Name: Advancing Allergic Rhinitis Research with Multicolor Immunofluorescence
Uploaded: 2023-09-22T14:50:00
Description: Watch this Scientific Journal Video about Advancing Allergic Rhinitis Research with Multicolor Immunofluorescence at JoVE.com

Questo lavoro è stato sostenuto dal Dipartimento provinciale di scienza e tecnologia del Sichuan (2021YJ0175).

Il protocollo e le procedure sperimentali hanno ricevuto l'approvazione del Comitato amministrativo e di ricerca sugli animali dell'Università di Medicina Tradizionale Cinese di Chengdu (Numero di registrazione: 2022DL-010). I ratti maschi di Sprague Dawley (SD) di otto settimane, del peso di 180-200 g, sono stati ottenuti commercialmente (vedi Tabella dei materiali) e alloggiati in un ciclo naturale luce/buio con temperatura controllata (23 ± 2 °C) e umidità relativa (55% ± 10%). Dodici ratti sono ...

Multicolor Immunofluorescence of the Nasal Tissue in an Allergic Rhinitis Rat Model

<ol>
	<li>Bro&#380;ek, J. 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=Allergic+rhinitis+and+its+impact+on+asthma+(ARIA)+Guidelines-2016+revision.">Allergic rhinitis and its impact on asthma (ARIA) Guidelines-2016 revision.</a> The Journal of Allergy and Clinical Immunology. 140 (4), 950-958 (2017).</li><li>Maurer, M., Zuberbier, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Undertreatment+of+rhinitis+symptoms+in+Europe:+findings+from+a+cross-sectional+questionnaire+survey.">Undertreatment of rhinitis symptoms in Europe: findings from a cross-sectional questionnaire survey.</a> Allergy. 62 (9), 1057-1063 (2007).</li><li>Ding, 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=Alpha-linolenic+acid+improves+nasal+mucosa+epithelial+barrier+function+in+allergic+rhinitis+by+arresting+CD4++T+cell+differentiation+via+IL-4R&#945;-JAK2-STAT3+pathway.">Alpha-linolenic acid improves nasal mucosa epithelial barrier function in allergic rhinitis by arresting CD4+ T cell differentiation via IL-4R&#945;-JAK2-STAT3 pathway.</a> Phytomedicine: International Journal of Phytotherapy and Phytopharmacology. 116, 154825 (2023).</li><li>Zhou, 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=Activation+of+NLRP3+inflammasome+contributes+to+the+inflammatory+response+to+allergic+rhinitis+via+macrophage+pyroptosis.">Activation of NLRP3 inflammasome contributes to the inflammatory response to allergic rhinitis via macrophage pyroptosis.</a> International Immunopharmacology. 110, 109012 (2022).</li><li>Watts, A. M., Cripps, A. W., West, N. P., Cox, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+allergic+inflammation+in+the+nasal+mucosa+of+allergic+rhinitis+sufferers+with+topical+pharmaceutical+agents.">Modulation of allergic inflammation in the nasal mucosa of allergic rhinitis sufferers with topical pharmaceutical agents.</a> Frontiers in pharmacology. 10, 294 (2019).</li><li>Patel, N., 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=Fungal+extracts+stimulate+solitary+chemosensory+cell+expansion+in+noninvasive+fungal+rhinosinusitis.">Fungal extracts stimulate solitary chemosensory cell expansion in noninvasive fungal rhinosinusitis.</a> International Forum of Allergy &amp; Rhinology. 9 (7), 730-737 (2019).</li><li>Granier, C., 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=Tim-3+expression+on+tumor-infiltrating+PD-1(+)CD8(+)+T+cells+correlates+with+poor+clinical+outcome+in+renal+cell+carcinoma.">Tim-3 expression on tumor-infiltrating PD-1(+)CD8(+) T cells correlates with poor clinical outcome in renal cell carcinoma.</a> Cancer Research. 77 (5), 1075-1082 (2017).</li><li>Badoua, C., 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=PD-1-ex+pressing+tumor-infiltrating+T+cells+are+a+favorable+prognostic+biomarker+in+HPV-associated+head+and+neck+cancer.">PD-1-ex pressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer.</a> Cancer Research. 73 (1), 128-138 (2013).</li><li>Fedchenko, N., Reifenrath, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+approaches+for+interpretation+and+reporting+of+immunohistochemistry+analysis+results+in+the+bone+tissue+-+a+review.">Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue - a review.</a> Diagnostic Pathology. 9, 221 (2014).</li><li>Faget, L., Hnasko, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tyramide+signal+amplification+for+immunofluorescent+enhancement.">Tyramide signal amplification for immunofluorescent enhancement.