Abstract：
BACKGROUND: Human immunodeficiency virus (HIV) infection is associated with pronounced oxidative stress, leading to the development of various virus-associated pathologies. A wealth of evidence suggests that, along with canonical enzymes of reactive oxygen species regulation, human blood contains antibodies with peroxidase, superoxide dismutase, and catalase activities. Here we show that the catalase activity of IgGs and their κκ-IgG, λλ-IgG, and κλ-IgG subfractions of HIV-infected individuals is significantly different compared to the healthy donors.
METHODS: Protein G-Sepharose sorbent was used to resolve IgG from blood of healthy donors and HIV-infected patients by affinity chromatography. Subfractions of κκ-IgG, λλ-IgG, and κλ-IgG were separated from IgGs samples of each group by affinity chromatography on sorbents containing immobilized antibodies to κ or λ light human chains. The IgG catalase activity level was measured spectrophotometrically by evaluating the decrease in optical density (A240) due to hydrogen peroxide decomposition.
RESULTS: The relative catalase activity of antibodies from HIV-infected patients (kcat = (1.41 ± 0.92) × 103 min-1, 95% CI: [1.01-1.81]) was statistically significant, 1.6 times higher (p = 0.014) compared to apparently healthy donors ((0.86 ± 0.49) × 103, 95% CI: [0.69-1.03]). The activity level of κκ-IgG HIV-infected patients ((0.44 ± 0.04) × 103 min-1) was 1.4 times higher than that of λλ-IgGs ((0.31 ± 0.025) × 103 min-1); the opposite was observed for κκ-IgGs from apparently healthy donors, which activity ((0.17 ± 0.015) × 103 min-1) was 3.1 times lower compared to λλ-IgGs ((0.53 ± 0.045) × 103 min-1).
CONCLUSIONS: Thus, the data obtained may indicate that IgG with increased catalase activity may prevent harmful processes arising from oxidative stress in HIV-infected patients, acting as an additional natural molecular mechanism of regulation of hydrogen peroxide level.
METHODS: Protein G-Sepharose sorbent was used to resolve IgG from blood of healthy donors and HIV-infected patients by affinity chromatography. Subfractions of κκ-IgG, λλ-IgG, and κλ-IgG were separated from IgGs samples of each group by affinity chromatography on sorbents containing immobilized antibodies to κ or λ light human chains. The IgG catalase activity level was measured spectrophotometrically by evaluating the decrease in optical density (A240) due to hydrogen peroxide decomposition.
RESULTS: The relative catalase activity of antibodies from HIV-infected patients (kcat = (1.41 ± 0.92) × 103 min-1, 95% CI: [1.01-1.81]) was statistically significant, 1.6 times higher (p = 0.014) compared to apparently healthy donors ((0.86 ± 0.49) × 103, 95% CI: [0.69-1.03]). The activity level of κκ-IgG HIV-infected patients ((0.44 ± 0.04) × 103 min-1) was 1.4 times higher than that of λλ-IgGs ((0.31 ± 0.025) × 103 min-1); the opposite was observed for κκ-IgGs from apparently healthy donors, which activity ((0.17 ± 0.015) × 103 min-1) was 3.1 times lower compared to λλ-IgGs ((0.53 ± 0.045) × 103 min-1).
CONCLUSIONS: Thus, the data obtained may indicate that IgG with increased catalase activity may prevent harmful processes arising from oxidative stress in HIV-infected patients, acting as an additional natural molecular mechanism of regulation of hydrogen peroxide level.
摘要：
背景：人类免疫缺陷病毒（HIV）感染与明显的氧化应激有关，导致各种病毒相关病理的发展。大量证据表明,以及活性氧调节的规范酶，人体血液中含有过氧化物酶抗体，超氧化物歧化酶,和过氧化氢酶活性。在这里，我们显示IgG的过氧化氢酶活性及其κκ-IgG，λλ-IgG，与健康供体相比，HIV感染个体的κλ-IgG亚组分显着不同。
方法：蛋白G-Sepharose吸附剂用于通过亲和层析从健康供体和HIV感染患者的血液中分离IgG。κκ-IgG亚组分,λλ-IgG，通过在含有固定的κ或λ轻链抗体的吸附剂上进行亲和层析，从每组的IgG样品中分离出κλ-IgG。通过评估由于过氧化氢分解导致的光密度(A240)的降低,用分光光度法测量IgG过氧化氢酶活性水平。
结果：HIV感染患者抗体的相对过氧化氢酶活性（kcat=（1.41±0.92）×103min-1，95％CI：[1.01-1.81]）具有统计学意义，与明显健康的供体（（0.86±0.49）×103，95％CI：[0.69-1.03]）相比，高出1.6倍（p=0.014）。HIV感染患者的κκκ-IgG活性水平（（0.44±0.04）×103min-1）是λλ-IgG的1.4倍（（0.31±0.025）×103min-1）；对于明显健康的供体，观察到相反的情况。与λλ-IgG（（0.53±0.045）×103min-1）相比，其活性（（0.17±0.015）×103min-1）低3.1倍。
结论：因此，获得的数据可能表明，过氧化氢酶活性增加的IgG可以预防HIV感染患者氧化应激引起的有害过程，作为调节过氧化氢水平的附加天然分子机制。
方法：蛋白G-Sepharose吸附剂用于通过亲和层析从健康供体和HIV感染患者的血液中分离IgG。κκ-IgG亚组分,λλ-IgG，通过在含有固定的κ或λ轻链抗体的吸附剂上进行亲和层析，从每组的IgG样品中分离出κλ-IgG。通过评估由于过氧化氢分解导致的光密度(A240)的降低,用分光光度法测量IgG过氧化氢酶活性水平。
结果：HIV感染患者抗体的相对过氧化氢酶活性（kcat=（1.41±0.92）×103min-1，95％CI：[1.01-1.81]）具有统计学意义，与明显健康的供体（（0.86±0.49）×103，95％CI：[0.69-1.03]）相比，高出1.6倍（p=0.014）。HIV感染患者的κκκ-IgG活性水平（（0.44±0.04）×103min-1）是λλ-IgG的1.4倍（（0.31±0.025）×103min-1）；对于明显健康的供体，观察到相反的情况。与λλ-IgG（（0.53±0.045）×103min-1）相比，其活性（（0.17±0.015）×103min-1）低3.1倍。
结论：因此，获得的数据可能表明，过氧化氢酶活性增加的IgG可以预防HIV感染患者氧化应激引起的有害过程，作为调节过氧化氢水平的附加天然分子机制。
