Name: intralymphatic immunotherapy and vaccination in mice
Uploaded: 2014-02-02T14:00:00
Description: Watch this Scientific Journal Video about Intralymphatic Immunotherapy and Vaccination in Mice at JoVE.com

Forfatterne er taknemmelige for den eksperimentelle hjælp til at udvikle metoden til intralymfatisk immunisering i mus fra Iris Erdmann, Barbara von Beust, og Julia Maria Martínez-Gómez. Tak også til Maggy Arras og Nikola Cesarovic for at lade os bruge deres kirurgiske teater for denne video produktion.

1.. Bedøvelse af mus <ol><li> Forbered bedøvelsesmidler ved at blande ketamin (dissociativ bedøvelse) og xylazin (beroligende og smertestillende middel) i saltvand. Koncentrationerne af ketamin og xylazin i den endelige opløsning er 12,5 og 2 mg / ml. </li><li> Injicer bedøvelsesmidler til mus ved intraperitoneal indgivelse under anvendelse af en sprøjte med en 25-30 G nål. Brug 0,1 ml/10 g legemsvægt. </li><li> Påfør et oftalmisk salve til musenes øjne for at forhindre udtørring af hornhinder. </li><li> S...

<ol>
	<li>Johansen, P., 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=Direct+intralymphatic+injection+of+peptide+vaccines+enhances+immunogenicity.">Direct intralymphatic injection of peptide vaccines enhances immunogenicity.</a> Eur. J. Immunol. 35, 568-574 (2005).</li><li>Maloy, K. 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=Intralymphatic+immunization+enhances+DNA+vaccination.">Intralymphatic immunization enhances DNA vaccination.</a> Proc. Natl. Acad. Sci. U.S.A. 98, 3299-3303 (2001).</li><li>Smith, K. 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=Enhancing+DNA+vaccination+by+sequential+injection+of+lymph+nodes+with+plasmid+vectors+and+peptides.">Enhancing DNA vaccination by sequential injection of lymph nodes with plasmid vectors and peptides.</a> Vaccine. 27, 2603-2615 (2009).</li><li>Kreiter, 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=Intranodal+vaccination+with+naked+antigen-encoding+RNA+elicits+potent+prophylactic+and+therapeutic+antitumoral+immunity.">Intranodal vaccination with naked antigen-encoding RNA elicits potent prophylactic and therapeutic antitumoral immunity.</a> Cancer Res. 70, 9031-9040 (2010).</li><li>Johansen, P., 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=Antigen+kinetics+determines+immune+reactivity.">Antigen kinetics determines immune reactivity.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 5189-5194 (2008).</li><li>Smith, K. 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=Lymph+node-targeted+immunotherapy+mediates+potent+immunity+resulting+in+regression+of+isolated+or+metastatic+human+papillomavirus-transformed+tumors.">Lymph node-targeted immunotherapy mediates potent immunity resulting in regression of isolated or metastatic human papillomavirus-transformed tumors.</a> Clin. Cancer Res. 15, 6167-6176 (2009).</li><li>Johansen, P., 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=Toll-like+receptor+ligands+as+adjuvants+in+allergen-specific+immunotherapy.">Toll-like receptor ligands as adjuvants in allergen-specific immunotherapy.</a> Clin. Exp. Allergy. 35, 1591-1598 (2005).</li><li>Johansen, P., 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=Heat+denaturation,+a+simple+method+to+improve+the+immunotherapeutic+potential+of+allergens.">Heat denaturation, a simple method to improve the immunotherapeutic potential of allergens.</a> Eur. J. Immunol. 35, 3591-3598 (2005).</li><li>Martinez-Gomez, J. 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=Intralymphatic+injections+as+a+new+administration+route+for+allergen-specific+immunotherapy.">Intralymphatic injections as a new administration route for allergen-specific immunotherapy.</a> Int. Arch. Allergy Immunol. 150, 59-65 (2009).</li><li>Martinez-Gomez, J. 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=Targeting+the+MHC+class+II+pathway+of+antigen+presentation+enhances+immunogenicity+and+safety+of+allergen+immunotherapy.">Targeting the MHC class II pathway of antigen presentation enhances immunogenicity and safety of allergen immunotherapy.</a> Allergy. 64, 172-178 (2009).</li><li>Mohanan, 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=Administration+routes+affect+the+quality+of+immune+responses:+A+cross-sectional+evaluation+of+particulate+antigen-delivery+systems.">Administration routes affect the quality of immune responses: A cross-sectional evaluation of particulate antigen-delivery systems.</a> J. Control Release. 147, 342-349 (2010).</li><li>Waeckerle-Men, 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=Lymph+node+targeting+of+BCG+vaccines+amplifies+CD4+and+CD8+T-cell+responses+and+protection+against+Mycobacterium+tuberculosis.">Lymph node targeting of BCG vaccines amplifies CD4 and CD8 T-cell responses and protection against Mycobacterium tuberculosis.</a> Vaccine. 31, 1057-1064 (2013).</li><li>von Beust, B. 