Name: human in vitro suppression as screening tool for the recognition of an early state of immune imbalance
Uploaded: 2011-07-22T12:00:00
Description: Watch this Scientific Journal Video about Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance at JoVE.com

इस अध्ययन अधिकतम McGee किशोर Diabetesat विस्कॉन्सिन और Wisconsin के बच्चों के अनुसंधान संस्थान के मेडिकल कॉलेज के लिए राष्ट्रीय अनुसंधान केंद्र द्वारा समर्थित किया गया. funders अध्ययन डिजाइन में कोई भूमिका नहीं थी, डेटा संग्रह और विश्लेषण, या पांडुलिपि की तैयारी.

1. एक दमन परख की स्थापना से पहले, सेल और बाद में उत्तेजना के लिए एक मानव विरोधी CD3 (UCHT1 क्लोन, अंतिम एकाग्रता 1μg/ml) के साथ कोट tosylactivated मोती की जरूरत है की जाँच करें कि क्या मोती कुशलता से इन विट्रो प्रसार परख ?...

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केवल Tregs के लिए अनूठी विशेषता के रूप में विषयों के बीच, दमनकारी समारोह मज़बूती से और समान परीक्षण किया जाना चाहिए एक ही के भीतर और विभिन्न अध्ययनों के बीच रोग के विकास के विभिन्न चरणों पर. हम दमन हमारे इस ?...

<table border="1"><tbody><tr><th> अभिकर्मक या साधन के नाम </th><th> कंपनी </th><th> सूचीपत्र संख्या </th><th> टिप्पणियां (वैकल्पिक) </th></tr><tr><td> Ficoll - Paque PLUS </td><td> Amersham Pharmacia बायोटेक </td><td> 17-1440-03 </td><td></td></tr><tr><td> DPBS-1X </td><td> Gibco </td><td> 14190-144 </td><td></td></tr><tr><td> Trypan ब्लू </td><td> Invitrogen </td><td> 15250-061 </td><td></td></tr><tr><td> विरोधी CD4 microbeads </td><td> Miltenyi </td><td> 130-045-101 </td><td></td></tr><tr><td> प्री - जुदाई फिल्टर </td><td> Miltenyi </td><td> 130-041-407 </td><td></td></tr><tr><td> रास स्तंभ </td><td> Miltenyi </td><td> 130-042-401 </td><td></td></tr><tr><td> EDTA </td><td> Invitrogen </td><td> 15575-020 </td><td></td></tr><tr><td> BSA </td><td> सिग्मा Aldrich </td><td> B4287 </td><td></td></tr><tr><td> विरोधी मानव CD4 APCCy7 (क्लोन RPA-टी -4) </td><td> बी.डी. Pharmingen </td><td> 557852 </td><td></td></tr><tr><td> विरोधी मानव CD25 पीई (क्लोन एम-A251, आईएल 2Rα) </td><td> बी.डी. Pharmingen </td><td> 555432 </td><td></td></tr><tr><td> विरोधी मानव CD8 FITC (क्लोन RPA-T8) </td><td> बी.डी. Pharmingen </td><td> 555366 </td><td></td></tr><tr><td> विरोधी मानव CD14 FITC (M5E2 क्लोन, LPS रिसेप्टर) </td><td> बी.डी. Pharmingen </td><td> 555397 </td><td></td></tr><tr><td> विरोधी मानव CD32 FITC (FLI8.26 क्लोन, द्वितीय FcγR - प्रकार) </td><td> बी.डी. Pharmingen </td><td> 555448 </td><td></td></tr><tr><td> विरोधी मानव CD116 FITC (M5D12 क्लोन, जीएम - CSFRα श्रृंखला) </td><td> बी.डी. Pharmingen </td><td> 554532 </td><td></td></tr><tr><td> Dynalbeads M-450 tosylactivated </td><td> Invitrogen </td><td> 140-13 </td><td></td></tr><tr><td> एंटी - मानव CD3 </td><td> Ancell </td><td> 144-024 </td><td></td></tr><tr><td> Buffer1 </td><td> घर का बना </td><td></td><td> 0.1M ना बी 2 4 7 हे pH7.6 </td></tr><tr><td> Buffer2 </td><td> घर का बना </td><td></td><td> PBS/2mM / 0.1 EDTA% pH7.4 BSA </td></tr><tr><td> Buffer3 </td><td> घर का बना </td><td></td><td> 0.2M Tris/0.1% pH8.5 BSA </td></tr><tr><td> पूरा RPMI मीडिया </td><td> घर का बना </td><td></td><td> 1640 मीडिया 2 मिमी एल glutamine 5 मिमी HEPES यू 100 / μg / मिलीग्राम / peni strept 0.5 मिमी सोडियम पाइरूवेट RPMI </td></tr><tr><td> [3 एच] thymidine </td><td> Perkin एल्मर </td><td> NET027Z005MC </td><td></td></tr><tr><td> मानव जमा अटल बिहारी सीरम </td><td> अटलांटा बायोलॉजिकल </td><td> S40110 </td><td></td></tr><tr><td> Multiscreen फसल प्लेट </td><td> Millipore </td><td> MAHFC1H60 </td><td></td></tr><tr><td> 20 Microscint </td><td> Perkin एल्मर </td><td> 6013621 </td><td></td></tr></tbody></table>

