Antibody isotype analysis of malaria-nematode co-infection: problems and solutions associated with cross-reactivity.
Antibody isotype analysis of malaria-nematode co-infection: problems and solutions associated with cross-reactivity.
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DOI：
10.1186/1471-2172-11-6
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发表时间：
2010-02-17
期刊：
影响因子：
3
通讯作者：
Allen JE
中科院分区：
文献类型：
--
作者：
Fairlie-Clarke KJ;Lamb TJ;Langhorne J;Graham AL;Allen JE
抗体同种型反应可用作感染期间免疫偏倚的指标。然而，在寄生虫合并感染的研究中，交叉反应性抗体的出现使免疫偏倚的解释变得复杂。为了自信地将免疫偏倚的变化归因于共感染寄生虫的存在，我们提出了实用的方法来解释抗体的交叉反应性。交叉反应性抗体的潜在影响疾病的结果也进行了讨论。利用两个小鼠模型的疟疾蠕虫共感染，我们分析了抗体反应的小鼠单独或共同感染疟原虫chabaudi chabaudi和日本圆线虫巴西或Litomosoides sigmodontis。我们观察到交叉反应的抗体反应，无论是否粗寄生虫抗原制剂或纯化的重组蛋白在ELISA中使用的抗原识别两种病原体。这些反应在对照小鼠中不明显。通过计算抗体滴度确定交叉反应性与抗原特异性应答的相对强度。此外，我们分析了抗体与高碘酸盐处理的抗原的结合，以区分针对蛋白质与碳水化合物部分的反应。高碘酸盐处理影响抗原特异性和交叉反应性反应。例如，发现疟疾诱导的交叉反应性IgG 1应答靶向蠕虫抗原的碳水化合物组分，因为在高碘酸盐处理后未检测到它们。有趣的是，重组疟疾抗原裂殖子表面蛋白-119（MSP-119）的高碘酸盐处理导致疟疾感染小鼠中抗原特异性IgG 2a应答的检测增加。这表明糖基化可能已经掩蔽蛋白质表位，并且高碘酸盐处理的MSP-119可能更接近地反映感染期间观察到的天然非糖基化抗原。为了在共感染研究期间利用抗体同种型作为免疫偏倚的量度，重要的是将抗原特异性与交叉反应性抗体应答分开。计算抗体滴度，而不是使用单一稀释的血清，作为反应相对强度的测量，在很大程度上实现了这一点。消除抗原的碳水化合物部分也被证明是有用的，所述碳水化合物部分通常可以是交叉反应性抗体的靶标。
Antibody isotype responses can be useful as indicators of immune bias during infection. In studies of parasite co-infection however, interpretation of immune bias is complicated by the occurrence of cross-reactive antibodies. To confidently attribute shifts in immune bias to the presence of a co-infecting parasite, we suggest practical approaches to account for antibody cross-reactivity. The potential for cross-reactive antibodies to influence disease outcome is also discussed. Utilising two murine models of malaria-helminth co-infection we analysed antibody responses of mice singly- or co-infected with Plasmodium chabaudi chabaudi and Nippostrongylus brasiliensis or Litomosoides sigmodontis. We observed cross-reactive antibody responses that recognised antigens from both pathogens irrespective of whether crude parasite antigen preparations or purified recombinant proteins were used in ELISA. These responses were not apparent in control mice. The relative strength of cross-reactive versus antigen-specific responses was determined by calculating antibody titre. In addition, we analysed antibody binding to periodate-treated antigens, to distinguish responses targeted to protein versus carbohydrate moieties. Periodate treatment affected both antigen-specific and cross-reactive responses. For example, malaria-induced cross-reactive IgG1 responses were found to target the carbohydrate component of the helminth antigen, as they were not detected following periodate treatment. Interestingly, periodate treatment of recombinant malaria antigen Merozoite Surface Protein-119 (MSP-119) resulted in increased detection of antigen-specific IgG2a responses in malaria-infected mice. This suggests that glycosylation may have been masking protein epitopes and that periodate-treated MSP-119 may more closely reflect the natural non-glycosylated antigen seen during infection. In order to utilize antibody isotypes as a measure of immune bias during co-infection studies, it is important to dissect antigen-specific from cross-reactive antibody responses. Calculating antibody titre, rather than using a single dilution of serum, as a measure of the relative strength of the response, largely accomplished this. Elimination of the carbohydrate moiety of an antigen that can often be the target of cross-reactive antibodies also proved useful.
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DOI：
10.4269/ajtmh.2005.73.1124
发表时间：
2005-12-01
影响因子：
3.3
作者：
Lyke, KE;Dicko, A;Doumbo, OK
通讯作者：
Doumbo, OK
影响因子：
3.1
作者：
Helmby, H;Kullberg, M;Troye-Blomberg, M
通讯作者：
Troye-Blomberg, M
影响因子：
2.1
作者：
Lawrence, RA;Gray, CA;Maizels, RM
通讯作者：
Maizels, RM
影响因子：
3
作者：
Hoeve MA;Mylonas KJ;Fairlie-Clarke KJ;Mahajan SM;Allen JE;Graham AL
通讯作者：
Graham AL
DOI：
10.1186/1475-2883-3-1
发表时间：
2004-01-22
期刊：
Filaria journal
影响因子：
--
作者：
Li, Ben-Wen;Rush, Amy;Weil, Gary J
通讯作者：
Weil, Gary J