Name: selective cell elimination from mixed 3d culture using a near infrared photoimmunotherapy technique
Uploaded: 2016-03-14T14:00:00.000+00:00
Duration: 5 min 18 s
Description: Watch this Scientific Journal Video about Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique at JoVE.com

この研究は、米国立衛生研究所、国立癌研究所、癌研究センターの学内研究プログラムによってサポートされていました。

注：以下のプロトコルは、NIR-PITを使用して、特定の細胞を排除するために必要な手順を説明します。コントロールとNIR-PITと細胞生存率に関するその他の詳細は、別の場所で18見つけることができます。 モノクローナル抗体にIR700の1コンジュゲートモノクローナル抗体（mAb） <ol><li> 0.1 MのNa 2 HPO 4（pHは8.6）溶液中の2-5 mg / mlとに関心のmAbを準備します。 </li><li>マイクロ遠心チューブ中の0.1MのNa 2 HPO 4溶液（pHは8.6）中の10mM IR700の30.8ナノモルとmAbの6.8ナノモルを混合し、アルミ箔で覆われて1時間、室温でインキュベートします。 </li><li>二回、15ミリリットルのPBSでPD-10カラム（材料/機器の表を参照）を洗浄します。ステップ1.2からサンプルをロードします。 </li><li>製造業者の指示に従ってPBS溶出により、PD-10カラムを介して混合物を精製します。 注：ここでは、溶出はIR700のバンド色に沿っていました。 300μlのサンプルについて、PBS溶出画分4.4 mlを典型的には2.5〜mlです。 </li><li>決定します分光光度計9で595nmの吸収を測定することによりクマシー染色でタンパク質濃度。各mAb分子9にコンジュゲート蛍光体分子の数を確認するために、689 nmでの吸収がIR700の濃度を決定します。 注：分光光度計で1モノクローナル抗体でIR700分子の最適なコンジュゲーション数を決定することが重要です。一般的に、1抗体分子中の3 IR700分子の周りには、両方のin vitroおよびin vivoの仕事に最適です。 HPLCおよびSDS-PAGE法は、mAbおよびIR700が結合しているか否かを確認するために使用することができます。 </li><li>タンパク質濃度を決定した後、4℃で保管してください。 </li></ol>混合3D細胞培養（混合スフェロイド）の調製<ol><li>ドロッププレートをぶら下げのプレート貯留部に無菌水（約1 ml）を適用します。 </li><li> A431-LUC-GFP細胞および3T3-RFP細胞 - 様々な目的の細胞タイプの比率を準備 -各サンプルは合計5,000個の細胞を含有する培養培地50μlに懸濁しました。 注：培地中の目的の細胞タイプの割合は、細胞型に応じてその他 100、100、10：100、25：100、50 1を準備します。 </li><li> 37℃、5％二酸化炭素の加湿インキュベーター内で96ウェル懸滴プレートにおいて5~7日間ミックスをインキュベートします。 2日ごとに培地を変更します。注：正確な3Dスフェロイドを作る静かにプレートを処理するために、細胞を含有する液滴は、3D形状を形成する前に、簡単に落ちるように。 </li><li> 40X倍率 - 10Xで反転明視野顕微鏡を用いてスフェロイドの形態と大きさを観察します。注：それは細胞の種類やハンギングドロップの大きさに依存するが、回転楕円体の直径は96ウェル懸滴プレート中でのインキュベーションの7日後の周りの400から600ミクロンであることを確認してください。 </li></ol>混合3D細胞培養のためのインビトロ NIR-PIT 3. <ol><li>ヘクタールのメディアを変更10μg/ mlの抗体光吸収複合体（APC）を含む培地にドロッププレートnging、37℃、5％二酸化炭素の加湿インキュベーター内で6時間インキュベートします。 </li><li> 6時間のインキュベーション後、新鮮な培地（フェノールレッドを含まない）で2回回転楕円体を洗浄します。先端がカットオフで優しく滅菌200μlのピペットチップを使用して、100μlの新鮮なフェノールレッドを含まない培地でガラス底の50ミリメートル皿にスフェロイドを転送します。各皿に1回転楕円体を配置します。 </li><li>形態の変化を検出するために倒立明視野顕微鏡でスフェロイドを守ってください。光学レポーター（ 例えば、GFPおよびRFP）を観察するには、以下のフィルタ設定で蛍光顕微鏡を使用する：GFP - 469 nmの励起フィルター、および525 nmの発光フィルター; RFP - 559 nmの励起フィルター、および630nmの発光フィルター。 </li><li>照射用ガラスボトムディッシュの上に発光ダイオード（LED）を配置します。 注：スフェロイドは、NIRに露出させることができます顕微鏡上または層流フード内のいずれかの光。 <ol><li>光パワーメータ9でNIR光のパワー密度を測定します。この測定によれば、2 J / cm 2で670〜710ナノメートルの波長で光を放射する発光ダイオード（LED）を介して近赤外光を照射します。 注：LEDの光は、約5インチの最大の深さに浸透することができます。 NIR-PITの細胞傷害性効果は関係なく、電力密度と暴露23の期間だけ与えられたエネルギーに依存しています。 </li></ol></li><li>照射後、新鮮な培地の50μlの新た懸滴プレートに回転楕円体を移し、37℃、5％二酸化炭素の加湿インキュベーター中で1日間インキュベートします。 </li><li>優しくカットオフチップで滅菌200μlのピペットチップを使用して、新鮮な培地（フェノールレッドを含まない）の100μlでガラス底の皿にスフェロイドを転送します。 NIR-PIT fiを使って後の蛍光顕微鏡1日でスフェロイドを守ってLTERの設定は、ステップ3.3に記載します。 <ol><li> 、または蛍光顕微鏡（ 図2B）14,18,22 を用いて、細胞質GFP蛍光の消失によって2μg/ mlの最終濃度で培地にヨウ化プロピジウムを添加することによって、死細胞を検出します。 </li></ol></li><li>標的細胞が完全に除去されていない場合、繰り返し3.1~3.6手順。 </li></ol>

