Name: contact hypersensitivity as a murine model of allergic contact dermatitis
Uploaded: 2022-09-26T15:00:00
Description: Watch this Scientific Journal Video about Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis at JoVE.com

この研究は、subvention SUBZによって支持された。ヴロツワフの医科大学のA020.22.060, ポーランド, 科学高等教育省からの助成金によって N N401 545940 MSとIP2012 0443 72 MMSに.

この記事で紹介したすべての実験は、クラクフの第1回動物実験に関する地方倫理委員会のガイドラインに従って実施されました。記載されているすべての手順は、特にケタミン/キシラジンを麻酔薬として使用すること、耳の両側を使用して物質/ハプテンを塗布すること、耳を切り落とすこと、および眼球除去による血液の採取に関して、地元の勧告に従って実施された。本研究には、生後6...

<ol>
	<li>Nosbaum, A., Vocanson, M., Rozieres, A., Hennino, A., Nicolas, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allergic+and+irritant+contact+dermatitis.">Allergic and irritant contact dermatitis.</a> European Journal of Dermatology. 19 (4), 325-332 (2009).</li><li>Hertl, 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=Predominance+of+epidermal+CD8++T+lymphocytes+in+bullous+cutaneous+reactions+caused+by+beta-lactam+antibiotics.">Predominance of epidermal CD8+ T lymphocytes in bullous cutaneous reactions caused by beta-lactam antibiotics.</a> The Journal of Investigative Dermatology. 101 (6), 794-799 (1993).</li><li>Martin, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T+lymphocyte-mediated+immune+responses+to+chemical+haptens+and+metal+ions:+implications+for+allergic+and+autoimmune+disease.">T lymphocyte-mediated immune responses to chemical haptens and metal ions: implications for allergic and autoimmune disease.</a> International Archives of Allergy and Immunology. 134 (3), 186-198 (2004).</li><li>Takeyoshi, M., Iida, K., Suzuki, K., Yamazaki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skin+sensitization+potency+of+isoeugenol+and+its+dimers+evaluated+by+a+non-radioisotopic+modification+of+the+local+lymph+node+assay+and+guinea+pig+maximization+test.">Skin sensitization potency of isoeugenol and its dimers evaluated by a non-radioisotopic modification of the local lymph node assay and guinea pig maximization test.</a> Journal of Applied Toxicology. 28 (4), 530-534 (2008).</li><li>Nakamura, K., Aizawa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+the+genetic+control+of+picryl+chloride+contact+hypersensitivity+reaction+in+inbred+rats.">Studies on the genetic control of picryl chloride contact hypersensitivity reaction in inbred rats.</a> Transplantation Proceedings. 13 (2), 1400-1403 (1981).</li><li>Asherson, G. L., Ptak, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contact+and+delayed+hypersensitivity+in+the+mouse.+I.+Active+sensitization+and+passive+transfer.">Contact and delayed hypersensitivity in the mouse. I. Active sensitization and passive transfer.</a> Immunology. 15 (3), 405-416 (1968).</li><li>Honda, T., Egawa, G., Grabbe, S., Kabashima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+of+immune+events+in+the+murine+contact+hypersensitivity+model:+toward+the+understanding+of+allergic+contact+dermatitis.">Update of immune events in the murine contact hypersensitivity model: toward the understanding of allergic contact dermatitis.</a> The Journal of Investigative Dermatology. 133 (2), 303-315 (2013).</li><li>Kaplan, D. H., Jenison, M. C., Saeland, S., Shlomchik, W. D., Shlomchik, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidermal+langerhans+cell-deficient+mice+develop+enhanced+contact+hypersensitivity.">Epidermal langerhans cell-deficient mice develop enhanced contact hypersensitivity.</a> Immunity. 23 (6), 611-620 (2005).</li><li>Wang, 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=Langerin+expressing+cells+promote+skin+immune+responses+under+defined+conditions.">Langerin expressing cells promote skin immune responses under defined conditions.</a> Journal of Immunology. 180 (7), 4722-4727 (2008).</li><li>Wang, B., 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=CD4++Th1+and+CD8++type+1+cytotoxic+T+cells+both+play+a+crucial+role+in+the+full+development+of+contact+hypersensitivity.">CD4+ Th1 and CD8+ type 1 cytotoxic T cells both play a crucial role in the full development of contact hypersensitivity.</a> Journal of Immunology. 165 (12), 6783-6790 (2000).</li><li>Mori, 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=Cutaneous+hypersensitivities+to+hapten+are+controlled+by+IFN-gamma-upregulated+keratinocyte+Th1+chemokines+and+IFN-gamma-downregulated+langerhans+cell+Th2+chemokines.">Cutaneous hypersensitivities to hapten are controlled by IFN-gamma-upregulated keratinocyte Th1 chemokines and IFN-gamma-downregulated langerhans cell Th2 chemokines.</a> The Journal of Investigative Dermatology. 128 (7), 1719-1727 (2008).</li><li>Peszkowski, M. J., Warfvinge, G., Larsson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allergic+and+irritant+contact+responses+to+DNFB+in+BN+and+LEW+rat+strains+with+different+TH1/TH2+profiles.">Allergic and irritant contact responses to DNFB in BN and LEW rat strains with different TH1/TH2 profiles.</a> Acta Dermato-Venereologica. 74 (5), 371-374 (1994).</li><li>Henningsen, S. J., Mickell, J., Zachariae, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+kinins+in+dinitrochlorobenzene+contact+dermatitis+of+guinea-pigs.">Plasma kinins in dinitrochlorobenzene contact dermatitis of guinea-pigs.</a> Acta Allergologica. 25 (5), 327-331 (1970).</li><li>Maibach, H. I., Maguire, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elicitation+of+delayed+hypersensitivity+(DNCB+contact+dermatitis)+in+markedly+panleukopenic+guinea+pigs.">Elicitation of delayed hypersensitivity (DNCB contact dermatitis) in markedly panleukopenic guinea pigs.</a> The Journal of Investigative Dermatology. 41, 123-127 (1963).</li><li>Martel, B. C., Lovato, P., B&#228;umer, W., Olivry, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+animal+models+of+atopic+dermatitis+for+preclinical+studies.">Translational animal models of atopic dermatitis for preclinical studies.</a> The Yale Journal of Biology and Medicine. 90 (3), 389-402 (2017).</li><li>Jin, H., He, R., Oyoshi, M., Geha, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+atopic+dermatitis.">Animal models of atopic dermatitis.</a> The Journal of Investigative Dermatology. 129 (1), 31-40 (2009).</li><li>Li, Y. Z., Lu, X. Y., Jiang, W., Li, L. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-inflammatory+effect+of+qingpeng+ointment+in+atopic+dermatitis-like+murine+model.">Anti-inflammatory effect of qingpeng ointment in atopic dermatitis-like murine model.</a> Evidence-Based Complementary and Alternative. 2013, 907016 (2013).</li><li>Hoggatt, J., Hoggatt, A. F., Tate, T. A., Fortman, J., Pelus, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bleeding+the+laboratory+mouse:+Not+all+methods+are+equal.">Bleeding the laboratory mouse: Not all methods are equal.</a> Experimental Hematology. 44 (2), 132-137 (2016).</li><li>Bedoya, S. K., Wilson, T. D., Collins, E. L., Lau, K., Larkin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+th17+differentiation+of+na&#239;ve+CD4+T+lymphocytes.">Isolation and th17 differentiation of na&#239;ve CD4 T lymphocytes.</a> Journal of Visualized Experiments. (79), e50765 (2013).</li><li>Hemocytometer - Counting of Cells. Amrita University Available from: <a target="_blank" href="https://www.youtube.com/watch?v=MKS0KM3lr90">https://www.youtube.com/watch?v=MKS0KM3lr90</a> (2011)</li><li>Majewska-Szczepanik, M., Strzepa, A., Marci&#324;ska, K., Wen, L., Szczepanik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epicutaneous+immunization+with+TNP-Ig+and+Zymosan+induces+TCR&#945;&#946;++CD4++contrasuppressor+cells+that+reverse+skin-induced+suppression+via+IL-17A.">Epicutaneous immunization with TNP-Ig and Zymosan induces TCR&#945;&#946;+ CD4+ contrasuppressor cells that reverse skin-induced suppression via IL-17A.</a> International Archives of Allergy and Immunology. 164 (2), 122-136 (2014).</li><li>Majewska-Szczepanik, M., Zemelka-Wi&#261;cek, M., Ptak, W., Wen, L., Szczepanik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epicutaneous+immunization+with+DNP-BSA+induces+CD4++CD25++Treg+cells+that+inhibit+Tc1-mediated+CS.">Epicutaneous immunization with DNP-BSA induces CD4+ CD25+ Treg cells that inhibit Tc1-mediated CS.</a> Immunology and Cell Biology. 90 (8), 784-795 (2012).</li><li>Directive 2010/63 / EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. Official Journal of the European Union Available from: <a target="_blank" href="https://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:En:PDF">https://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:En:PDF</a> (2010)</li><li>Colvin, R. B., Dvorak, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+clotting+system+in+cell-mediated+hypersensitivity.+II.+Kinetics+of+fibrinogen/fibrin+accumulation+and+vascular+permeability+changes+in+tuberculin+and+cutaneous+basophil+hypersensitivity+reactions.">Role of the clotting system in cell-mediated hypersensitivity. II. Kinetics of fibrinogen/fibrin accumulation and vascular permeability changes in tuberculin and cutaneous basophil hypersensitivity reactions.</a> Journal of Immunology. 114, 377-387 (1975).</li><li>Szczepanik, 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=Regulation+of+contact+sensitivity+in+non-obese+diabetic+(NOD)+mice+by+innate+immunity.">Regulation of contact sensitivity in non-obese diabetic (NOD) mice by innate immunity.</a> Contact Dermatitis. 79 (4), 197-207 (2018).</li><li>Askenase, P. W., Majewska-Szczepanik, M., Kerfoot, S., Szczepanik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Participation+of+iNKT+cells+in+the+early+and+late+components+of+Tc1-mediated+DNFB+contact+sensitivity:+Cooperative+role+of+&#947;&#948;-T+cells.">Participation of iNKT cells in the early and late components of Tc1-mediated DNFB contact sensitivity: Cooperative role of &#947;&#948;-T cells.</a> Scandinavian Journal of Immunology. 73 (5), 465-477 (2011).</li><li>Zemelka-Wi&#261;cek, M., Majewska-Szczepanik, M., Ptak, W., Szczepanik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epicutaneous+immunization+with+protein+antigen+induces+antigen-non-specific+suppression+of+CD8+T+cell+mediated+contact+sensitivity.">Epicutaneous immunization with protein antigen induces antigen-non-specific suppression of CD8 T cell mediated contact sensitivity.</a> Pharmacological Reports. 64 (6), 1485-1496 (2012).</li><li>Van Loveren, 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=Use+of+micrometers+and+calipers+to+measure+various+components+of+delayed-type+hypersensitivity+ear+swelling+reactions+in+mice.">Use of micrometers and calipers to measure various components of delayed-type hypersensitivity ear swelling reactions in mice.</a> Journal of Immunological Methods. 67 (2), 311-319 (1984).</li><li>Tsuji, R. F., 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=B+cell-dependent+T+cell+responses:+IgM+antibodies+are+required+to+elicit+contact+sensitivity.">B cell-dependent T cell responses: IgM antibodies are required to elicit contact sensitivity.</a> The Journal of Experimental Medicine. 196 (10), 1277-1290 (2002).</li><li>Campos, R. 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=Cutaneous+immunization+rapidly+activates+liver+invariant+Valpha14+NKT+cells+stimulating+B-1+B+cells+to+initiate+T+cell+recruitment+for+elicitation+of+contact+sensitivity.">Cutaneous immunization rapidly activates liver invariant Valpha14 NKT cells stimulating B-1 B cells to initiate T cell recruitment for elicitation of contact sensitivity.</a> The Journal of Experimental Medicine. 198 (12), 1785-1796 (2003).</li><li>R&#252;hl-Muth, A. C., Maler, M. D., Esser, P. R., Martin, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+of+a+fat-enriched+diet+causes+the+loss+of+resistance+to+contact+hypersensitivity.">Feeding of a fat-enriched diet causes the loss of resistance to contact hypersensitivity.</a> Contact Dermatitis. 85 (4), 398-406 (2021).</li><li>Bour, 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=histocompatibility+complex+class+I-restricted+CD8++T+cells+and+class+II-restricted+CD4++T+cells,+respectively,+mediate+and+regulate+contact+sensitivity+to+dinitrofluorobenzene.">histocompatibility complex class I-restricted CD8+ T cells and class II-restricted CD4+ T cells, respectively, mediate and regulate contact sensitivity to dinitrofluorobenzene.</a> European Journal of Immunology. 25 (11), 3006-3010 (1995).</li><li>Majewska-Szczepanik, 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=Obesity+aggravates+contact+hypersensitivity+reaction+in+mice.">Obesity aggravates contact hypersensitivity reaction in mice.</a> Contact Dermatitis. 87 (1), 28-39 (2022).</li><li>Katagiri, K., Arakawa, S., Kurahashi, R., Hatano, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+contact+hypersensitivity+in+diet-induced+obese+mice.">Impaired contact hypersensitivity in diet-induced obese mice.</a> Journal of Dermatological Science. 46 (2), 117-126 (2007).</li><li>Bouloc, A., Cavani, A., Katz, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contact+hypersensitivity+in+MHC+class+II-deficient+mice+depends+on+CD8+T+lymphocytes+primed+by+immunostimulating+Langerhans+cells.">Contact hypersensitivity in MHC class II-deficient mice depends on CD8 T lymphocytes primed by immunostimulating Langerhans cells.</a> The Journal of Investigative Dermatology. 111 (1), 44-49 (1998).</li><li>Martin, 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=Peptide+immunization+indicates+that+CD8++T+cells+are+the+dominant+effector+cells+in+trinitrophenyl-specific+contact+hypersensitivity.">Peptide immunization indicates that CD8+ T cells are the dominant effector cells in trinitrophenyl-specific contact hypersensitivity.</a> The Journal of Investigative Dermatology. 115 (2), 260-266 (2000).</li><li>Vennegaard, M. 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=Epicutaneous+exposure+to+nickel+induces+nickel+allergy+in+mice+via+a+MyD88-dependent+and+interleukin-1-dependent+pathway.">Epicutaneous exposure to nickel induces nickel allergy in mice via a MyD88-dependent and interleukin-1-dependent pathway.</a> Contact Dermatitis. 71 (4), 224-232 (2014).</li></ol>

