The authors acknowledge the help from the staff of Laboratory Animal Medicine, University of Nevada, Reno. MM received funding from Nevada INBRE.

下記に概説プロトコル（数-00682）は、ネバダ大学リノ校で施設内動物管理使用委員会（IACUC）によって承認され、ネバダ大学、リノの動物研究倫理委員会のガイドラインに従っています。手短に言えば、マウスをイソフルランで麻酔し、ノーズコーンを20分間連続イソフルランレベルを維持するために使用しました。獣医軟膏は、麻酔下で、目の乾燥を防ぐために使用されました。つま先のピンチは、麻酔のレベルを確認するために使用され、呼吸数は、プロシージャ全体で測定しました。マウスは、個々のケージに保管し、意識を取り戻したまで監視しました。マウスは、ダニの供給のために一度だけ使用し、ポストティック回復を安楽死させました。安楽死は、動物医療スタッフの人員の研究室で行いました。 CO 2及び頸椎脱臼は、動物を安楽死させるために使用されました。 注：ティックで作業が完了個人用保護EQを使用する必要がありますuipment。未成熟ダニをカウントするため、ゴムバンド、及び閉鎖つま先靴でキャップ白衣のスリーブ開口部を覆うように白白衣、長いニトリル手袋を着用します。ダニをマウスに寄生するために、ヘアネット、使い捨てカバーオール、袖オーバー長袖手袋、フットカバーを使用しています。わがままなダニを検出するために、白や明るい色のカバーオールを使用してください。定期的にダニ用手袋と袖を確認してください。 動物住宅用1.準備室<ol><li>ダニが寄生し、家のマウスに別の部屋を指定します。偶発チック脱出を防ぐために、外部のドアの内側に粘着マットまたは両面カーペットテープを置きます。 </li></ol> 2.カウントはマウス加害ティック<ol><li>すべての7&quot; ×5&quot; ×14&quot; プラスチック容器の上縁の周りに両面テープを入れて、約2cmまで水で満たします。 </li><li> ＆容器の中心に別の小さなボックスまたはペトリ皿を置き、作成1センチメートルまで水で満たします＃34;堀 &quot; </li><li>ペトリ皿の中に幼虫やニンフを含むバイアルを保管します。 </li><li>バイアルから幼虫または幼虫を取り除き、顕微鏡下で数えるために、細かいペイントブラシを使用してください。 50匹の幼虫または25匹のニンバを別々のシンチレーションバイアルに数える。バイアルをナイロンメッシュまたはオルガンディッシュのスクリーンで直ちに覆い、ゴムバンドで閉めます。 </li></ol> 3.未成熟なティックでマウスに感染する<ol><li>白または薄い色の作業台を使用し、両面テープを作業エリアの周辺に貼り付けます。 </li><li>イソフルランでマウスを麻酔する。つま先をつまんで麻酔のレベルを確認してください。麻酔を終えたら、マウスをペーパータオルで覆われた加熱パッドに移し、継続的なイソフルラン供給のためにノーズコーンに取り付けます。 </li><li>乾燥を避けるために石油ゼリーベースの眼軟膏を塗布する。イソフルランレベルを調節するためのマウスの呼吸パターンに注意してください（毎分80~230回の呼吸が正常です）。もしbrであればイソフルランレベルを下げてくださいeathing率）は、動物を殺す避けるために毎分80未満の呼吸です。 </li><li> 50幼虫または若虫25の1つのバイアルを取り、場所はペイントブラシ（ 図1）と耳の間の頭部に毛下ティック。ダニは添付するのに十分な時間を与えるために配置した後20分間麻酔下でマウスを保管してください。 </li><li>静的蓋と白の寝具標準、プラスチック製の靴箱型の使い捨てマウスケージにマウスを移動します。正常なマウスケアに従っておもちゃ、水、および食品を自由に提供します。 </li><li>トップエッジの周りに両面粘着テープと並ぶ大きなラットやスナネズミケージ内にマウスケージを保管してください。水3センチ（ 図2）と外側のケージを埋めます。 </li></ol> 4.マウスからダニの収集<ol><li>未熟ティックは日3及び摂食の6の間で切り離します。 3日後に毎日充血ダニのためのケージと水の堀を確認してください。 </li><li>日4および6.痛みの間にケージから取り外したダニを集めますtブラシまたは軟らかい鉗子を使用してゴムバンドで固定されたナイロンメッシュ布でキャップされたきれいなシンチレーションバイアルに保管してください。 </li><li> 7日目にマウスに麻酔をかけ、残りの付着したダニを確認する。 </li><li> 7日目に詰まったダニについて寝具、フードトラフ、水筒を点検する。上記の手順に従ってマウスを安楽死させる。使い捨てケージ、寝具、ウォーターボトル、フードトラフをオートクレーブして、脱落しないようにします。 </li></ol> 5. Fed Ticksの保管<ol><li> 90％の湿度で虫歯を防ぎ、20 湿潤および温度制御されたインキュベーター中で脱皮が起こるまで、12：12の明暗サイクルを繰り返す。これには約12〜18週間かかります。 </li><li>湿度が高いとダニの生存が促進されるが、ダニは真菌の過増殖を起こしやすい。少なくとも1週間に1回、顕微鏡下で飼育されたダニを金型検査する。検出された場合、生存者を70％エタノールで5回洗浄する分、乾燥するためにろ紙に移し、水ですすぎ、そして新しい、きれいなバイアルに移します。 </li></ol>

