Name: near infrared photoimmunotherapy for mouse models of pleural dissemination
Uploaded: 2021-02-09T12:30:00
Description: Watch this Scientific Journal Video about Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination at JoVE.com

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所有活体实验均按照名古屋大学动物护理和使用委员会实验室动物资源的护理和使用指南（批准 #2017-29438，#2018-30096，#2019-31234，#2020-20104）进行。在名古屋大学动物中心购买并饲养了六周大的同源性裸体小鼠。在小鼠进行手术时，他们麻醉时会用异黄素（介绍：4-5%，维持2-3%）：爪子被用钳子压住， 以确认麻醉的深度。
1. IR700 与 mAb 的结合
<ol><li>孵化 mAb ...

<ol>
	<li>Xu, X., Lu, H., Lee, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+Infrared+Light+Triggered+Photo/Immuno-Therapy+Toward+Cancers.">Near Infrared Light Triggered Photo/Immuno-Therapy Toward Cancers.</a> Frontiers in Bioengineering and Biotechnology. 8, (2020).</li><li>Mitsunaga, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+cell-selective+in+vivo+near+infrared+photoimmunotherapy+targeting+specific+membrane+molecules.">Cancer cell-selective in vivo near infrared photoimmunotherapy targeting specific membrane molecules.</a> Nature Medicine. 17, 1685-1691 (2011).</li><li>Kobayashi, H., Choyke, P. 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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy+and+biodistribution+with+an+OC125-chlorin+immunoconjugate+in+an+in+vivo+murine+ovarian+cancer+model.">Photoimmunotherapy and biodistribution with an OC125-chlorin immunoconjugate in an in vivo murine ovarian cancer model.</a> British Journal of Cancer. 70, 474-480 (1994).</li><li>Sato, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoinduced+Ligand+Release+from+a+Silicon+Phthalocyanine+Dye+Conjugated+with+Monoclonal+Antibodies:+A+Mechanism+of+Cancer+Cell+Cytotoxicity+after+Near-Infrared+Photoimmunotherapy.">Photoinduced Ligand Release from a Silicon Phthalocyanine Dye Conjugated with Monoclonal Antibodies: A Mechanism of Cancer Cell Cytotoxicity after Near-Infrared Photoimmunotherapy.</a> ACS Central Science. 4, 1559-1569 (2018).</li><li>Sato, K., Nagaya, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+in+the+treatment+of+pleural+disseminated+NSCLC:+Preclinical+experience.">Near infrared photoimmunotherapy in the treatment of pleural disseminated NSCLC: Preclinical experience.</a> Theranostics. 5, 698-709 (2015).</li><li>Nishinaga, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+Phototherapy+for+Malignant+Pleural+Mesothelioma:+Near-Infrared+Photoimmunotherapy+Targeting+Podoplanin.">Targeted Phototherapy for Malignant Pleural Mesothelioma: Near-Infrared Photoimmunotherapy Targeting Podoplanin.</a> Cells. 9, 1019 (2020).</li><li>Sato, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+prevents+lung+cancer+metastases+in+a+murine+model.">Near infrared photoimmunotherapy prevents lung cancer metastases in a murine model.</a> Oncotarget. 6, 19747-19758 (2015).</li><li>Sato, K., Nagaya, T., Mitsunaga, M., Choyke, P. 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L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy+of+Gastric+Cancer+Peritoneal+Carcinomatosis+in+a+Mouse+Model.">Photoimmunotherapy of Gastric Cancer Peritoneal Carcinomatosis in a Mouse Model.</a> PLoS One. 9, 113276 (2014).</li><li>McLemore, T. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+intrapulmonary,+percutaneous+intrathoracic,+and+subcutaneous+models+for+the+propagation+of+human+pulmonary+and+nonpulmonary+cancer+cell+lines+in+athymic+nude+mice.">Comparison of intrapulmonary, percutaneous intrathoracic, and subcutaneous models for the propagation of human pulmonary and nonpulmonary cancer cell lines in athymic nude mice.</a> Cancer Research. 48, 2880-2886 (1988).</li><li>Manzotti, C., Audisio, R. A., Pratesi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Importance+of+orthotopic+implantation+for+human+tumors+as+model+systems:+relevance+to+metastasis+and+invasion.">Importance of orthotopic implantation for human tumors as model systems: relevance to metastasis and invasion.</a> Clinical &amp; Experimental Metastasis. 11, 5-14 (1993).</li><li>Lwin, T. M., Hoffman, R. M., Bouvet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advantages+of+patient-derived+orthotopic+mouse+models+and+genetic+reporters+for+developing+fluorescence-guided+surgery.">Advantages of patient-derived orthotopic mouse models and genetic reporters for developing fluorescence-guided surgery.</a> Journal of Surgical Oncology. 118, 253-264 (2018).</li><li>Sordat, B. C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> From Ectopic to Orthotopic Tumor Grafting Sites: Evidence for a Critical Role of the Host Tissue Microenvironment for the Actual Expression of the Malignant Phenotype. , 43-53 (2017).