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我々は、NIR-PITを用いて、非標的細胞に損傷を与えることなく、混合三次元細胞培養物から特定の細胞除去の方法を示します。これまでのところ、実用的な細胞除去の方法組織が確立されるか、移植後はありません。したがって、NIR-PITは、これを達成するための有望な方法です。 APCはモノクローナル抗体自体と同様の薬物動態を示すので、この技術は、 インビボ 18,22 <em...

他の細胞に損傷を与えることなく、特定の細胞を除去することは、特に確立された組織に、非常に困難であり、組織工学の分野における細胞除去方法に対する緊急の必要性があります。 3 -最近再生医療の分野では、胚性幹細胞（ES）、多能性幹細胞（のPSC）、または誘導多能性幹細胞（IPS）を使用して、組織培養、材料1が期待されています。 この組織再生が有望であるが、望ましくない細胞による汚染が主要な関心事です。また、移植4,5後の腫瘍形成能の安全性の懸念があります。多くの研究は、これらの問題に焦点を当ててきたが、特に再生医療6で、特定の細胞を排除するために- 8、実用的な方法が開発されていません。 近赤外photoimmunotherapy（NIR-PIT）は、抗体の光吸収conjugatに基づく治療法ですE（APC）。 APCは、細胞特異的なモノクローナル抗体（mAb）と光吸収、IR700で構成されています。 IR700は、親水性シリカ、フタロシアニン誘導体であり、それ自体9によって光毒性を誘発しません。 IR700は、共有リジン分子の側鎖のアミド残基を介して抗体に結合されます。 APCは、細胞膜上の標的分子に結合し、その後、690 nmの近赤外光への曝露後にほぼ即時の細胞壊死を誘導します。 14 - NIR光への曝露の間に、細胞膜の破裂は、細胞死に9をリード。 21 - NIR-PITは、抗EGFR、抗HER2、抗PSMA、抗CD25、抗メソテリン、抗GPC3、および抗CEA 15を含む複数の抗体または抗体断片と効果的であることが証明されています。したがって、NIR-PITは、標的分子の広範囲に対して使用することができます。また、NIR-PITは、NIR-ガーゼを制限することによって、特定の領域の選択的処理を可能にする十分に制御された治療でありますトン照射18,22。 ここでは、混合3D培養物からNIR-PITを使用して、特定の細胞除去の方法を提示します。

