Dextrans
Centrifugation, Density Gradient
Fluorescein-5-isothiocyanate
Cell Separation
Glomerular Basement Membrane
Centrifugation, Isopycnic
Diatrizoate
Deficiency in Msh2 affects the efficiency and local sequence specificity of immunoglobulin class-switch recombination: parallels with somatic hypermutation. (1/226)
During maturation of the immune response, IgM+ B cells switch to expression of one of the downstream isotypes (IgG, A or E). This class switching occurs by region-specific recombination within the IgH locus through an unknown mechanism. A lack of switch recombination in mice deficient in components of the DNA-dependent protein kinase (DNA-PK)-Ku complex has pointed to a role for non-homologous end joining. Here we characterize a switching defect in mice lacking a protein involved in DNA mismatch recognition. Mice deficient in Msh2 give diminished IgG (but not IgM) responses following challenge with both T cell-dependent and T cell-independent antigens. This appears to reflect a B cell-intrinsic defect since B cells from Msh2-deficient mice also exhibit impaired switching (but not blasting or proliferation) on in vitro culture with lipopolysaccharide. Furthermore, those switches that do occur in Msh2-deficient B cells reveal a shift in the distribution of recombination sites used: the breakpoints are more likely to occur in consensus motifs. These results, which intriguingly parallel the effects of Msh2 deficiency on hypermutation, suggest a role for Msh2 in the mechanics of class-switch recombination. (+info)Cutting edge: CD40 ligand is a limiting factor in the humoral response to T cell-dependent antigens. (2/226)
CD40 ligand (CD40L) plays a crucial role in T cell-dependent B cell responses, but whether its abundance is a limiting factor in their development is unclear. This question was addressed in transgenic mice expressing the murine CD40L gene under the control of the IL-2-promoter (CD40Ltg+). The fraction of activated T cells from the CD40Ltg+ mice with detectable levels of surface CD40L was modestly greater (1.1- to 2-fold) than littermate controls and paralleled an approximately 1.8-fold increase in CD40L mRNA abundance. In response to trinitrophenol (TNP)-keyhole limpet hemocyanin and tetanus/diphtheria vaccine, CD40Ltg+ mice developed higher titers of high-affinity IgG and IgG1 Ab than wild-type mice. In contrast, the Ab response of CD40Ltg+ and control mice was similar in response to the T-independent Ag TNP-Ficoll. These results suggest that a modest increment in expression of CD40L accelerates the development of T-dependent responses, and that CD40L plays a limiting role in the induction of high-affinity Ab and Ab-class switching. (+info)A comparison of the direct and reporter antigen popliteal lymph node assay for the detection of immunomodulation by low molecular weight compounds. (3/226)
The direct popliteal lymph node assay (PLNA) is a predictive test used to detect the immune-stimulating potential of pharmaceuticals and other low molecular weight compounds (LMWCs) with known autoimmunogenic or sensitizing properties. Two limitations in the PLNA are the existence of false negatives and the inability of the assay to provide mechanistic information. Recently the direct PLNA was modified by incorporating reporter antigens (RA), either TNP-Ficoll or TNP-OVA. In the RA-PLNA, immune stimulation is detected by measuring IgM or IgG TNP-specific antibody-forming cells (AFC) using an enzyme-linked immunospot (ELISPOT) assay. The RA-PLNA, when using potent, known autoimmunogenic compounds, may provide greater sensitivity compared to the direct PLNA and might distinguish LMWCs that have intrinsic adjuvant activity from those that create neo-antigens, using TNP-OVA and TNP-Ficoll, respectively. The purpose of this study was to rigorously compare the two assays. Our first objective was to investigate the interlaboratory reproducibility of the RA-PLNA using four autoimmunogenic LMWC models, plus one negative control LMWC. Subsequently, we tested seven LMWCs with known sensitizing properties and compared the results from the direct and modified assay. The test group included LMWCs thought to be mechanistically distinct and similar to compounds typically encountered in preclinical safety assessment. All control and treatment AFC plaques were collected (76 total), pooled, coded to conceal their source, and counted. The interlaboratory reproducibility of the RA-PLNA was demonstrated with the model autoimmunogenic compounds HgCl2, diphenylhydantoin, D-penicillamine, and