我々は、この作品[R33 CA174616-01A1]を資金調達NIHから国立癌研究所のIMATプログラムに感謝します。

1.ターゲット細胞の調製<ol><li> 37°Cの時に（10％の熱不活性化ウシ胎児血清（FBS）、4nmのL-グルタミン、500 IU / mlペニシリン/ストレプトマイシンを、RPMI 1640培地）増殖培地中の標的細胞（ 例えば 、Raji細胞リンパ腫細胞）を維持5％CO 2インキュベーター中。 </li><li>遠心分離機15ミリリットルチューブに標的細胞をペレット化するためのサンプル。遠心分離時間は、異なる細胞型によって変化する。 Raji細胞のための468×gで3分間遠心する。 </li><li>最初の上清の表面に形成された気泡を吸引し、全上清を除去。 </li><li>チューブに10mlのPBSを追加します。再懸濁細胞塊を分割するには、上下に数回ソリューションをピペッティングすることにより細胞を。 </li><li>遠心分離機468×gで3分間のサンプル。 </li><li>上清の表面に形成された気泡を吸引し、全上清を除去。 </li></ol>細胞マイクロアレイの作製<ol><li> Obtain細胞付着の96ウェルプレートキットまたは単一セルアレイ8ウェルチャンバースライド（材料/試薬の表を参照）。 </li><li>キットからのDNA試薬にアクティベーターの1.2μlのを適用します。バイアルにキャップを、それを反転し、優しく溶液を混合する振る。 </li><li>ソリューションは、RTで20分間反応させる。 </li><li> （15mlチューブ中で）標的細胞ペレットを混合した溶液を塗布する。 </li><li> 500 nMの最終濃度緑色蛍光細胞質ゾル染料を追加します。 </li><li>オービタルシェーカー上でRTで30分間混合した溶液および色素で標的細胞をインキュベートする。 </li><li> 5mLのPBSで細胞を2回洗浄し、10mlの増殖培地中で細胞を再懸濁する。 </li><li>各ウェルにステップ2.7から溶液100μlを適用します。血球計算盤を用いて細胞数を数える。細胞数はウェル当たり1×10 6の細胞に5×10 4の範囲であることを確認してください。 </li><li> RTでインキュベーションの10〜15分後、静かに200から30で二回のサンプルを洗う未結合の細胞を除去する0μlの増殖培地。 注：約5-10μlの増殖培地は、洗浄後に各ウェルに残っています。 </li></ol> 3.全血細胞毒性アッセイ<ol><li>各サンプルウェルにヒト全血180μlのを追加します。 </li><li>ミリリットル/ 5mgのに抗CD20抗体溶液を得、そして作るために連続希釈を行い50、20、10、5、2,1、1.5 mlチューブ中の抗CD20抗体溶液を0.5μg/ mlであった。よく三重を作る各サンプルに抗CD20抗体の各濃度の20μlのを追加します。 </li><li>各ウェルに500 nMでの最終濃度でヨウ化プロピジウムを追加する。 </li><li> 16時間、5％CO 2、37℃で、得られたプレートをインキュベート。 </li><li>画像スライドまたは8X倍率に自動撮像システム96ウェルプレート上の細胞。 </li><li>結果を定量化するために自動撮像システムの細胞計数プログラムを使用します。 NOTE：自動撮像システムのソフトウェアは、単純な提供細胞計数のためのステップバイステップの操作。このソフトウェアを使用するプロトコルは、プロバイダのウェブサイトで見つけることができます。 </li><li>ソフトウェアのメインページから[レビュー結果]を選択します。 </li><li> [サンプルのために保存したファイル]を選択します。 </li><li>最初のゲートは、FITCその後ゲートが強度&gt; 20を意味TxRed、&gt; 50強さを意味します。 </li><li> [プレートの概要]を選択します。 </li><li> [エクスポートテーブル]を選択します。 注：プログラムのパーセンテージ細胞殺傷式を以下に示す： ％の標的細胞の死滅=（緑と赤の両方の染色された細胞の数/細胞の＃は緑色に染色）* 100％ </li></ol>