<table><tbody><tr><td>IRDye 700DX Ester Infrared Dye</td><td>LI-COR Bioscience (Lincoln, NE, USA)</td><td>929-70011</td><td></td></tr><tr><td>Na&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt;</td><td>SIGMA-ALDRICH (St. Louis, MO, USA)</td><td>S9763</td><td></td></tr><tr><td>Sephadex G25 column (PD-10)&amp;nbsp;</td><td>GE Healthcare (Piscataway, NJ, USA)</td><td>17-0851-01</td><td></td></tr><tr><td>Coomassie (bradford) Plus protein assay</td><td>Thermo Fisher Scientific Inc (Waltham, MA, USA)</td><td>PI-23200</td><td></td></tr><tr><td>Perfecta3D 96-Well hanging Drop Plates</td><td>3D Biomatrix Inc (Ann Arbor, MI, USA)</td><td>HDP1096-8</td><td></td></tr><tr><td>Optical power meter</td><td>Thorlabs (Newton, NJ, USA)</td><td>PM100</td><td></td></tr><tr><td>LED: L690-66-60</td><td>Marubeni America Co. (Santa Clara, CA, USA)</td><td>L690-66-60</td><td></td></tr><tr><td>Vectibix (panitumumab)</td><td>Amgen (Thousand Oaks, CA, USA)</td><td></td><td></td></tr><tr><td>35&amp;nbsp;mm glass bottom dish, dish size 35&amp;nbsp;mm, well size 10&amp;nbsp;mm</td><td>Cellvis (Mountain View, CA, USA)</td><td>D35-10-0-N</td><td></td></tr></tbody></table>

selective cell elimination from mixed 3d culture using a near infrared photoimmunotherapy technique

Recent developments in tissue engineering offer innovative solutions for many diseases. For example, tissue engineering using induced pluripotent stem cell (iPS) emerged as a new method in regenerative medicine. Although this tissue regeneration is promising, contamination with unwanted cells during tissue cultures is a major concern. Moreover, there is a safety concern regarding tumorigenicity after transplantation. Therefore, there is an urgent need for eliminating specific cells without damaging other cells that need to be protected, especially in established tissue. Here, we present a method for specific cell elimination from a mixed 3D cell culture in vitro with near infrared photoimmunotherapy (NIR-PIT) without damaging non-targeted cells. This technique enables the elimination of specific cells from mixed cell cultures or tissues.

光学NIR-PITの効果を監視するには、EGFRを過剰発現するA431細胞株は、遺伝的にもGFPおよびルシフェラーゼ（A431-LUC-GFP）を発現するように変更されました。 NIR-PITの非標的として、のBalb / 3T3細胞株を光学的にRFP（3T3-RFP）を発現するように改変しました。 APC、パニツムマブ-IR700（パン-IR700）を合成しました。細胞（A431-LUC-GFPおよび3T3-RFP）の種々の比率で構成された混合ス?...

Watch this Scientific Journal Video about Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique at JoVE.com

Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique

Eliminating specific cells without damaging other cells is extremely difficult, especially in established tissue, yet there is an urgent need for a cell elimination method in the tissue engineering field. Here, we present a method for specific cell elimination from a mixed 3D cell culture using near infrared photoimmunotherapy (NIR-PIT).