CHSは、皮膚中の自己タンパク質抗原に結合し、ネオ抗原を生成するハプテン を介して 誘導される。CHSは、循環抗原特異的CHSエフェクターT細胞の局所的な血管外動員によって媒介され、これは、チャレンジ組織において腫脹をもたらし、二次皮膚を同じハプテンに曝露してから24時間後にピークに達する6。組織の腫脹は、主に、白血球および白血球依存性フィブリン?...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64329">Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis</a>. The Protocol&#160;and Representative Results sections were updated.
Step 3.2.1 of the Protocol has been updated from:
On day &#34;4&#34;, prepare 0.4% hapten: TNCB in an acetone-ethanol mixture (ratio 1:1). Prepare the solution just before use in a glass vial and protect it from light by covering the vial with aluminum foil.
to:
On day &#34;4&#34;, prepare 0.4% hapten: TNCB in an acetone: olive oil&#160;mixture (ratio 1:1). Prepare the solution just before use in a glass vial and protect it from light by covering the vial with aluminum foil.
Figure 1 of the Representative Results has been updated from:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64329/64329fig01.jpg" /> 
Figure 1: Induction of CHS. Sensitization, challenge, and ear measurement. Abbreviations: CHS = contact hypersensitivity reaction; TNCB = 2,4,6-trinitrochlorobenzene. <a href="https://www.jove.com/files/ftp_upload/64329/64329fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
to:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64329/64329fig1v2.jpg" /> 
Figure 1: Induction of CHS. Sensitization, challenge, and ear measurement. Abbreviations: CHS = contact hypersensitivity reaction; TNCB = 2,4,6-trinitrochlorobenzene. <a href="https://www.jove.com/files/ftp_upload/64329/64329fig1v2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
Figure 2 of the Representative Results has been updated from:
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/64329/64329fig02.jpg" /> 
Figure 2: Adoptive transfer of the CHS-effector cells. Abbreviations: ALNs =axillary and inguinal lymph nodes; CHS = contact hypersensitivity reaction; i.v. = intravenously; SPLs = spleens; TNCB = 2,4,6-trinitrochlorobenzene. <a href="https://www.jove.com/files/ftp_upload/64329/64329fig02large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
to:
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/64329/64329fig2v2.jpg" /> 
Figure 2: Adoptive transfer of the CHS-effector cells. Abbreviations: ALNs =axillary and inguinal lymph nodes; CHS = contact hypersensitivity reaction; i.v. = intravenously; SPLs = spleens; TNCB = 2,4,6-trinitrochlorobenzene. <a href="https://www.jove.com/files/ftp_upload/64329/64329fig2v2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64329">Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis</a>. The Protocol&#160;and Representative Results sections were updated.

