Delivery of Antibodies into the Brain Using Focused Scanning Ultrasound

Gerhard Leinenga; Liviu-Gabriel Bodea; Wee  Kiat Koh; Rebecca  M. Nisbet; J&#252;rgen G&#246;tz

doi:10.3791/61372

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Delivery of Antibodies into the Brain Using Focused Scanning Ultrasound

집중 스캐닝 초음파를 사용하여 뇌로 항체 전달

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07:34 min

July 18, 2020

DOI: 10.3791/61372-v

Gerhard Leinenga¹, Liviu-Gabriel Bodea¹, Wee Kiat Koh¹, Rebecca M. Nisbet¹, Jürgen Götz¹

¹Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute,The University of Queensland

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Overview

This study presents a method for transiently opening the blood-brain barrier (BBB) in mouse models to facilitate the delivery of fluorescently-labeled antibodies and to activate microglia. The method allows for noninvasive delivery and monitoring of antibody uptake in the brain through histological techniques.

Key Study Components

Area of Science

Neuroscience
Immunology
Therapeutic delivery

Background

The blood-brain barrier limits the uptake of therapeutic antibodies to the brain.
Microglia play a critical role in brain immune responses.
Current methodologies for BBB opening and antibody delivery are limited.
Fluorescent labeling aids visualization of antibody uptake and microglia activation.

Purpose of Study

To develop a protocol to transiently open the BBB for antibody delivery.
To activate microglia in a controlled experimental setting.
To evaluate the effectiveness of this delivery method through histology.

Methods Used

The methodology involves focused ultrasound and microbubble injection into the mouse brain.
The main biological model utilized is anesthetized mice.
Key steps include the preparation of microbubbles and antibodies, and careful ultrasound targeting.
Fluorescent antibodies are purified and visualized to measure interaction with microglial cells.

Main Results

The technique successfully demonstrated antibody delivery to the brain.
Microglia exhibited increased phagocytic activity following antibody administration.
Visualization techniques confirmed effective localization and uptake of antibodies.
Scanning patterns enabled targeting of specific brain regions, broadening potential therapeutic applications.

Conclusions

This study provides a reliable method for enhancing therapeutic antibody delivery across the BBB.
It opens avenues for future research in drug development and understanding brain mechanisms.
The findings highlight the potential for this method in studying neuroimmunological interactions.

Frequently Asked Questions

What are the advantages of this antibody delivery method?

The method allows for noninvasive delivery of antibodies into the brain, which enhances therapeutic potential while minimizing surgical risks associated with traditional methods.

How is the biological model implemented in this study?

Anesthetized mice are used as the biological model, allowing researchers to conduct precise interventions without distress to the animals.

What types of data are obtained using this method?

The study provides data on the concentration and localization of antibody uptake in the brain, alongside changes in microglial activity indicated by histological staining.

How can this method be adapted for other studies?

This technique can be adjusted to target different brain regions or to investigate other therapeutic agents, thus broadening its application in neuroscience research.

Are there limitations to this approach?

While effective, the method requires careful control of ultrasound parameters to avoid potential damage to surrounding brain tissue and must be used with caution in studies.

여기에 제시된 프로토콜은 대소 또는 마우스 뇌 전체에 걸쳐 혈액-뇌 장벽 (BBB)을 일시적으로 열어 형광 표지 항체를 전달하고 미세 아교세포를 활성화시키는 프로토콜입니다. 또한 조직학에 의한 항체 전달 및 미세아교세포 활성화를 검출하는 방법이 제시된다.

치료용 항체 중 극히 일부만이 뇌에 흡수됩니다. 우리의 방법을 사용하면 항체가 혈액-뇌 장벽을 일시적으로 열어 뇌로 전달됩니다. 이 기술을 사용하면 비침습적 방식으로 쥐의 뇌에 항체를 전달할 수 있습니다.

세포 계수기를 사용하여 마이크로 거품의 품질 관리를 실행하기 위하여는, 합병기에서 마이크로 거품 해결책을 제거하고 마이크로 거품 해결책 작은 유리병의 격막을 관통하기 위하여 19의 계기 바늘을 이용하십시오. 19 게이지 바늘이 장착된 1밀리리터 주사기를 사용하여 100마이크로리터의 마이크로 버블을 5밀리리터의 여과된 유동 용액에 희석하고 100마이크로리터의 희석된 마이크로 버블 용액을 큐벳의 여과된 유동 용액 10밀리리터에 피펫으로 묽니다. 세포 계수기 플랫폼의 큐벳을 제자리에 고정하고 샘플 수집에 30미크론 조리개가 사용되는지 확인합니다.

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