</a> Methods in Molecular Biology. 1318, 161-172 (2015).</li><li>Parra, E. R., Villalobos, P., Rodriguez-Canales, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+multiple+faces+of+programmed+cell+death+ligand+1+expression+in+malignant+and+nonmalignant+cells.">The multiple faces of programmed cell death ligand 1 expression in malignant and nonmalignant cells.</a> Applied Immunohistochemistry &amp; Molecular Morphology.: AIMM. 27 (4), 287-294 (2019).</li><li>Welsh, A. W., 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=Standardization+of+estrogen+receptor+measurement+in+breast+cancer+suggests+false-negative+results+are+a+function+of+threshold+intensity+rather+than+percentage+of+positive+cells.">Standardization of estrogen receptor measurement in breast cancer suggests false-negative results are a function of threshold intensity rather than percentage of positive cells.</a> Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology. 29 (22), 2978-2984 (2011).</li><li>Parra, E. R., 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=Validation+of+multiplex+immunofluorescence+panels+using+multispectral+microscopy+for+immune-profiling+of+formalin-fixed+and+paraffin-embedded+human+tumor+tissues.">Validation of multiplex immunofluorescence panels using multispectral microscopy for immune-profiling of formalin-fixed and paraffin-embedded human tumor tissues.</a> Scientific Reports. 7 (1), 13380 (2017).</li><li>Miao, M., Xiang, L., Miao, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specification+for+preparation+of+allergic+rhinitis+animal+model+(draft).">Specification for preparation of allergic rhinitis animal model (draft).</a> Chinese Traditional and Herbal Drugs. 49 (1), 50-57 (2018).</li><li>Xu, J., 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=Astragalus+polysaccharides+attenuate+ovalbumin-induced+allergic+rhinitis+in+rats+by+inhibiting+nlrp3+inflammasome+activation+and+nod2-mediated+nf-+&#954;+b+activation.">Astragalus polysaccharides attenuate ovalbumin-induced allergic rhinitis in rats by inhibiting nlrp3 inflammasome activation and nod2-mediated nf- &#954; b activation.</a> Journal of Medicinal Food. 24 (1), 1-9 (2021).</li><li>Bousquet, J., 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=Allergic+rhinitis.">Allergic rhinitis.</a> Nature reviews. 6 (1), 95 (2020).</li><li>Huang, C., 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=Expression+of+activated+signal+transducer+and+activator+of+transcription-3+as+a+predictive+and+prognostic+marker+in+advanced+esophageal+squamous+cell+carcinoma.">Expression of activated signal transducer and activator of transcription-3 as a predictive and prognostic marker in advanced esophageal squamous cell carcinoma.</a> World Journal of Surgical Oncology. 13, 314 (2015).</li><li>Leynaert, B., Bousquet, J., Neukirch, C., Liard, R., Neukirch, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perennial+rhinitis:+An+independent+risk+factor+for+asthma+in+nonatopic+subjects:+results+from+the+European+Community+Respiratory+Health+Survey.">Perennial rhinitis: An independent risk factor for asthma in nonatopic subjects: results from the European Community Respiratory Health Survey.</a> The Journal of Allergy and Clinical Immunology. 104 (2), 301-304 (1999).</li><li>Guerra, S., 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=Rhinitis+is+an+independent+risk+factor+for+developing+cough+apart+from+colds+among+adults.">Rhinitis is an independent risk factor for developing cough apart from colds among adults.</a> Allergy. 60 (3), 343-349 (2005).</li><li>Greisner, W. A., Settipane, R. J., Settipane, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Co-existence+of+asthma+and+allergic+rhinitis:+a+23-year+follow-up+study+of+college+students.">Co-existence of asthma and allergic rhinitis: a 23-year follow-up study of college students.</a> Allergy and Asthma Proceedings. 19 (4), 185-188 (1998).</li><li>Pawankar, R., Mori, S., Ozu, C., Kimura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overview+on+the+pathomechanisms+of+allergic+rhinitis.">Overview on the pathomechanisms of allergic rhinitis.</a> Asia Pacific Allergy. 1 (3), 157-167 (2011).