विशिष्ट प्रतिजन नियमन के लिए ट्रांस विवो विलंबित प्रकार अतिसंवेदनशीलता परख
Summary
हम संभावित भ्रष्टाचार अस्वीकृति का ऊंचा जोखिम के बिना प्रत्यारोपण के बाद प्रतिरक्षादमन की वापसी तय करने के लिए इस्तेमाल किया जा सकता है कि एक मूल्यवान नैदानिक परख का वर्णन है. परख विलंबित प्रकार अतिसंवेदनशीलता के सिद्धांतों का उपयोग करता है और प्राप्तकर्ता द्वारा घुड़सवार दोनों दाता विशिष्ट प्रेरक और विनियामक प्रतिरक्षा प्रतिक्रिया का सटीक आकलन प्रदान करता है.
Abstract
Delayed-type hypersensitivity response (DTH) is a rapid in vivo manifestation of T cell-dependent immune response to a foreign antigen (Ag) that the host immune system has experienced in the recent past. DTH reactions are often divided into a sensitization phase, referring to the initial antigen experience, and a challenge phase, which usually follows several days after sensitization. The lack of a delayed-type hypersensitivity response to a recall Ag demonstrated by skin testing is often regarded as an evidence of anergy. The traditional DTH assay has been effectively used in diagnosing many microbial infections.
Despite sharing similar immune features such as lymphocyte infiltration, edema, and tissue necrosis, the direct DTH is not a feasible diagnostic technique in transplant patients because of the possibility of direct injection resulting in sensitization to donor antigens and graft loss. To avoid this problem, the human-to-mouse “trans-vivo” DTH assay was developed 1,2. This test is essentially a transfer DTH assay, in which human peripheral blood mononuclear cells (PBMCs) and specific antigens were injected subcutaneously into the pinnae or footpad of a naïve mouse and DTH-like swelling is measured after 18-24 hr 3. The antigen presentation by human antigen presenting cells such as macrophages or DCs to T cells in highly vascular mouse tissue triggers the inflammatory cascade and attracts mouse immune cells resulting in swelling responses. The response is antigen-specific and requires prior antigen sensitization. A positive donor-reactive DTH response in the Tv-DTH assay reflects that the transplant patient has developed a pro-inflammatory immune disposition toward graft alloantigens.
The most important feature of this assay is that it can also be used to detect regulatory T cells, which cause bystander suppression. Bystander suppression of a DTH recall response in the presence of donor antigen is characteristic of transplant recipients with accepted allografts 2,4-14. The monitoring of transplant recipients for alloreactivity and regulation by Tv-DTH may identify a subset of patients who could benefit from reduction of immunosuppression without elevated risk of rejection or deteriorating renal function.
A promising area is the application of the Tv-DTH assay in monitoring of autoimmunity15,16 and also in tumor immunology 17.
Protocol
Representative Results
Discussion
ट्रांस विवो डीटीएच परख प्रत्यारोपण, कैंसर और autoimmune रोगियों में सेल की मध्यस्थता प्रतिक्रियाओं का आकलन करने में एक संभावित नैदानिक आवेदन के साथ एक उपन्यास नैदानिक परीक्षण है. यह याद टी प्रेरक प्रति…
Declarações
The authors have nothing to disclose.
Acknowledgements
लेखकों प्रत्यारोपण प्राप्तकर्ताओं में विनियमित डीटीएच प्रतिक्रिया के बारे में हमारी समझ के विकास के लिए कर रहा हूँ Vanbuskirk के योगदान को स्वीकार करना होगा. इस काम एनआईएच अनुदान PO1AI084853 और R01AI066219-06 समर्थित, और यूरोपीय संघ द्वारा प्रायोजित एक अध्ययन के द्वारा किया गया था.
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
| ACD tube for blood collection | BD | 02-684-26 | |
| Lymphocyte Separation Medium | Cellgro | 25-072-CV | |
| Dulbecco’s Phosphate-Buffered Saline | Cellgro | 21-031-CM | Without calcium & magnesium |
| ACK Lysis Buffer | BioWhittaker | 10-548E | |
| TT/DT or EBV | Sanofi Pasteur Inc./ Meridian Life Science, Inc. | TT/DT 25 μg/injection EBV 8 μg/injection | |
| Protease inhibitor PMSF | Sigma-Aldrich | 78830 | |
| Eosine for cell count | Sigma-Aldrich | E-6003 | |
| Alloantigen | Purified HLA antigens, synthetic allopeptides can be used instead of donor cell-free lysates | ||
| 50 ml sterile centrifuge tubes | Fisher Scientific | 06-443-18 | |
| 10 ml pipettes and pipettor | BD Falcon | 13-675-20 | |
| 2 ml safe-lock tubes | Costar | 3213 | |
| 1000 μl, 100 μl , 10 μl pipettes with sterile tips | |||
| Hemocytometer | Fisher | 02-671-10 | |
| Full size centrifuge and microfuge | Beckman Coulter/Eppendorf | ||
| 1/2cc or 1cc insulin syringes | Becton Dickinson | 14-826-79 | 28 gauge |
| Vibracell sonicator | Divtech Equipment Co. Sonocs& Materials Inc | 2 mm probe | |
| Dial thickness gauge | Mitutoyo, Japan | ||
| SCID mice | Harlan | ||
| Isoflurane | Piramal Healthcare | Inhalant anesthesia |
Referências
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- VanBuskirk, A. M., et al. Human allograft acceptance is associated with immune regulation. J. Clin. Invest. 106, 145-155 (2000).
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