Erratum: Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis

アレルギー性接触皮膚炎(ACD)は、ハプテンと呼ばれる低分子量化学物質への曝露に起因するIV型過敏反応によって引き起こされる先進国における一般的な皮膚炎症性疾患である。ヒトにおける接触感作を引き起こす物質には、重金属イオン(クロム、ニッケル、鉄、コバルト)、テルペンチン、化粧品中に存在する香料、染料、および防腐剤(例えば、パラフェニレンジアミン)、いくつかの薬物(例えば、ネオマイシン、ベンゾカイン)、βラクタム系抗生物質(すなわち、ペニシリン)、植物によって産生される化学物質(ペンタデカカテコール、ポイズンツタに存在する物質)、ならびに写真産業で使用されるヒドロキノンが含まれる1,2.ACD病因薬は、産業だけで100,000以上の化学物質が使用され、毎年2,000の新しい化学物質が合成されているため、非常に高いです。現在までに、ハプテン/アレルゲンに接触している可能性のある3,700以上の分子が同定されています3。接触過敏反応(CHS)は、マウス、モルモット、ラットで研究することができ、有機溶媒に溶解した反応性化学ハプテンの局所皮膚適用によって誘発され得るACDの実験モデルである4,5,6。この研究は、様々な試験によって測定および定量化することができるマウスにおけるCHS反応を研究するのに役立つかもしれない客観的な実験室法を記述することを目的としている。
CHSは感作(誘導)相とエフェクター(チャレンジ)相からなる。動物モデルでは、ハプテンはまず体内のタンパク質に共有結合してネオ抗原を作り出します。感作期の間、活性化ケラチノサイトは、炎症誘発性サイトカイン腫瘍壊死因子α(TNF-α)およびインターロイキン1β(IL-1β)7を産生することにより、皮膚樹状細胞(sDC)の遊走および成熟を促進する7。表皮ランゲルハンス細胞(LC)は、CHS誘導およびエフェクター相8の間に抗原を提示する。感作中にハプテンに曝露されたLCは、調節細胞およびエフェクター細胞の両方の誘導を促進する9。いくつかの研究からの証拠の増加は、CHS応答が、CD4+ MHCクラスII制限性Th1細胞、特徴的な炎症性浸潤物を採用するためにインターフェロンγ(IFN-γ)を局所的に放出すること、IFN-γも放出することができるが、ほとんどがケラチノサイトへの細胞傷害性損傷を媒介するCD8+ MHCクラスI制限性Tc1リンパ球、および現在インターロイキン17(IL-17)産生Th17細胞10によって媒介され得ることを示唆している。11.
様々な種1 2、13、14およびハプテンを用いたいくつかの異なるCHSモデルが開発されている(異なるハプテン、溶媒、および適用時間の詳細な比較は表1に要約されている)。頻繁に使用される実験種であるマウスは、CHSの研究においていくつかの利点を提供する。マウスには他の種に比べて系統、ノックアウト(KO)、トランスジェニック動物が多く、非常に魅力的な動物となっています15。さらに、CHSモデルには多くの動物が必要であり、マウスはここでより経済的です。動物モデルは、すべての面でACDを模倣するわけではない。特に、彼らは人間には一般的ではない痂皮と落屑を示しています16,17。慢性疾患の特徴は、主に記載されたモデルが長期間にわたってハプテンの適用を想定していないため、再現が困難である。しかし、ACDの重要な側面の多くが再現されていることがここで確認されています。また、ヒトと同様に、これらの特徴は局所アレルギー反応と関連していることも示されている。このプロトコルで概説されているハプテン、その溶媒、およびその用途の選択は、結果が多数のインビトロ試験によって確認されており、現在のバージョンが確立されるまで長年にわたって実験室で試験および修正されたという事実によって決定された。マウスモデルは、ACDの発症に関与し、新しい治療法の前臨床評価に不可欠な細胞サブセットまたはサイトカインの分析を可能にする。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>70% ethanol</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>65350-M</td>
			<td>for surface disinfection</td>
		</tr>
		<tr>
			<td>96-well flat-bottom plates, polypropylene</td>
			<td>Greiner Bio-One GmbH, Kremsmunster, Austria</td>
			<td>655101</td>
			<td>for MPO and Evans blue measurement - plates should be made of polypropylene, that has a lower binding capacity so proteins or DNA will not bind</td>
		</tr>
		<tr>
			<td>Acetone (ACS reagent, &#8805;99.5%)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>179124</td>
			<td></td>
		</tr>
		<tr>
			<td>Aluminum foil</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>Z185140</td>
			<td></td>
		</tr>
		<tr>
			<td>Analytical balance</td>
			<td>Sartorius Weighing Technology GmbH, Goettingen, Germany</td>
			<td>PRACTUM224-1s, 29105177</td>
			<td></td>
		</tr>
		<tr>
			<td>Automated tissue processor</td>
			<td>MediMeas Instruments, Sarsehri, Haryana, India</td>
			<td>MTP-E-212</td>
			<td>automatically prepare tissue samples by fixing, dehydrating, clearing, and infiltrating them with paraffin</td>
		</tr>
		<tr>
			<td>BD Vacutainer SST II Advance (tube with gel for obtaining serum)</td>
			<td>Becton Dickinson (BD), Franklin Lakes, NJ, USA</td>
			<td>BD 366882</td>
			<td></td>
		</tr>
		<tr>
			<td>Bicinchoninic acid kit for protein determination</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>BCA1-1KT</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotin Rat Anti-Mouse IgG1</td>
			<td>Becton Dickinson (BD Biosciences), Franklin Lakes, NJ, USA</td>
			<td>553441</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA (bovine serum albumine)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>A9418</td>
			<td>protein assays &#38; analysis, 2 mg/mL</td>
		</tr>
		<tr>
			<td>Cell strainer, pore size 70 &#956;m</td>
			<td>BIOLOGIX, China</td>
			<td>15-1070</td>
			<td>suitable for 50 mL tubes</td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>VWR, Radnor, Pennsylvania, United States</td>
			<td>631-1583</td>
			<td>24 mm, but it possible to use different size</td>
		</tr>
		<tr>
			<td>Disposable pipettes capacity: 5 mL, 10 mL, 25 mL</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>Z740301, Z740302, Z740303</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS (Dulbecco&#8242;s phosphate buffered saline)</td>
			<td>ThermoFisher Scientific,&#160; Waltham, Massachusetts, USA</td>
			<td>14190144</td>
			<td>no calcium, no magnesium, mammalian cell culture</td>
		</tr>
		<tr>
			<td>DPX Mountant for histology</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>6522</td>
			<td>mounting media for H-E, might be used some other e.g, Canada balsam</td>
		</tr>
		<tr>
			<td>Dumont 5 tweezers - straight</td>
			<td>Animalab, Poznan, Poland</td>
			<td>11251-10FST</td>
			<td>surgical instruments for procedures on mice (should be steriled)</td>
		</tr>
		<tr>
			<td>Dumont 7 tweezers - bent</td>
			<td>Animalab, Poznan, Poland</td>
			<td>11272-50FST</td>
			<td>surgical instruments for procedures on mice (should be steriled)</td>
		</tr>
		<tr>
			<td>Eosin Y solution, alcoholic</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>HT110116</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf Safe-Lock Tubes 1.5 mL</td>
			<td>Eppenforf, Germany</td>
			<td>3,01,20,086</td>
			<td>polypropylene</td>
		</tr>
		<tr>
			<td>Eppendorf Safe-Lock Tubes, 2.0 mL</td>
			<td>Eppenforf, Germany</td>
			<td>3,01,20,094</td>
			<td>polypropylene, round bottom (the homogenization beads can easily move)</td>
		</tr>
		<tr>
			<td>Ethanol 100% (absolute alkohol)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>1.07017</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol 96%</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>1.59010</td>
			<td></td>
		</tr>
		<tr>
			<td>Evans blue</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>E2129</td>
			<td></td>
		</tr>
		<tr>
			<td>FBS (fetal bovine serum)</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>A3160802</td>
			<td></td>
		</tr>
		<tr>
			<td>Formalin solution, neutral buffered, 10%</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>HT501128</td>
			<td></td>
		</tr>
		<tr>
			<td>Formamide 99.5% (GC)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>F7503</td>
			<td></td>
		</tr>
		<tr>
			<td>Freezer -20 &#176;C</td>
			<td>Bosch, Germany</td>
			<td>GSN54AW30</td>
			<td></td>
		</tr>
		<tr>
			<td>Fridge +4 &#176;C / freezer -20 &#176;C</td>
			<td>Bosch, Germany</td>
			<td>KGV36V10</td>
			<td>mammalian Cell Culture, qualified, Brazil, 10 x 50 mL</td>
		</tr>
		<tr>
			<td>Glass microskope slides, SuperFrost Plus</td>
			<td>VWR, Radnor, Pennsylvania, United States</td>
			<td>631-0108, 631-0446, 631-0447, 631-0448, 631-0449</td>
			<td>Slides that eliminates the need to apply adhesive materials or protein coatings, to preventing any tissue sections loss during staining.</td>
		</tr>
		<tr>
			<td>Graph Pad Prism</td>
			<td>GraphPad Software Inc.</td>
			<td>v. 9.4.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Grey soap</td>
			<td>Pollena Ostrzesz&#243;w, Producent Chemii Gospodarczej Sp. Z.o.o. , Sp. K., Poland</td>
			<td>Bialy jelen soap bar</td>
			<td>grey Soap Bar Natural Hypoallergenic. Generally available product</td>
		</tr>
		<tr>
			<td>H2SO4 (sulfuric acid) 1 mol/l (1 M)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>1.60313</td>
			<td></td>
		</tr>
		<tr>
			<td>Harris hematoxylin solution</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>HHS16</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemocytometer</td>
			<td>VWR, Avantor, U.S.A</td>
			<td>612-5719</td>
			<td>manual counting chamber is recommend, which is more accurate</td>
		</tr>
		<tr>
			<td>Hexadecyltrimethylammonium bromide</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>H5882</td>
			<td></td>
		</tr>
		<tr>
			<td>Homogenizer</td>