近赤外Photoimmunotherapy法を用いた混合3D文化からの選択的な細胞除去

Recent developments in tissue engineering offer innovative solutions for many diseases. For example, tissue engineering using induced pluripotent stem ...

selective-cell-elimination-from-mixed-3d-culture-using-near-infrared

Research

JoVE Journal

Bioengineering

7.8K ビュー.  National Cancer Institute. Eliminating specific cells without damaging other cells is extremely difficult, especially in established tissue, yet there is an urgent need for a cell elimination method in the tissue engineering field. Here, we present a method for specific cell elimination from a mixed 3D cell culture using near infrared photoimmunotherapy (NIR-PIT). 

ビデオ: Selective Cell Elimination from Mixed 3D Culture Using a Near Infrared Photoimmunotherapy Technique

Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery

Monoclonal antibody-based immunotherapy targeting tumor antigens is now a mainstay of cancer treatment. One of the clinically relevant mechanisms of action of the antibodies is antibody-dependent cellular cytotoxicity (ADCC), where the antibody binds to the cancer cells and engages the cellular component of the immune system, e.g., natural killer (NK) cells, to kill the tumor cells. The effectiveness of these therapies could be improved by identifying adjuvant compounds that increase the sensitivity of the cancer cells or the potency of the immune cells. In addition, undiscovered drug interactions in cancer patients co-medicated for previous conditions or cancer-associated symptoms may determine the success of the antibody therapy; therefore, such unwanted drug interactions need to be eliminated. With these goals in mind, we created a cancer ADCC model and describe here a simple protocol to find ADCC-modulating drugs. Since 3D models such as cancer cell spheroids are superior to 2D cultures in predicting in vivo responses of tumors to anticancer therapies, spheroid co-cultures of EGFP-expressing HER2+ JIMT-1 breast cancer cells and the NK92.CD16 cell lines were set up and induced with Trastuzumab, a monoclonal antibody clinically approved against HER2-positive breast cancer. JIMT-1 spheroids were allowed to form in cell-repellent U-bottom 96-well plates. On day 3, NK cells and Trastuzumab were added. The spheroids were then stained with Annexin V-Alexa 647 to measure apoptotic cell death, which was quantitated in the peripheral zone of the spheroids with an automated microscope. The applicability of our assay to identify ADCC-modulating molecules is demonstrated by showing that Sunitinib, a receptor tyrosine kinase inhibitor approved by the FDA against metastatic cancer, almost completely abolishes ADCC. The generation of the spheroids and image acquisition and analysis pipelines are compatible with high-throughput screening for ADCC-modulating compounds in cancer cell spheroids.

Here, we present a method to identify compounds that modulate the ADCC mechanism, an important cancer cell-killing mechanism of antitumor antibodies. The cytotoxic effect of NK cells is measured in breast cancer cell spheroids in the presence of Trastuzumab. Image analysis identifies live and dead killer and target cells in spheroids.

腫瘍スフェロイドモデルにおける抗体依存性細胞傷害性の定量化:創薬への応用

Monoclonal antibody-based immunotherapy targeting tumor antigens is now a mainstay of cancer treatment. One of the clinically relevant mechanisms of ...

がん研究

An In-House-Built and Light-Emitting-Diode-Based Photodynamic Therapy Device for Enhancing Verteporfin Cytotoxicity in a 2D Cell Culture Model