</li><li>Sato, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy:+comparative+effectiveness+of+two+monoclonal+antibodies+targeting+the+epidermal+growth+factor+receptor.">Photoimmunotherapy: comparative effectiveness of two monoclonal antibodies targeting the epidermal growth factor receptor.</a> Molecular Oncology. 8, 620-632 (2014).</li><li>Nakajima, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+conjugate+and+light+dose+on+photo-immunotherapy+induced+cytotoxicity.">The effects of conjugate and light dose on photo-immunotherapy induced cytotoxicity.</a> BMC Cancer. 14, 389 (2014).</li><li>Nagaya, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+of+B-cell+lymphoma.">Near infrared photoimmunotherapy of B-cell lymphoma.</a> Molecular Oncology. 10, 1404-1414 (2016).</li><li>Sato, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+in+the+treatment+of+disseminated+peritoneal+ovarian+cancer.">Near infrared photoimmunotherapy in the treatment of disseminated peritoneal ovarian cancer.</a> Molecular Cancer Therapeutics. 14, 141-150 (2015).</li><li>Colin, D. J., Bejuy, O., Germain, S., Triponez, F., Serre-Beinier, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantation+and+monitoring+by+pet/ct+of+an+orthotopic+model+of+human+pleural+mesothelioma+in+athymic+mice.">Implantation and monitoring by pet/ct of an orthotopic model of human pleural mesothelioma in athymic mice.</a> Journal of Visualized Experiments. 2019, (2019).</li><li>Opitz, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+recurrence+model+of+malignant+pleural+mesothelioma+for+investigation+of+intrapleural+treatment.">Local recurrence model of malignant pleural mesothelioma for investigation of intrapleural treatment.</a> European Journal of Cardio-Thoracic Surgery. 31, 772-778 (2007).</li><li>Bunn, P. A., Kelly, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+chemotherapeutic+agents+prolong+survival+and+improve+quality+of+life+in+non-small+cell+lung+cancer:+a+review+of+the+literature+and+future+directions.">New chemotherapeutic agents prolong survival and improve quality of life in non-small cell lung cancer: a review of the literature and future directions.</a> Clinical Cancer Research. 4, 1087-1100 (1998).</li><li>Astoul, P., Wang, X., Hoffman, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-like+nude-mouse+and+scid-mouse+models+of+human+lung+and+pleural+cancer+(review).">Patient-like nude-mouse and scid-mouse models of human lung and pleural cancer (review).</a> International Journal of Oncology. 3, 713-718 (1993).</li><li>Yamaguchi, H., Pantarat, N., Suzuki, T., Evdokiou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+photoimmunotherapy+using+a+small+protein+mimetic+for+HER2-overexpressing+breast+cancer.">Near-infrared photoimmunotherapy using a small protein mimetic for HER2-overexpressing breast cancer.</a> International Journal of Molecular Sciences. 20, (2019).</li><li>Jing, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+and+selective+elimination+of+glioblastoma+stem+cells+with+theranostic+Near-Infrared-Labeled+CD133-Specific+antibodies.">Imaging and selective elimination of glioblastoma stem cells with theranostic Near-Infrared-Labeled CD133-Specific antibodies.</a> Theranostics. 6, 862-874 (2016).</li><li>Burley, T. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+photoimmunotherapy+targeting+EGFR-Shedding+new+light+on+glioblastoma+treatment.">Near-infrared photoimmunotherapy targeting EGFR-Shedding new light on glioblastoma treatment.</a> International Journal of Cancer. 142, 2363-2374 (2018).</li><li>Nagaya, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+using+a+fiber+optic+diffuser+for+treating+peritoneal+gastric+cancer+dissemination.">Near infrared photoimmunotherapy using a fiber optic diffuser for treating peritoneal gastric cancer dissemination.</a> Gastric Cancer. 22, 463-472 (2019).</li><li>Nagaya, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoscopic+near+infrared+photoimmunotherapy+using+a+fiber+optic+diffuser+for+peritoneal+dissemination+of+gastric+cancer.">Endoscopic near infrared photoimmunotherapy using a fiber optic diffuser for peritoneal dissemination of gastric cancer.</a> Cancer Science. 109, 1902-1908 (2018).</li><li>Harada, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+photoimmunotherapy+with+galactosyl+serum+albumin+in+a+model+of+diffuse+peritoneal+disseminated+ovarian+cancer.">Near-infrared photoimmunotherapy with galactosyl serum albumin in a model of diffuse peritoneal disseminated ovarian cancer.</a> Oncotarget. 7, 79408-79416 (2016).</li><li>Journals, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=JNCI+Journal+of+the+National+Cancer+Institute+Way+to+Better+DNA.">JNCI Journal of the National Cancer Institute Way to Better DNA.</a> Annals of Internal Medicine. 37, 1-9 (2008).</li></ol>