			<td>QIAGEN Hilden, Germany</td>
			<td>Tissue Lyser LT, SN 23.1001/05234</td>
			<td>homogenizer with stainless steel beads (diameter 5 mm) for 2 mL centrifuge tubes</td>
		</tr>
		<tr>
			<td>Horseradish peroxidase streptavidin (HRP streptavidin)</td>
			<td>Vector Laboratories, Inc., Burlingame, CA, USA</td>
			<td>SA-5004-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrogen peroxide solution (H202)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>H1009</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator Heracell 150i</td>
			<td>Thermo Electron LED Gmbh, Germany</td>
			<td>41071068</td>
			<td>37 oC in the atmosphere of 5 % CO2, and 65 0C for deparaffinization the sections for histology</td>
		</tr>
		<tr>
			<td>Ketamine 100 mg/mL, solution for injection</td>
			<td>Biowet Pulawy Sp. z o.o., Pulawy, Poland</td>
			<td>cat.# not avaliable</td>
			<td></td>
		</tr>
		<tr>
			<td>KH2PO4 (potassium dihydrogen phosphate) 99.995% anhydrous basis</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>1.05108</td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory Centrifuge</td>
			<td>Heraeus Megafuge 1.0R, Thermo Scientific, Germany</td>
			<td>B00013899</td>
			<td>speed to 300 x g, with cooling to 4 0C</td>
		</tr>
		<tr>
			<td>Laboratory Centrifuge</td>
			<td>Heraeus Fresco 21, Thermo Scientific, Germany</td>
			<td>75002425</td>
			<td>speed to 3,000 x g, with cooling to 4 0C</td>
		</tr>
		<tr>
			<td>Mask (FFP2)</td>
			<td>VWR, Radnor, Pennsylvania, United States</td>
			<td>111-0917</td>
			<td>for working with ortho-dianisine dihydrochloride</td>
		</tr>
		<tr>
			<td>Mice</td>
			<td>Breeding unit of the Chair of Biomedical Sciences, Faculty of Health Sciences, Jagiellonian University Medical College, Krakow, Poland</td>
			<td>CBA/J, C57BL/6</td>
			<td></td>
		</tr>
		<tr>
			<td>Micrometer</td>
			<td>Mitutoyo, Tokyo, Japan</td>
			<td>193-111</td>
			<td>digit Outside Micrometer, Ratchet Stop, 0-25mm Range, 0.001mm Graduation, +/-0.002mm Accuracy, https://shop.mitutoyo.pl/web/mitutoyo/pl_PL/all/all/Mikrometr%20analogowy%20/PR/193-111/index.xhtml&#160;&#160;</td>
		</tr>
		<tr>
			<td>microplate, 96 well, microlon, high binding (for ELISA test)</td>
			<td>Greiner Bio-One GmbH, Kremsmunster, Austria</td>
			<td>655061</td>
			<td>with maxi-sorp binding surfaces for reliable and reproducible results in colormetric assays</td>
		</tr>
		<tr>
			<td>Microscope with objectives</td>
			<td>Leica Microsystems CMS GmbH, Germany</td>
			<td>DM1000, 294011-082007</td>
			<td>histology presented in the paper was performed under ThermoFisher Scientific EVOS M5000 Imaging System, with objectives: FL 20X LWDPH, 0.40NA/3.1WD and FL 40X LWDPH 0.65NA/1.79WD</td>
		</tr>
		<tr>
			<td>Myeloperoxidase from human leukocytes (MPO standard)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>M6908</td>
			<td></td>
		</tr>
		<tr>
			<td>Na2HPO4 x 7 H2O (sodium phosphate dibasic heptahydrate)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>S9390</td>
			<td></td>
		</tr>
		<tr>
			<td>Olive-oil</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>75343</td>
			<td>pure, natural</td>
		</tr>
		<tr>
			<td>OptEIA Mouse IFN-&#947; ELISA Set</td>
			<td>Becton Dickinson (BD Biosciences), Franklin Lakes, NJ, USA</td>
			<td>555138</td>
			<td></td>
		</tr>
		<tr>
			<td>Ortho-dianisine dihydrochloride</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>D3252</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraffin wax</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>76242</td>
			<td>beads, waxy solid</td>
		</tr>
		<tr>
			<td>PBS (phosphate buffered saline)</td>
			<td>ThermoFisher Scientific,&#160; Waltham, Massachusetts, USA</td>
			<td>20012027</td>
			<td>pH 7.2, mammalian cell culture</td>
		</tr>
		<tr>
			<td>ph meter</td>
			<td>Elmetron, Poland</td>
			<td>CP-401</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipettes, variable volume with tips</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>EP3123000900-1EA</td>
			<td>3-pack, Option 1, 0.5-10 uL/10-100 uL/100-1000 uL, includes epT.I.P.S.</td>
		</tr>
		<tr>
			<td>Razor blade</td>
			<td>VWR, Radnor, Pennsylvania, United States</td>
			<td>PERS94-0462</td>
			<td>scraper and cutter blades, single edge, aluminium spine, 100 blades per box, individually wrapped</td>
		</tr>
		<tr>
			<td>Rotary microtome</td>
			<td>MRC Laboratory-Instruments, Essex, CM20 2HU UK</td>
			<td>HIS-202A</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors - straight, sharp / sharp</td>
			<td>Animalab, Poznan, Poland</td>
			<td>14060-10FST</td>
			<td>Surgical instruments for procedures on mice (should be steriled)</td>
		</tr>
		<tr>
			<td>Screw cap (open top)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>27056</td>
			<td>black polypropylene hole cap, for use with 22&#160;mL vial with 20-400 thread</td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>BioTek Instruments, U.S.A</td>
			<td>201446</td>
			<td>universal microplate spectrophotometer: &#955; range: 200 - 999 nm, absorbance measurement range: 0.000 - 4.000 Abs</td>
		</tr>
		<tr>
			<td>Staining dish 20 slides with rack</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>S6141</td>
			<td>e.g. 20 slide staining dishes complete with covers, slide rack and handle</td>
		</tr>
		<tr>
			<td>Sterile Disposable Biopsy Punch 6mm</td>
			<td>Integra LifeSciences, Princeton, NJ, USA</td>
			<td>33-36</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td>Animalab, Poznan, Poland</td>
			<td>52138-46</td>
			<td>surgical instruments for procedures on mice (should be steriled)</td>
		</tr>
		<tr>
			<td>Tissue processing cassettes</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>Z672122</td>
			<td>tissue processing/ embedding cassettes with lid</td>
		</tr>
		<tr>
			<td>TMB Substrate Reagent Set</td>
			<td>Becton Dickinson (BD Biosciences), Franklin Lakes, NJ, USA</td>
			<td>555214</td>
			<td></td>
		</tr>
		<tr>
			<td>TNCB (2,4,6-trinitrochlorobenzene)</td>
			<td>Tokyo Chemical Industry CO., LTD, Japan</td>
			<td>C0307</td>
			<td></td>
		</tr>
		<tr>
			<td>TNP-BSA (bovine serum albumin conjugated with 2,4,6-trinitrophenyl)</td>
			<td>Biosearch Technologies LGC, Petaluma, CA, USA</td>
			<td>T-5050</td>
			<td></td>
		</tr>
		<tr>
			<td>T-PER (tissue protein extration reagent)</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>78510</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubes 15 mL sterile</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>CLS430055 (Corning)</td>
			<td>polypropylene, conical bottom</td>
		</tr>
		<tr>
			<td>Tubes 50 mL, sterile</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>CLS430290 (Corning)</td>
			<td>polypropylene, conical bottom</td>
		</tr>
		<tr>
			<td>Tween 20</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>P1379</td>
			<td></td>
		</tr>
		<tr>
			<td>Vials, screw top, clear glass (vial only) 22 mL</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>27173</td>
			<td>for the preparation of hapten, screwed on so that it does not evaporate</td>
		</tr>
		<tr>
			<td>Water bath</td>
			<td>AJL Electronic, Poland</td>
			<td>LW102</td>
			<td></td>
		</tr>
		<tr>
			<td>Wax (paraffin) dispenser</td>
			<td>VWR, Radnor, Pennsylvania, United States</td>
			<td>114-8737</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine (xylapan 20 mg/mL) solution for injection</td>
			<td>Vetoquinol Biowet Sp. z o.o., Gorzow Wielkopolski, Poland</td>
			<td>cat.# not avaliable</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene (histological grade)</td>
			<td>Merck KGaA, Darmstadt, Germany</td>
			<td>534056</td>
			<td></td>
		</tr>
	</tbody>
</table>