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著者は、競合する経済的利益を持っていません。

WCAは、理想的にはそのようなCDCおよびADCCアッセイ9-11、などと前臨床動物試験の前に、伝統的なターゲットの上映の後に利用され、単一細胞の解像度12-16、を用いたin vitro抗がんスクリーニングツールにおいて重要である。現在、CDCまたはADCCアッセイの一次標的スクリーニングアッセイは、すべての単純化されたメディアまたは緩衝液系中で実施される。?...

製薬業界における最近の進歩は、腫瘍細胞抗体パーソナライズ癌治療の特定の識別を実現する上で関心が高まってきた。しかし、いくつかの障害は、プロセスで遭遇している。前臨床開発中の抗がん活性を有する薬剤のわずか5％がフェーズII-III試験の1,2で十分な有効性を示した後にライセンスされています。多くの例では抗腫瘍薬3-5主としてそれらの異なる血液成分にヒトおよび実験動物では異なる挙動を示すことが示されているように（in vitroおよびin vivoの両方で ）前臨床戦略は最適以下である。 抗腫瘍薬物スクリーニングプラットフォームの必要性に対処するために費用のかかる動物実験および臨床試験前のチェックポイントを提供するために、ヒト全血の細胞毒性アッセイ（WCA）は、より関連性の生物学的環境において抗腫瘍薬の有効性を評価するために提案されている。ザ·全血の細胞毒性アッセイは、抗体およびヒト全血の他の薬物候補への個々の細胞の応答を評価するために用いることができる。 WCAは、ハイスループットおよびハイコンテントイメージング6でハイスループット細胞測位技術を組み込むことによって現像される。自動化されたイメージングシステムを利用することにより、生きている細胞および死細胞の両方の数を高精度に判定することができる。による標的細胞は、同一の焦点面上に固定されているという事実のため、WCAは、赤血球を除去することなく、リアルタイムでの定量的細胞毒性分析を提供することができる。また、自動イメージングシステムは、その指定した基準に達した場合にのみ、標的細胞（ 例えば 、蛍光標識された細胞、および細胞形態）などのいくつかの利点ゲートされ、処理さを提供する。また、一日あたり144のプレートを製造することができる。したがって、この撮像能力とスループットが高いcの実行を可能にするontentハイスループット実験を同時に。 WCAおよび自動画像化システムを組み合わせることにより、高スループット定量的細胞毒性分析は、複数の生物学的関連性環境内で達成することができる。

<table><tbody><tr><td>Cell attachment 96 well plate kits&amp;nbsp;</td><td>Adheren</td><td>AP9601</td><td></td></tr><tr><td>Suspension Single cell array 8 well chamber slide</td><td>Adheren</td><td>SS0801</td><td></td></tr><tr><td>Adherent Single cell array 8 well chamber slide</td><td>Adheren</td><td>SS0802</td><td></td></tr><tr><td>Lymphoma cell line CD20+</td><td>ATCC</td><td>CCL-86</td><td>Raji cells</td></tr><tr><td>Lymphoma cell line CD20-</td><td>ATCC</td><td>CRL-8083</td><td>MC/CAR</td></tr><tr><td>RPMI 1640 with L-glutamine</td><td>Life Technologies</td><td>11875-119</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Thermo</td><td>SH30070.01HI</td><td></td></tr><tr><td>Peni/Strep</td><td>Life Technologies</td><td>15070063</td><td></td></tr><tr><td>Cytosolic dye</td><td>Life Technologies</td><td>C7025</td><td>Cell Tracker Green</td></tr><tr><td>Rituxan (Biosimilar)&amp;nbsp;</td><td>Eureka Therapeutics</td><td></td><td></td></tr><tr><td>Human whole blood</td><td>Allcells</td><td>WB001</td><td></td></tr><tr><td>Propidium Iodide</td><td>Sigma</td><td>P4170-10MG</td><td></td></tr><tr><td>Automatic imaging system</td><td>Molecular Devices</td><td>Contact Vendor</td><td>Cell Reporter</td></tr><tr><td>Cell counting program</td><td>Molecular Devices</td><td>Contact Vendor</td><td>Cell Reporter</td></tr></tbody></table>

