وأيد هذه الدراسة ماكجي ماكس المركز القومي للبحوث الطبية لكلية Diabetesat الأحداث ويسكونسن وطفولة معهد البحوث ويسكونسن. كان الممولين أي دور في تصميم الدراسة وجمع البيانات وتحليلها ، أو في الإعداد للمخطوطة.

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الإعلان عن أي تضارب في المصالح.

والميزة الفريدة فقط لTregs ، يجب اختبار الوظيفة القمعية وموثوق بها بشكل موحد بين الموضوعات في مختلف مراحل تطور المرض داخل نفسه ، وبين الدراسات المختلفة. ونحن نقدم تفاصيل قمع مقايسة وضعت في مختبرنا ومساهمتنا في توحيد هذا الاختبار. في دراستنا التحسين واسعة ، ونحن مصممون ...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><th> اسم كاشف أو صك </th><th> شركة </th><th> فهرس العدد </th><th> تعليقات (اختياري) </th></tr><tr><td> Ficoll - PLUS Paque </td><td> أمرشام فارماسيا في مجال التكنولوجيا الحيوية </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> التريبان الأزرق </td><td> Invitrogen </td><td> 15250-061 </td><td></td></tr><tr><td> مكافحة CD4 microbeads </td><td> Miltenyi </td><td> 130-045-101 </td><td></td></tr><tr><td> قبل فصل المرشحات </td><td> Miltenyi </td><td> 130-041-407 </td><td></td></tr><tr><td> LS العمود </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> سيغما الدريخ </td><td> B4287 </td><td></td></tr><tr><td> مكافحة الإنسان CD4 - APCCy7 (استنساخ RPA - T4) </td><td> دينار بحريني Pharmingen </td><td> 557852 </td><td></td></tr><tr><td> مكافحة الإنسان CD25 - PE (M - A251 استنساخ ؛ IL - 2Rα) </td><td> دينار بحريني Pharmingen </td><td> 555432 </td><td></td></tr><tr><td> مكافحة الإنسان CD8 - FITC (استنساخ RPA - T8) </td><td> دينار بحريني Pharmingen </td><td> 555366 </td><td></td></tr><tr><td> مكافحة الإنسان CD14 - FITC (استنساخ M5E2 ؛ مستقبلات LPS) </td><td> دينار بحريني Pharmingen </td><td> 555397 </td><td></td></tr><tr><td> مكافحة الإنسان CD32 - FITC (استنساخ FLI8.26 ؛ FcγR من النوع الثاني) </td><td> دينار بحريني Pharmingen </td><td> 555448 </td><td></td></tr><tr><td> مكافحة الإنسان CD116 - FITC (استنساخ M5D12 ؛ GM - CSFRα السلسلة) </td><td> دينار بحريني Pharmingen </td><td> 554532 </td><td></td></tr><tr><td> tosylactivated Dynalbeads M - 450 </td><td> Invitrogen </td><td> 140-13 </td><td></td></tr><tr><td> مكافحة الإنسان CD3 </td><td> أنسل </td><td> 144-024 </td><td></td></tr><tr><td> Buffer1 </td><td> محلية الصنع </td><td></td><td> 0.1M نا 2 B 4 O 7 pH7.6 </td></tr><tr><td> Buffer2 </td><td> محلية الصنع </td><td></td><td> PBS/2mM BSA EDTA / 0.1 ٪ pH7.4 </td></tr><tr><td> Buffer3 </td><td> محلية الصنع </td><td></td><td> 0.2M Tris/0.1 ٪ BSA pH8.5 </td></tr><tr><td> استكمال وسائل الاعلام RPMI </td><td> محلية الصنع </td><td></td><td> RPMI 1640 وسائل الإعلام 2 مم L - الجلوتامين HEPES ملي 5 100 U / ميكروغرام / مل peni / strept الصوديوم 0.5 مم البيروفات </td></tr><tr><td> [3 H] ثيميدين </td><td> بيركن إلمر </td><td> NET027Z005MC </td><td></td></tr><tr><td> الإنسان المجمعة AB المصل </td><td> اتلانتا البيولوجية </td><td> S40110 </td><td></td></tr><tr><td> لوحة متعددة الشاشات الحصاد </td><td> ميليبور </td><td> MAHFC1H60 </td><td></td></tr><tr><td> Microscint 20 </td><td> بيركن إلمر </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

