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61.
Human cytotoxic T cell clones directed against herpes simplex virus-infected cells. IV. Recognition and activation by cloned glycoproteins gB and gD 总被引:20,自引:0,他引:20
J M Zarling P A Moran R L Burke C Pachl P W Berman L A Lasky 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(12):4669-4673
Results of studies in mice and clinical observations in man indicate that T cell-mediated immunity is important in resistance to herpes simplex virus (HSV) infections. This study was undertaken to elucidate the viral antigen specificity of human HSV-immune T cells. Purified HSV-1 glycoproteins gB-1 and gD-1, cloned and expressed in mammalian cells, were found to stimulate proliferation of, and interleukin 2 (IL 2) production by, peripheral blood lymphocytes (PBL) of HSV seropositive individuals, indicating the presence of memory T cells to gB-1 and gD-1 in individuals with serologic evidence of immunity to HSV. Second, T cell clones, generated by stimulation of PBL with HSV-1, were found to recognize gB-1 or gD-1, as evidenced by the ability of the clones to proliferate in response to stimulation with gB-1 or gD-1 in the absence of exogenous IL 2. Third, HSV-specific T cell clones, lytic for HSV-1 or both HSV-1- and HSV-2-infected autologous target cells, were generated after stimulation of PBL with purified cloned gB-1 or gD-1. Our findings, that human HSV-specific T cells can recognize and be activated by HSV subunit antigens gB-1 or gD-1, imply that these glycoproteins play a role in human T cell-mediated immunity to HSV and support the contention that a gB-1 or gD-1 subunit vaccine may be protective in man. 相似文献
62.
63.
The functional domains of coagulation factor VIII:C 总被引:11,自引:0,他引:11
R L Burke C Pachl M Quiroga S Rosenberg N Haigwood O Nordfang M Ezban 《The Journal of biological chemistry》1986,261(27):12574-12578
A lack of factor VIII:C, manifested as a bleeding disorder due to the absence of clot formation, is known as hemophilia A, an X chromosome-linked inherited disease afflicting 1-2 males/10,000. To determine the minimum functional domain(s) essential for factor VIII:C activity, we have expressed the amino-terminal (92-kDa) and carboxyl-terminal (80-kDa) proteolytic cleavage products as individual, secreted polypeptides in monkey cells without the 909-residue central region. We have found that neither terminal domain alone is able to promote coagulation in factor VIII:C-deficient plasma. However, when the 92- and 80-kDa peptides are co-expressed, clotting activity is readily detected. Thus, these two chains alone constitute an active or activatable complex. The central domain is required neither for activity nor for the assembly of an active complex from two chains expressed in trans. These results suggest that a truncated derivative of factor VIII:C may be useful in coagulation therapy. 相似文献
64.
65.
This study explores, by kinetic analysis, the movement of calcium among cellular pools of target cells in which cytotoxicity is induced by human lymphotoxin, and evaluates the requirement for calcium in this reaction. We employed the kinetic model of Borle to quantitate flux rates and pool sizes. It was found that the rate of flux between the surface (plasma membrane-glycocalyx) compartment and the intracellular compartment was greatly increased. The size of the total exchangeable intracellular calcium pool was not altered, but there was an apparent decrease in the size of the surface calcium pool. This latter phenomenon may be related to the blebbing and exfoliation of plasma membranes under the influence of the lymphokine. Lymphotoxin-induced cytotoxicity is observed in calcium-free medium and over a range of calcium concentrations. These results argue against cell death due to a massive in rush of calcium into the cell under these circumstances. 相似文献
66.