</li><li>Van Nguyen, 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=Anti-allergic+rhinitis+activity+of+&#945;-lipoic+acid+via+balancing+th17/treg+expression+and+enhancing+nrf2/ho-1+pathway+signaling.">Anti-allergic rhinitis activity of &#945;-lipoic acid via balancing th17/treg expression and enhancing nrf2/ho-1 pathway signaling.</a> Scientific Reports. 10 (1), 12528 (2020).</li><li>Ivanov, I. 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=The+orphan+nuclear+receptor+ror&#947;t+directs+the+differentiation+program+of+proinflammatory+IL-17++T+helper+cells.">The orphan nuclear receptor ror&#947;t directs the differentiation program of proinflammatory IL-17+ T helper cells.</a> Cell. 126 (6), 1121-1133 (2006).</li><li>Fontenot, J. D., Gavin, M. A., Rudensky, A. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Foxp3+programs+the+development+and+function+of+CD4+CD25++regulatory+T+cells.">Foxp3 programs the development and function of CD4+CD25+ regulatory T cells.</a> Nature Immunology. 4 (4), 330-336 (2003).</li><li>Zhou, X., 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=Instability+of+the+transcription+factor+Foxp3+leads+to+the+generation+of+pathogenic+memory+T+cells+in+Vivo.">Instability of the transcription factor Foxp3 leads to the generation of pathogenic memory T cells in Vivo.</a> Nature Immunology. 10 (9), 1000-1007 (2009).</li><li>Ziegler, S. F., Buckner, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Foxp3+and+the+regulation+of+Treg/Th17+differentiation.">Foxp3 and the regulation of Treg/Th17 differentiation.</a> Microbes and Infection. 11 (5), 594-598 (2009).</li><li>Yaling, Y., Gui, Y., Shuqi, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+research+advances+of+toll-like+receptors+and+allergic+rhinitis.">Recent research advances of toll-like receptors and allergic rhinitis.</a> Medical Recapitulate. 23 (7), 1273-1276 (2017).</li><li>Shan, H., Jie, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+between+TLR4+gene+polymorphisms+and+allergic+rhinitis.">Correlation between TLR4 gene polymorphisms and allergic rhinitis.</a> Journal of Clinical Otorhinolaryngology, Head, and Neck Surgery. 31 (13), 991-994 (2017).</li><li>Xu, O., 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=Immunoregulatory+mechanism+of+Treg+cells+and+TICAM-1+patients+with+severe+allergic+rhinitis.">Immunoregulatory mechanism of Treg cells and TICAM-1 patients with severe allergic rhinitis.</a> Chinese Journal of Coal Industry Medicine. 23 (3), 303-306 (2020).</li><li>Granier, C., 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=Tim-3+expression+on+tumor-infiltrating+PD-1+CD8++T+cells+correlates+with+poor+clinical+outcome+in+renal+cell+carcinoma.">Tim-3 expression on tumor-infiltrating PD-1+CD8+ T cells correlates with poor clinical outcome in renal cell carcinoma.</a> Cancer Research. 77 (5), 1075-1082 (2017).</li><li>Badoual, C., 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=PD-1+expressing+tumor-infiltrating+T+cells+are+a+favorable+prognostic+biomarker+in+hpv-associated+head+and+neck+cancer.">PD-1 expressing tumor-infiltrating T cells are a favorable prognostic biomarker in hpv-associated head and neck cancer.</a> Cancer Research. 73 (1), 128-138 (2013).</li><li>Gustavson, M. D., 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=Development+of+an+unsupervised+pixel-based+clustering+algorithm+for+compartmentalization+of+immunohistochemical+expression+using+automated+quantitative+analysis.">Development of an unsupervised pixel-based clustering algorithm for compartmentalization of immunohistochemical expression using automated quantitative analysis.</a> Applied Immunohistochemistry &amp; Molecular Morphology. 17 (4), 329-337 (2009).</li><li>Lu, S., 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+biomarker+modalities+for+predicting+response+to+PD-1/PD-L1+checkpoint+blockade:+a+systematic+review+and+meta-analysis.">Comparison of biomarker modalities for predicting response to PD-1/PD-L1 checkpoint blockade: a systematic review and meta-analysis.</a> JAMA Oncology. 5 (8), 1195-1204 (2019).</li><li>Hernandez, S., 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=Multiplex+immunofluorescence+tyramide+signal+amplification+for+immune+cell+profiling+of+paraffin-embedded+tumor+tissues.">Multiplex immunofluorescence tyramide signal amplification for immune cell profiling of paraffin-embedded tumor tissues.</a> Frontiers in Molecular Biosciences. 8, 667067 (2021).</li></ol>

