ELISA for Aging Biomarkers Induced by Telomere Dysfunction in Human Plasma

Hong Jiang,Wenqing Chen,Jianyong Wu,Ying Chen,Zhangfei Shou,Jianghua Chen,Lihui Qu,Zhenyu Ju,Qiang He,Huiping Wang

doi:10.1155/2010/121947

Abstract

Background. We identified cathelicidin related antimicrobial protein (CRAMP) secreted from telomere dysfunctional bone marrow cells of late generation telomerase knockout mice (G4mTerc−/−), increased in blood and various tissues. It can represented human aging and disease. The main aim of this study is to investigate the sensitive direct enzyme-linked immunosorbent assay (ELISA) method to analyze the human aging and disease in plasma and the detailed methods to quantify the direct ELISA of these aging biomarkers. Methods. Telomere lengths of 50 healthy persons are measured with real-time PCR in blood cells. Plasma samples from all subjects are analyzed using direct ELISA. Results. From 25 years old person to 78 years, the telomere length becomes shorter during aging. In blood plasma, the expression levels of CRAMP increases during human aging. There is the reverse correspondence between the telomere length and the plasma CRAMP level. We also find that the fresh plasma, the frozen plasma which thawed less than 3 times, and the plasma kept in the room temperature less than 3 hours are better for the ELISA analyze of CRAMP in the plasma. Conclusion. This CRAMP ELISA could become a powerful tool for investigating the relationship between human aging and telomere length shortening.

Highlights

Telomeres cap the chromosome ends and prevent the activation of DNA damage checkpoints inducing cell cycle arrest or apoptosis [1, 2]
From 25 years old person to 78 years, the telomere length becomes shorter during aging (Figure 1)
We identified cathelicidin related antimicrobial protein (CRAMP) secreted from telomeredysfunctional bone-marrow cells of late-generation telomerase knockout mice (G4mTerc−/−), increased in blood and in various tissues of aging G4mTerc−/− mice and represented human aging and disease (PNAS)

Summary

Introduction

Telomeres cap the chromosome ends and prevent the activation of DNA damage checkpoints inducing cell cycle arrest (senescence) or apoptosis [1, 2]. In aging telomerase knockout mice (G4mTerc−/−), telomere dysfunction is associated with a decline in stem cell function, impaired organ maintenance, and a shortened lifespan [8,9,10,11]. We identified cathelicidin related antimicrobial protein (CRAMP) secreted from telomere dysfunctional bone marrow cells of late generation telomerase knockout mice (G4mTerc−/−), increased in blood and various tissues. It can represented human aging and disease. This CRAMP ELISA could become a powerful tool for investigating the relationship between human aging and telomere length shortening

Objectives

Results

Conclusion