the negative control compound phenobarbital, by detecting TNP-specific IgM and polyclonal IgG production to both reporter antigens. Additionally, the sensitizing effects of streptozotocin were identified using an IgG2a ELISPOT with both TNP-OVA and TNP-Ficoll. With the extended test group, the sensitizing effects of aniline, a false negative LMWC in the direct PLNA, was not detected in this study when using the direct PLNA. However, there was an increase of IgG1 AFCs using TNP-OVA, when compared to control (508 +/- 113 vs. 12 +/- 4 respectively). Glafenine, diclofenac, and ibuprofen, all associated with drug-induced anaphylaxis in humans, produced significant increases in IgG1 production to TNP-OVA. Of these three LMWCs, only diclofenac, which has been documented to induce neo-antigen formation, was detected with TNP-Ficoll. Hydralazine immunomodulation could be detected only with the direct PLNA although significant increases in IgM were identified with the co-injection of either reporter antigen. Isoniazid and methyldopa consistently produced negative responses in both assays. In summary, this study has demonstrated acceptable interlaboratory reproducibility of the RA-PLNA, using model autoimmunogenic LMWCs. Additionally, it demonstrated that an advantage of the RA-PLNA was that it identified all anaphylactic-associated LMWCs tested, detected the false negative compound aniline, and revealed what is thought to be the mechanism(s) associated with diclofenac-induced immunostimulation. (+info)Anti-4-1BB monoclonal antibodies abrogate T cell-dependent humoral immune responses in vivo through the induction of helper T cell anergy. (4/226)
The 4-1BB receptor (CDw137), a member of the tumor necrosis factor receptor superfamily, has been shown to costimulate the activation of T cells. Here we show that anti-mouse 4-1BB monoclonal antibodies (mAbs) inhibit thymus-dependent antibody production by B cells. Injection of anti-4-1BB mAbs into mice being immunized with cellular or soluble protein antigens induced long-term anergy of antigen-specific T cells. The immune response to the type II T cell-independent antigen trinintrophenol-conjugated Ficoll, however, was not suppressed. Inhibition of humoral immunity occurred only when anti-4-1BB mAb was given within 1 wk after immunization. Anti-4-1BB inhibition was observed in mice lacking functional CD8(+) T cells, indicating that CD8(+) T cells were not required for the induction of anergy. Analysis of the requirements for the anti-4-1BB-mediated inhibition of humoral immunity revealed that suppression could not be adoptively transferred with T cells from anti-4-1BB-treated mice. Transfer of BALB/c splenic T cells from sheep red blood cell (SRBC)-immunized and anti-4-1BB-treated mice together with normal BALB/c B cells into C.B-17 severe combined immunodeficient mice failed to generate an anti-SRBC response. However, B cells from the SRBC-immunized, anti-4-1BB-treated BALB/c mice, together with normal naive T cells, exhibited a normal humoral immune response against SRBC after transfer, demonstrating that SRBC-specific B cells were left unaffected by anti-4-1BB mAbs. (+info)Crowding effects on EcoRV kinetics and binding. (5/226)
The cytosol of the cell contains high concentrations of small and large macromolecules, but experimental data are often obtained in dilute solutions that do not reflect in vivo conditions. We have studied the crowding effect that large macromolecules have on EcoRV cleavage by adding high-molecular-weight Ficoll 70 to reaction solutions. Results indicate that Ficoll has surprisingly little effect on overall EcoRV reaction velocity because of offsetting increases in V(max) and K(m), and stronger nonspecific binding. The changes in measured parameters can largely be attributed to the excluded volume effects on reactant activities and the slowing of protein diffusion. Covolume reduction upon binding appears to reinforce nonspecific binding strength, and k(cat) appears to be slowed by stronger nonspecific binding, which slows product release. The data also suggest that effective Ficoll particle volume decreases as its concentration increases above a few weight percent, which may be due to Ficoll interpenetration or compression. (+info)Torque-speed relationship of the flagellar rotary motor of Escherichia coli. (6/226)