在这项研究中，我们展示了一种测量NIR-PIT对MPM胸膜传播模型的治疗效果的方法。用NIR-PIT进行了高度选择性的细胞杀戮;因此，正常组织几乎没有损坏23，24，25。通过这种选择性细胞杀伤，NIR-PIT在传播的9、26型中被证明是安全的。但是，某些步骤可能采用替代...

尽管进行了数十年的研究，癌症仍然是导致死亡的主要原因之一。原因之一是放射治疗和化疗是高侵入性技术，这可能限制其治疗效果。细胞或分子靶向疗法，这是侵入性较小的技术，正受到越来越多的关注。光免疫疗法是一种通过结合免疫治疗和光疗协同提高治疗效果的治疗方法。免疫疗法通过增加肿瘤微环境的免疫原性，减少免疫调节抑制，提高肿瘤免疫力，导致肿瘤在体内的破坏。光疗通过光敏剂和光线的组合破坏原发性肿瘤，从肿瘤细胞中释放出的肿瘤特异性抗原增强肿瘤免疫力。肿瘤可以选择性地使用光敏剂治疗，因为它们是特定和选择性的目标细胞。光疗模式包括光动力疗法（PDT）、光热疗法（PTT）和光化学疗法1。
近红外光免疫疗法（NIR-PIT）是一种最近开发的抗肿瘤光疗方法，结合了光化学疗法和免疫疗法1，2。NIR-PIT 是一种分子靶向疗法，通过将近红外硅邻苯二甲酸酯染料 IRdye 700DX （IR700） 与单克隆抗体 （mAb） 结合，针对特定细胞表面分子。目标细胞的细胞膜在用NIR光（690纳米）3照射时被破坏。
使用靶向光疗法的概念，结合传统的光敏剂和抗体或靶向PDT是超过30年的历史4，5。先前的研究试图通过将常规PDT制剂与抗体结合来瞄准它们。然而，由于光敏剂6，7的疏水性，这些结合体被困在肝脏中，因此成功有限。此外，NIR-PIT的机制与传统的PDT机制完全不同。传统的光敏剂产生氧化应激，这种应激来自能量转换，这种能量转换吸收光能，脱位到兴奋状态，过渡到地面状态，并导致凋亡。然而，NIR-PIT通过光化学反应8将光敏剂聚合在膜上，直接破坏细胞膜，从而导致快速坏死。NIR-PIT 在很多方面优于传统的目标 PDT。传统光敏剂的消亡系数较低，需要将大量光敏剂附着在单个抗体分子上，从而可能降低结合亲和力。大多数传统光敏剂是疏水剂，因此很难将光敏剂与抗体结合，而不影响其免疫反应或体内目标积累。传统光敏剂通常吸收可见范围内的光线，减少组织渗透。
关于NIR-PIT针对内胸肿瘤，如肺癌和恶性胸膜间皮瘤（MPM）细胞的几项研究已经报告了9，10，11，12，13，14，15，16，17。然而，只有少数报告描述了NIR-PIT在胸膜传播MPM或肺癌模型9，10，11，12的疗效。皮下肿瘤异种模型被认为是标准肿瘤模型，目前广泛用于评估新疗法18的抗肿瘤效果。然而，皮下肿瘤微环境是不允许发展一个适当的组织结构或条件，适当地回顾一个真正的恶性表型19，20，21，22。理想情况下，应建立正位疾病模型，以便更精确地评估抗肿瘤效应。
在这里，我们展示了一种在小鼠模型的胸膜传播肺癌，这是使用NIR-PIT治疗的功效评估方法。通过将肿瘤细胞注射到胸腔中，并使用荧光酶发光进行确认，生成胸腔传播小鼠模型。小鼠接受静脉注射，注射与IR700和NIR照射到胸部的mAb。使用荧光酶发光评估治疗效果。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.25w/v% Trypsin-1mmol/l EDTA 4Na Solution with Phenol Red</td>
			<td>Wako</td>
			<td>209-016941</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>1mL syringe</td>
			<td>TERUMO</td>
			<td>SS-01T</td>
			<td>for mice experiment</td>
		</tr>
		<tr>
			<td>30G needle</td>
			<td>Nipro</td>
			<td>1907613</td>
			<td>for mice experiment</td>
		</tr>
		<tr>
			<td>BALB/cSlc-nu/nu</td>
			<td>Japan SLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Collidal Blue Staining Kit</td>
			<td>Invitrogen</td>
			<td>LC6025</td>
			<td>use for gel protein staining</td>
		</tr>
		<tr>
			<td>Coomassie (bradford) Plus protein assay</td>
			<td>Thermo Fisher Scientific Inc (Waltham, MA, USA)</td>
			<td>PI-23200</td>
			<td>for measuring the APC concentration</td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide (DMSO)</td>
			<td>Wako</td>
			<td>043-07216</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>D-Luciferin (potassium salt)</td>
			<td>Cayman Chemical</td>
			<td>14681</td>
			<td>for bioluminescence imaging and DLIT</td>
		</tr>
		<tr>
			<td>GraphPad Prism7</td>
			<td>GraphPad software</td>