<ol>
	<li>Gleim, E. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+associated+with+tick+bites+and+pathogen+prevalence+in+ticks+parasitizing+humans+in+Georgia,+USA.">Factors associated with tick bites and pathogen prevalence in ticks parasitizing humans in Georgia, USA.</a> Parasites &amp;amp; Vectors. 9 (125), 1-13 (2016).</li><li>Krober, T., Guerin, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+feeding+assays+for+hard+ticks.">In vitro feeding assays for hard ticks.</a> Trends Parasitol. 23 (9), 445-449 (2007).</li><li>Kuhnert, 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+Hard+Ticks+In+Vitro:+New+Perspectives+for+Rearing+and+for+the+Identification+of+Systemic+Acaricides.">Feeding of Hard Ticks In Vitro: New Perspectives for Rearing and for the Identification of Systemic Acaricides.</a> ALTEX. 13 (2), 76-87 (1996).</li><li>Voigt, W. 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=In+vitro+feeding+of+instars+of+the+ixodid+tick+Amblyomma+variegaturn+on+skin+membranes+and+its+application+to+the+transmission+of+Theileria+mutans+and+Cowdria+ruminantium.">In vitro feeding of instars of the ixodid tick Amblyomma variegaturn on skin membranes and its application to the transmission of Theileria mutans and Cowdria ruminantium.</a> Parasitol. 107, 257-263 (1993).</li><li>Gregson, J. D., Smith, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ticks.">Ticks.</a> Insect Colonization and Mass Production. , 49-72 (1966).</li><li>Sonenshine, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biology of Ticks. 2, (1993).</li><li>Bouchard, K. R., Wikel, S. K., Marquaedt, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Care,+maintenance,+and+experimental+infestation+of+ticks+in+the+laboratory+setting.">Care, maintenance, and experimental infestation of ticks in the laboratory setting.</a> Biology of Disease Vectors. , 705-711 (2005).</li><li>Schumaker, T. S., Barros, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+cycle+of+Ornithodoros+(Alectorobius)+talaje.+(Acari:Argasidae)+in+laboratory.">Life cycle of Ornithodoros (Alectorobius) talaje. (Acari:Argasidae) in laboratory.</a> J Med Entomol. 32, 249-254 (1995).</li><li>Banks, C. W., Oliver, J. H., Hopla, C. E., Dotson, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+life+cycle+of+Ixodes+woodi+(Acari:Ixodidae).">Laboratory life cycle of Ixodes woodi (Acari:Ixodidae).</a> J. Med. Entomol. 35, 177-179 (1998).</li><li>James, A. M., Oliver, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+and+host+preference+of+immature+Ixodes+dammini,I.scapularis,and+I.pacificus.(Acari:Ixodidae).">Feeding and host preference of immature Ixodes dammini,I.scapularis,and I.pacificus.(Acari:Ixodidae).</a> J. Med. Entomol. 27, 324-330 (1990).</li></ol>

The authors have nothing to disclose.