real-time cytotoxicity assays in human whole blood

全体的な細胞毒性の時間情報へのアクセスを可能にする生細胞ベースの全血の細胞毒性アッセイ（WCA）は、ハイスループット細胞測位技術で開発されている。標的腫瘍細胞集団は、最初の配列形式に固定化予めプログラム、および緑色蛍光の細胞質ゾル色素で標識されている。セルアレイを形成した後、抗体医薬品は、ヒト全血との組合せで添加される。ヨウ化プロピジウム（PI）は、細胞死を評価するために添加される。セル配列は、自動画像化システムを用いて分析される。細胞質ゾルの色素が標的腫瘍細胞集団にラベルを付けながら、PIは死んだ腫瘍細胞集団にラベルを付けます。従って、標的癌細胞の殺傷の割合は死んだ標的細胞の数の標的細胞の生存数を計算することによって定量することができる。この方法では、研究者は、時間依存性および用量依存性の細胞毒性の情報にアクセスすることができる。驚くべきことに、有害ラジオません化学物質が使用されている。 WCAは、ここで提示リンパ腫、白血病、および固形腫瘍細胞株を用いて試験された。したがって、WCAは研究者が、関連性の高いex vivoの状態で薬効を評価することができます。

抗CD20抗体およびリンパ腫細胞（Raji細胞およびMC / CAR）は、全血の細胞毒性アッセイ（WCA）7,8を実証するためのモデル系として選択した。 MC / CAR細胞がその膜上のCD20の低コピー数を有していたRaji細胞では、細胞表面上のCD20の高いコピー数を有していた。標的細胞は、最初に緑色蛍光細胞質色素で緑色に染色し、96ウェルプレート上に配列した。万 - 5万標的リンパ腫細胞を、各ウェル?...

Watch this Scientific Journal Video about Real-time Cytotoxicity Assays in Human Whole Blood at JoVE.com

Real-time Cytotoxicity Assays in Human Whole Blood

全血細胞毒性アッセイ（WCA）は、蛍光顕微鏡および自動化された画像処理をハイスループット細胞測位技術を組み込むことによって開発された細胞毒性アッセイである。ここでは、抗CD20抗体で処置したリンパ腫細胞の定量的な細胞傷害性分析を提供するために、ヒト全血中でリアルタイムに分析することができる方法について説明します。

ヒト全血でのリアルタイム細胞毒性アッセイ

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

real-time-cytotoxicity-assays-in-human-whole-blood

Research

JoVE Journal

Medicine

12.1K ビュー.  Adheren, Inc. 全血細胞毒性アッセイ（WCA）は、蛍光顕微鏡および自動化された画像処理をハイスループット細胞測位技術を組み込むことによって開発された細胞毒性アッセイである。ここでは、抗CD20抗体で処置したリンパ腫細胞の定量的な細胞傷害性分析を提供するために、ヒト全血中でリアルタイムに分析することができる方法について説明します。 

ビデオ: Real-time Cytotoxicity Assays in Human Whole Blood

Determining Optimal Cytotoxic Activity of Human Her2neu Specific CD8 T cells by Comparing the Cr51 Release Assay to the xCELLigence System