human in vitro suppression as screening tool for the recognition of an early state of immune imbalance

Regulatory T cells (Tregs) are critical mediators of immune tolerance to self-antigens. In addition, they are crucial regulators of the immune response following an infection. Despite efforts to identify unique surface marker on Tregs, the only unique feature is their ability to suppress the proliferation and function of effector T cells. While it is clear that only in vitro assays can be used in assessing human Treg function, this becomes problematic when assessing the results from cross-sectional studies where healthy cells and cells isolated from subjects with autoimmune diseases (like Type 1 Diabetes-T1D) need to be compared. There is a great variability among laboratories in the number and type of responder T cells, nature and strength of stimulation, Treg:responder ratios and the number and type of antigen-presenting cells (APC) used in human in vitro suppression assays. This variability makes comparison between studies measuring Treg function difficult. The Treg field needs a standardized suppression assay that will work well with both healthy subjects and those with autoimmune diseases. We have developed an in vitro suppression assay that shows very little intra-assay variability in the stimulation of T cells isolated from healthy volunteers compared to subjects with underlying autoimmune destruction of pancreatic &beta;-cells. The main goal of this piece is to describe an in vitro human suppression assay that allows comparison between different subject groups. Additionally, this assay has the potential to delineate a small loss in nTreg function and anticipate further loss in the future, thus identifying subjects who could benefit from preventive immunomodulatory therapy1. Below, we provide thorough description of the steps involved in this procedure. We hope to contribute to the standardization of the in vitro suppression assay used to measure Treg function. In addition, we offer this assay as a tool to recognize an early state of immune imbalance and a potential functional biomarker for T1D.

Watch this Scientific Journal Video about Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance at JoVE.com

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

Tregs प्रतिरक्षा प्रणाली की प्रबल दमन कर रहे हैं. अद्वितीय सतह मार्करों की एक कमी के लिए उन्हें परिभाषित है, इसलिए, Tregs की परिभाषा मुख्य रूप से कार्य कर रहे हैं. यहाँ हम एक अनुकूलित वर्णन<em&gt; इन विट्रो में</em&gt; परख जोखिम में विषयों में प्रतिरक्षा असंतुलन की पहचान के लिए T1D विकसित करने में सक्षम.

मानवीय इन विट्रो में इम्यून असंतुलन का एक प्रारंभिक राज्य की मान्यता के लिए स्क्रीनिंग उपकरण के रूप में दमन

Regulatory T cells (Tregs) are critical mediators of immune tolerance to self-antigens. In addition, they are crucial regulators of the immune response ...

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JoVE Journal

Immunology and Infection

Human In Vitro Suppression as Screening Tool for the Recognition of an Early State of Immune Imbalance

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

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एक का प्रयोग न्यूमोकोकल औपनिवेशीकरण पर प्रोबायोटिक्स के प्रभाव की जांच<em&gt; इन विट्रो</em&gt; पालन परख

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An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

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एक<em&gt; इन विट्रो</em&gt; एचआईवी सह-संक्रमण के संदर्भ में मलेरिया के लिए इम्यून प्रतिक्रियाओं को मापने के लिए मॉडल

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Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

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छोटे अणुओं मिलाना सी-di-जीएमपी में संकेत है कि जांच के लिए एक उच्च throughput सेटअप की स्थापना<em&gt; Pseudomonas aeruginosa</em

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