contact hypersensitivity as a murine model of allergic contact dermatitis

接触過敏症(CHS)は、マウスで研究することができるアレルギー性接触皮膚炎(ACD)の実験モデルである。この研究は、様々な試験によって測定および定量化することができるマウスにおけるCHS反応を研究するのに役立つかもしれない客観的な実験室法を提示することを目的としている。CHSを誘導するために、「0」日目に、マウスをアセトン - エタノール混合物中のハプテン2,4,6−トリニトロクロロベンゼン(TNCB)による腹部皮膚塗装によって予め剃毛したスポット上で感作し、一方、陰性対照マウスは、ビヒクル単独 - アセトン - エタノール混合物で偽感作した。「4」日目に、試験群および対照群の両方で希釈TNCBで両耳を塗ることによってCHSの惹起(チャレンジ)の前にベースライン耳厚をマイクロメーターで測定した。24時間後、耳の腫れをマイクロメーターで測定した。CHSは、炎症を起こした組織に腫脹を引き起こすT細胞媒介性免疫応答の一例であり、同じハプテンによる皮膚チャレンジの24時間後にピークに達する。耳水腫の増加は、耳の体重の増加、ミエロペルオキシダーゼ(MPO)活性、耳抽出物中の前炎症性サイトカイン濃度、組織学的検査における浮腫性真皮の肥厚の増加、および耳の血管透過性と相関した。対照マウスと比較した場合、試験群の血清中のTNP特異的IgG1抗体の濃度も増加した。さらに、CHSは、以前にTNCBで感作されたドナーから得られたCHSエフェクター細胞を用いて首尾よく転写することができる。CHSエフェクター細胞をナイーブレシピエントマウスに静脈内投与し、その後、同じ希釈ハプテンでチャレンジした。耳の腫れは、24時間後にマイクロメーターで測定した。

CHS誘導のために、動物を、150μLの5%TNCBまたは偽物を用いて、ビヒクル単独で感作した皮膚塗装(腹部) を介して 感作した。「4」日目に、TNCBで以前に接触感作したマウス(試験群)および対照群マウス(偽感作)の両マウスにおいて、0.4%TNCBの10 μLによる接触塗装(チャレンジ)により両耳の耳膨潤応答を誘導した。提示されたデータは、TNCBで感作し、4日後にチャレンジしたマウスが、同様にチ...

Watch this Scientific Journal Video about Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis at JoVE.com

Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis

接触過敏症(CHS)は、アレルギー性接触皮膚炎(ACD)のマウス実験モデルである。CHSは、胸部および腹部の剃毛された皮膚を塗ることによって反応性ハプテンによる感作に基づいており、その後の耳の皮膚の課題を希釈ハプテンで、様々な方法で評価される腫脹反応を引き起こす。

アレルギー性接触皮膚炎のマウスモデルとしての接触過敏症

Contact hypersensitivity (CHS) is an experimental model of allergic contact dermatitis (ACD) that can be studied in mice. This study aims to present an ...