プロトコル内の重要なステップ偶発的な逃げを避けるためにダニを飼育するときは、いくつかのレベルの安全対策が必要です。粘着テープと水面を使用することは、安全性を確保する上で重要です。体温を一定に保つために、麻酔したマウスをヒーティングパッド上に置くことが重要です。我々はまた、マウスをシェービングしても、ダニ付着のための特別な利益は得られ...

ティックは、脊椎動物の必須の赤血球性外部寄生虫であり、世界中に分布している。米国では、少なくとも11種のダニは公衆衛生上重要な病原菌のベクターである1 。 Ixodes肩甲介は、ライム病（ Borrelia burgdorferi ）再発熱（ B. miyamotoi）、ヒト顆粒球アナプラズマ症（ Anaplasma phagocytophilum ）、およびバベシア症（Babesia spp。）の原因物質のようないくつかの病原体の伝達に関与する。疾病ベクターとしての肩甲介の重要性にもかかわらず、研究室での研究のためにこれらのクモ形類を野生から豊富に収集することは、必ずしも実現可能ではない。したがって、健全な実験室で飼育されたダニの生産は、ダニ生物学およびダニ病原体の相互作用に関する研究にとって不可欠である。 I. scapularisを含むすべての硬ダニ（Ixodidae科）の生活環は、卵と3つの活性成分eステージ：幼虫、ニンフ、および成体。各活性段階は、脊椎動物の宿主に供給される。付着と摂食の数日にわたってダニとその宿主との間で起こる複雑な相互作用は人工フィーダーを用いて複製することはほとんど不可能であり、大量飼育のために十分な数の飼養されたダニを提供する可能性は低い2、3、4 。したがって、生きたマウスおよびウサギは、未成熟（幼虫および幼虫）およびダニの成熟段階（成虫）をそれぞれ飼育するための宿主として最も頻繁に使用される。各発達段階の間に複数の宿主が血液供給のために必要であるため、ダニの飼育が複雑になり、時間とコストが掛かる5,6,7。ほとんどのダニ飼育プロトコルでは、マウスを吊り下げられたワイヤーグリッドの床ケージ7,8または円筒状のカード動物は自由に移動し、それ自体6、9、10をグルーミングすることができないような寸法のGE。 これらの円筒形のケージは、後のワイヤグリッドで靴箱ケージに転送されます。充血し、剥離ダニは、次いで下水から収集されます。しかし、連邦準備制度を露光におけるこの方法の結果は、真菌の増殖を増加させ、死亡率9をチェックすることができ、尿及び糞便で汚染された水ティック。また、ダニ水トラフから脱出、ならびにマウスにストレスを引き起こす可能性が高くなります。これらの問題を回避するために、我々はここではプラスチック製の靴箱型の使い捨てケージ内でマウスの幼虫ダニの摂食を示しています。この方法は、マウスの正常な動作を可能に充血ダニ回復を増加し、汚染によるダニの死亡率を減少させます。

<table><tbody><tr><td>Puralube vet ointment</td><td>Amazon</td><td></td><td>Available from any Pet store or online store</td></tr><tr><td>Disposable mouse cage</td><td>&amp;nbsp;Innovive, San Diego, CA&amp;nbsp;</td><td>MV 2</td><td>Set of bottom and lid</td></tr><tr><td>White Alpha dri bedding&amp;nbsp;</td><td>Lab Supply, Fort Worth, TX</td><td>&amp;nbsp;ALPHA-Dri&amp;trade;</td><td></td></tr></tbody></table>

rearing ixodes scapularis, the black-legged tick: feeding immature stages on mice