Cytotoxic CD8 T cells constitute a subgroup of T cells that are capable of inducing the death of infected or malignant host cells1. These cells express a specialized receptor, called the T cell receptor (TCR), which can recognize a specific antigenic peptide bound to HLA class I molecules2. Engagement of infected cells or tumor cells through their HLA class I molecule results in production of lytic molecules such as granzymes and perforin resulting in target cell death. While it is useful to determine frequencies of antigen-specific CD8 T cells using assays such as the ELIspot or flow cytometry, it is also helpful to ascertain the strength of CD8 T cell responses using cytotoxicity assays3. The most recognizable assay for assessing cytotoxic function is the Chromium Release Assay (CRA), which is considered a standard assay 4. The CRA has several limitations, including exposure of cells to gamma radiation, lack of reproducibility, and a requirement for large numbers of cells. Over the past decade, there has been interest in adopting new strategies to overcome these limitations. Newer approaches include those that measure caspase 
release 4, BLT esterase activity 5 and surface expression of CD107 6. The impedance-based assay, using the Roche xCelligence system, was examined in the present paper for its potential as an alternative to the CRA. Impedance or opposition to an electric current occurs when adherent tumor cells bind to electrode plates. Tumor cells detach following killing and electrical impedance is reduced which can be measured by the xCelligence system. The ability to adapt the impedance-based approach to assess cell-mediated killing rests on the observation that T cells do not adhere tightly to most surfaces and do not appear to have much impact on impedance thus diminishing any concern of direct interference of the T cells with the measurement. Results show that the impedance-based assay can detect changes in the levels of antigen-specific cytotoxic CD8 T cells with increased sensitivity relative to the standard CRA. Based on these results, impedance-based approaches may be good alternatives to CRAs or other approaches that aim to measure cytotoxic CD8 T cell functionality.

The chromium release assay, a common assay for detecting cytotoxic T cell activity, has several limitations. Using antigen-specific CD8 T cells and the human breast cancer tumor line, SKBR3, in the present article, an impedance-based approach was examined for the capability of detecting cell killing.

xCELLigenceシステムにCR51リリースアッセイを比較することによって、人間Her2neu特異的CD8 T細胞の最適な細胞傷害活性を決定する

Cytotoxic CD8 T cells constitute a subgroup of T  cells that are capable of inducing the death of infected or malignant host  cells1. These cells express ...

免疫・感染学

Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells

Cancer immunotherapy can harness the specificity of immune response to target and eliminate tumors. Adoptive cell therapy (ACT) based on the adoptive transfer of T cells genetically modified to express a chimeric antigen receptor (CAR) has shown considerable promise in clinical trials1-4. There are several advantages to using CAR+ T cells for the treatment of cancers including the ability to target non-MHC restricted antigens and to functionalize the T cells for optimal survival, homing and persistence within the host; and finally to induce apoptosis of CAR+ T cells in the event of host toxicity5.
Delineating the optimal functions of CAR+ T cells associated with clinical benefit is essential for designing the next generation of clinical trials. Recent advances in live animal imaging like multiphoton microscopy have revolutionized the study of immune cell function in vivo6,7. While these studies have advanced our understanding of T-cell functions in vivo, T-cell based ACT in clinical trials requires the need to link molecular and functional features of T-cell preparations pre-infusion with clinical efficacy post-infusion, by utilizing in vitro assays monitoring T-cell functions like, cytotoxicity and cytokine secretion. Standard flow-cytometry based assays have been developed that determine the overall functioning of populations of T cells at the single-cell level but these are not suitable for monitoring conjugate formation and lifetimes or the ability of the same cell to kill multiple targets8.
Microfabricated arrays designed in biocompatible polymers like polydimethylsiloxane (PDMS) are a particularly attractive method to spatially confine effectors and targets in small volumes9. In combination with automated time-lapse fluorescence microscopy, thousands of effector-target interactions can be monitored simultaneously by imaging individual wells of a nanowell array. We present here a high-throughput methodology for monitoring T-cell mediated cytotoxicity at the single-cell level that can be broadly applied to studying the cytolytic functionality of T cells.

