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C DeLisi 《Journal of theoretical biology》1975,51(2):337-345
Equations are presented which can be used to describe the inhibition of plaques by multifunctional antigen which binds γG antibody bivalently. The interaction is treated as a bimolecular reaction which is irreversible within the time of the experiment. It is shown that under these conditions the characteristics of the inhibition curve, and their relationship to kinetic and thermodynamic parameters are strongly dependent upon how antibody interacts with RBCs. When the epitope coating is dense, multivalent attachment of antibody is likely and the interaction is considered irreversible. When the epitope coating is sparse, only rapid, reversible univalent attachment is considered and local equilibrium is assumed to hold.The first case leads to an abrupt inhibition curve whose position is determined by the forward rate constant and RBC density. The second case leads to broad asymmetric curves. For this situation the relation between the extent of inhibition and the affinity of the population is generally complicated and reliable affinity information is difficult to obtain. This is contrasted to results obtained previously for unifunctional inhibitors from which reliable affinity information can, in principle, be obtained. The results emphasize the need for carefully designed experiments if affinity information is to be obtained from inhibition studies. 相似文献
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C DeLisi 《Journal of theoretical biology》1975,51(2):313-335
We present a mathematical theory of hapten inhibition of hemolytic plaque formation. The treatment is based upon the mathematical model for plaque growth presented by DeLisi &; Bell (1974). The lymphocyte under consideration is embedded in an infinite three-dimensional medium, and is secreting antibodies isotropically at a constant rate. As the antibodies diffuse from the source they can bind reversibly to hapten, and in the most general case reversibly to red blood cell (RBC) epitope. The model leads to a non-linear diffusion equation coupled to a set of first order differential equations. The system must, in general, be solved numerically. However, in many cases of experimental interest simplifications arise which permit closed form solutions to be obtained. In this paper we have developed solutions for three special cases.In the first example antibodies can bind only univalently to RBCs, as would be expected if the epitopes are sparsely distributed. In this case reaction between antibody site and RBC epitope is rapid ( ⪆ 1 sec) and reversible and local equilibrium is assumed. This leads to a “pure” diffusion equation in the free antibody concentration, but with a reduced diffusion coefficient.In another example univalent attachment of an antibody site to a RBC epitope is followed by a rapid irreversible intramolecular reaction. This might be expected for example if the epitope density is large. An exact solution to the resulting diffusion equation was also found in this case. In order to assess an intermediate situation, we also solved the equations for a model in which intramolecular reaction is slow and irreversible.The theory predicts that the type of information one can obtain from inhibition experiments depends critically upon the preparation of the RBC. If the cell is sufficiently haptenated so that rapid irreversible multivalent attachment is favorable, a differential plot of the inhibition curve will reflect the affinity distribution of antibody sites for free hapten. If only univalent attachment with RBCs is possible, so that antibody sites bind to RBC hapten in the same way they bind to free hapten, then a differential plot of the inhibition curve will reflect the secretion rate distribution. 相似文献
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The hemolytic plaque assay: theory for finite layers. 总被引:1,自引:0,他引:1
We extend the mathematical theory of hemolytic plaque growth to include plaques produced by cells secreting antibodies in layers of finite thickness. Previous theories have assumed that the layer was either two-dimensional or of infinite thickness. By using the method of images we derive an equation for the plaque radius as a function of time for layers of any thickness. We show that at short times and at long times the equation reduces to the appropriate infinite three-dimensional and two-dimensional limiting forms, and obtain expressions for estimating the range of times for which these limiting results are valid. For the liquid monolayer technique we obtain a new limiting result. The equation for the plaque radius is a transcendental equation which we solve numerically for a number of cases of interest. These results illustrate a variety of different features of plaque growth associated with the finite thickness of the layer. Experimental studies are usually carried out in layers whose thicknesses are not standardized. In the assays commonly used the thickness h can vary more than six hundred fold, i.e. 1 × 10?3 cm ?h? 6.5 × 10?1 cm. Such variation in h will cause widely different kinetics of plaque growth. For typical plaque experiments of one hour duration the two-dimensional limit is valid when h ? 3 × 10?3 cm while the infinite thickness limit is valid when h? 10?1 cm. For thicknesses in between these values the finite layer results must be used. 相似文献
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We use the mathematical theory of plaque growth to determine if there is merit in performing a hemolytic plaque assay in the presence of an external electric field. In particular, we study the effects of an electric field on the transport of anti-bodies secreted by a single lymphocyte and on the size and shape of the plaques they produce. Our results indicate that in the presence of an applied electric field: (1) The mobility of the antibodies produced by the antibody forming cell can be determined from the plaque shape. (In the electric field the plaques are no longer circular, but cigar shaped.) (2) By changing the magnitude or direction of the applied electric field more than one plaque can be generated by a single AFC. Thus changes in mobility or the rate of antibody secretion can be assayed. (3) Plaques will reach a steady state size; for good emitters (cells that secrete antibodies at a high rate or that secrete high affinity antibodies) this steady state will be achieved rapidly.Equations are given which describe both the temporal development and steady state plaque size and shape. From the equations, computer generated plots of plaques produced by typical antibody farming cells are presented. These plots are then used to show how pictures of plaques formed in an electric field can be analyzed to determine the antibody mobility. 相似文献
