreases were observed, with changes ranging from 2.4{\%} per day to 325{\%} per day. Proteins with significant changes in one cohort did not always show significant changes in the other group. These results support the need to separate plasma or serum from whole blood as quickly as possible and/or to standardize the length of time to processing within a given study of peripheral blood protein concentrations. When this is not possible, care should be taken to account for differences due to sample age.",
author = "Jackman, {Rachael P.} and Utter, {Garth H} and Heitman, {John W.} and Hirschkorn, {Dale F.} and Law, {Jacqueline P.} and Nelly Gefter and Busch, {Michael P.} and Norris, {Philip J.}",
year = "2011",
month = "2",
doi = "10.1128/CVI.00465-10",
language = "English (US)",
volume = "18",
pages = "318--326",
journal = "Clinical and Vaccine Immunology",
issn = "1556-6811",
publisher = "American Society for Microbiology",
number = "2",

}

TY - JOUR

T1 - Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma

AU - Jackman, Rachael P.

AU - Utter, Garth H

AU - Heitman, John W.

AU - Hirschkorn, Dale F.

AU - Law, Jacqueline P.

AU - Gefter, Nelly

AU - Busch, Michael P.

AU - Norris, Philip J.

PY - 2011/2

Y1 - 2011/2

N2 - Measurement of peripheral blood cytokines and other immunomodulatory proteins is a useful and popular tool for assessing human immune responses to a wide range of assaults. A common challenge in this work is obtaining fresh, high-quality samples and limiting the time between blood collection and the separation of plasma or serum from cells. In this study we sought to determine the effect of sample age at the time of processing on the measured levels of 41 soluble immune mediators. Two cohorts were examined: healthy lab donors and trauma patients, who have significant immune perturbation. Whole-blood samples were aliquoted, and plasma was isolated, at days 0, 1, 2, and 3 after collection. Multiplexing techniques were used to measure protein concentrations, and general estimating equations were used to determine if there was a significant change over time. Over the 3-day period examined, only 15 of the 41 proteins showed no significant change in either cohort. Among the remaining proteins both increases and decreases were observed, with changes ranging from 2.4% per day to 325% per day. Proteins with significant changes in one cohort did not always show significant changes in the other group. These results support the need to separate plasma or serum from whole blood as quickly as possible and/or to standardize the length of time to processing within a given study of peripheral blood protein concentrations. When this is not possible, care should be taken to account for differences due to sample age.

AB - Measurement of peripheral blood cytokines and other immunomodulatory proteins is a useful and popular tool for assessing human immune responses to a wide range of assaults. A common challenge in this work is obtaining fresh, high-quality samples and limiting the time between blood collection and the separation of plasma or serum from cells. In this study we sought to determine the effect of sample age at the time of processing on the measured levels of 41 soluble immune mediators. Two cohorts were examined: healthy lab donors and trauma patients, who have significant immune perturbation. Whole-blood samples were aliquoted, and plasma was isolated, at days 0, 1, 2, and 3 after collection. Multiplexing techniques were used to measure protein concentrations, and general estimating equations were used to determine if there was a significant change over time. Over the 3-day period examined, only 15 of the 41 proteins showed no significant change in either cohort. Among the remaining proteins both increases and decreases were observed, with changes ranging from 2.4% per day to 325% per day. Proteins with significant changes in one cohort did not always show significant changes in the other group. These results support the need to separate plasma or serum from whole blood as quickly as possible and/or to standardize the length of time to processing within a given study of peripheral blood protein concentrations. When this is not possible, care should be taken to account for differences due to sample age.

UR - http://www.scopus.com/inward/record.url?scp=79551494483&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79551494483&partnerID=8YFLogxK

U2 - 10.1128/CVI.00465-10

DO - 10.1128/CVI.00465-10

M3 - Article

C2 - 21159926

AN - SCOPUS:79551494483

VL - 18

SP - 318

EP - 326

JO - Clinical and Vaccine Immunology

JF - Clinical and Vaccine Immunology

SN - 1556-6811

IS - 2

ER -