We describe a single-cell high-throughput assay to measure cytotoxicity of T cells when incubated with tumor target cells. This method employs a dense, elastomeric array of sub-nanoliter wells (~100,000 wells/array) to spatially confine the T cells and target cells at defined ratios and is coupled to fluorescence microscopy to monitor effector-target conjugation and subsequent apoptosis.

T細胞の定量的なハイスループットシングルセル細胞毒性アッセイ

Cancer immunotherapy can harness the specificity of  immune response to target and eliminate tumors. Adoptive cell therapy (ACT)  based on the adoptive ...

An Automated Differential Nuclear Staining Assay for Accurate Determination of Mitocan Cytotoxicity

The contribution of mitochondria to oncogenic transformation is a subject of wide interest and active study. As the field of cancer metabolism becomes more complex, the goal of targeting mitochondria using various compounds that inflict mitochondrial damage (so-called mitocans) is becoming quite popular. Unfortunately, many existing cytotoxicity assays, such as those based on tetrazolium salts or resazurin require functional mitochondrial enzymes for their performance. The damage inflicted by compounds that target mitochondria often compromises the accuracy of these assays. Here, we describe a modified protocol based on differential staining with two fluorescent dyes, one of which is cell-permeant (Hoechst 33342) and the other of which is not (propidium iodide). The difference in staining allows living and dead cells to be discriminated. The assay is amenable to automated microscopy and image analysis, which increases throughput and reduces bias. This also allows the assay to be used in high-throughput fashion using 96-well plates, making it a viable option for drug discovery efforts, particularly when the drugs in question have some level of mitotoxicity. Importantly, results obtained by Hoechst/PI staining assay show increased consistency, both with trypan blue exclusion results and between biological replicates when the assay is compared to other methods.

The protocol describes a rapid, high-throughput, reliable, inexpensive, and unbiased assay for efficiently determining cellular viability. This assay is particularly useful when cells&#39; mitochondria have been damaged, which interferes with other assays. The assay uses automated counting of cells stained with two nuclear dyes &#8211; Hoechst 33342 and propidium iodide.

マイトカン細胞毒性の正確な決定のための自動差動核染色アッセイ

The contribution of mitochondria to oncogenic transformation is a subject of wide interest and active study. As the field of cancer metabolism becomes ...

がん研究

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Cytolytic activity of CD8+ T cells is rarely evaluated. We describe here a new cell-based assay to measure the capacity of antigen-specific CD8+ T cells to kill CD4+ T cells loaded with their cognate peptide. Target CD4+ T cells are divided into two populations, labeled with two different concentrations of CFSE. One population is pulsed with the peptide of interest (CFSE-low) while the other remains un-pulsed (CFSE-high). Pulsed and un-pulsed CD4+ T cells are mixed at an equal ratio and incubated with an increasing number of purified CD8+ T cells. The specific killing of autologous target CD4+ T cells is analyzed by flow cytometry after coculture with CD8+ T cells containing the antigen-specific effector CD8+ T cells detected by peptide/MHCI tetramer staining. The specific lysis of target CD4+ T cells measured at different effector versus target ratios, allows for the calculation of lytic units, LU30/106 cells. This simple and straightforward assay allows for the accurate measurement of the intrinsic capacity of CD8+ T cells to kill target CD4+ T cells.

This protocol describes a sensitive, cell-based cytotoxicity assay. By enumerating the decrease in frequency of live target CD4+ T cells in the presence of an increasing number of effector CD8+ T cells, this assay allows for the direct assessment of cytolytic activity of antigen-specific CD8+ T cells.

Cytolytic activity of CD8+ T cells is rarely evaluated. We describe here a new cell-based assay to measure the capacity of antigen-specific CD8+ T cells ...