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=Improving+the+therapeutic+index+of+CpG+oligodeoxynucleotides+by+intralymphatic+administration.">Improving the therapeutic index of CpG oligodeoxynucleotides by intralymphatic administration.</a> Eur. J. Immunol. 35, 1869-1876 (2005).</li><li>Senti, G., 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=Intralymphatic+immunotherapy+for+cat+allergy+induces+tolerance+after+only+3+injections.">Intralymphatic immunotherapy for cat allergy induces tolerance after only 3 injections.</a> J. Allergy Clin. Immunol. 129, 1290-1296 (2012).</li><li>Senti, G., 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=Intralymphatic+allergen+administration+renders+specific+immunotherapy+faster+and+safer:+a+randomized+controlled+trial.">Intralymphatic allergen administration renders specific immunotherapy faster and safer: a randomized controlled trial.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 17908-17912 (2008).</li><li>Lesterhuis, W. 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=Route+of+administration+modulates+the+induction+of+dendritic+cell+vaccine-induced+antigen-specific+T+cells+in+advanced+melanoma+patients.">Route of administration modulates the induction of dendritic cell vaccine-induced antigen-specific T cells in advanced melanoma patients.</a> Clin. Cancer Res. 17, 5725-5735 (2011).</li><li>Fadul, C. E., 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=Immune+response+in+patients+with+newly+diagnosed+glioblastoma+multiforme+treated+with+intranodal+autologous+tumor+lysate-dendritic+cell+vaccination+after+radiation+chemotherapy.">Immune response in patients with newly diagnosed glioblastoma multiforme treated with intranodal autologous tumor lysate-dendritic cell vaccination after radiation chemotherapy.</a> J. Immunother. 34, 382-389 (2011).</li><li>Eizenberg, P., 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=Acceptance+of+Intanza(R)+9+mug+intradermal+influenza+vaccine+in+routine+clinical+practice+in+Australia+and+Argentina.">Acceptance of Intanza(R) 9 mug intradermal influenza vaccine in routine clinical practice in Australia and Argentina.</a> Adv. Ther. 28, 640-649 (2011).</li><li>Durando, P., 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=Adjuvants+and+alternative+routes+of+administration+towards+the+development+of+the+ideal+influenza+vaccine.">Adjuvants and alternative routes of administration towards the development of the ideal influenza vaccine.</a> Hum. Vaccin. 7, 29-40 (2011).</li><li>Barth, R. 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=A+randomized+trial+of+ex+vivo+CD40L+activation+of+a+dendritic+cell+vaccine+in+colorectal+cancer+patients:+tumor-specific+immune+responses+are+associated+with+improved+survival.">A randomized trial of ex vivo CD40L activation of a dendritic cell vaccine in colorectal cancer patients: tumor-specific immune responses are associated with improved survival.</a> Clin. Cancer Res. 16, 5548-5556 (2010).</li><li>Schwaab, 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=Clinical+and+immunologic+effects+of+intranodal+autologous+tumor+lysate-dendritic+cell+vaccine+with+Aldesleukin+(Interleukin+2)+and+IFN-{alpha}2a+therapy+in+metastatic+renal+cell+carcinoma+patients.">Clinical and immunologic effects of intranodal autologous tumor lysate-dendritic cell vaccine with Aldesleukin (Interleukin 2) and IFN-{alpha}2a therapy in metastatic renal cell carcinoma patients.</a> Clin. Cancer Res. 15, 4986-4992 (2009).</li><li>Senti, G., Johansen, P., Kundig, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intralymphatic+immunotherapy.">Intralymphatic immunotherapy.</a> Curr. Opin. Allergy Clin. Immunol. 9, 537-543 (2009).</li><li>Catron, D. M., Itano, A. A., Pape, K. A., Mueller, D. L., Jenkins, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+the+first+50+hr+of+the+primary+immune+response+to+a+soluble+antigen.">Visualizing the first 50 hr of the primary immune response to a soluble antigen.</a> Immunity. 21, 341-347 (2004).</li><li>Itano, A. A., Jenkins, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigen+presentation+to+naive+CD4+T+cells+in+the+lymph+node.">Antigen presentation to naive CD4 T cells in the lymph node.</a> Nat. Immunol. 4, 733-739 (2003).</li><li>Johansen, P., Mohanan, D., Martinez-Gomez, J. M., Kundig, T. 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Intralymfatisk immunisering og immunterapi har vist sig at være egnet til stimulering af både antistofrespons og T-celle responser. Som påvist i denne video-artikel, intralymfatisk procedure ved vaccination er en hurtig og nem metode til at stimulere stærke immunreaktioner i mus. En uddannet kirurg kan udføre proceduren i løbet af 3-4 min. Sessionen kan også deles mellem to kirurger, hvor man typisk gør anæstesi og suturering og den anden kirurg gør snittet og injektionen. Effektiviteten af ​​proceduren er...