التنظيمية خلايا T (Tregs) هي الحاسمة وسطاء التسامح المناعي في تقرير المصير ، مستضدات. بالإضافة إلى ذلك ، هم المنظمين الحاسمة للاستجابة مناعية بعد العدوى. على الرغم من الجهود المبذولة لتحديد علامة فريدة من نوعها على سطح Tregs ، الميزة الفريدة فقط هو قدرتها على منع انتشار الأسلحة النووية وظيفة الخلايا التائية المستجيب. في حين أنه من الواضح أن المقايسات فقط في المختبر يمكن استخدامها في تقييم وظيفة Treg الإنسان ، وهذا يصبح مشكلة عند تقييم النتائج من دراسات مستعرضة فيها الخلايا السليمة والخلايا المعزولة من الموضوعات مع أمراض المناعة الذاتية (مثل مرض السكري نوع 1 - T1D) الحاجة ويمكن مقارنة. هناك تفاوت كبير بين المختبرات في عدد ونوع من الخلايا التائية المستجيب وطبيعة وقوة التحفيز ، Treg : نسب المستجيب وعدد ونوع من خلايا مولدة تقديم (APC) المستخدمة في قمع الإنسان في فحوصات المختبر. هذا التباين يجعل المقارنة بين الدراسات قياس وظيفة Treg صعبة. الحقل Treg يحتاج الى فحص قمع موحدة من شأنها أن تعمل بشكل جيد مع كل من الاشخاص الاصحاء والمصابين بأمراض المناعة الذاتية. طورنا في الفحص المختبري القمع الذي يظهر القليل جدا من داخل مقايسة التباين في تحفيز خلايا تي المعزولة من المتطوعين الأصحاء مقارنة مع الموضوعات الأساسية تدمير المناعة الذاتية للخلايا البنكرياس β. الهدف الرئيسي لهذه القطعة لوصف تجربة في المختبر قمع الإنسان أن يسمح بإجراء مقارنة بين مجموعات مختلفة الموضوع. بالإضافة إلى ذلك ، هذا الاختبار لديه القدرة على رسم خسارة صغيرة في nTreg وظيفة وتوقع مزيد من الخسائر في المستقبل ، ومن ثم تحديد الموضوعات التي يمكن أن تستفيد من المعالجة المناعية الوقائية 1. أدناه ، ونحن نقدم وصف دقيق للخطوات التي ينطوي عليها هذا الإجراء. نأمل أن تسهم في توحيد في المختبر فحص قمع المستخدمة لقياس وظيفة Treg. بالإضافة إلى ذلك ، فإننا نقدم هذا الاختبار كأداة ليتعرف على الدولة في وقت مبكر من الخلل المناعي والعلامات البيولوجية المحتملة وظيفية للT1D.

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 والمكثفات قوية من الجهاز المناعي. هناك عدم وجود علامات سطح فريدة لتعريف بهم ، وبالتالي ، تعريفات Tregs وظيفية في المقام الأول. نحن هنا تصف الأمثل<em&gt; في المختبر</em&gt; مقايسة قادرة على تحديد الخلل المناعي في المواد المعرضة للخطر لتطوير T1D.

الإنسان في المختبر قمع وأداة الكشف عن الاعتراف للدولة في وقت مبكر من الخلل المناعي

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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Immunology and Infection

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

14.4K Views.  Medical College of Wisconsin. Tregs والمكثفات قوية من الجهاز المناعي. هناك عدم وجود علامات سطح فريدة لتعريف بهم ، وبالتالي ، تعريفات Tregs وظيفية في المقام الأول. نحن هنا تصف الأمثل في المختبر مقايسة قادرة على تحديد الخلل المناعي في المواد المعرضة للخطر لتطوير T1D.

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

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.

إدخال إجهاد القص في دراسة الالتصاق الجرثومي

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

Quantification of the Respiratory Burst Response as an Indicator of Innate Immune Health in Zebrafish

The phagocyte respiratory burst is part of the innate immune response to pathogen infection and involves the production of reactive oxygen species (ROS). ROS are toxic and function to kill phagocytized microorganisms. In vivo quantification of phagocyte-derived ROS provides information regarding an organism's ability to mount a robust innate immune response. Here we describe a protocol to quantify and compare ROS in whole zebrafish embryos upon chemical induction of the phagocyte respiratory burst. This method makes use of a non-fluorescent compound that becomes fluorescent upon oxidation by ROS. Individual zebrafish embryos are pipetted into the wells of a microplate and incubated in this fluorogenic substrate with or without a chemical inducer of the respiratory burst. Fluorescence in each well is quantified at desired time points using a microplate reader. Fluorescence readings are adjusted to eliminate background fluorescence and then compared using an unpaired t-test. This method allows for comparison of the respiratory burst potential of zebrafish embryos at different developmental stages and in response to experimental manipulations such as protein knockdown, overexpression, or treatment with pharmacological agents. This method can also be used to monitor the respiratory burst response in whole dissected kidneys or cell preparations from kidneys of adult zebrafish and some other fish species. We believe that the relative simplicity and adaptability of this protocol will complement existing protocols and will be of interest to researchers who seek to better understand the innate immune response.

