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Diagnostic advantage of double immunohistochemistry using two mutation-specific anti-IDH antibodies (HMab-1 and MsMab-1) in gliomas

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Isocitrate dehydrogenase (IDH) mutation is a valuable prognostic marker and a tool for decision-making for glioma treatment. An algorithm for IDH mutation screening was recently proposed—it consists of a two-step process of an initial search for the most common IDH1-R132H mutation using immunohistochemistry (IHC)-based assay, followed by DNA-based analysis of IHC-negative or -equivocal cases. Here, we report that immunohistochemistry using two mutation-specific anti-IDH monoclonal antibodies (mAbs)—an IDH1-R132H-specific mAb (clone HMab-1) and a multi-specific mAb (clone MsMab-1)—is easy and reliable for IDH mutation screening. We investigated the IDH status of 54 grade III gliomas. For the first screening, we used HMab-1 IHC and for the second, (of HMab-1-negative cases) we used MsMab-1 IHC. The double IHC screening results were confirmed using sequence analysis (100 % specificity and 100 % sensitivity). Thirty of 54 cases (55.6 %) had IDH mutations and the remaining 24 were of the IDH wild type; moreover, the screening results predicted grade III glioma prognosis. IDH sequencing procedures are popular but inconsistent across laboratories. By contrast, double IHC screening using HMab-1 and MsMab-1 is very reliable for detecting IDH1/2 mutations and can predict survival in grade III glioma patients.

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We thank Yoshiko Tsukada, Makiko Miyakawa, Takuro Nakamura, Noriko Saidoh, and Kanae Yoshida for their excellent technical assistance. This work was supported in part by Grants-in-Aid for Scientific Research (to S.T., No. 24390339; to Y.K., No. 25462242) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan; by the Platform for Drug Discovery, Informatics, and Structural Life Science from MEXT, Japan (to Y.K.); by the Basic Science and Platform Technology Program for Innovative Biological Medicine from MEXT, Japan (to Y.K.); by the Regional Innovation Strategy Support Program from MEXT, Japan (to Y.K.); by a Health Labour Sciences Research Grant from the Ministry of Health, Labour, and Welfare of Japan (to Y.K.); and by the Japan Brain Foundation (to S.T.) and the Japanese Foundation for Multidisciplinary Treatment of Cancer (to S.T.).

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Correspondence to Shingo Takano.

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Supplementary Figure 1. Background staining with MsMab-1. IDH1-R132H case (A, B), IDH wild-type case (C, D). HMab-1 stained nicely IDH1-R132H (A) and revealed no stain against IDH wild type (C) using LSAB2 kit. By contrast, MsMab-1 revealed background staining both against IDH1-R132H (B) and IDH wild type (D) using LSAB2 kit. However, MsMab-1 revealed no background staining against the same case using Envision + kit (see Supplementary Fig. 2F) (PPTX 393 kb)


Supplementary Figure 2. Comparison between HMab-1 and MsMab-1 against IDH1-R132H. HMab-1 (A, C, E) detected IDH1-R132H more strongly and frequently than MsMab-1 (B, D, F) (PPTX 689 kb)

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Takano, S., Kato, Y., Yamamoto, T. et al. Diagnostic advantage of double immunohistochemistry using two mutation-specific anti-IDH antibodies (HMab-1 and MsMab-1) in gliomas. Brain Tumor Pathol 32, 169–175 (2015).

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