Vacciner er typisk subkutant eller intramuskulært, til stimulering af immunresponser. Succesen for denne procedure kræver effektiv dræning af vaccine til lymfeknuder, hvor antigen-præsenterende celler kan interagere med lymfocytter til generering af T-og B-celle respons. Desuden styrke og type af immunrespons afhænger også af tætheden eller hyppigheden af ​​sådanne interaktioner samt mikromiljø selv, især indholdet af cytokiner. Da kun en lille brøkdel af en vaccine injiceres i en perifer væv når en lymfeknude, vaccination af mus og mennesker ved direkte indsprøjtning af vaccinen i lymfeknuder, stedet blev immunresponser genereres, er blevet udført. Hos mennesker er proceduren styret af ultralyd, en procedure også anvendes til administration af imaging agenter for visualisering og diagnose i lymfesystemet. Hos mus, er proceduren invasiv. Her blev en lille (5-10 mm) incision er lavet i lyske-regionen i bedøvede dyr 1, er lymfeknude lokaliseret og immobiliseres med en pincet, og et volumen på 10-20 ul af vaccinen injiceres under visuel kontrol 10 pi anvendes til første injektioner og unge mus med små lymfeknuder, hvorimod 20 pi kan injiceres i lymfeknuder ældre eller allerede immuniserede mus, der har større lymfeknuder. Snittet kan lukkes med et enkelt sting hjælp kirurgiske suturer. Ved denne fremgangsmåde har mus vaccineret med plasmid-DNA 2,3, messenger-RNA 4, peptid 1,3,5,6, protein 7-10, partikler 11, bakterier 12 samt adjuvanser 7,13 og stærk forbedring af immun responser mod alle typer af vacciner er blevet overholdt. Den intralymfatisk metode til vaccination er især relevant i situationer, hvor konventionel vaccination producerer utilstrækkelig immunitet, eller hvor mængden af ​​tilgængelig vaccine er begrænset eller very dyrt. Hos mennesker har intralymfatisk immuniserings blevet anvendt på allergipatienter 14,15 eller til patienter med cancer 16-21. Selvom den nuværende opfattelse er, at den intralymfatisk metode er mere invasiv end andre injicerbare metoder såsom intramuskulære og subkutane injektioner smerteopfattelsen ikke er højere end efter en venepunktur 15. Det forventes, at intralymfatisk vaccination vil blive et alternativ eller supplement til andre metoder til profylaktiske og især terapeutiske vaccinationer. Denne artikel beskriver i detaljer, hvordan proceduren for intralymfatisk vaccination udføres i mus. Alle procedurer, der er beskrevet, blev godkendt af Cantonal Veterinary Agency Zürich og udføres ifølge schweiziske føderale retningslinjer og direktiver om beskyttelse af dyr, der anvendes til videnskabelige formål.

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			<td height="42" style="height:42px;width:233px;">Ketamine (Ketasol-100)</td>
			<td style="width:135px;">Graeub AG, Switzerland</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Anesthetics</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:233px;">Xylazine (Rompun)</td>
			<td style="width:135px;">Bayer, Germany</td>
			<td style="width:119px;"></td>
			<td>Anesthetics</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Viscotears Eye-Gel</td>
			<td style="width:135px;">Novartis, Switzerland</td>
			<td style="width:119px;"></td>
			<td>To keep eyes from drying out during anesthesia.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">BD Micro-Fine 0.5 ml</td>
			<td style="width:135px;">BD Medical, France</td>
			<td style="width:119px;"></td>
			<td>29 G Insulin syringes with permanently attached needles</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">6-0 Dermalon Monofilament nylon</td>
			<td style="width:135px;">Covidien, MA, USA</td>
			<td style="width:119px;"></td>
			<td>For sutures (0.7 metric, 18G, 45 cm, Blue)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Curved forceps, 4.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>For incision and holding of lymph node</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Straight surgical scissors, 4.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>For incision</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:233px;">Needle holder, 5.5 inch</td>
			<td style="width:135px;">Polymed, Switzerland</td>
			<td style="width:119px;"></td>
			<td>To close incision with suture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:233px;"></td>
			<td style="width:135px;"></td>
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			<td></td>
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intralymphatic immunotherapy and vaccination in mice