The innate immune response protects organisms against pathogen infection. A critical component of the innate immune response, the phagocyte respiratory burst, generates reactive oxygen species that kill invading microorganisms. We describe a respiratory burst assay that quantifies reactive oxygen species produced when the innate immune response is chemically induced.

الكمي للاستجابة انفجر الجهاز التنفسي باعتبارها مؤشر الفطري المناعي الصحة في الزرد

The phagocyte respiratory burst is part of the innate immune response to  pathogen infection and involves the production of reactive oxygen species (ROS). ...

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

Human respiratory syncytial virus (HRSV) infections present a broad spectrum of disease severity, ranging from mild infections to life-threatening bronchiolitis. An important part of the pathogenesis of severe disease is an enhanced immune response leading to immunopathology.	Here, we describe a protocol used to investigate the immune response of human immune cells to an HRSV infection. First, we describe methods used for culturing, purification and quantification of HRSV. Subsequently, we describe a human in vitro model in which peripheral blood mononuclear cells (PBMCs) are stimulated with live HRSV. This model system can be used to study multiple parameters that may contribute to disease severity, including the innate and adaptive immune response. These responses can be measured at the transcriptional and translational level. Moreover, viral infection of cells can easily be measured using flow cytometry. Taken together, stimulation of PBMC with live HRSV provides a fast and reproducible model system to examine mechanisms involved in HRSV-induced disease.

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

Human respiratory syncytial virus (HRSV) can cause severe bronchiolitis in young infants. Part of the pathogenesis of severe HRSV disease is caused by the host immune response. Stimulation of primary human immune cells with HRSV provides a fast and reproducible model system to study activation of inflammatory pathways and infection.

Human respiratory syncytial  virus (HRSV) infections present a broad spectrum of disease severity, ranging  from mild infections to life-threatening ...

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. In vitro adherence assays can be used to study the attachment of pneumococci to epithelial cell monolayers and to investigate potential interventions, such as the use of probiotics, to inhibit pneumococcal&#160;colonization.&#160;The protocol described here is used to investigate the effects of the probiotic Streptococcus salivarius on the adherence of pneumococci to the human epithelial cell line CCL-23 (sometimes referred to as HEp-2 cells). The assay involves three main steps: 1) preparation of epithelial and bacterial cells, 2) addition of bacteria to epithelial cell monolayers, and 3) detection of adherent pneumococci by viable counts (serial dilution and plating) or quantitative real-time PCR (qPCR). This technique is relatively straightforward and does not require specialized equipment other than a tissue culture setup. The assay can be used to test other probiotic species and/or potential inhibitors of pneumococcal colonization and can be easily modified to address other scientific questions regarding pneumococcal adherence and invasion.

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

In vitro adherence assays can be used to study the attachment of Streptococcus pneumoniae to epithelial cell monolayers and to investigate potential interventions such as the use of probiotics for inhibiting pneumococcal colonization.

التحقيق في آثار البروبيوتيك على المكورات الرئوية الاستعمار باستخدام<em&gt; في المختبر</em&gt; الالتزام الفحص

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

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 mice. Recently, myeloid differentiation factor 88 (MyD88) signaling was shown to be important for initial T cell priming and memory T cell development during WNV NS4B-P38G mutant infection. In this study, two flow cytometry-based methods &#8211; an in vitro T cell priming assay and an intracellular cytokine staining (ICS) &#8211; were utilized to assess dendritic cells (DCs) and T cell functions. In the T cell priming assay, cell proliferation was analyzed by flow cytometry following co-culture of DCs from both groups of mice with carboxyfluorescein succinimidyl ester (CFSE) - labeled CD4+ T cells of OTII transgenic mice. This approach provided an accurate determination of the percentage of proliferating CD4+ T cells with significantly improved overall sensitivity than the traditional assays with radioactive reagents. A microcentrifuge tube system was used in both cell culture and cytokine staining procedures of the ICS protocol. Compared to the traditional tissue culture plate-based system, this modified procedure was easier to perform at biosafety level (BL) 3 facilities. Moreover, WNV- infected cells were treated with paraformaldehyde in both assays, which enabled further analysis outside BL3 facilities. Overall, these in vitro immunological assays can be used to efficiently assess cell-mediated immune responses during WNV infection.