ここで紹介する客観的な実験室の方法は、ヒトのアレルギー性接触皮膚炎を模倣するマウスの接触過敏反応を研究するのに役立つ可能性があります。接触過敏反応は、耳浮腫だけでなく、この反応に関与する細胞メカニズムの研究を可能にする様々な実験室試験によっても測定することができる。より接触性のある過敏症モデルは、さまざまな環境要因と新しい物質をテストして、テストされた要因がT細胞依存性免疫応答を調節するかどうかを示すことができ、新しい治療法の実施に使用できます。かみそりの刃でマウスの皮膚を剃り、足にラベルを付けることから始めます。接触過敏症(CHS)の誘発の6時間前に、灰色の石鹸を水で塗り、マウスの胸部と腹部の2センチメートル四方の領域を剃ります。同じ日に、以前に剃った場所に150マイクロリットルの新しく調製した5%ハプテンを適用してマウスを感作します。コントロールマウスでは、車両のみを適用します。動物をケージに戻す前に、ハプテンスポットを30秒間乾燥させます。4日目に、実験群を知らない観察者がゼロ時間でマイクロメーターを使用して麻酔をかけたマウスの耳の厚さを測定します。次に、テストグループとコントロールグループの耳の両側に10マイクロリットルの作りたての04%ハプテンを塗布し、30秒間乾燥させます。前回の0.4%ハプテン塗布から24時間後の5日目に、24時間測定のために耳の厚さの測定を繰り返します。耳の厚さ測定の24時間後、マウスがまだ深い麻酔下にあるときに、ハサミを使用して頭蓋骨のできるだけ近い耳を切り落とします。耳の遠位側で、生検パンチを使用して直径6ミリメートルのパンチを作ります。分析天びん上で各耳生検の重量を測定し、ミリグラムで表します。ミエロペルオキシダーゼ(MPO)アッセイでは、耳生検をステンレス鋼ビーズを備えた2ミリリットルの微量遠心チューブに入れ、500マイクロリットルの調製バッファーを追加します。生検をホモジナイザーで10分間ホモジナイズし、次にサンプルを摂氏4度で15分間冷却してから、再び10分間ホモジナイズします。生検が均質化されたらビーズを取り出します。ホモジネートを摂氏マイナス20度で30分間凍結します。サンプルを解凍してボルテックスし、均質化と凍結の手順を3回繰り返します。完了したら、ホモジネートを3, 000 Gで摂氏4度で30分間遠心分離します。上清をピペットで回収してMPO活性を測定し、タンパク質1ミリグラムあたりの単位で表します。血管透過性試験を行うには、0日目にマウスを感作し、4日目に、前述の手順を使用して耳にハプテンを適用して直接挑戦します。23時間後、ダルベッコのリン酸緩衝生理食塩水(DPBS)に1%エバンスブルー染料をグラム体重あたり8.3マイクロリットルを麻酔したマウスに静脈内注射します。1時間後、マウスに再度麻酔をかけ、前述のように耳生検を採取する。組織から色素を抽出するには、1ミリリットルのホルムアミドを含むチューブ内で生検を摂氏37度、5%二酸化炭素の雰囲気中で18時間インキュベートします。耳の生検を収集した後、マウスがまだ深い麻酔下にあるときに、ピンセットで眼球を取り除きます。マウスに穏やかな圧力をかけ、後眼窩洞から血清採取のためにチューブに血液を採取します。チューブを6回反転させ、血液が凝固するまで30分待ちます。次に、チューブを1, 300〜2, 000 Gで室温で10分間遠心分離します。前述の手順を使用して、0日目にTNCBハプテンでドナーマウスを感作します 4日目に、皮膚を消毒した後、鉗子を使用して麻酔をかけたマウスから腋窩リンパ節と鼠径リンパ節(ALN)を分離し、続いて脾臓を分離します。2つの顕微鏡スライドのつや消しの端の間で組織をマッシュし、細胞懸濁液を孔径70マイクロメートルのセルストレーナーに通します。次に、1%ウシ胎児血清を添加したDPBSで細胞を洗浄し、摂氏4度で10分間300Gで遠心分離します。上清をデカントし、残りの細胞ペレットを1〜5ミリリットルのDPBSに再懸濁します。ALNと脾臓の1:1の混合物を調製して、200マイクロリットルのDBPSで10〜7番目の細胞の7.0倍までの濃度を達成してから、麻酔をかけたマウスにCHSエフェクター細胞の混合物を静脈内注射します。ゼロ時間とチャレンジの24時間後に耳の厚さを測定します。データは、TNCBに感作し、4日後に耳に挑戦したマウスは、偽感作され、同様に挑戦された対照マウスと比較して、耳の腫れが有意に増加したことを示しました。試験群における耳浮腫の増加は、耳重量、MPO活性、耳抽出物中のインターフェロンガンマ濃度、組織学的検査における真皮浮腫の変化、および耳血管透過性の増加をもたらし、対照動物と比較した場合、試験マウスの血清においてTNP特異的IgG1抗体が上昇した。以前にTNCBで感作したドナーからCHSエフェクター細胞を投与された動物は、細胞を受け取らなかった動物と比較して、有意に増加した耳の腫れを示しました。最も重要な瞬間は、接触過敏症を引き起こすことです。ハプテン溶液は非常に揮発性で光に敏感であるため、動物の皮膚にすばやく塗布する必要があります。単離されたエフェクター細胞に対して追加の試験を実施することができる。例えば、細胞培養物は、抗原の存在下で増殖する能力を評価するために確立され得る。

contact-hypersensitivity-as-murine-model-allergic-contact

Research

JoVE Journal

Immunology and Infection

Recognition of Epidermal Transglutaminase by IgA and Tissue Transglutaminase 2 Antibodies in a Rare Case of Rhesus Dermatitis

Tissue transglutaminase 2 (tTG2) is an intestinal digestive enzyme which deamidates already partially digested dietary gluten e.g. gliadin peptides. In genetically predisposed individuals, tTG2 triggers autoimmune responses that are characterized by the production of tTG2 antibodies and their direct deposition into small intestinal wall 1,2. The presence of such antibodies constitutes one of the major hallmarks of the celiac disease (CD). Epidermal transglutaminase (eTG) is another member of the transglutaminase family that can also function as an autoantigen in a small minority of CD patients. In these relatively rare cases, eTG triggers an autoimmune reaction (a skin rash) clinically known as dermatitis herpetiformis (DH). Although the exact mechanism of CD and DH pathogenesis is not well understood, it is known that tTG2 and eTG share antigenic epitopes that can be recognized by serum antibodies from both CD and DH patients 3,4.
In this study, the confocal microscopy examination of biopsy samples from skin lesions of two rhesus macaques (Macaca mulatta) with dermatitis (Table 1, Fig. 1 and 2) was used to study the affected tissues. In one animal (EM96) a spectral overlap of IgA and tTG2 antibodies (Fig. 3) was demonstrated. The presence of double-positive tTG2+IgA+ cells was focused in the deep epidermis, around the dermal papillae. This is consistent with lesions described in DH patients 3. When EM96 was placed on a gluten-free diet, the dermatitis, as well as tTG2+IgA+ deposits disappeared and were no longer detectable (Figs. 1-3). Dermatitis reappeared however, based on re-introduction of dietary gluten in EM96 (not shown). In other macaques including animal with unrelated dermatitis, the tTG2+IgA+ deposits were not detected. Gluten-free diet-dependent remission of dermatitis in EM96 together with presence of tTG2+IgA+ cells in its skin suggest an autoimmune, DH-like mechanism for the development of this condition. This is the first report of DH-like dermatitis in any non-human primate.

Recognition of Epidermal Transglutaminase by IgA and Tissue Transglutaminase 2 Antibodies in a Rare Case of Rhesus Dermatitis

Dermatitis herpetiformis (DH) is a chronic inflammatory condition characterized by an autoimmune reaction between IgA and epidermal transglutaminase (eTG). DH develops in a very small portion of gluten-sensitive and/or celiac patients. The results of this study indicate that DH can also develop in a rhesus monkey host with symptoms of idiopatic dermatitis.

の稀なケースでIgAおよび組織トランスグルタミナーゼ2抗体による表皮トランスグルタミナーゼの認識アカゲザル炎

Tissue transglutaminase 2 (tTG2) is an intestinal digestive enzyme which deamidates already partially digested dietary gluten e.g. gliadin peptides. In ...