Vacciner er typisk subkutant eller intramuskulært, til stimulering af immunresponser. Succesen for dette kræver en effektiv dræning af vaccine til lymfeknuder, hvor antigen-præsenterende celler kan interagere med lymfocytter til generering af de ønskede immunrespons. Styrke og type af immunrespons afhænger også af tætheden eller hyppigheden af ​​interaktioner samt mikromiljø, især indholdet af cytokiner. Da kun en brøkdel af perifert injicerede vacciner når lymfeknuder blev vaccination af mus og mennesker udføres ved direkte injektion af vaccine i lysken lymfeknuder, dvs intralymfatisk injektion. Hos mennesker er proceduren styret af ultralyd. I mus er en lille (5-10 mm) incision i lyske region bedøvede dyr, lymfeknude er lokaliseret og immobiliseres med en pincet, og et volumen på 10-20 ul af vaccinen injiceres under visuel kontrol.Snittet lukkes med en enkelt søm ved hjælp af kirurgiske suturer. Mus blev vaccineret med plasmid DNA, RNA, peptid, protein, partikler og bakterier såvel som adjuvanser og stærk forbedring af immunresponser mod alle typer af vacciner blev observeret. Den intralymfatisk metode til vaccination er især relevant i situationer, hvor konventionel vaccination producerer utilstrækkelig immunitet, eller hvor mængden af ​​tilgængelig vaccine er begrænset.

Proceduren for intralymfatisk injektioner i mus, på trods af kirurgisk art, er ligetil og relativt hurtigt. En uddannet person kan udføre proceduren i 3-4 min. Det snit, der er lukket med en søm typisk heler inden to dage (figur 1) Intralymfatisk vaccination eller immunisering er blevet udført med mRNA, plasmid-DNA, peptider, proteiner, virus og bakterier Figur. 2 illustrerer antistof-produktion efter immunisering med proteinet phospholipase A2 (PLA2), d...

Watch this Scientific Journal Video about Intralymphatic Immunotherapy and Vaccination in Mice at JoVE.com

Intralymphatic Immunotherapy and Vaccination in Mice

Intralymfatisk Immunterapi og Vaccination i mus

Profylaktisk og terapeutisk vaccination ofte undlader at stimulere stærke immunreaktioner på grund uge dræning af vaccinen til lymfeknuder og dermed dårlig inddragelse af immunceller. Ved direkte injektion af vaccine til lymfeknuder, såkaldt intralymfatisk indsprøjtning, effekten af ​​vaccinen kan være stærkt forbedret, og kan reduceres vaccinedoser.

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While clinical studies have established that antigen-loaded DC vaccines are safe and promising therapy for tumors 1, their clinical efficacy remains to be ...

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

High-throughput Screening for Bredspektret Chemical hæmmere af RNA virus

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

Measuring Local Anaphylaxis in Mice

Måling Lokale anafylaksi hos mus

Allergic responses are the result of the activation of mast cells and basophils, and the subsequent release of vasoactive and proinflammatory mediators. ...

Generation of CAR T Cells for Adoptive Therapy in the Context of Glioblastoma Standard of Care

Generation af CAR T-celler til adoptiv terapi i forbindelse med glioblastom standard for pleje

Adoptive T cell immunotherapy offers a promising strategy for specifically targeting and eliminating malignant gliomas. T cells can be engineered ex vivo ...

In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model

In Vivo Assay for Detection of Antigen-specific T-cell Cytolytic Function Using a Vaccination Model

In Vivo Assay til påvisning af Antigen-specifikke T-celle cytolytisk funktion ved hjælp af en Vaccination Model

Current methodologies for antigen-specific killing are limited to in vitro use or utilized in infectious disease models. However, there is not a protocol ...

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Generation af diskriminerende menneskelige monoklonale antistoffer fra sjældne Antigen-specifikke B celler cirkulerer i blodet

Monoclonal antibodies (mAbs) are powerful tools useful for both fundamental research and in biomedicine. Their high specificity is indispensable when the ...

Stereotactic Adoptive Transfer of Cytotoxic Immune Cells in Murine Models of Orthotopic Human Glioblastoma Multiforme Xenografts

Stereotactic adoptiv overførsel af cytotoksiske immunceller i Murine modeller af Orthotopic menneskelige glioblastom Multiforme Xenografts

Glioblastoma multiforme (GBM), the most frequent and aggressive primary brain cancer in adults, is generally associated with a poor prognosis, and scarce ...

Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination

Optimeret Interferon-gamma ELISpot Assay til foranstaltning T-celle respons i Guinea Gris Model efter vaccinationen

The guinea pig has played a pivotal role as a relevant small animal model in the development of vaccines for infectious diseases such as tuberculosis, ...