La rinite allergica (AR) è una malattia infiammatoria non infettiva della mucosa nasale derivante da una combinazione di fattori ambientali e genetici. È diventato un problema di salute globale, che ha un impatto sull'efficienza lavorativa, diminuisce la qualità della vita, compromette il sonno, la funzione cognitiva e causa irritabilità e affaticamento. L'AR colpisce il 10%-20% della popolazione mondiale¹ e comporta costi economici considerevoli, causando perdite annuali fino a 30-50 miliardi di euro nei paesi dell...

La rinite allergica (AR) è una malattia infiammatoria cronica non infettiva della mucosa nasale, mediata principalmente da immunoglobuline E (IgE) specifiche1,2. È caratterizzata da sintomi come starnuti, naso che cola, congestione nasale e prurito nasale. Con l'industrializzazione e l'urbanizzazione, la prevalenza dell'AR sta gradualmente aumentando, colpendo circa il 10%-20% della popolazione mondiale1. La tecnica dell'immunofluorescenza (IF) è un metodo di colorazione fluorescente che utilizza una reazione di legame anticorpo-antigene. Può essere impiegato per rilevare e quantificare la distribuzione e i livelli di espressione di proteine specifiche nei tessuti biologici o nelle cellule. Nella ricerca AR, l'IF può rilevare simultaneamente più bersagli, tra cui citochine correlate all'AR, cellule infiammatorie, recettori e altro ancora, facilitando l'esplorazione della patogenesi dell'AR e degli effetti dei farmaci 3,4,5,6.
Il processo di colorazione a immunofluorescenza multicolore (mIF) assomiglia molto all'IF, con l'aggiunta di una fase di eluizione dell'anticorpo durante ogni ciclo di colorazione. Questa modifica consente il rilevamento simultaneo di più biomarcatori sulla stessa sezione di tessuto attraverso una singola marcatura sequenziale e più cicli di ricolorazione. mIF si basa sull'amplificazione del segnale della tiramide (TSA), che consente cicli ripetuti di colorazione fluorescente TSA e l'uso del riscaldamento a microonde per rimuovere gli anticorpi mantenendo i segnali fluorescenti 7,8. Rispetto all'IF convenzionale, l'mIF offre diversi vantaggi: (1) è in grado di rilevare antigeni debolmente espressi che sono difficili da identificare con l'IFconvenzionale 9,10; (2) fornisce una colorazione di alta qualità con un migliore rapporto segnale/rumore; (3) consente di quantificare le strutture e le regioni di interesse tessuto-specifiche11; (4) il multiplexing di percorsi multipli utilizza in modo efficiente i tessuti e conserva risorse patologiche limitate12; (5) l'analisi multiparametrica attraverso mIF offre informazioni più approfondite sui tessuti, scoprendo informazioni biologiche nascoste13.
Nel complesso, mIF consente l'osservazione di diverse espressioni e distribuzioni dell'antigene all'interno dello stesso campione, facilitando lo studio delle proteine bersaglio. In futuro, i ricercatori che cercano di comprendere l'espressione e la distribuzione di più proteine bersaglio troveranno questa tecnica una scelta preziosa. Questo studio dimostra l'applicazione di mIF per la colorazione di campioni di mucosa nasale di ratti con AR e valuta la creazione di un modello di AR nel ratto.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Al(OH)3</td>
			<td>Sollerbauer Biotechnology Co., Ltd</td>
			<td>A7130</td>
			<td></td>
		</tr>
		<tr>
			<td>75% ethanol</td>
			<td>Anhui Yiren An Co., Ltd</td>
			<td>20210107</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonia</td>
			<td>Chengdu Kolon Chemical Co., Ltd</td>
			<td>2021070101</td>
			<td></td>
		</tr>
		<tr>
			<td>Anhydrous ethanol</td>
			<td>Chengdu Kolon Chemical Co., Ltd</td>
			<td>2022070501</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-fluorescence quenching sealer</td>
			<td>SouthernBiotech</td>
			<td>0100-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic dyeing machine</td>
			<td>Thermo scientific</td>
			<td>Varistain Gemini ES</td>
			<td></td>
		</tr>
		<tr>
			<td>Carrier slides</td>
			<td>Nantong Mei Wei De Experimental Equipment Co., Ltd</td>
			<td>220518001</td>
			<td></td>
		</tr>
		<tr>
			<td>Citrate-phosphate buffer&#160;</td>
			<td>Servicebio biotechnology co., Ltd</td>
			<td>G1201</td>
			<td></td>
		</tr>
		<tr>
			<td>Citric acid antigen repair solution (PH 6.0)</td>
			<td>Xavier Biotechnology Co., Ltd</td>
			<td>G1201</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Nantong Mei Wei De Experimental Equipment Co.</td>
			<td>220518001</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Nantong Mewtech Life Science Co., Ltd</td>
			<td>CS01-2450</td>
			<td></td>
		</tr>
		<tr>
			<td>CY3-Tyramide</td>
			<td>Sawell Biotechnology Co., Ltd</td>
			<td>G1223-50UL</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sawell Biotechnology Co., Ltd</td>
			<td>G1012</td>
			<td></td>
		</tr>
		<tr>
			<td>Decoloring shaker</td>
			<td>SCILOGEX</td>
			<td>S1010E</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA decalcification solution</td>