The output of a rotary motor is characterized by its torque and speed. We measured the torque-speed relationship of the flagellar rotary motor of Escherichia coli by a new method. Small latex spheres were attached to flagellar stubs on cells fixed to the surface of a glass slide. The angular speeds of the spheres were monitored in a weak optical trap by back-focal-plane interferometry in solutions containing different concentrations of the viscous agent Ficoll. Plots of relative torque (viscosity x speed) versus speed were obtained over a wide dynamic range (up to speeds of approximately 300 Hz) at three different temperatures, 22.7, 17.7, and 15.8 degrees C. Results obtained earlier by electrorotation (, Biophys. J. 65:2201-2216) were confirmed. The motor operates in two dynamic regimes. At 23 degrees C, the torque is approximately constant up to a knee speed of nearly 200 Hz, and then it falls rapidly with speed to a zero-torque speed of approximately 350 Hz. In the low-speed regime, torque is insensitive to changes in temperature. In the high-speed regime, it decreases markedly at lower temperature. These results are consistent with models in which torque is generated by a powerstroke mechanism (, Biophys. J. 76:580-587). (+info)Germinal centers without T cells. (7/226)
Germinal centers are critical for affinity maturation of antibody (Ab) responses. This process allows the production of high-efficiency neutralizing Ab that protects against virus infection and bacterial exotoxins. In germinal centers, responding B cells selectively mutate the genes that encode their receptors for antigen. This process can change Ab affinity and specificity. The mutated cells that produce high-affinity Ab are selected to become Ab-forming or memory B cells, whereas cells that have lost affinity or acquired autoreactivity are eliminated. Normally, T cells are critical for germinal center formation and subsequent B cell selection. Both processes involve engagement of CD40 on B cells by T cells. This report describes how high-affinity B cells can be induced to form large germinal centers in response to (4-hydroxy-3-nitrophenyl) acetyl (NP)-Ficoll in the absence of T cells or signaling through CD40 or CD28. This requires extensive cross-linking of the B cell receptors, and a frequency of antigen-specific B cells of at least 1 in 1,000. These germinal centers abort dramatically at the time when mutated high-affinity B cells are normally selected by T cells. Thus, there is a fail-safe mechanism against autoreactivity, even in the event of thymus-independent germinal center formation. (+info)A dual role for Src homology 2 domain-containing inositol-5-phosphatase (SHIP) in immunity: aberrant development and enhanced function of b lymphocytes in ship -/- mice. (8/226)
In this report, we demonstrate that the Src homology 2 domain-containing inositol-5-phosphatase (SHIP) plays a critical role in regulating both B cell development and responsiveness to antigen stimulation. SHIP(-/-) mice exhibit a transplantable alteration in B lymphoid development that results in reduced numbers of precursor B (fraction C) and immature B cells in the bone marrow. In vitro, purified SHIP(-/)- B cells exhibit enhanced proliferation in response to B cell receptor stimulation in both the presence and absence of Fcgamma receptor IIB coligation. This enhancement is associated with increased phosphorylation of both mitogen-activated protein kinase and Akt, as well as with increased survival and cell cycling. SHIP(-/)- mice manifest elevated serum immunoglobulin (Ig) levels and an exaggerated IgG response to the T cell-independent type 2 antigen trinitrophenyl Ficoll. However, only altered B cell development was apparent upon transplantation into nonobese diabetic-severe combined immunodeficient (NOD/SCID) mice. The in vitro hyperresponsiveness, together with the in vivo findings, suggests that SHIP regulates B lymphoid development and antigen responsiveness by both intrinsic and extrinsic mechanisms. (+info)Ficoll is not a medical term itself, but it is a type of synthetic polymer that is often used in laboratory settings for various medical and scientific purposes. Ficoll is a high-molecular-weight coopolymer of sucrose and epichlorohydrin, which forms a highly flexible and soluble structure with unique physical properties.
In medicine and research, Ficoll is commonly used as a component in density gradient media for the separation and purification of biological cells, viruses, and other particles based on their size, density, or sedimentation rate. The most common application of Ficoll is in the preparation of peripheral blood mononuclear cells (PBMCs) from whole blood samples.