免疫・感染学

Introducing Shear Stress in the Study of Bacterial Adhesion

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the initial and determining step of the pathogenesis. Although experimentally adhesion is mostly studied in static conditions adhesion actually takes place in the presence of flowing liquid. First encounters between bacteria and their host often occur at the mucosal level, mouth, lung, gut, eye, etc. where mucus flows along the surface of epithelial cells. Later in infection, pathogens occasionally access the blood circulation causing life-threatening illnesses such as septicemia, sepsis and meningitis. A defining feature of these infections is the ability of these pathogens to interact with endothelial cells in presence of circulating blood. The presence of flowing liquid, mucus or blood for instance, determines adhesion because it generates a mechanical force on the pathogen. To characterize the effect of flowing liquid one usually refers to the notion of shear stress, which is the tangential force exerted per unit area by a fluid moving near a stationary wall, expressed in dynes/cm2. Intensities of shear stress vary widely according to the different vessels type, size, organ, location etc. (0-100 dynes/cm2). Circulation in capillaries can reach very low shear stress values and even temporarily stop during periods ranging between a few seconds to several minutes 1. On the other end of the spectrum shear stress in arterioles can reach 100 dynes/cm2 2. The impact of shear stress on different biological processes has been clearly demonstrated as for instance during the interaction of leukocytes with the endothelium 3. To take into account this mechanical parameter in the process of bacterial adhesion we took advantage of an experimental procedure based on the use of a disposable flow chamber 4. Host cells are grown in the flow chamber and fluorescent bacteria are introduced in the flow controlled by a syringe pump. We initially focused our investigations on the bacterial pathogen Neisseria meningitidis, a Gram-negative bacterium responsible for septicemia and meningitis. The procedure described here allowed us to study the impact of shear stress on the ability of the bacteria to: adhere to cells 1, to proliferate on the cell surface 5and to detach to colonize new sites 6 (Figure 1). Complementary technical information can be found in reference 7. Shear stress values presented here were chosen based on our previous experience1 and to represent values found in the literature. The protocol should be applicable to a wide range of pathogens with specific adjustments depending on the objectives of the study.

During the infection process, a key step is the adhesion of pathogens with host cells. In most instances this adhesion step occurs in the presence of mechanical stress generated by flowing liquid. We describe a technique that introduces shear stress as an important parameter in the study of bacterial adhesion.

細菌付着の研究ではせん断応力を導入する

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like allergic reactions 1. In clinical practice, diagnosis of drug hypersensitivity is mainly performed by patient history, as skin testing is not reliable and oral provocation testing bears life-threatening risks for the patient 2. Hence, evidence for an underlying IgE-mediated pathomechanism is hard to obtain. 
Here, we present an in vitro method based on the use of human basophils derived from drug-hypersensitive patients that mimics the allergic effector reaction in vivo. As basophils of drug-allergic patients carry IgE molecules specific for the culprit drug, they become activated upon IgE receptor crosslinking and release allergic effector molecules. The activation of basophils can be monitored by the determination of the upregulation of CD63 surface expression using flow cytometry 3. 
In the case of low molecular weight drugs, conjugates are designed to enable IgE receptor crosslinking on basophils. As depicted in Figure 1, two representatives of NSAIDs, PP and DF, are covalently bound to human serum albumin (HSA) via a carboxyl group reacting with the primary amino group of lysine residues. DF carries an intrinsic carboxyl group and, thus, can be used directly 4, whereas a carboxyl group-containing derivative of PP had to be organochemically synthesized prior to the study 1.
The coupling degree of the low molecular weight compounds on the protein carrier molecule and their spatial distribution is important to guarantee crosslinking of two IgE receptor molecules. The here described protocol applies high performance-size exclusion chromatography (HPSEC) equipped with a sequential refractive index (RI) and ultra violet (UV) detection system for determination of the coupling degree.
As the described methodology may be applied for other drugs, the basophil activation test (BAT) bears the potential to be used for the determination of IgE-mediated mechanisms in drug hypersensitivity. Here, we determine PP hypersensitivity as IgE-mediated and DF hypersensitivity as non-IgE-mediated by BAT.

Basophil activation test is a potent tool for the detection of IgE-dependent allergies in vitro. Here, an optimized protocol for basophil activation test is used to investigate drug hypersensitivity. A method for the efficient production of covalent drug-protein conjugates and their physicochemical characterization is described.

薬物過敏症におけるIgEが介在する機序の解明のための好塩基球の活性化試験

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like ...

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Graft-versus-host disease (GVHD) is the limiting barrier to the broad use of bone marrow transplant as a curative therapy for a variety of hematological deficiencies. GVHD is caused by mature alloreactive T cells present in the bone marrow graft that are infused into the recipient and cause damage to host organs. However, in mice, T cells must be added to the bone marrow inoculum to cause GVHD. Although extensive work has been done to characterize T cell responses post transplant, bioluminescent imaging technology is a non-invasive method to monitor T cell trafficking patterns in vivo. 
Following lethal irradiation, recipient mice are transplanted with bone marrow cells and splenocytes from donor mice. T cell subsets from L2G85.B6 (transgenic mice that constitutively express luciferase) are included in the transplant. By only transplanting certain T cell subsets, one is able to track specific T cell subsets in vivo, and based on their location, develop hypotheses regarding the role of specific T cell subsets in promoting GVHD at various time points. At predetermined intervals post transplant, recipient mice are imaged using a Xenogen IVIS CCD camera. Light intensity can be quantified using Living Image software to generate a pseudo-color image based on photon intensity (red = high intensity, violet = low intensity). 
Between 4-7 days post transplant, recipient mice begin to show clinical signs of GVHD. Cooke et al.1 developed a scoring system to quantitate disease progression based on the recipient mice fur texture, skin integrity, activity, weight loss, and posture. Mice are scored daily, and euthanized when they become moribund. Recipient mice generally become moribund 20-30 days post transplant. 
Murine models are valuable tools for studying the immunology of GVHD. Selectively transplanting particular T cell subsets allows for careful identification of the roles each subset plays. Non-invasively tracking T cell responses in vivo adds another layer of value to murine GVHD models.

Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model

Murine bone marrow transplantation is a widely used technique to study immunological mechanisms governing graft-versus-host disease in humans. The ability to monitor T cell trafficking patterns in vivo allows for detailed analysis of the development and perpetuation of T cell responses during graft-versus-host disease.

移植片対宿主病の誘導および<emインビボで&gt;</emMHCマッチしたマウスモデルを使用する&gt; T細胞のモニタリング

Graft-versus-host disease (GVHD) is the limiting barrier to the broad use of bone marrow transplant as a curative therapy for a variety of hematological ...

Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently shown that UPEC have an important role in the initiation of chronic pelvic pain2, a feature of Chronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS)3,4. Infection of the prostate by clinically relevant UPEC can initiate and establish chronic pain through mechanisms that may involve tissue damage and the initiation of mechanisms of autoimmunity5.
A challenge to understanding the pathogenesis of UPEC in the prostate is the relative inaccessibility of the prostate gland to manipulation. We utilized a previously described intraurethral infection method6 to deliver a clinical strain of UPEC into male mice thereby establishing an ascending infection of the prostate. Here, we describe our protocols for standardizing the bacterial inoculum7 as well as the procedure for catheterizing anesthetized male mice for instillation of bacteria.
CP/CPPS is primarily characterized by the presence of tactile allodynia4. Behavior testing was based on the concept of cutaneous hyperalgesia resulting from referred visceral pain8-10. An irritable focus in visceral tissues reduces cutaneous pain thresholds allowing for an exaggerated response to normally non-painful stimuli (allodynia). Application of normal force to the skin result in abnormal responses that tend to increase with the intensity of the underlying visceral pain. We describe methodology in NOD/ShiLtJ mice that utilize von Frey fibers to quantify tactile allodynia over time in response to a single infection with UPEC bacteria.

