Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers

A K Allan, G M Hawksworth, L R Woodhouse, B Sutherland, J C King, J H Beattie

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)-polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13.7 mg/d for 5 weeks (baseline) followed by 4.6 mg/d for 10 weeks (marginal intake) and then 13.7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to beta -actin mRNA which was also measured by competitive RT-PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10.3 (se 3.7) fg MT-2A mRNA/pg beta -actin mRNA, which then decreased by 64 % during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.

Original languageEnglish
Pages (from-to)747-756
Number of pages10
JournalBritish Journal of Nutrition
Volume84
Publication statusPublished - 2000

Keywords

  • metallothionein
  • RT-PCR
  • zinc deficiency
  • capillary electrophoresis
  • POLYMERASE CHAIN-REACTION
  • INTERNAL STANDARDS
  • PRESCHOOL-CHILDREN
  • COMPETITIVE PCR
  • MESSENGER-RNA
  • POOR GROWTH
  • BLOOD
  • DEFICIENCY
  • EXPRESSION
  • ANOREXIA

Cite this

Allan, A. K., Hawksworth, G. M., Woodhouse, L. R., Sutherland, B., King, J. C., & Beattie, J. H. (2000). Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers. British Journal of Nutrition, 84, 747-756.

Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers. / Allan, A K ; Hawksworth, G M ; Woodhouse, L R ; Sutherland, B ; King, J C ; Beattie, J H .

In: British Journal of Nutrition, Vol. 84, 2000, p. 747-756.

Research output: Contribution to journalArticle

Allan, AK, Hawksworth, GM, Woodhouse, LR, Sutherland, B, King, JC & Beattie, JH 2000, 'Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers' British Journal of Nutrition, vol. 84, pp. 747-756.
Allan, A K ; Hawksworth, G M ; Woodhouse, L R ; Sutherland, B ; King, J C ; Beattie, J H . / Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers. In: British Journal of Nutrition. 2000 ; Vol. 84. pp. 747-756.
@article{7c057354898644f3a505b16f6dab32f4,
title = "Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers",
abstract = "Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)-polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13.7 mg/d for 5 weeks (baseline) followed by 4.6 mg/d for 10 weeks (marginal intake) and then 13.7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to beta -actin mRNA which was also measured by competitive RT-PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10.3 (se 3.7) fg MT-2A mRNA/pg beta -actin mRNA, which then decreased by 64 {\%} during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.",
keywords = "metallothionein, RT-PCR, zinc deficiency, capillary electrophoresis, POLYMERASE CHAIN-REACTION, INTERNAL STANDARDS, PRESCHOOL-CHILDREN, COMPETITIVE PCR, MESSENGER-RNA, POOR GROWTH, BLOOD, DEFICIENCY, EXPRESSION, ANOREXIA",
author = "Allan, {A K} and Hawksworth, {G M} and Woodhouse, {L R} and B Sutherland and King, {J C} and Beattie, {J H}",
year = "2000",
language = "English",
volume = "84",
pages = "747--756",
journal = "British Journal of Nutrition",
issn = "0007-1145",
publisher = "Cambridge Univ. Press.",

}

TY - JOUR

T1 - Lymphocyte metallothionein mRNA responds to marginal zinc intake in human volunteers

AU - Allan, A K

AU - Hawksworth, G M

AU - Woodhouse, L R

AU - Sutherland, B

AU - King, J C

AU - Beattie, J H

PY - 2000

Y1 - 2000

N2 - Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)-polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13.7 mg/d for 5 weeks (baseline) followed by 4.6 mg/d for 10 weeks (marginal intake) and then 13.7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to beta -actin mRNA which was also measured by competitive RT-PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10.3 (se 3.7) fg MT-2A mRNA/pg beta -actin mRNA, which then decreased by 64 % during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.

AB - Marginal Zn deficiency is thought to be prevalent in both developed and developing countries. However, the extent of Zn deficiency is not known, due to the lack of a reliable diagnostic indicator. Blood plasma and erythrocyte concentrations of metallothionein (MT) reflect Zn status, but measurement of MT is dependent on the availability of sensitive immunoassays. Our aim was to show whether measurement of T lymphocyte MT-2A mRNA, using a competitive reverse transcriptase (RT)-polymerase chain reaction (PCR) assay, could indicate Zn status in human subjects in a residential Zn-depletion study. In the study, the Zn intake of seven volunteers was maintained at 13.7 mg/d for 5 weeks (baseline) followed by 4.6 mg/d for 10 weeks (marginal intake) and then 13.7 mg/d (repletion) for 5 weeks. The quantitative assay was developed using standard techniques and concentrations of MT-2A mRNA were normalized by reference to beta -actin mRNA which was also measured by competitive RT-PCR assay. An alternative method of measuring the PCR product using capillary electrophoresis with laser-induced fluorescence detection was also evaluated. There was considerable inter-individual variation in MT-2A mRNA concentration and the mean level at the end of the baseline period was 10.3 (se 3.7) fg MT-2A mRNA/pg beta -actin mRNA, which then decreased by 64 % during the low Zn intake period. After repletion, MT-2A mRNA returned to baseline concentrations. In contrast, plasma Zn was unchanged by marginal Zn intake or repletion. The effect of low Zn in all individuals was consistent. We conclude that this assay is a sensitive method of evaluating marginal changes in dietary Zn intake.

KW - metallothionein

KW - RT-PCR

KW - zinc deficiency

KW - capillary electrophoresis

KW - POLYMERASE CHAIN-REACTION

KW - INTERNAL STANDARDS

KW - PRESCHOOL-CHILDREN

KW - COMPETITIVE PCR

KW - MESSENGER-RNA

KW - POOR GROWTH

KW - BLOOD

KW - DEFICIENCY

KW - EXPRESSION

KW - ANOREXIA

M3 - Article

VL - 84

SP - 747

EP - 756

JO - British Journal of Nutrition

JF - British Journal of Nutrition

SN - 0007-1145

ER -