血红蛋白对中性粒细胞细胞色素b - 245的光谱干扰及其被一氧化碳消除的情况。

Spectroscopic interference of hemoglobin with neutrophil cytochrome b-245 and its elimination by carbon monoxide.

作者信息

Pasquier C, Marquetty C, Chollet-Martin S, Hakim J

出版信息

J Immunol Methods. 1985 Feb 28;77(1):147-53. doi: 10.1016/0022-1759(85)90192-9.

Abstract

A cytochrome b, designated as cytochrome b-245, exists in neutrophils and is probably involved in their stimulated oxidative burst. As a rule, its concentration is spectroscopically measured by the height of its alpha-peak at 558-559 nm (dithionite-reduced minus oxidized). Hemoglobin (Hb), which usually contaminates neutrophils isolated from blood, interferes with the spectroscopic measurement of the cytochrome. Hb contamination from less than 0.4 red blood cells per 100 neutrophils leads to over-estimation of the cytochrome by approximately 50%. This interference can be overcome by bubbling CO through neutrophil homogenates heavily contaminated by Hb, prior to the conventional spectroscopic procedure. The cytochrome B-245 concentration obtained in neutrophils by CO bubbling is 7.2 +/- 1.28 pmoles per 10(6) polymorphonuclear neutrophils.

摘要

一种被称为细胞色素b - 245的细胞色素存在于中性粒细胞中，可能参与其受刺激后的氧化爆发过程。通常，其浓度通过在558 - 559nm处其α峰的高度（连二亚硫酸盐还原减去氧化态）进行光谱测定。血红蛋白（Hb）通常会污染从血液中分离出的中性粒细胞，干扰细胞色素的光谱测量。每100个中性粒细胞中少于0.4个红细胞的Hb污染会导致细胞色素的估计值高估约50%。在传统光谱测定程序之前，通过向被Hb严重污染的中性粒细胞匀浆中鼓入CO，可以克服这种干扰。通过鼓入CO在中性粒细胞中获得的细胞色素B - 245浓度为每10⁶个多形核中性粒细胞7.2 +/- 1.28皮摩尔。