Ficoll-Paque is a commercially available density gradient medium that contains Ficoll and a high-density solution of sodium diatrizoate. When a blood sample is layered onto the Ficoll-Paque solution and centrifuged, the various cell types in the blood separate into distinct bands based on their densities. The PBMCs, which include lymphocytes, monocytes, and other immune cells, collect at the interface between the Ficoll layer and the plasma layer, allowing for easy isolation and further analysis.
Therefore, while not a medical term itself, Ficoll plays an essential role in many laboratory procedures used in medical research and diagnostics.
Trinitrobenzenes are not typically referred to in the context of medical definitions, as they are chemical compounds that do not have direct relevance to medical fields. However, I can provide you with some general information about trinitrobenzenes.
Trinitrobenzenes are a group of chemical compounds consisting of a benzene ring (a cyclic aromatic hydrocarbon) with three nitro groups (-NO2) attached to it. The most common and well-known compound in this group is 1,3,5-trinitrobenzene (TNB). These compounds are highly explosive and have been used in military and industrial applications for their energetic properties.
In a medical or clinical context, trinitrobenzenes may be encountered only if someone is exposed to these chemicals in an occupational or accidental setting. Exposure can lead to local skin irritation, eye damage, or respiratory issues. Chronic exposure or high-dose acute exposure might cause more severe health problems, including damage to the liver and kidneys. However, trinitrobenzenes are not used as therapeutic agents or diagnostic tools in medicine.
Dextrans are a type of complex glucose polymers that are formed by the action of certain bacteria on sucrose. They are branched polysaccharides consisting of linear chains of α-1,6 linked D-glucopyranosyl units with occasional α-1,3 branches.
Dextrans have a wide range of applications in medicine and industry. In medicine, dextrans are used as plasma substitutes, volume expanders, and anticoagulants. They are also used as carriers for drugs and diagnostic agents, and in the manufacture of immunoadsorbents for the removal of toxins and pathogens from blood.
Dextrans can be derived from various bacterial sources, but the most common commercial source is Leuconostoc mesenteroides B-512(F) or L. dextranicum. The molecular weight of dextrans can vary widely, ranging from a few thousand to several million Daltons, depending on the method of preparation and purification.
Dextrans are generally biocompatible and non-toxic, but they can cause allergic reactions in some individuals. Therefore, their use as medical products requires careful monitoring and testing for safety and efficacy.
Centrifugation, Density Gradient is a medical laboratory technique used to separate and purify different components of a mixture based on their size, density, and shape. This method involves the use of a centrifuge and a density gradient medium, such as sucrose or cesium chloride, to create a stable density gradient within a column or tube.
The sample is carefully layered onto the top of the gradient and then subjected to high-speed centrifugation. During centrifugation, the particles in the sample move through the gradient based on their size, density, and shape, with heavier particles migrating faster and further than lighter ones. This results in the separation of different components of the mixture into distinct bands or zones within the gradient.
This technique is commonly used to purify and concentrate various types of biological materials, such as viruses, organelles, ribosomes, and subcellular fractions, from complex mixtures. It allows for the isolation of pure and intact particles, which can then be collected and analyzed for further study or use in downstream applications.
In summary, Centrifugation, Density Gradient is a medical laboratory technique used to separate and purify different components of a mixture based on their size, density, and shape using a centrifuge and a density gradient medium.
Fluorescein-5-isothiocyanate (FITC) is not a medical term per se, but a chemical compound commonly used in biomedical research and clinical diagnostics. Therefore, I will provide a general definition of this term:
Fluorescein-5-isothiocyanate (FITC) is a fluorescent dye with an absorption maximum at approximately 492-495 nm and an emission maximum at around 518-525 nm. It is widely used as a labeling reagent for various biological molecules, such as antibodies, proteins, and nucleic acids, to study their structure, function, and interactions in techniques like flow cytometry, immunofluorescence microscopy, and western blotting. The isothiocyanate group (-N=C=S) in the FITC molecule reacts with primary amines (-NH2) present in biological molecules to form a stable thiourea bond, enabling specific labeling of target molecules for detection and analysis.