			<td></td>
			<td>for statistical analysis</td>
		</tr>
		<tr>
			<td>Image Studio</td>
			<td>Li-Cor Biosciences</td>
			<td></td>
			<td>for analyzing 700 nm fluorescent image</td>
		</tr>
		<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>isoflurane</td>
			<td>Wako</td>
			<td>095-06573</td>
			<td>for mice anesthesia</td>
		</tr>
		<tr>
			<td>IVIS Spectrum CT</td>
			<td>PerkinElmer</td>
			<td></td>
			<td>for capturing bioluminescent image and DLIT</td>
		</tr>
		<tr>
			<td>Living Image</td>
			<td>PerkinElmer</td>
			<td></td>
			<td>for analyzing bioluminescent image and DLIT</td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>SIGMA-ALDRICH (St. Louis, MO, USA)</td>
			<td>S9763</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>NIR Laser</td>
			<td>Changchun New Industries Optoelectronics Technology</td>
			<td>MRL-III-690R</td>
			<td>for NIR irradiation</td>
		</tr>
		<tr>
			<td>Novex WedgeWell 4 to 20%, Tris-Glycine, 1.0 mm, Mini Protein Gel, 12 well</td>
			<td>Invitrogen</td>
			<td>XP04202BOX</td>
			<td>use for SDS-PAGE</td>
		</tr>
		<tr>
			<td>NuPAGE LDS Sample Buffer (x4)</td>
			<td>Invitrogen</td>
			<td>NP0007</td>
			<td>use for SDS-PAGE</td>
		</tr>
		<tr>
			<td>Optical power meter</td>
			<td>Thorlabs (Newton, NJ, USA)</td>
			<td>PM100</td>
			<td>for measuring the output of the NIR laser&#160;</td>
		</tr>
		<tr>
			<td>PBS(-)</td>
			<td>Wako</td>
			<td>166-23555</td>
			<td></td>
		</tr>
		<tr>
			<td>Pearl Trilogy imaging system</td>
			<td>Li-Cor Biosciences</td>
			<td></td>
			<td>for capturing 700 nm fluorecent image</td>
		</tr>
		<tr>
			<td>Penicilin-Streptomycin Solution (x100)</td>
			<td>Wako</td>
			<td>168-23191</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>Puromycin Dihydrochloride</td>
			<td>ThermoFisher</td>
			<td>A1113803</td>
			<td>for luciferase transfection</td>
		</tr>
		<tr>
			<td>RediFect Red-Fluc-Puromycin Lentiviral Prticles</td>
			<td>PerkinElmer</td>
			<td>CLS960002</td>
			<td>for luciferase transfection</td>
		</tr>
		<tr>
			<td>RPMI-1640 with L-glutamine and Phenol Red</td>
			<td>Wako</td>
			<td>189-02025</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>Sephadex G25 column (PD-10)&#160;</td>
			<td>GE Healthcare (Piscataway, NJ, USA)</td>
			<td>17-0851-01</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>UV-1900i</td>
			<td>Shimadzu</td>
			<td></td>
			<td>for measuring the APC concentration</td>
		</tr>
	</tbody>
</table>