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

Two flow cytometry-based methods &#8211; an in vitro T cell priming assay and intracellular cytokine staining were utilized to measure antigen presenting capacity of dendritic cells and antigen-specific T cell responses to a West Nile virus mutant infection in mice.

في تحليل المختبر من الاستجابة المناعية الخلوية بوساطة Myd88 لفيروس غرب النيل المسخ سلالة العدوى

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

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. Although understanding the disease dynamics for each of these pathogens independently is vital, it is also important that the interactions between these pathogens are investigated and understood. 
We have developed a versatile in vitro model of HIV-malaria co-infection to study host immune responses to malaria in the context of HIV infection. Our model allows the study of secreted factors in cellular supernatants, cell surface and intracellular protein markers, as well as RNA expression levels. The experimental design and methods used limit variability and promote data reliability and reproducibility. 
All pathogens used in this model are natural human pathogens (Plasmodium falciparum and HIV-1), and all infected cells are naturally infected and used fresh. We use human erythrocytes parasitized with P. falciparum and maintained in continuous in vitro culture. We obtain freshly isolated peripheral blood mononuclear cells from chronically HIV-infected volunteers. Every condition used has an appropriate control (P. falciparum parasitized vs. normal erythrocytes), and every HIV-infected donor has an HIV uninfected control, from which cells are harvested on the same day. This model provides a realistic environment to study the interactions between malaria parasites and human immune cells in the context of HIV infection.

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

Human co-infection is difficult to replicate in vitro. However, human malaria parasites can readily be cultured in vitro, as can freshly isolated human peripheral blood mononuclear cells naturally infected with HIV. This provides an excellent model for studying early immune responses to malaria parasites in the context of HIV co-infection.

ل<em&gt; في المختبر</em&gt; نموذج لقياس الاستجابات المناعية لمكافحة الملاريا في سياق الفيروس المسبب للإيدز

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. ...

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

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. Regulatory systems that utilize intracellular cyclic di-GMP (c-di-GMP) as a second messenger are one such class of target. c-di-GMP is a signaling molecule found in almost all bacteria that acts to regulate an extensive range of processes including antibiotic resistance, biofilm formation and virulence. The understanding of how c-di-GMP signaling controls aspects of antibiotic resistant biofilm development has suggested approaches whereby alteration of the cellular concentrations of the nucleotide or disruption of these signaling pathways may lead to reduced biofilm formation or increased susceptibility of the biofilms to antibiotics. We describe a simple high-throughput bioreporter protocol, based on green fluorescent protein (GFP), whose expression is under the control of the c-di-GMP responsive promoter cdrA, to rapidly screen for small molecules with the potential to modulate c-di-GMP cellular levels in Pseudomonas aeruginosa (P. aeruginosa). This simple protocol can screen upwards of 3,500 compounds within 48 hours and has the ability to be adapted to multiple microorganisms.

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

This article describes the high-throughput assay that has been successfully established to screen large libraries of small molecules for their potential ability to manipulate cellular levels of cyclic di-GMP in Pseudomonas aeruginosa, providing a new powerful tool for antibacterial drug discovery and compound testing.

إنشاء إعداد إنتاجية عالية لفحص الجزيئات الصغيرة التي تعدل ج-دي-GMP في اشارة<em&gt; الزائفة الزنجارية</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...

Determining Immune System Suppression versus CNS Protection for Pharmacological Interventions in Autoimmune Demyelination

A major hallmark of the autoimmune demyelinating disease multiple sclerosis (MS) is immune cell infiltration into the brain and spinal cord resulting in myelin destruction, which not only slows conduction of nerve impulses, but causes axonal injury resulting in motor and cognitive decline. Current treatments for MS focus on attenuating immune cell infiltration into the central nervous system (CNS). These treatments decrease the number of relapses, improving quality of life, but do not completely eliminate relapses so long-term disability is not improved. Therefore, therapeutic agents that protect the CNS are warranted. In both animal models as well as human patients with MS, T cell entry into the CNS is generally considered the initiating inflammatory event. In order to assess if a drug protects the CNS, any potential effects on immune cell infiltration or proliferation in the periphery must be ruled out. This protocol describes how to determine whether CNS protection observed after drug intervention is a consequence of attenuating CNS-infiltrating immune cells or blocking death of CNS cells during inflammatory insults. The ability to examine MS treatments that are protective to the CNS during inflammatory insults is highly critical for the advancement of therapeutic strategies since current treatments reduce, but do not completely eliminate, relapses (i.e., immune cell infiltration), leaving the CNS vulnerable to degeneration.