			<td>Wuhan Xavier Biotechnology Co., Ltd</td>
			<td>CR2203047</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric heating blast dryer&#160;</td>
			<td>Shanghai Yiheng Scientific Instruments Co., Ltd</td>
			<td>DHG-9240A</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding box marking machine</td>
			<td>Thermo scientific</td>
			<td>&#160;PrintMate AS</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding machine</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-P5</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast tissue dewatering machine</td>
			<td>Thermo scientific</td>
			<td>STP420 ES</td>
			<td></td>
		</tr>
		<tr>
			<td>Film sealer</td>
			<td>Thermo scientific</td>
			<td>Autostainer 360</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC-Tyramide</td>
			<td>Sawell Biotechnology Co., Ltd</td>
			<td>G1222-50UL</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>Sunny Optical Technology Co.Ltd</td>
			<td>CX40</td>
			<td></td>
		</tr>
		<tr>
			<td>Foxp3</td>
			<td>Affinity Biosciences Co., Ltd</td>
			<td>bs-10211R</td>
			<td></td>
		</tr>
		<tr>
			<td>Freezing table</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-L5</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat Anti-Rabbit IgG H&#38;L (HRP)</td>
			<td>Liankebio Co., Ltd</td>
			<td>GAR0072</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat serum</td>
			<td>Biosharp</td>
			<td>BL210A</td>
			<td></td>
		</tr>
		<tr>
			<td>H&#38;E staining kit</td>
			<td>Leagene</td>
			<td>DH0020</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic forceps</td>
			<td>Shanghai Medical Devices Co., Ltd</td>
			<td>J31010</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric acid</td>
			<td>Sichuan Xilong Science Co., Ltd</td>
			<td>210608</td>
			<td></td>
		</tr>
		<tr>
			<td>Immunohistochemical pen</td>
			<td>Biosharp</td>
			<td>BC004</td>
			<td></td>
		</tr>
		<tr>
			<td>Microwave oven</td>
			<td>Midea</td>
			<td>M1-L213B</td>
			<td></td>
		</tr>
		<tr>
			<td>Neutral gum</td>
			<td>Sinopharm Group Chemical Reagent Co., Ltd</td>
			<td>10004160</td>
			<td></td>
		</tr>
		<tr>
			<td>Ovalbumin&#160;</td>
			<td>Sollerbauer Biotechnology Co., Ltd</td>
			<td>A804010</td>
			<td></td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Shanghai Yiheng Scientific Instruments Co., Ltd</td>
			<td>DHG-9240A</td>
			<td></td>
		</tr>
		<tr>
			<td>Palm centrifuge</td>
			<td>SCILOGEX</td>
			<td>D1008E</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Beyotime Biotechnology Co., Ltd</td>
			<td>P0099-100ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Pathology section scanner</td>
			<td>3DHISTECH Kft</td>
			<td>Pannoramic SCAN&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS buffer</td>
			<td>Biosharp</td>
			<td>G4202</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette&#160;</td>
			<td>Dragon</td>
			<td>KE0003087/KA0056573</td>
			<td></td>
		</tr>
		<tr>
			<td>Ror&#947;t</td>
			<td>Affinity Biosciences Co., Ltd</td>
			<td>DF3196</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Quanzhou Excellence Medical Co., Ltd</td>
			<td>20170022</td>
			<td></td>
		</tr>
		<tr>
			<td>Self-fluorescent quenching agent Sudan Black B</td>
			<td>Bioengineering Co., Ltd</td>
			<td>A602008-0025</td>
			<td></td>
		</tr>
		<tr>
			<td>Slicer</td>
			<td>Thermo scientific</td>
			<td>HM325</td>
			<td></td>
		</tr>
		<tr>
			<td>Slicing machine</td>
			<td>Thermo scientific</td>
			<td>HM325</td>
			<td></td>
		</tr>
		<tr>
			<td>Slide</td>
			<td>Nantong Mewtech Life Science Co., Ltd</td>
			<td>PC2-301</td>
			<td></td>
		</tr>
		<tr>
			<td>Sprague Dawley rats</td>
			<td>Sichuan Academy of Traditional Chinese Medicine</td>
			<td>SYX <img alt="Equation 1" src="/files/ftp_upload/65937/65937eq01.jpg" style="margin: auto;" /> 2023-0100</td>
			<td></td>
		</tr>
		<tr>
			<td>TICAM-1</td>
			<td>Affinity Biosciences Co., Ltd</td>
			<td>DF6289</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue scissors</td>
			<td>Shanghai Medical Devices Co., Ltd</td>
			<td>J22120</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue spreading baking sheet machine</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JK-6</td>
			<td></td>
		</tr>
		<tr>
			<td>TYR-690 fluorescent dyes</td>
			<td>Shanghai Rutron Biotechnology Co., Ltd</td>
			<td>RC0086-34RM</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex mixer</td>
			<td>SCILOGEX</td>
			<td>SLK-O3000-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Water bath-slide drier</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JK-6</td>
			<td></td>
		</tr>
		<tr>
			<td>Wax trimmer</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JXL-818</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Chengdu Kolon Chemical Co., Ltd</td>
			<td>2022051901</td>
			<td></td>
		</tr>
	</tbody>
</table>

