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

The authors are grateful for the experimental help in developing the method of intralymphatic immunization in mice from Iris Erdmann, Barbara von Beust, and Julia Maria Mart&#237;nez-G&#243;mez. Thanks also to Maggy Arras and Nikola Cesarovic for letting us use their surgical theatre for this video production.&#160;

1. Anesthesia of Mice
<ol>
	<li>Prepare the anesthetics by mixing ketamine (dissociative anesthetic) and xylazine (sedative and analgesic) in buffered saline. The concentrations of ketamine and xylazine in the final solution are 12.5 and 2 mg/ml, respectively.</li>
	<li>Inject the anesthetics to the mice by intraperitoneal administration using a syringe with a 25-30 G needle. Use 0.1 ml/10 g of body weight.</li>
	<li>Apply an ophthalmic ointment to the mice&#8217;s eyes in order to prevent drying-out of the corneas.</l...

<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. M., Gander, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lympho-geographical+concepts+in+vaccine+delivery.">Lympho-geographical concepts in vaccine delivery.</a> J. Control Release. 148, 56-62 (2010).</li><li>Johansen, P., von Moos, S., Mohanan, D., Kundig, T. M., Senti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+routes+for+allergen+immunotherapy.">New routes for allergen immunotherapy.</a> Hum. Vacc. Immunother. 8, 1525-1533 (2012).</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:+from+the+rationale+to+human+applications.">Intralymphatic immunotherapy: from the rationale to human applications.</a> Curr. Top. Microbiol. Immunol. 352, 71-84 (2011).</li><li>Duthie, M. S., Windish, H. P., Fox, C. B., Reed, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+defined+TLR+ligands+as+adjuvants+within+human+vaccines.">Use of defined TLR ligands as adjuvants within human vaccines.</a> Immunol. Rev. 239, 178-196 (2011).</li><li>Ribas, 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=Intra-lymph+node+prime-boost+vaccination+against+Melan+A+and+tyrosinase+for+the+treatment+of+metastatic+melanoma:+results+of+a+phase+1+clinical+trial.">Intra-lymph node prime-boost vaccination against Melan A and tyrosinase for the treatment of metastatic melanoma: results of a phase 1 clinical trial.</a> Clin. Cancer Res. 17, 2987-2996 (2011).</li><li>Bedrosian, 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=Intranodal+administration+of+peptide-pulsed+mature+dendritic+cell+vaccines+results+in+superior+CD8++T-cell+function+in+melanoma+patients.">Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8+ T-cell function in melanoma patients.</a> J. Clin. Oncol. 21, 3826-3835 (2003).</li><li>Lesimple, 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=Injection+by+various+routes+of+melanoma+antigen-associated+macrophages:+biodistribution+and+clinical+effects.">Injection by various routes of melanoma antigen-associated macrophages: biodistribution and clinical effects.</a> Cancer Immunol. Immunother. 52, 438-444 (2003).</li><li>Brown, 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=Adenovirus-transduced+dendritic+cells+injected+into+skin+or+lymph+node+prime+potent+simian+immunodeficiency+virus-specific+T+cell+immunity+in+monkeys.">Adenovirus-transduced dendritic cells injected into skin or lymph node prime potent simian immunodeficiency virus-specific T cell immunity in monkeys.</a> J. Immunol. 171, 6875-6882 (2003).</li><li>Fong, L., Brockstedt, D., Benike, C., Wu, L., Engleman, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendritic+cells+injected+via+different+routes+induce+immunity+in+cancer+patients.">Dendritic cells injected via different routes induce immunity in cancer patients.</a> J. Immunol. 166, 4254-4259 (2001).</li></ol>

Intralymphatic immunization and immunotherapy have been shown to be appropriate for stimulation of both antibody responses and T-cell responses. As demonstrated in this video-article, the intralymphatic procedure of vaccination is a quick and easy method for stimulating strong immune responses in mice. A trained surgeon can perform the procedure during 3-4 min. The session can also be shared between two surgeons where one is typically doing anesthesia and the suturing and the second surgeon is doing the incision and the ...