This paper describes a novel, simple, and low-cost device to perform in vitro photodynamic therapy (PDT) assays, named the PhotoACT. The device was built using a set of conventional programmable light-emitting diodes (LEDs), a liquid crystal display (LCD) module, and a light sensor connected to a commercial microcontroller board. The box-based structure of the prototype was made with medium-density fiberboards (MDFs). The internal compartment can simultaneously allocate four cell culture multiwell microplates.
As a proof of concept, we studied the cytotoxic effect of the photosensitizer (PS) verteporfin against the HeLa cell line in two-dimensional (2D) culture. HeLa cells were treated with increasing concentrations of verteporfin for 24 h. The drug-containing supernatant medium was discarded, the adherent cells were washed with phosphate-buffered saline (PBS), and drug-free medium was added. In this study, the effect of verteporfin on cells was examined either without light exposure or after exposure for 1 h to light using red-green-blue (RGB) values of 255, 255, and 255 (average fluence of 49.1 &#177; 0.6 J/cm2). After 24 h, the cell viability was assessed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay.
Experimental results showed that exposure of cells treated with verteporfin to the light from the device enhances the drug&#39;s cytotoxic effect via a mechanism mediated by reactive oxygen species (ROS). In addition, the use of the prototype described in this work was validated by comparing the results with a commercial PDT device. Thus, this LED-based photodynamic therapy prototype represents a good alternative for in vitro studies of PDT.

Here, we describe a novel, simple, and low-cost device to successfully perform in vitro photodynamic therapy (PDT) assays using two-dimensional HeLa cell culture and verteporfin as a photosensitizer.

2D細胞培養モデルにおけるベルテポルフィン細胞毒性を高めるための自作発光ダイオードベースの光線力学治療装置

This paper describes a novel, simple, and low-cost device to perform in vitro photodynamic therapy (PDT) assays, named the PhotoACT. The device was built ...

生化学

In Vitro and In Vivo Evaluation of Photocontrolled Biologically Active Compounds - Potential Drug Candidates for Cancer Photopharmacology

Photocontrolled, biologically active compounds are an emerging class of &#34;smart&#34; drug candidates. They provide additional safety in systemic chemotherapy due to their precise spatiotemporal activation by directing a benign, non-ionizable light to a specific location within the patient&#39;s body. This paper presents a set of methods to evaluate the in vitro potency and ex vivo efficiency of the photoactivation of photocontrolled, biologically active compounds as well as the in vivo efficacy at early stages of drug development. The methodology is applied to anticancer cytotoxic peptides, namely, the diarylethene-containing analogs of a known antibiotic, gramicidin S. The experiments are performed using 2D (adherent cells) and 3D (spheroids) cell cultures of a cancer cell line (Lewis lung carcinoma, LLC), live tissue surrogates (pork meat mince), and an allograft cancer model (subcutaneous LLC) in immunocompetent mice. The selection of the most effective compounds and estimation of realistic phototherapeutic windows are performed via automated fluorescence microscopy. The photoactivation efficiency at varying illumination regimens is determined at different depths in a model tissue, and the optimal light dosage is applied in the final therapeutic in vivo experiment.

In Vitro and In Vivo Evaluation of Photocontrolled Biologically Active Compounds - Potential Drug Candidates for Cancer Photopharmacology

This protocol presents a set of experiments adopted for the evaluation of photoswitchable anticancer peptides, that can be used in the preclinical screening of such compounds. This includes cytotoxicity assessment in 2D and 3D cell cultures, the evaluation of ex vivo (model tissue) photoisomerization efficiency, and in vivo efficacy.

光制御された生物学的に活性な化合物のin vitroおよびin vivo評価-癌光薬理学の潜在的な薬剤候補

Photocontrolled, biologically active compounds are an emerging class of &#34;smart&#34; drug candidates. They provide additional safety in systemic ...

化学

LED-Based In Vitro Screening for Assessing Photoactivable Molecules in Bacterial Photodynamic Inactivation

The rise of multi-drug-resistant infections and the lack of new antibiotic classes have renewed interest in alternative therapies like photodynamic inactivation of bacteria (aPDI). This process involves the administration of a photosensitizer (PS) activated by a suitable visible light source, producing exacerbated levels of reactive oxygen species (ROS) that damage crucial cellular biomolecules, ultimately causing bacterial cell death. It is crucial to create standardized, easy-to-use, and reproducible initial tests to assess and compare the effectiveness of light-induced phototoxicity of potential photosensitizers. This study introduces a simple and efficient in vitro technique for assessing photodynamic activity against planktonic bacterial cells. By employing a 96-well microplate format along with a large LED panel, the system facilitates the systematic evaluation of various compounds. Such a configuration allows for high-throughput screening of potential photosensitizers in a controlled and consistent environment, simplifying the process of identifying promising candidates for further development. This flexible platform serves as an important step in advancing the development of innovative photodynamic therapies for managing antibiotic-resistant infections.