Flow Cytometric Analysis of Natural Killer Cell Lytic Activity in Human Whole Blood

NK cell cytotoxicity is a widely used measure to determine the effect of outside intervention on NK cell function. However, the accuracy and reproducibility of this assay can be considered unstable, either because of user&#39;s errors or because of the sensitivity of NK cells to experimental manipulation. To eliminate these issues, a workflow that reduces them to a minimum was established and is presented here. To illustrate, we obtained blood samples, at various time points, from runners (n = 4) that were submitted to an intense bout of exercise. First, NK cells are simultaneously identified and isolated through CD56 tagging and magnetic-based sorting, directly from whole blood and from as little as one milliliter. The sorted NK cells are removed of any reagent or capping antibodies. They can be counted in order to establish an accurate NK cell count per milliliter of blood. Secondly, the sorted NK cells (effectors cells or E) can be mixed with 3,3&#39;-Diotadecyloxacarbocyanine Perchlorate (DiO) tagged K562 cells (target cells or T) at an assay-optimal 1:5 T:E ratio, and analyzed using an imaging flow-cytometer that allows for the visualization of each event and the elimination of any false positive or false negatives (such as doublets or effector cells). This workflow can be completed in about 4 h, and allows for very stable results even when working with human samples. When available, research teams can test several experimental interventions in human subjects, and compare measurements across several time points without compromising the data&#39;s integrity.

This work describes an advanced workflow for the accurate and fast determination of NK (Natural Killer) cell count and NK cell cytotoxicity in human blood samples.

ヒト全血中のナチュラルキラー細胞溶解活性のフローサイトメトリー分析

NK cell cytotoxicity is a widely used measure to determine the effect of outside intervention on NK cell function. However, the accuracy and ...

A Flow Cytometry-Based Cytotoxicity Assay for the Assessment of Human NK Cell Activity

Within the innate immune system, effector lymphocytes known as natural killer (NK) cells play an essential role in host defense against aberrant cells, specifically eliminating tumoral and virally infected cells. Approximately 30 known monogenic defects, together with a host of other pathological conditions, cause either functional or classic NK cell deficiency, manifesting in reduced or absent cytotoxic activity. Historically, cytotoxicity has been investigated with radioactive methods, which are cumbersome, expensive and potentially hazardous. This article describes a streamlined, clinically applicable flow cytometry-based method to quantify NK cell cytotoxic activity. In this assay, peripheral blood mononuclear cells (PBMCs) or purified NK cell preparations are co-incubated at different ratios with a target tumor cell line known to be sensitive to NK cell-mediated cytotoxicity (NKCC). The target cells are pre-labeled with a fluorescent dye to allow their discrimination from the effector cells (NK cells). After the incubation period, killed target cells are identified by a nucleic acid stain, which specifically permeates dead cells. This method is amenable to both diagnostic and research applications and, thanks to the multi-parameter capabilities of flow cytometry, has the added advantage of potentially enabling a deeper analysis of NK cell phenotype and function.

A flow cytometry-based method to quantitatively determine the cytotoxic activity of human natural killer cells is shown here.

ヒト NK 細胞活性の評価の流れのフローサイトメトリーを用いた細胞毒性の試金

Within the innate immune system, effector lymphocytes known as natural killer (NK) cells play an essential role in host defense against aberrant cells, ...

A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells

Chimeric antigen receptor (CAR) T-cell therapy for cancer has achieved significant clinical benefit for resistant and refractory hematological malignancies such as childhood acute lymphocytic leukemia. Efforts are currently underway to extend this promising therapy to solid tumors in addition to other hematological cancers. Here, we describe the development and production of potent CAR T cells targeting antigens with unique or preferential expression on solid and liquid tumor cells. The in vitro potency of these CAR T cells is then evaluated in real-time using the highly sensitive impedance-based xCELLigence assay. Specifically, the impact of different costimulatory signaling domains, such as glucocorticoid-induced tumor necrosis factor receptor (TNFR)-related protein (GITR), on the in vitro potency of CAR T cells is examined. This report includes protocols for: generating CAR T cells for preclinical studies using lentiviral gene transduction, expanding CAR T cells, validating CAR expression, and running and analyzing xCELLigence potency assays.