マダニのscapularis、ライム病のベクトルは、東部と中西部米国で最も重要な病気のベクトルのうちの1つです。この種は、各開発段階のための脊椎動物宿主からの血液の食事を必要とする3つのホストダニで、成人女性は、再生のための血液の食事を必要とします。給餌のための5日間、完全に充血したときにホストを降ろし - 幼虫は3のために自分のホストに接続ティック。いくつかの異なるホスト上のこの依存性や充血のための長い添付ファイルの時間は、実験室の設定で飼育ダニを複雑にします。しかし、生物学をダニやダニ - 病原体相互作用を理解するために、健康、研究室飼育の生産が不可欠であるダニ。ここでは、マウスを餌未熟ダニのための簡単な、費用対効果の高いプロトコルを示しています。我々は、マウスで減少し、ストレスのための既存のプロトコルを変更し、ダニのwiの接触を避けるために、メッシュボトムなし使い捨てケージを使用して、成功と生存を供給増加ダニマウスの尿や糞で汚染された水。

我々は、改善された給電効率を既存のダニ飼育プロトコル6、10を変更し、マウス宿主へのストレスを低減します。結果は、標準的なシューボックススタイルのマウスのケージは、ダニの飼育に適していることを示しています。白い寝具は、供給されたダニの容易な収集のための良好なコントラストを提供します。ほとんど?...

Watch this Scientific Journal Video about Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice at JoVE.com

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

健康的な実験室飼育の生産は、ダニの生物学の研究、およびダニ病原体相互作用に不可欠であるダニ。ここでは、費用対効果およびマウスにストレスの少ないです未熟ダニ供給のための単純なプロトコルを示しています。

飼育<em&gt; Ixodes肩甲骨、</em&gt;ブラック・ティック・ティック：未熟期をマウスに与える

Ixodes scapularis, the vector of Lyme disease, is one of the most important disease vectors in the eastern and Midwestern United States. This species is a ...

rearing-ixodes-scapularis-black-legged-tick-feeding-immature-stages

Research

JoVE Journal

Immunology and Infection

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

11.9K ビュー.  University of Nevada, Reno. 健康的な実験室飼育の生産は、ダニの生物学の研究、およびダニ病原体相互作用に不可欠であるダニ。ここでは、費用対効果およびマウスにストレスの少ないです未熟ダニ供給のための単純なプロトコルを示しています。 

ビデオ: Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut

Lyme disease is caused by infection with the spirochete pathogen Borrelia burgdorferi, which is maintained in nature by a tick-rodent infection cycle 1. A tick-borne murine model 2 has been developed to study Lyme disease in the laboratory. While na&iacute;ve ticks can be infected with B. burgdorferi by feeding them on infected mice, the molting process takes several weeks to months to complete. Therefore, development of more rapid and efficient tick infection techniques, such as a microinjection-based procedure, is an important tool for the study of Lyme disease 3,4. The procedure requires only hours to generate infected ticks and allows control over the delivery of equal quantities of spirochetes in a cohort of ticks. This is particularly important as the generation of B. burgdorferi infected ticks by the natural feeding process using mice fails to ensure 100% infection rate and potentially results in variation of pathogen burden amongst fed ticks. Furthermore, microinjection can be used to infect ticks with B. burgdorferi isolates in cases where an attenuated strain is unable to establish infection in mice and thus can not be naturally acquired by ticks 5. This technique can also be used to deliver a variety of other biological materials into ticks, for example, specific antibodies or double stranded RNA 6. In this article, we will demonstrate the microinjection of nymphal ticks with in vitro-grown B. burgdorferi. We will also describe a method for localization of Lyme disease pathogens in the tick gut using confocal immunofluorescence microscopy.

Lyme disease research studies often require generation of ticks infected with the pathogen Borrelia burgdorferi, a process that typically takes several weeks. Here we demonstrate a microinjection-based tick infection procedure that can be accomplished within hours. We also demonstrate an immunofluorescence method for in situ localization of B. burgdorferi within ticks.

ティック腸内ライム病の病原体の迅速な伝達とローカリゼーションのための方法

Lyme disease is caused by infection with the spirochete pathogen Borrelia burgdorferi, which is maintained in nature by a tick-rodent infection cycle 1.  ...