immunofluorescent labeling in nasal mucosa tissue sections of allergic rhinitis rats via multicolor immunoassay

La rinite allergica (AR) è una malattia infiammatoria cronica e non infettiva della mucosa nasale, mediata principalmente da immunoglobuline E specifiche (IgE), che colpisce circa il 10%-20% della popolazione mondiale. Mentre la colorazione a immunofluorescenza (IF) è stata a lungo una tecnica standard per rilevare l'espressione proteica specifica della malattia, le tecniche convenzionali di IF sono limitate nella loro capacità di rilevare i livelli di espressione di tre o più proteine nello stesso campione. Di conseguenza, negli ultimi anni sono state sviluppate tecniche di IF multicolore, che consentono la marcatura simultanea di più bersagli in cellule o tessuti.
Questo protocollo fornisce una panoramica completa del processo per stabilire un modello di ratto di AR, ottenere campioni di mucosa nasale e le procedure tecniche per l'immunofluorescenza multicolore. Tutti i ratti del gruppo AR hanno mostrato sintomi tipici come starnuti, naso che cola e prurito al naso, con osservazioni comportamentali che hanno ottenuto un punteggio di ≥5 punti. La colorazione con ematossilina ed eosina (H&E) ha rivelato un aumento della conta delle cellule infiammatorie e un'integrità della mucosa nasale interrotta nel gruppo AR. L'immunofluorescenza multicolore (mIF) ha dimostrato un aumento dell'espressione di RORγt e TICAM-1, mentre l'espressione di Foxp3 è diminuita nel tessuto della mucosa nasale dei ratti AR.