We describe a quantitative real-time in vitro cytolysis assay system to evaluate the potency of chimeric antigen receptor T cells targeting liquid and solid tumor cells. This protocol can be extended to assess other immune effector cells, as well as combination treatments.

固体および造形癌細胞を標的とするキメラ抗原受容体T細胞のリアルタイム効力アッセイ

Chimeric antigen receptor (CAR) T-cell therapy for cancer has achieved significant clinical benefit for resistant and refractory hematological ...

<meta charset="utf8"></head><body>真菌およびウイルス免疫の分析に最適化された全血イムノアッセイ

Whole Blood Assay with Dual Co-Stimulation for Antigen-Specific Analysis of Host Immunity to Fungal and Viral Pathogens

Rapid and resource-efficient sample processing, high throughput, and high robustness are critical for effective scientific and clinical application of advanced antigen-specific immunoassays. Traditionally, such immunoassays, especially antigen-specific T-cell analysis by flow cytometry or enzyme-linked immunosorbent spot assays, often rely on the isolation of peripheral blood mononuclear cells. This process is time-consuming, subject to many pre-analytic confounders, and requires large blood volumes. Whole blood-based assays provide a facile alternative with increased pre-analytic robustness and lower blood volume requirements. Furthermore, whole blood-based assays allow for the preservation of inter-cellular interactions that are not captured by assays using isolated cell subsets. Recently, a refined whole blood immunoassay with dual anti-CD28 and anti-CD49d co-stimulation for comprehensive analysis of both antigen-specific T-cell functions and complex intercellular interactions in response to various fungal and viral antigens has been proposed. This protocol provides guidance for the preparation of stimulation tubes, blood stimulation, and downstream sample processing for flow cytometry, cytokine secretion assays, and transcriptional analyses. This includes a validated and functionally equivalent, previously unpublished, low-volume protocol (250 &#181;L) to make flow cytometric and cytokine-based T-cell monitoring more accessible for studies in pediatric patients or preclinical studies in small animals (e.g., mice). Altogether, these protocols provide a versatile toolbox for complex antigen-specific immune analysis in both clinical and translational research settings.

<meta charset="utf8"></head><body>著者スポットライト:全血アッセイを使用した救命救急患者における免疫モニタリングの進歩

Whole blood-based immunoassays provide a facile and resource-efficient tool to analyze antigen-specific immunity for diagnostic and research purposes. This article provides an optimized whole blood-based protocol with dual co-stimulation for comprehensive analysis of host immunity to fungal and viral pathogens, including a low-volume version for pediatric patients and small animals.

真菌およびウイルス性病原体に対する宿主免疫の抗原特異的解析のための二重共刺激による全血アッセイ

Rapid and resource-efficient sample processing, high throughput, and high robustness are critical for effective scientific and clinical application of ...

ヒト全血でのリアルタイム細胞毒性アッセイ

要約

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xCELLigenceシステムにCR51リリースアッセイを比較することによって、人間Her2neu特異的CD8 T細胞の最適な細胞傷害活性を決定する

T細胞の定量的なハイスループットシングルセル細胞毒性アッセイ

マイトカン細胞毒性の正確な決定のための自動差動核染色アッセイ

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

ヒト全血中のナチュラルキラー細胞溶解活性のフローサイトメトリー分析

ヒト NK 細胞活性の評価の流れのフローサイトメトリーを用いた細胞毒性の試金

固体および造形癌細胞を標的とするキメラ抗原受容体T細胞のリアルタイム効力アッセイ

真菌およびウイルス性病原体に対する宿主免疫の抗原特異的解析のための二重共刺激による全血アッセイ

真菌およびウイルス性病原体に対する宿主免疫の抗原特異的解析のための二重共刺激による全血アッセイ

真菌およびウイルス性病原体に対する宿主免疫の抗原特異的解析のための二重共刺激による全血アッセイ