1,635 research outputs found
Clonal analysis of a human antibody response. Quantitation of precursors of antibody-producing cells and generation and characterization of monoclonal IgM, IgG, and IgA to rabies virus.
We quantitated and characterized the changes in the human B cell repertoire, at the clonal level, before and after immunization with rabies virus. Moreover, we generated 10 monoclonal cell lines producing IgM, IgG, and IgA antibodies to the virus. We found that in healthy subjects, not previously exposed to the virus, nearly 2% of the circulating B lymphocytes were committed to the production of antibodies that bound the virus. These B cells expressed the surface CD5 molecule. The antibodies they produced were polyreactive IgM that displayed a relatively low affinity for the virus components (Kd, 1.0-2.4 x 10(-6) g/microliters). After immunization, different anti-virus (IgG and IgA) antibody-producing cells consistently appeared in the circulation and increased from less than 0.005% to greater than 10% of the total B cells committed to the production of IgG and IgA, respectively. Most of such B cells do not express CD5 and produce monoreactive antibodies of high affinity for rabies virus (Kd, 6.5 x 10(-9) to 1.2 x 10(-10) g/microliters). One of these IgG mAbs efficiently neutralized rabies virus in vitro and in vivo, as detailed elsewhere (Dietzschold, B., P. Casali, Y. Ueki, M. Gore, C. E. Rupprecht, A. L. Notkins, and H. Koprowski, manuscript submitted for publication). Hybridization experiments using probes specific for the different human V gene segment families revealed that cell precursors producing low affinity IgM binding to rabies virus utilized a restricted number of VH gene segments (i.e., only members of the VHIIIb subfamily), whereas cell precursors producing high affinity IgG and IgA to rabies virus utilized an assortment of different VH gene segments (i.e., members of the VHI, VHIII, VHIV, and VHVI families and VHIIIb subfamily). In conclusion, our studies show that EBV transformation in conjunction with limiting dilution technology and somatic cell hybridization techniques are useful methods for quantitating, at the B cell clonal level, the human antibody response to foreign Ags and for generating human mAbs of predetermined specificity and high affinity
Synthesis and properties of novel bis(1,3-benzodithiolium)-type dications containing a biaryl unit: New redox systems undergoing reversible structural changes by electron transfer
In order to develop new redox systems which undergo reversible structural changes by electron transfer, bis(1,3-benzodithiolium)-type dications (5 21) containing a biaryl unit have been synthesized by hydride abstraction of the corresponding bis(1,3-benzodithiol-2-yl)biaryis (8). Reduction of 5 2, with zinc gave the corresponding intramolecular cyclization products (6), which reverted to 5(2+) by oxidation. Cyclic voltammetry also showed the efficient interconversion between both states. X-Ray analyses revealed that the twist angle of the biaryl unit decreases largely by reductive intramolecular cyclization.ArticleHETEROCYCLES. 69(1): 365 (2006)journal articl
A New Experimental Approach to Evaluate Plasma-induced Damage in Microcantilever
Plasma etching, during micro-fabrication processing is indispensable for fabricating MEMS structures. During the plasma processes, two major matters, charged ions and vacuum–ultraviolet (VUV) irradiation damage, take charge of reliability degradation. The charged ions induce unwanted sidewall etching, generally called as “notching”, which causes degradation in brittle strength. Furthermore, the VUV irradiation gives rise to crystal defects on the etching surface. To overcome the problem, neutral beam etching (NBE), which use neutral particles without the VUV irradiation, has been developed. In order to evaluate the effect of the NBE quantitatively, we measured the resonance property of a micro-cantilever before and after NBE treatment. The thickness of damage layer (δ) times the imaginary part of the complex Young's modulus (Eds) were then compared, which is a parameter of surface damage. Although plasma processes make the initial surface of cantilevers damaged during their fabrication, the removal of that damage by NBE was confirmed as the reduction in δEds. NBE will realize a damage-free surface for microstructures
Potential Role of Protein Kinase B in Insulin-induced Glucose Transport, Glycogen Synthesis, and Protein Synthesis
Various biological responses stimulated by insulin
have been thought to be regulated by phosphatidylinosi-tol
3-kinase, including glucose transport, glycogen syn-thesis,
and protein synthesis. However, the molecular
link between phosphatidylinositol 3-kinase and these
biological responses has been poorly understood. Re-cently,
it has been shown that protein kinase B (PKB/c-Akt/
Rac) lies immediately downstream from phosphati-dylinositol
3-kinase. Here, we show that expression of a
constitutively active form of PKB induced glucose up-take,
glycogen synthesis, and protein synthesis in L6
myotubes downstream of phosphatidylinositol 3-kinase
and independent of Ras and mitogen-activated protein
kinase activation. Introduction of constitutively active
PKB induced glucose uptake and protein synthesis but
not glycogen synthesis in 3T3L-1 adipocytes, which lack
expression of glycogen synthase kinase 3 different from
L6 myotubes. Furthermore, we show that deactivation
of glycogen synthase kinase 3 and activation of rapamy-cin-
sensitive serine/threonine kinase by PKB in L6 myo-tubes
might be involved in the enhancement of glycogen
synthesis and protein synthesis, respectively. These re-sults
suggest that PKB acts as a key enzyme linking
phosphatidylinositol 3-kinase activation to multiple bi-ological
functions of insulin through regulation of
downstream kinases in skeletal muscle, a major target
tissue of insulin
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