Immunofluorescent Labeling in Nasal Mucosa Tissue Sections of Allergic Rhinitis Rats via Multicolor Immunoassay

Sei ratti SD sono stati indotti con successo nel modello AR attraverso l'iniezione intraperitoneale OVA e la sfida nasale. L'AR è stata indotta in tutti i ratti del gruppo AR, che rappresentavano il 100% del gruppo. Tutti i ratti del gruppo AR hanno mostrato sintomi tipici come starnuti, naso che cola e prurito al naso. Tutte le osservazioni comportamentali hanno ottenuto un punteggio di ≥5 punti (Tabella 2).
I risultati della colorazione H&E al 21° giorno di mo...

Watch this Scientific Journal Video about Advancing Allergic Rhinitis Research with Multicolor Immunofluorescence at JoVE.com

Advancing Allergic Rhinitis Research with Multicolor Immunofluorescence

Questo protocollo descrive una tecnica di immunofluorescenza multicolore per la valutazione del modello di rinite allergica nel ratto.

Marcatura immunofluorescente in sezioni di tessuto della mucosa nasale di ratti con rinite allergica tramite test immunologico multicolore

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), ...

La nostra ricerca principale si basa su uno studio sulla rinite allergica. La nostra ricerca intende analizzare quali proteine sono associate alla rinite allergica. L'immunofluorescenza multicolore è attualmente utilizzata per far progredire la ricerca nel nostro campo.In studi futuri, ci concentreremo sui possibili meccanismi della MTC nel trattamento della rinite allergica.