Infection of the prostate may be a contributing factor in mediating pelvic pain in chronic prostatitis. We describe the procedure for preparation of standardized bacterial inoculum, instillation of bacteria into the urethra of male mice and methodology for measuring tactile allodynia in mice over time.

細菌性前立腺炎のマウスモデルにおける接触性アロディニアの測定

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently ...

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

Legionella pneumophila, the causative agent of a severe pneumonia named Legionnaires&#39; disease, is an important human pathogen that infects and replicates within alveolar macrophages. Its virulence depends on the Dot/Icm type IV secretion system (T4SS), which is essential to establish a replication permissive vacuole known as the Legionella containing vacuole (LCV). L. pneumophila infection can be modeled in mice however most mouse strains are not permissive, leading to the search for novel infection models. We have recently shown that the larvae of the wax moth Galleria mellonella are suitable for investigation of L. pneumophila infection. G. mellonella is increasingly used as an infection model for human pathogens and a good correlation exists between virulence of several bacterial species in the insect and in mammalian models. A key component of the larvae&#39;s immune defenses are hemocytes, professional phagocytes, which take up and destroy invaders. L. pneumophila is able to infect, form a LCV and replicate within these cells. Here we demonstrate protocols for analyzing L. pneumophila virulence in the G. mellonella model, including how to grow infectious L. pneumophila, pretreat the larvae with inhibitors, infect the larvae and how to extract infected cells for quantification and immunofluorescence microscopy. We also describe how to quantify bacterial replication and fitness in competition assays. These approaches allow for the rapid screening of mutants to determine factors important in L. pneumophila virulence, describing a new tool to aid our understanding of this complex pathogen.

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

The larva of the wax moth Galleria mellonella was recently established as an in vivo model to study Legionella pneumophila infection. Here, we demonstrate fundamental techniques to characterize the pathogenesis of Legionella in the larvae, including inoculation, measurement of bacterial virulence and replication as well as extraction and analysis of infected hemocytes.

使用<em&gt;ハチノスツヅリガ</em&gt;モデル生物として研究する<em&gt;レジオネラ·ニューモフィラ</em&gt;感染症

Legionella pneumophila, the causative agent of a severe pneumonia named Legionnaires&#39; disease, is an important human pathogen that infects and ...

Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori

Recently infection biologists have employed stem cell derived cultures to answer the need for new and better models to study host-pathogen interactions. Three cellular sources have been used: Embryonic stem cells (ESC), induced pluripotent stem cells (iPSC) or adult stem cells. Here, culture of mouse and human gastric organoids derived from adult stem cells is described and used for infection with the gastric pathogen Helicobacter pylori. Human gastric glands are isolated from resection material, seeded in a basement matrix and embedded in medium containing growth factors epidermal growth factor (EGF), R-spondin, Noggin, Wnt, fibroblast growth factor (FGF) 10, gastrin and transforming growth factor (TGF) beta inhibitor. In these conditions, gastric glands grow into 3-dimensional organoids containing 4 lineages of the stomach. The organoids expand indefinitely and can be frozen and thawed similarly as cell lines. For infection studies, bacteria are microinjected into the lumen of the organoids. Infected organoids are processed for imaging. The described methods can be adapted to other organoids and infections with other bacteria, viruses or parasites. This allows the study of infection-induced changes in primary cells.

Stem cell derived cultures harbor tremendous potential to model infectious diseases. Here, the culture of mouse and human gastric organoids derived from adult stem cells is described. The organoids are microinjected with the gastric pathogen Helicobacter pylori.

感染症のためのモデルとしてのオルガノイド：人間の文化とマウス胃オルガノイドとピロリ菌のマイクロインジェクション

Recently infection biologists have employed stem cell derived cultures to answer the need for new and better models to study host-pathogen interactions. ...

Zebrafish as a Model to Assess the Teratogenic Potential of Nitrite

High nitrate levels in the environment may&#160;result in congenital defects or miscarriages in humans. Presumably, this is due to the conversion of nitrate to nitrite by gut and salivary bacteria. However, in other mammalian studies, high nitrite levels do not cause birth defects, although they can lead to poor reproductive outcomes. Thus, the teratogenic potential of nitrite is not clear. It would be useful to have a vertebrate model system to easily assess teratogenic effects of nitrite or any other chemical of interest. Here, we demonstrate the utility of zebrafish (Danio rerio) to screen compounds for toxicity and embryonic defects. Zebrafish embryos are fertilized externally and have rapid development, making them a good model for teratogenic studies. We show that increasing the time of exposure to nitrite negatively affects survival. Increasing the concentration of nitrite also adversely affects survival, whereas nitrate does not. For embryos that survive nitrite exposure, various defects can occur, including pericardial and yolk sac&#160;edema, swim bladder noninflation, and craniofacial malformation. Our results indicate that the zebrafish is a convenient system for studying the teratogenic potential of nitrite. This approach can easily be adapted to test other chemicals for their effects on early vertebrate development.

Exposure to teratogens may cause birth defects. Zebrafish are useful for determining the teratogenic potential of chemicals. We demonstrate the utility of zebrafish by exposing embryos to various levels of nitrite and also at different times of exposure. We show that nitrite can be toxic and cause severe developmental defects.

亜硝酸塩の催奇形性を評価するためのモデルとしてのゼブラフィッシュ

High nitrate levels in the environment may&#160;result in congenital defects or miscarriages in humans. Presumably, this is due to the conversion of ...

A Murine Model of Group B Streptococcus Vaginal Colonization

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of 10 - 30% of adults. In immune-compromised individuals, including neonates, pregnant women, and the elderly, GBS may switch to an invasive pathogen causing sepsis, arthritis, pneumonia, and meningitis. Because GBS is a leading bacterial pathogen of neonates, current prophylaxis is comprised of late gestation screening for GBS vaginal colonization and subsequent peripartum antibiotic treatment of GBS-positive mothers. Heavy GBS vaginal burden is a risk factor for both neonatal disease and colonization. Unfortunately, little is known about the host and bacterial factors that promote or permit GBS vaginal colonization. This protocol describes a technique for establishing persistent GBS vaginal colonization using a single &#946;-estradiol pre-treatment and daily sampling to determine bacterial load. It further details methods to administer additional therapies or reagents of interest and to collect vaginal lavage fluid and reproductive tract tissues. This mouse model will further the understanding of the GBS-host interaction within the vaginal environment, which will lead to potential therapeutic targets to control maternal vaginal colonization during pregnancy and to prevent transmission to the vulnerable newborn. It will also be of interest to increase our understanding of general bacterial-host interactions in the female vaginal tract.

A Murine Model of Group B Streptococcus Vaginal Colonization

The purpose of this protocol is to imitate human group B Streptococcus (GBS) vaginal colonization in a murine model. This method may be used to investigate host immune responses and bacterial factors contributing to GBS vaginal persistence, as well as to test therapeutic strategies.

グループBのマウスモデル<em&gt;連鎖球菌</em&gt;膣植民地化

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of ...

Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment

Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between genetic and environmental factors are known to play an important role in the pathophysiology of atopic dermatitis. However, we still lack a detailed picture of the pathogenesis of this disease. Thus, it is of importance to develop appropriate animal models for elucidating the progression of atopic dermatitis. Moreover, investigating the effect of environmental factors such as air pollutants on atopic dermatitis expands understanding of the disease. Here, we describe a method for inducing atopic dermatitis in rats with neonatal capsaicin treatment and a protocol for exposure of a constant concentration of formaldehyde to rats to reveal effects on the development of atopic dermatitis in infantile and adolescent periods. These protocols have been successfully applied to several experiments and can be used for other substances.

We describe here methods for neonatal capsaicin treatment to induce atopic dermatitis in rats and exposure of vaporized formaldehyde to investigate the effects of formaldehyde inhalation on atopic dermatitis.

新生児カプサイシン処置によるアトピー性皮膚炎のモデルラットにホルムアルデヒドの暴露の影響

Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between ...