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this procedure requires efficient drainage of vaccine to lymph nodes where antigen presenting cells can interact with lymphocytes for generation of T- and B-cell responses. In addition, the strength and the type of immune responses induced also depend on the density or frequency of such interactions as well as the microenvironment itself, especially the content of cytokines. As only a small fraction of a vaccine injected into a peripheral tissue reaches a lymph node, vaccinations of mice and humans by direct injection of the vaccine into lymph nodes, the site were immune responses are generated, have been performed. In man, the procedure is guided by ultrasound, a procedure also used for the administration of imaging agents for visualization and diagnosis in the lymphatic system. In mice, the procedure is invasive. Here, a small (5-10 mm) incision is made in the inguinal region of anesthetized animals1, the lymph node is localized and immobilized with forceps, and a volume of 10-20 &#956;l of the vaccine is injected under visual control; 10 &#956;l is used for first injections and in young mice with small lymph nodes, whereas 20 &#956;l can be injected into lymph nodes of older or already primed mice, which have larger lymph nodes. The incision can be closed with a single stitch using surgical sutures. By this method, mice have vaccinated with plasmid DNA2,3, messenger RNA4, peptide1,3,5,6, protein7-10, particles11, bacteria12 as well as adjuvants7,13, and strong improvement of immune responses against all type of vaccines have been observed. The intralymphatic method of vaccination is especially appropriate in situations where conventional vaccination produces insufficient immunity or where the amount of available vaccine is limited or very costly. In human, the intralymphatic method of immunization has been applied to allergy patients14,15 or to patients with cancer16-21. Although the current notion is that the intralymphatic method is more invasive than other injectable methods such as intramuscular and subcutaneous injections, the pain perception is not higher than after a venous puncture15. It is expected that intralymphatic vaccination will become an alternative or complement to other methods of prophylactic and especially therapeutic vaccinations. This article describes in details how the procedure of intralymphatic vaccination is performed in mice. All procedures described, were approved by the Cantonal Veterinary Agency of Zurich and performed according Swiss federal guidelines and directives on the protection of animals used for scientific purposes.

<table border="0" cellpadding="0" cellspacing="0" width="653">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="42">
			<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>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

intralymphatic immunotherapy and vaccination in mice

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this requires efficient drainage of vaccine to lymph nodes where antigen presenting cells can interact with lymphocytes for generation of the wanted immune responses. The strength and the type of immune responses induced also depend on the density or frequency of interactions as well as the microenvironment, especially the content of cytokines. As only a minute fraction of peripherally injected vaccines reaches the lymph nodes, vaccinations of mice and humans were performed by direct injection of vaccine into inguinal lymph nodes, i.e.&#160;intralymphatic injection. In man, the procedure is guided by ultrasound. In mice, a small (5-10 mm) incision is made in the inguinal region of anesthetized animals, the lymph node is localized and immobilized with forceps, and a volume of 10-20 &#956;l of the vaccine is injected under visual control. The incision is closed with a single stitch using surgical sutures. Mice were vaccinated with plasmid DNA, RNA, peptide, protein, particles, and bacteria as well as adjuvants, and strong improvement of immune responses against all type of vaccines was observed. The intralymphatic method of vaccination is especially appropriate in situations where conventional vaccination produces insufficient immunity or where the amount of available vaccine is limited.

The procedure of intralymphatic injections in mice, despite the surgical nature, is straight forward and relatively fast. A trained person can perform the procedure in 3-4 min. The incision that is closed with one stitch typically heals within two days (Figure 1)
Intralymphatic vaccination or immunization has been performed with mRNA, plasmid DNA, peptides, proteins, virus and bacteria. Figure 2 illustrates the antibody production after immunization with the p...

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

Intralymphatic Immunotherapy and Vaccination in Mice

Prophylactic and therapeutic vaccination often fails to stimulate strong immune responses due to week drainage of the vaccine to lymph nodes and consequently poor involvement of immune cells. By direct injection of vaccine to lymph nodes, so-called intralymphatic injection, vaccine efficacy can be strongly improved and vaccine doses can be reduced.

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this requires efficient drainage of ...

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