This protocol describes how to determine whether pharmacological treatments for experimental autoimmune encephalomyelitis show CNS protection as a consequence of suppressing immune cell infiltration or are neuroprotective during the onslaught of immune cell infiltration.

تحديد قمع نظام المناعة مقابل حماية الجهاز العصبي المركزي للالدوائية التدخلات في إزالة الميالين المناعة الذاتية

A major hallmark of the autoimmune demyelinating disease multiple sclerosis (MS) is immune cell infiltration into the brain and spinal cord resulting in ...

Analysis of Lymphocyte Extravasation Using an In Vitro Model of the Human Blood-brain Barrier

Lymphocyte extravasation into the central nervous system (CNS) is critical for immune surveillance. Disease-related alterations of lymphocyte extravasation might result in pathophysiological changes in the CNS. Thus, investigation of lymphocyte migration into the CNS is important to understand inflammatory CNS diseases and to develop new therapy approaches. Here we present an in vitro model of the human blood-brain barrier to study lymphocyte extravasation. Human brain microvascular endothelial cells (HBMEC) are confluently grown on a porous polyethylene terephthalate transwell insert to mimic the endothelium of the blood-brain barrier. Barrier function is validated by zonula occludens immunohistochemistry, transendothelial electrical resistance (TEER) measurements as well as analysis of evans blue permeation. This model allows investigation of the diapedesis of rare lymphocyte subsets such as CD56brightCD16dim/- NK cells. Furthermore, the effects of other cells, cytokines and chemokines, disease-related alterations, and distinct treatment regimens on the migratory capacity of lymphocytes can be studied. Finally, the impact of inflammatory stimuli as well as different treatment regimens on the endothelial barrier can be analyzed.

Analysis of Lymphocyte Extravasation Using an In Vitro Model of the Human Blood-brain Barrier

Here, we describe a human blood-brain barrier model enabling to investigate lymphocyte transmigration into the central nervous system in vitro.

تحليل الخلايا الليمفاوية التسرب باستخدام<em&gt; في المختبر</em&gt; نموذج من الدم في الدماغ الإنسان الحاجز

Lymphocyte extravasation into the central nervous system (CNS) is critical for immune surveillance. Disease-related alterations of lymphocyte ...

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

Mycobacterium tuberculosis remains the number one infectious agent in the world today. With the emergence of antibiotic resistant strains, new clinically relevant methods are needed that evaluate the disease process and screen for potential antibiotic and vaccine treatments. Positron Emission Tomography/Computed Tomography (PET/CT) has been established as a valuable tool for studying a number of afflictions such as cancer, Alzheimer&#39;s disease, and inflammation/infection. Outlined here are a number of strategies that have been employed to evaluate PET/CT images in cynomolgus macaques that are infected intrabronchially with low doses of M. tuberculosis. Through evaluation of lesion size on CT and uptake of 18F-fluorodeoxyglucose (FDG) in lesions and lymph nodes in PET images, these described methods show that PET/CT imaging can predict future development of active versus latent disease and the propensity for reactivation from a latent state of infection. Additionally, by analyzing the overall level of lung inflammation, these methods determine antibiotic efficacy of drugs against M. tuberculosis in the most clinically relevant existing animal model. These image analysis methods are some of the most powerful tools in the arsenal against this disease as not only can they evaluate a number of characteristics of infection and drug treatment, but they are also directly translatable to a clinical setting for use in human studies.

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

Here, we present a protocol to describe the analysis of 18F-FDG PET/CT imaging in non-human primates that have been infected with M. tuberculosis to study disease process, drug treatment, and disease reactivation.

تحليل 18"فدج التصوير" PET/CT كأداة لدراسة بكتريا السل العدوى والعلاج في الإنسان عدم الرئيسات

Mycobacterium tuberculosis remains the number one infectious agent in the world today. With the emergence of antibiotic resistant strains, new clinically ...