Cell separation is a process used to separate and isolate specific cell types from a heterogeneous mixture of cells. This can be accomplished through various physical or biological methods, depending on the characteristics of the cells of interest. Some common techniques for cell separation include:
1. Density gradient centrifugation: In this method, a sample containing a mixture of cells is layered onto a density gradient medium and then centrifuged. The cells are separated based on their size, density, and sedimentation rate, with denser cells settling closer to the bottom of the tube and less dense cells remaining near the top.
2. Magnetic-activated cell sorting (MACS): This technique uses magnetic beads coated with antibodies that bind to specific cell surface markers. The labeled cells are then passed through a column placed in a magnetic field, which retains the magnetically labeled cells while allowing unlabeled cells to flow through.
3. Fluorescence-activated cell sorting (FACS): In this method, cells are stained with fluorochrome-conjugated antibodies that recognize specific cell surface or intracellular markers. The stained cells are then passed through a laser beam, which excites the fluorophores and allows for the detection and sorting of individual cells based on their fluorescence profile.
4. Filtration: This simple method relies on the physical size differences between cells to separate them. Cells can be passed through filters with pore sizes that allow smaller cells to pass through while retaining larger cells.
5. Enzymatic digestion: In some cases, cells can be separated by enzymatically dissociating tissues into single-cell suspensions and then using various separation techniques to isolate specific cell types.
These methods are widely used in research and clinical settings for applications such as isolating immune cells, stem cells, or tumor cells from biological samples.
The Glomerular Basement Membrane (GBM) is a part of the filtration barrier in the nephron of the kidney. It is a thin, porous sheet of extracellular matrix that lies between the glomerular endothelial cells and the visceral epithelial cells (podocytes). The GBM plays a crucial role in the process of ultrafiltration, allowing the passage of water and small molecules while preventing the loss of larger proteins into the urine. It is composed mainly of type IV collagen, laminin, nidogen, and heparan sulfate proteoglycans. Certain kidney diseases, such as Goodpasture's disease and some forms of glomerulonephritis, can involve damage to the GBM.
Isopycnic centrifugation is a type of centrifugation technique used in medical and scientific research. The term "isopycnic" refers to the process of separating particles based on their density, where the density of the particles is equal to that of the surrounding medium. In this technique, a sample containing particles of different densities is placed in a gradient medium within a centrifuge tube and then subjected to high-speed centrifugation.
During centrifugation, the particles move through the gradient medium until they reach a layer where their density matches that of the surrounding medium. Once the particles reach this point, they will no longer continue to move, even if the centrifugation continues for an extended period. This results in the separation of particles based on their densities, with denser particles settling at lower levels and less dense particles settling at higher levels.
Isopycnic centrifugation is a useful technique for separating and purifying various types of biological particles, such as viruses, organelles, and subcellular structures. It can also be used to study the properties of these particles, including their density, size, and shape.
Diatrizoate is a type of contrast medium that is used during X-ray examinations, such as CT scans and urography, to help improve the visibility of internal body structures. It is a type of iodinated compound, which means it contains iodine atoms. Diatrizoate works by blocking the absorption of X-rays, causing the areas where it is injected or introduced to appear white on X-ray images. This can help doctors to diagnose a variety of medical conditions, including problems with the urinary system and digestive tract.
Like all medications and contrast agents, diatrizoate can have side effects, including allergic reactions, kidney damage, and thyroid problems. It is important for patients to discuss any potential risks and benefits of using this agent with their healthcare provider before undergoing an X-ray examination.
Bovine Serum Albumin (BSA) is not a medical term per se, but a biochemical term. It is widely used in medical and biological research. Here's the definition:
Bovine Serum Albumin is a serum albumin protein derived from cows. It is often used as a stabilizer, an emulsifier, or a protein source in various laboratory and industrial applications, including biochemical experiments, cell culture media, and diagnostic kits. BSA has a high solubility in water and can bind to many different types of molecules, making it useful for preventing unwanted interactions between components in a solution. It also has a consistent composition and is relatively inexpensive compared to human serum albumin, which are factors that contribute to its widespread use.
Ficoll - Wikipedia
Ficoll Density Gradient Media, Other Supplements - Creative Bioarray
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