Abstract
Monoclonal immunoglobulin-G (IgG) antibodies are now emerging as therapeutic tools to tackle various disorders, including those affecting the brain. However, little is known about how these IgG molecules behave in the brain. To better understand the potential behavior of IgG molecules in the brain, here we established a specific protocol to immunolocalize rat IgG injected into mouse striatum with an anti-rat IgG antibody. Using double immunolabeling, IgG-like immunoreactivity (IR) was mainly found in neurons but scarcely observed in glia 1 h after intrastriatal injection of IgG, whereas some surrounding glia contained IgG-like IR 24 h after injection. However, preabsorption with a large excess of rat IgG to confirm the authenticity of this labeling failed to eliminate this neuronal IgG-like IR but rather exhibited nuclear staining in glial cells. Because this unexpected nuclear staining escalated with increasing amount of absorbing IgG, we postulated that this nuclear staining is due to formation of immune complex IgG–anti-IgG, which can be removed by centrifugal filtration. As expected, this nuclear staining in glial cells was eliminated after centrifugal filtration of the IgG/anti-IgG mixture, and authentic IgG-like IR was chiefly detected in the cytoplasm of neurons around the injection channel. This study is the first demonstration of neuronal redistribution of injected IgG in the mouse brain. Neuronal internalization of exogenous IgG may be advantageous especially when the therapeutic targets of monoclonal IgG are intraneuronal such as neurofibrillary tangles or Lewy bodies.
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Abbreviations
- CNS:
-
Central nervous system
- CSF:
-
Cerebrospinal fluid
- FcγR:
-
Fc-gamma receptor
- GFAP:
-
Glial fibrillary acidic protein
- IBA1:
-
Ionized calcium-binding adaptor molecule 1
- IF:
-
Immunofluorescence
- IgG:
-
Immunoglobulin-G
- IHC:
-
Immunohistochemistry
- IR:
-
Immunoreactivity
- PBS:
-
Phosphate-buffered saline
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Acknowledgements
The authors are grateful to Dr. Tetsuya Nagata and Takashi Ishii for generous support and technical assistance, respectively. TB. appreciates the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) (Tokyo) for kindly providing a scholarship, Super Global University Category. This work was also supported by a grant (grant number 13423765) from Center of Open Innovation Network for Smart Health; COINS (Kanagawa, Japan), organized by Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) (Tokyo) to TY.
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All authors had full access to all the data in the study and took responsibility for the integrity of the data and the accuracy of the data analysis. Conceptualization, TU; methodology, AY, AN, and TU; investigation, TB; formal analysis, TB, AY, and TU; writing—original draft, TB, AY; writing—review and editing, TU; visualization, TB, AY, and TU; supervision, TU and TY; funding acquisition, TY.
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The animal protocols for this study were approved by the ethics committee of Tokyo Medical and Dental University (A2021-191A).
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Bannawongsil, T., Yamada, A., Nakamura, A. et al. Neuronal internalization of immunoglobulin G injected into the mouse brain by a novel absorption strategy to avoid unwanted interaction with immune complex using centrifugal filtration. Histochem Cell Biol 158, 159–168 (2022). https://doi.org/10.1007/s00418-022-02107-y
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DOI: https://doi.org/10.1007/s00418-022-02107-y