near infrared photoimmunotherapy for mouse models of pleural dissemination

光免疫疗法的疗效可以更准确地评估与矫名小鼠模型比皮下模型。可用于评估肺癌或恶性胸膜间皮瘤等胸内疾病的治疗方法。
近红外光免疫疗法（NIR-PIT）是最近开发的癌症治疗策略，它结合了肿瘤靶向抗体的特异性与光吸收物（IR700Dye）暴露在NIR光下后引起的毒性。NIR-PIT的功效已报告使用各种抗体：然而，只有少数报告在矫名模型中显示了这种策略的治疗效果。在本研究中，我们展示了一个使用NIR-PIT治疗的胸膜传播肺癌模型的疗效评估示例。

抗多普兰素抗体NZ-1与IR700结合产生NZ-1-IR700。我们确认了 NZ-1 和 IR700 在 SDS-PAGE（图 8）上的绑定。Luciferase表达H2373（H2373-luc）是由通过用荧光酶基因10转染恶性间皮瘤细胞（H2373）来制备的。
我们麻醉了8-12周大的雌性同源性同源性裸鼠，并将1×10个H2373-luc细胞注射到胸腔中。将肿瘤细胞注射到小鼠体内的当天被指示为第1天。
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Watch this Scientific Journal Video about Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination at JoVE.com

Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination

近红外光免疫疗法（NIR-PIT）是一种新兴的癌症治疗策略，利用抗体光吸收剂（IR700Dye）结合光和NIR光来破坏癌细胞。在这里，我们提出了一种方法，评估NIR-PIT的抗肿瘤作用，在小鼠模型的胸膜传播肺癌和恶性胸膜间皮瘤使用生物发光成像。