Structural basis of anthracycline selectivity for unilamellar phosphatidylcholine vesicles: an equilibrium binding study 总被引:2,自引:0,他引:2
Fluorescence anisotropy titration was used to determine the equilibrium binding affinities of several anthracycline antitumor antibiotics for sonicated dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) vesicles at 27.5 degrees C. Eight daunomycin analogues, all differing from the parent by one structural change in the aglycon portion of the molecule, as well as four anthracycline congeners modified in the amino sugar were studied. Double-reciprocal plots were used to determine overall binding affinities (K). It was shown that structural changes in both the aglycon and amino sugar portions of the daunomycin molecule strongly modulated K values for DMPC and DPPC bilayers. For modifications in the aglycon portion of an anthracycline, a correlation between drug hydrophobicity and membrane affinity was observed. The number of binding sites per phospholipid molecule (n) and the apparent association constant (Kapp) where K = nKapp, were determined at several temperatures for adriamycin, daunomycin, and carminomycin. The n values were found to be independent of temperature for fluid-phase DMPC or solid-phase DPPC bilayers. The Kapp values (25 degrees C) ranged from (0.82-4.4) X 10(5) M-1 for DMPC vesicles to (4.4-7.3) X 10(5) M-1 for DPPC vesicles. Although the Kapp values for the three drugs were similar for a particular bilayer, major differences were noted in the values of n and, therefore, in the overall vesicle affinities (nKapp). van't Hoff plots showed that anthracycline binding was exothermic; in all cases but one binding was accompanied by a decrease in entropy.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
67.
Reduction potential of iron in transferrin 总被引:1,自引:0,他引:1
D C Harris A L Rinehart D Hereld R W Schwartz F P Burke A P Salvador 《Biochimica et biophysica acta》1985,838(3):295-301
The reduction potential of Fe3+ in transferrin was measured spectrophotometrically by equilibration with methyl viologen in the presence of sodium dithionite. For an ionic strength near 0.1 M at 25 degrees C and pH 7.3 under 0.048 atm. CO2, half of the iron is reduced at a potential near -0.40 V (vs. standard hydrogen electrode). At least one disulfide bond of the protein is partially reduced at a potential of -0.44 V, as evidenced by reaction with [14C]iodoacetate. 相似文献
68.
Prenatal and lactational transmission of Toxocara canis and Ancylostoma caninum: experimental infection of the bitch before pregnancy 总被引:1,自引:0,他引:1
Breeding of five parasite-free and five experimentally infected (6000 Toxocara canis eggs orally and 2500 Ancylostoma caninum larvae subcutaneously) beagle bitches was done so that pairs of bitches (1 uninfected, 1 infected) whelped simultaneously. Pups born to an infected bitch were removed at birth and nursed by the paired uninfected bitch until 4 weeks of age when pups were necropsied to determine the number of parasites they had acquired prenatally from their infected mother. Pups born to the parasite-free bitch were nursed by the infected bitch until necropsied at 4 weeks of age to determine the number of parasites passed via the lactational route. Of 680 ascarids transmitted to pups by either route, 98.5% were transmitted prenatally and 1.5% lactationally. Transmission of 2746 hookworms to 22 pups occurred solely by the lactational route; prenatal transmission of this parasite did not occur in any of the 25 pups born to infected bitches. 相似文献
69.
Studies on the maintenance of cytochromes P-450 and b5, monooxygenases and cytochrome reductases in primary cultures of rat hepatocytes 总被引:2,自引:0,他引:2
The cytochrome P-450 content of rat hepatocytes declined rapidly over 72 h in culture, due primarily to denaturation to cytochrome P-420. Six different media were investigated for their ability to conserve cytochrome P-450 during culture, and the most successful was a modified Earle's medium. After 72 h culture in this medium, cytochromes P-450 and b5, NADH-cytochrome b5- and NADPH-cytochrome c-reductases were maintained at 40, 100, 35 and 52% of fresh cell values, respectively. Cytochrome P-450 showed differential functional stability during culture with ethoxyresorufin O-deethylation being more stable than either pentoxyphenoxazone O-depentylation or biphenyl 4-hydroxylation. Monooxygenase than did cytochrome P-450 content. This discrepancy was not explained by loss of flavin nucleotides, FMN or FAD. 相似文献
70.
Phytotoxins from Alternaria helianthi: radicinin,and the structures of deoxyradicinol and radianthin
A novel compound, radianthin, with phytotoxic activity was isolated from liquid cultures of Alternaria helianthi and identified as a pyrone related to radicinin. A second metabolite was identified as radicinin itself while deoxyradicinol is described for the first time as a natural product. 相似文献