免疫・感染学

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne relapsing fever (Borrelia spp.), Rocky Mountain Spotted fever (Rickettsia rickettsii), ehrlichiosis (Ehrlichia chaffeensis and E. equi), anaplasmosis (Anaplasma phagocytophilum), encephalitis (tick-borne encephalitis virus), babesiosis (Babesia spp.), Colorado tick fever (Coltivirus), and tularemia (Francisella tularensis) 1-8. To be properly transmitted into the host these infectious agents differentially regulate gene expression, interact with tick proteins, and migrate through the tick 3,9-13. For example, the Lyme disease agent, Borrelia burgdorferi, adapts through differential gene expression to the feast and famine stages of the tick's enzootic cycle 14,15. Furthermore, as an Ixodes tick consumes a bloodmeal Borrelia replicate and migrate from the midgut into the hemocoel, where they travel to the salivary glands and are transmitted into the host with the expelled saliva 9,16-19.
As a tick feeds the host typically responds with a strong hemostatic and innate immune response 11,13,20-22. Despite these host responses, I. scapularis can feed for several days because tick saliva contains proteins that are immunomodulatory, lytic agents, anticoagulants, and fibrinolysins to aid the tick feeding 3,11,20,21,23. The immunomodulatory activities possessed by tick saliva or salivary gland extract (SGE) facilitate transmission, proliferation, and dissemination of numerous tick-borne pathogens 3,20,24-27. To further understand how tick-borne infectious agents cause disease it is essential to dissect actively feeding ticks and collect tick saliva. This video protocol demonstrates dissection techniques for the collection of hemolymph and the removal of salivary glands from actively feeding I. scapularis nymphs after 48 and 72 hours post mouse placement. We also demonstrate saliva collection from an adult female I. scapularis tick.

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

The collection of infected tick hemolymph, salivary glands, and saliva is important to study how tick-borne pathogens cause disease. In this protocol we demonstrate how to collect hemolymph and salivary glands from feeding Ixodes scapularis nymphs. We also demonstrate saliva collection from female I. scapularis adults.

マダニScapularisダニの唾液、唾液腺、および体液コレクション

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne ...

Rearing and Injection of Manduca sexta Larvae to Assess Bacterial Virulence

Manduca sexta, commonly known as the tobacco hornworm, is considered a significant agricultural pest, feeding on solanaceous plants including tobacco and tomato. The susceptibility of M. sexta larvae to a variety of entomopathogenic bacterial species1-5, as well as the wealth of information available regarding the insect's immune system6-8, and the pending genome sequence9 make it a good model organism for use in studying host-microbe interactions during pathogenesis. In addition, M. sexta larvae are relatively large and easy to manipulate and maintain in the laboratory relative to other susceptible insect species. Their large size also facilitates efficient tissue/hemolymph extraction for analysis of the host response to infection.
The method presented here describes the direct injection of bacteria into the hemocoel (blood cavity) of M. sexta larvae. This approach can be used to analyze and compare the virulence characteristics of various bacterial species, strains, or mutants by simply monitoring the time to insect death after injection. This method was developed to study the pathogenicity of Xenorhabdus and Photorhabdus species, which typically associate with nematode vectors as a means to gain entry into the insect. Entomopathogenic nematodes typically infect larvae via natural digestive or respiratory openings, and release their symbiotic bacterial contents into the insect hemolymph (blood) shortly thereafter10. The injection method described here bypasses the need for a nematode vector, thus uncoupling the effects of bacteria and nematode on the insect. This method allows for accurate enumeration of infectious material (cells or protein) within the inoculum, which is not possible using other existing methods for analyzing entomopathogenesis, including nicking11 and oral toxicity assays12. Also, oral toxicity assays address the virulence of secreted toxins introduced into the digestive system of larvae, whereas the direct injection method addresses the virulence of whole-cell inocula.
The utility of the direct injection method as described here is to analyze bacterial pathogenesis by monitoring insect mortality. However, this method can easily be expanded for use in studying the effects of infection on the M. sexta immune system. The insect responds to infection via both humoral and cellular responses. The humoral response includes recognition of bacterial-associated patterns and subsequent production of various antimicrobial peptides7; the expression of genes encoding these peptides can be monitored subsequent to direct infection via RNA extraction and quantitative PCR13. The cellular response to infection involves nodulation, encapsulation, and phagocytosis of infectious agents by hemocytes6. To analyze these responses, injected insects can be dissected and visualized by microscopy13, 14.