This protocol describes a straightforward and economical method for evaluating the effectiveness of potential photosensitizers in antibacterial photodynamic inactivation (aPDI), using a 96-well plate format combined with an LED panel light source. This approach enables the simultaneous testing of multiple experimental conditions, including different concentrations, compounds, and bacterial strains.

細菌の光力学的不活性化における光活性分子を評価するためのLEDベース In Vitro スクリーニング

The rise of multi-drug-resistant infections and the lack of new antibiotic classes have renewed interest in alternative therapies like photodynamic ...

Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has been challenged. Here, we fabricate iron oxide nanoparticle shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform. Iron oxide nanoparticles serve as both magnetic and photothermal agents. Once intravenously injected, NSMs can be magnetically guided to the tumor site. Ultrasound triggers the release of iron oxide nanoparticles, facilitating the penetration of nanoparticles deep into the tumor due to the cavitation effect of microbubbles. Thereafter, magnetic hyperthermia and photothermal therapy can be performed on the tumor for combinational cancer therapy, a solution for cancer resistance due to the tumor heterogeneity. In this protocol, the synthesis and characterization of NSMs including structural, chemical, magnetic and acoustic properties were performed. In addition, the anti-cancer efficacy by thermal therapy was investigated using in vitro cell cultures. The proposed delivery strategy and combination therapy holds great promise in cancer treatment to improve both delivery and anticancer efficacies.

Presented here is a protocol for the fabrication of iron oxide nanoparticle-shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform for magnetic hyperthermia and photothermal combination cancer therapy.

磁気温熱療法とポト温熱併用がん治療のための磁気、音響、光三重応答性マイクロバブル

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has ...

生物工学

An In Vitro Approach to Photodynamic Therapy

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light photoactivation to induce cell apoptosis. The Federal Drug Administration has approved PDT for the treatment of actinic keratosis, and clinical guidelines recommend PDT as a treatment for certain non-melanoma skin cancers and acne vulgaris. PDT is an advantageous therapeutic modality as it is low cost, non-invasive, and associated with minimal adverse events and scaring. In the first step of PDT, a PS is applied and allowed to accumulate intracellularly. Subsequent light irradiation induces reactive oxygen species formation, which may ultimately lead to cell apoptosis, membrane disruption, mitochondrial damage, immune modulation, keratinocyte proliferation, and collagen turnover. Herein, we present an in vitro method to study PDT in an adherent cell line. This treatment protocol is designed to simulate PDT and may be adjusted to studying the use of PDT with various cell lines, photosensitizers, incubation temperatures, or photoactivation wavelengths. Squamous cell carcinoma cells were incubated with 0, 0.5, 1.0, and 2 mM 5-aminolevulinic acid (5-ALA) for 30 min and photoactivated with 417 nm blue light for 1,000 s. The primary outcome measure was apoptosis and necrosis, as measured by annexin-V and 7-aminoactinomycin D flow cytometry. There was a dose-dependent increase in cell apoptosis following thirty-minute incubation of 5-ALA. To achieve high inter-test validity, it is important to maintain consistent incubation and light parameters when performing in vitro PDT experiments. PDT is a useful clinical procedure and in vitro research may allow for the development of novel PSs, optimization of protocols, and new indications for PDT.

An In Vitro Approach to Photodynamic Therapy

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light photoactivation to induce apoptosis. We present an in vitro PDT protocol designed to simulate PDT that may be used to study differences in PS incubation and light treatment parameters.

体外光線力学的治療へのアプローチ

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light ...