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Plant and Soil - Aquatic plants, including rice, develop iron (Fe) plaques on their roots due to radial oxygen loss (ROL), and these plaques accumulate both beneficial and toxic elements. Silicon... 相似文献
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Potentiation of hemolytic plaque formation by incubation of immunized spleen cells in phenothiazine derivatives 总被引:2,自引:0,他引:2
G A Becker A V Pisciotta 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1967,124(3):764-767
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A mechanistic kinetic model of gel firmness development during milk gel formation is presented. The model correctly accounts for the influence of enzymatic kappa-casein hydrolysis on the rate of firmness development in renneted milk gels. The model used is based on two first-order reactions occurring in series. The first reaction is enzymatically controlled and corresponds to the formation of gel crosslink sites by kappa-casein hydrolysis. The second reaction is nonenzymatic and corresponds to the process of crosslink formation and depletion of active sites. The model successfully predicts gel firmness development in the temperature range 31-45 degrees C for a variety of initial enzyme concentrations. 相似文献
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The direct hemolytic plaque response to hemocyanin in the rabbit 总被引:1,自引:0,他引:1
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The pathogenic Neisseriae Neisseria meningitidis and Neisseria gonorrhoeae, initiate colonization by attaching to host cells using type IV pili. Subsequent adhesive interactions are mediated through the binding of other bacterial adhesins, in particular the Opa family of outer membrane proteins. Here, we have shown that pilus-mediated adhesion to host cells by either meningococci or gonococci triggers the rapid, localized formation of dramatic cortical plaques in host epithelial cells. Cortical plaques are enriched in both components of the cortical cytoskeleton and a subset of integral membrane proteins. These include: CD44v3, a heparan sulphate proteoglycan that may serve as an Opa receptor; EGFR, a receptor tyrosine kinase; CD44 and ICAM-1, adhesion molecules known to mediate inflammatory responses; f-actin; and ezrin, a component that tethers membrane components to the actin cytoskeleton. Genetic analyses reveal that cortical plaque formation is highly adhesin specific. Both pilE and pilC null mutants fail to induce cortical plaques, indicating that neisserial type IV pili are required for cortical plaque induction. Mutations in pilT, a gene required for pilus-mediated twitching motility, confer a partial defect in cortical plaque formation. In contrast to type IV pili, many other neisserial surface structures are not involved in cortical plaque induction, including Opa, Opc, glycolipid GgO4-binding adhesins, polysialic acid capsule or a particular lipooligosaccharide variant. Furthermore, it is shown that type IV pili allow gonococci to overcome the inhibitory effect of heparin, a soluble receptor analogue, on gonococcal invasion of Chang and A431 epithelial cells. These and other observations strongly suggest that type IV pili play an active role in initiating neisserial infection of the mucosal surface in vivo. The functions of type IV pili and other neisserial adhesins are discussed in the specific context of the mucosal microenvironment, and a multistep model for neisserial colonization of mucosal epithelia is proposed. 相似文献
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The effects of a series of n-alcohols and n-carboxylic acids on lipoxygenase activity was studied. It was shown that to a large extent the effects of these compounds could be ascribed to physiochemical interaction with the substrate solution rather than a direct action on the enzyme itself. The effect of better substrate analogues such as stearate and oleate could also be ascribed to this effect. A type-2 lipoxygenase was found to have a very unusual velocity-substrate relationship which could be normalized by addition of calcium chloride in amounts stoichiometric with the substrate. An excess of calcium inhibited the enzyme. By comparison of results with linoleoyl sulphate/linoleoyl alcohol mixed micelles, an explanation for this unusual velocity-substrate activity is presented. 相似文献
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Restriction of bacteriophage plaque formation in Streptomyces spp. 总被引:4,自引:11,他引:4
Several Streptomyces species that produce restriction endonucleases were characterized for their ability to propagate 10 different broad host range bacteriophages. Each species displayed a different pattern of plaque formation. A restrictionless mutant of S. albus G allowed plaque formation by all 10 phages, whereas the wild-type strain showed plaques with only 2 phages. DNA isolated from three of the phages was analyzed for the presence of restriction sites for Streptomyces species-encoded enzymes, and a very strong correlation was established between the failure to form plaques on Streptomyces species that produced particular restriction enzymes and the presence of the corresponding restriction sites in the phage DNA. Also, the phages that lacked restriction sites in their DNA generally formed plaques on the corresponding restriction endonuclease-producing hosts at high efficiency. The DNAs from the three phages analyzed also generally contained either many or no restriction sites for the Streptomyces species-produced enzymes, suggesting a strong evolutionary trend to either eliminate all or tolerate many restriction sites. The data indicate that restriction plays a major role in host range determination for Streptomyces phages. Analysis of bacteriophage host ranges of many other uncharacterized Streptomyces hosts has identified four relatively nonrestricting hosts, at least two of which may be suitable hosts for gene cloning. The data also suggest that several restriction systems remain to be identified in the genus Streptomyces. 相似文献