Rearing and Injection of Manduca sexta Larvae to Assess Bacterial Virulence

The method described here utilizes direct injection of entomopathogenic bacteria into the hemocoel of Manduca sexta insect larvae. M. sexta is a commercially available and well-studied insect. Thus, this method represents a simple approach to analyzing host-bacterial interactions from the perspective of one or both partners.

の飼育とインジェクション<em&gt;たばこスズメガ</em細菌毒性を評価するための&gt;幼虫

Manduca sexta, commonly known as the tobacco hornworm, is considered a significant agricultural pest, feeding on solanaceous plants including tobacco and ...

Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the attachment and blood feeding of Ixodes species ticks on mammalian hosts. In nature, this zoonotic bacterial pathogen may use a variety of reservoir hosts, but the white-footed mouse (Peromyscus leucopus) is the primary reservoir for larval and nymphal ticks in North America. Humans are incidental hosts most frequently infected with B. burgdorferi by the bite of ticks in the nymphal stage. B. burgdorferi adapts to its hosts throughout the enzootic cycle, so the ability to explore the functions of these spirochetes and their effects on mammalian hosts requires the use of tick feeding. In addition, the technique of xenodiagnosis (using the natural vector for detection and recovery of an infectious agent) has been useful in studies of cryptic infection. In order to obtain nymphal ticks that harbor B. burgdorferi, ticks are fed live spirochetes in culture through capillary tubes. Two animal models, mice and nonhuman primates, are most commonly used for Lyme disease studies involving tick feeding. We demonstrate the methods by which these ticks can be fed upon, and recovered from animals for either infection or xenodiagnosis.

Lyme disease is the most commonly-reported vector-borne disease in North America. The causative agent, Borrelia burgdorferi is a spirochete bacterium transmitted by Ixodid ticks. Transmission and detection of infection in animal models is optimized by the use of tick feeding, which we describe here.

ライム病研究における送信および外因診断法のために動物にダニの摂食

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the  attachment and blood feeding of Ixodes species ticks on ...

Measuring Local Anaphylaxis in Mice

Allergic responses are the result of the activation of mast cells and basophils, and the subsequent release of vasoactive and proinflammatory mediators. Exposure to an allergen in a sensitized individual can result in clinical symptoms that vary from minor erythema to life threatening anaphylaxis. In the laboratory, various animal models have been developed to understand the mechanisms driving allergic responses. Herein, we describe a detailed method for measuring changes in vascular permeability to quantify localized allergic responses. The local anaphylaxis assay was first reported in the 1920s, and has been adapted from the technique published by Kojima et al. in 20071. In this assay, mice sensitized to OVA are challenged in the left ear with vehicle and in the right ear with OVA. This is followed by an intravenous injection of Evans Blue dye. Ten min after injecting Evans Blue, the animal is euthanized and the dye that has extravasated into the ears is extracted overnight in formamide. The absorbance of the extracted dye is then quantified with a spectrophotometer. This method reliably results in a visual and quantifiable manifestation of a local allergic response.

Allergic responses, characterized by the activation of mast cells and basophils, are driven by the cross-linking of IgE and release of proinflammatory mediators. A quantitative assessment of allergic responses can be achieved by using Evans Blue dye to monitor changes in vascular permeability after allergen challenge.

マウスにおけるローカルアナフィラキシーの測定

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

Generation of Immature, Mature and Tolerogenic Dendritic Cells with Differing Metabolic Phenotypes

Immune response results from a complex interplay between the antigen non-specific innate immune system and the antigen specific adaptive immune system. The immune system is a constant balance in maintaining tolerance to self-molecules and reacting rapidly to pathogens. Dendritic cells (DCs) are powerful professional antigen presenting cells that link the innate immune system to the adaptive immune system and balance the adaptive response between self and non-self. Depending on the maturation signals, immature dendritic cells can be selectively stimulated to differentiate into immunogenic or tolerogenic DCs. Immunogenic dendritic cells provide proliferation signals to antigen-specific T cells for clonal expansion; while tolerogenic dendritic cells regulate tolerance by antigen-specific T-cell deletion or clonal expansion of regulatory T-cells. Due to this unique property, dendritic cells are highly sought after as therapeutic agents for cancer and autoimmune diseases. Dendritic cells can be loaded with specific antigens in vitro and injected into the human body to mount a specific immune response both immunogenic and tolerogenic. This work presents a means to generate in vitro from monocytes, immature monocyte derived dendritic cells (moDCs), tolerogenic and mature moDCs that differ in surface marker expression, function and metabolic phenotypes.