医学

Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination model can be used for the evaluation of treatment methods for intrathoracic diseases such as lung cancer or malignant pleural mesothelioma.
Near-infrared photoimmunotherapy (NIR-PIT) is a recently developed cancer treatment strategy that combines the specificity of tumor-targeting antibodies with toxicity caused by a photoabsorber (IR700Dye) after exposure to NIR light. The efficacy of NIR-PIT has been reported using various antibodies; however, only a few reports have shown the therapeutic effect of this strategy in an orthotopic model. In the present study, we demonstrate an example of efficacy evaluation of the pleural disseminated lung cancer model, which was treated using NIR-PIT.

Near-infrared photoimmunotherapy (NIR-PIT) is an emerging cancer therapeutic strategy that utilizes an antibody-photoabsorber (IR700Dye) conjugate and NIR light to destroy cancer cells. Here, we present a method to evaluate the antitumor effect of NIR-PIT in a mouse model of pleural disseminated lung cancer and malignant pleural mesothelioma using bioluminescence imaging.

胸膜普及のマウスモデルに対する近赤外光免疫療法

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination ...

Candida glabrataにおけるプチコロニー形成と薬剤耐性の解析

Induction of Petite Colonies in Candida glabrate via Rose Bengal-Mediated Photodynamic Therapy

Facing a 40% mortality rate in candidemia patients, drug-resistant Candida and their petite mutants remain a major treatment challenge. Antimicrobial photodynamic therapy (aPDT) targets multiple fungal structures, unlike antibiotics/antifungals, potentially thwarting resistance. Traditional methods for inducing petite colonies rely on ethidium bromide or fluconazole, which can influence drug susceptibility and stress responses. This study investigated the application of green light (peak 520 nm) and rose bengal (RB) photosensitizer to combat a drug-resistant Candida glabrata isolate. The findings revealed that aPDT treatment significantly inhibited cell growth (&#8805;99.9% reduction) and effectively induced petite colony formation, as evidenced by reduced size and loss of mitochondrial redox indicator staining. This study provides initial evidence that aPDT can induce petite colonies in a multidrug-resistant C. glabrata strain in vitro, offering a potentially transformative approach for combating resistant fungal infections.

著者スポットライト:抗真菌研究のための薬剤耐性 カンジ ダのプチコロニーを誘導するための新しいアプローチとしての光線力学療法

The significance of petite colonies in Candida spp. drug resistance has not been fully explored. Antimicrobial photodynamic therapy (aPDT) offers a promising strategy against drug-resistant fungal infections. This study demonstrates that rose bengal-mediated aPDT effectively deactivates Candida glabrata and induces petite colonies, presenting a unique procedure.

ローズベンガル媒介光線力学療法によるカンジダ炾粒体におけるプチコロニーの誘導

Facing a 40% mortality rate in candidemia patients, drug-resistant Candida and their petite mutants remain a major treatment challenge. Antimicrobial ...

近赤外Photoimmunotherapy法を用いた混合3D文化からの選択的な細胞除去

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腫瘍スフェロイドモデルにおける抗体依存性細胞傷害性の定量化:創薬への応用

2D細胞培養モデルにおけるベルテポルフィン細胞毒性を高めるための自作発光ダイオードベースの光線力学治療装置

光制御された生物学的に活性な化合物のin vitroおよびin vivo評価-癌光薬理学の潜在的な薬剤候補

細菌の光力学的不活性化における光活性分子を評価するためのLEDベース In Vitro スクリーニング

磁気温熱療法とポト温熱併用がん治療のための磁気、音響、光三重応答性マイクロバブル

体外光線力学的治療へのアプローチ

胸膜普及のマウスモデルに対する近赤外光免疫療法

ローズベンガル媒介光線力学療法によるカンジダ炾粒体におけるプチコロニーの誘導

ローズベンガル媒介光線力学療法によるカンジダ炾粒体におけるプチコロニーの誘導

ローズベンガル媒介光線力学療法によるカンジダ炾粒体におけるプチコロニーの誘導