近红外光免疫疗法为小鼠模型的胸膜传播

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

该方法可用于评估NIR-PIT在临床相关肿瘤模型中对胸肿瘤的治疗效果。使用胸膜传播癌症模型的NIR-PIT程序易于理解和执行。要建立鼠标传播模型，请使用聚苯乙烯泡沫制作阻塞剂，并用 70% 乙醇进行消毒。将针头连接到塞子上，尖端从塞子中伸出五毫米。用干净的钳子弯曲一个30米针的尖端，或用70%乙醇清洁的硬物，用PBS的100微升中10倍于6个目标肿瘤细胞填充注射器。确认麻醉后，19至21克，8至12周大，雌性，同源性，同源性裸体鼠标，对踏板反射缺乏反应。将针头通过腹间空间插入胸部，上下移动针头以避免接触肋骨。当尖端穿过间歇空间时，将注射器放置，使其压在小鼠身上，并注入目标细胞的整个体积。注射后，将鼠标卷起两到三次，将细胞扩散到胸腔内，并将鼠标放回笼中进行监测，直到完全恢复。细胞注射后24小时，此后每天给麻醉肿瘤细胞注射小鼠注射200微升，每毫升15毫克D-露西弗林。10 分钟后，将鼠标放入生物发光成像仪中，并在成像仪软件中打开采集控制面板。选择发光、照片和覆盖。将曝光时间设置为自动、从宾宁到小、F/Stop 到一到一用于发光和八到照片，将视场设置为 C.Click 获取以成像生物发光。将显示格式设置为 Radians，并在工具调色板面板中从兴趣区域工具中选择圆圈。单击"感兴趣区域"以测量表面生物发光强度，并使用配置测量选择与实验相关的值。将此数据表导出为 csv 文件。然后使用文件中的生物发光强度量化的总通量值。在对肿瘤注射小鼠进行近红外光疗之前，使用功率计测量690纳米波长激光的光剂量，并将输出调整为每平方厘米100毫瓦。治疗前24小时，通过肿瘤注射动物的尾静脉，静脉注射50至200微升PBS的抗体光敏剂。在光疗治疗当天，将麻醉结合注射的肿瘤小鼠置于苏平位置，用铝箔保护非目标部位。当所有防护罩都放置完毕后，使用每平方厘米 100 焦耳的激光用近红外光照射胸腔约 30 秒。当辐照完成，老鼠醒来后，将动物送回笼子，并测量每天的生物发光。在这项具有代表性的分析中，SDS-PAGE分析证实了抗多普兰素抗体与IR700的结合。肿瘤细胞注射后，应进行生物发光成像和扩散发光成像断层扫描，以确定哪些小鼠在胸腔中表现出足够的荧光酶活性，以便进一步研究。注射后的第五天，抗多普兰素抗体IR700注射小鼠在胸肿瘤中表现出高IR700荧光和荧光酶活性，表明静脉注射的IR700结合抗体到达传播的胸膜肿瘤部位。值得注意的是，接受近红外光免疫疗法治疗的胸膜传播小鼠的荧光素活性降低，而对照组的相对光单元表现出强度的逐渐增加。由于所需的NIR辐照能量取决于细胞系和抗体，请务必在体外提前检查情况。

Research

JoVE Journal

Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

体内近红外荧光（NIRF）血管内分子成像炎性斑块，多式联运方针成像动脉粥样硬化

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

科研

医学

Near Infrared Optical Projection Tomography for Assessments of &beta;-cell Mass Distribution in Diabetes Research

Near Infrared Optical Projection Tomography for Assessments of ÃƒÅ½Ã‚Â²-cell Mass Distribution in Diabetes Research

近红外光学投影层析成像技术在糖尿病研究β细胞质量分布的评估

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of ...

Mouse Models for Graft Arteriosclerosis

嫁接动脉硬化的小鼠模型

Graft arteriosclerois (GA),  also called allograft vasculopathy, is a pathologic lesion that develops over  months to years in transplanted organs ...

Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery

组织鬼怪模拟，以评估乳癌手术潜在的近红外荧光成像中的应用

Inaccuracies in intraoperative tumor localization and evaluation of surgical margin status result in suboptimal outcome of breast-conserving surgery ...

Mouse Kidney Transplantation: Models of Allograft Rejection

鼠肾移植：移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

鼠标颈淋巴结高频超声成像

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

结合近红外荧光成像和微计算机断层扫描直接可视化脑血栓

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

多发性硬化症的小鼠模型EAE视神经炎及脑部炎症的生物发光和近红外成像

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

"一站式" 近红外光谱对骨骼肌神经血管的耦合、氧化能力及微血管功能的作用

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

近红外标记剂对大鼠尾静脉注射的新的最佳验证

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...