Immature dendritic cells can be selectively differentiated into tolerogenic or mature dendritic cells to regulate the balance between immunity and tolerance. This work presents a means to generate from immature monocyte derived dendritic cells (moDCs), in vitro tolerogenic and mature moDCs that differ in metabolic phenotypes.

代謝表現型が異なる未熟、熟女と寛容原性樹状細胞の生成

Immune response results from a complex interplay between the antigen non-specific innate immune system and the antigen specific adaptive immune system. ...

ダニ媒介性疾患研究のためのダニ幼虫へのヒト曝露

A Model for Experimental Exposure of Humans to Larval Ixodes scapularis Ticks

Tickborne diseases are a significant public health problem in the United States and worldwide. Ticks are obligate blood-feeding arthropods; an ixodid tick must remain attached to the skin of the host and complete its multi-day feeding process to acquire its blood meal. Exposing animals to ticks is a common practice for studying host responses to tick bites and tickborne diseases. We developed the procedure, conducted the first human research study, and published the findings on exposing human volunteers to uninfected larval Ixodes scapularis ticks. This article describes the methodology used to construct the containment dressing, how to apply and secure the ticks to the host, how to maintain the dressing, and how to remove the ticks from the host. Exposing volunteers to tick bites is an experimental procedure and must be performed under a clinical research protocol approved by the appropriate regulatory authorities. This method allows for translational research to better understand the human response to tick bites and foster the development of diagnostics, prevention, and therapies for tickborne diseases.

著者スポットライト:マダニ研究およびライム病研究のための制御されたヒト曝露モデル

This article presents the methodology for exposing humans to larval Ixodes scapularis for clinical research. The technique is relatively simple, tolerable by the research volunteers, and can be modified according to experimental needs. Such research involving human subjects must be conducted under clinical study protocols approved by the appropriate regulatory authorities.

幼虫 Ixodes scapularis Ticsへのヒトの実験的曝露モデル

Tickborne diseases are a significant public health problem in the United States and worldwide. Ticks are obligate blood-feeding arthropods; an ixodid tick ...

A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of mouse models in tick research is critical for understanding their biology and tick-host-pathogen interactions. Here we demonstrate a non-laborious technique for the feeding of immature stages of hard ticks on laboratory mice. The benefit of the method is its simplicity, short duration, and the ability to monitor or collect ticks at different time points of an experiment. In addition, the technique allows attachment of two individual capsules on the same mouse, which is beneficial for a variety of experiments where two different groups of ticks are required to feed on the same animal. The non-irritating and flexible capsule is made from easily accessible materials and minimizes the discomfort of the experimental animals. Furthermore, euthanasia is not necessary, mice recover completely after the experiment and are available for re-use.

In this study, a feeding system for nymphal and larval stages of hard tick was developed using a capsule attached to laboratory mouse. The feeding capsule is made from flexible materials and remains firmly attached to the mouse for at least one week and allows comfortable monitoring of tick feeding.

実験室マウスにおける未熟なハードティックステージのカプセルベースモデル

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of ...

飼育

要約

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ティック腸内ライム病の病原体の迅速な伝達とローカリゼーションのための方法

マダニScapularisダニの唾液、唾液腺、および体液コレクション

の飼育とインジェクション

ライム病研究における送信および外因診断法のために動物にダニの摂食

マウスにおけるローカルアナフィラキシーの測定

代謝表現型が異なる未熟、熟女と寛容原性樹状細胞の生成

幼虫 Ixodes scapularis Ticsへのヒトの実験的曝露モデル

幼虫 Ixodes scapularis Ticsへのヒトの実験的曝露モデル

幼虫 Ixodes scapularis Ticsへのヒトの実験的曝露モデル

実験室マウスにおける未熟なハードティックステージのカプセルベースモデル