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Biomedical subjects

K Leung

Publications and source records attributed to K Leung.

108 records · Page 6Linked to original sources

Selective suppression of the cytotoxic T cell response to influenza virus in mice.

Mice injected with inactivated (UV light-irradiated) influenza virus produce specific antibody, become sensitized for a delayed-type hypersensitivity reaction, but do not generate specific cytotoxic T (Tc) cells. If injected 4-5 days later with infectious virus, the formation of Tc cells is suppressed by > 90%. If A strain viruses are used, the suppression observed is cross-reactive within A strain viruses but does not extend to B/LEE or to Sendai virus. Serum from mice injected with UV-irradiated virus contains antibodies which on adoptive transfer can inhibit Tc cell formation when infectious homologous virus is used to challenge the recipients. Spleen cells from the same mice, upon adoptive transfer, also inhibit (50-70%) Tc cell formation if transferred within 24 h of injection of infectious virus, and the specificity pattern observed is cross-reactive within A strains. The activity of the cells mediating suppression is destroyed by monospecific anti-Thy-1.2 antibody and complement. The immune cells require I region sharing between donor and recipient mice for their suppressor activity to be effective. (There is also a partial requirement for K, D region sharing, but the possible rejection of transferred cells is not excluded.) Dilution assays in which clonal expansion of Tc precursors is used to estimate their frequency and the presence of T helper (Th) cells indicate that suppressed mice possess Tc precursors and primed cells which, upon restimulation, act as Th cells. Furthermore, injection of irradiated Th cells with inactivated virus does not significantly reduce the ensuing suppression.

Animals↗

Bovine prolactin, TSH, T and T concentrations as affected by tall fescue summer toxicosis and temperature.

Serum concentrations of prolactin (PRL), thyrotropin (TSH), thyroxine (T4) and triiodothyronine (T3) were compared between Holstein calves fed a toxic fescue (G1-307) and Holstein calves fed a less toxic fescue (G1-306) at one of three environmental temperatures (low, 10 to 13 C; medium, 21 to 23 C; high, 34 to 35 C). Three calves were assigned to each treatment combination, and hormone concentrations were determined before (basal) and after thyrotropin-releasing hormone (TRH) was injected at 15 microgram/100 kg body weight via jugular cannula. Calves fed the toxic G1-307 had lower basal PRL concentrations than those fed G1-306 (1.8 +/- .1 vs 6.0 +/- 1.2 ng/ml). Increasing ambient temperature increased basal PRL levels in calves fed G1-306 (i.e., low [2.3 +/- .3] < medium [3.6 +/- .9] < high [12.1 +/- 1.3 ng/ml]) but not in calves fed G1-307 (i.e. low [1.8 +/- .1] = medium [2.0 +/- .1] = high [1.6 +/- .1]). Similarly, after TRH injection, PRL release was greater in calves fed the less toxic G1-306 (35.7 +/- 5.2 ng/ml) than in those fed G1-307 (5.6 +/- .6 ng/ml) and was positively affected by temperature (i.e., low [6.9 +/- 1.2] < medium [18.8 +/- 5.6] < high [36.3 +/- 5.9 ng/ml]). In contrast, basal and TRH-induced TSH concentrations were unaffected by temperature or fescue. However, T3 (time 0) was inversely related to temperature (1.06 +/- .1, 1.40 +/- .1 and 1.87 +/- .2 ng/ml for high, medium and low temperature, respectively). After TRH injections, T3 increased linearly and at the same rate (.007 ng/ml/min) at all temperatures, but T4 concentrations increased only in low temperature groups. Neither T4 nor T3 concentrations were affected by forage fed. We conclude that some factor(s) in toxic fescue inhibits PRL secretion, especially at high temperature, an environmental condition closely associated with fescue summer toxicosis in cattle. Increased body temperature and respiration rates associated with fescue summer toxicosis apparently are not mediated by TSH or thyroid hormone secretion.

Animals↗

Serum thyrotropin, thyroxine, and tri-iodothyronine in dairy cows fed varying amounts of iodine.

The objective was to determine whether dairy cows fed supplemental iodine daily at 2.5 or 5.0 mg per kg body weight for 49 wk had altered thyroid status relative to controls fed no supplemental iodine. Average daily dose of iodine was 1.6 and 3.3 g. Iodine as ethylene diamine dihydriodide was fed to Holstein cows beginning at 8 to 10 wk of lactation, for the remainder of that lactation, through the dry period, and into the next lactation. On day 341 after onset of iodine feeding, each cow was injected intravenously with 15 microgram thyrotropin releasing hormone per 100 kg body weight. Thyrotropin in jugular blood collected before injection of releasing hormone averaged 7.6, 6.1, and 8.2 ng/ml for cows given 0, 2.5, and 5.0 mg iodine/kg body weight; there were no differences between means. Releasing hormone increased thyrotropin, thyroxine, and tri-iodothyronine in serum of all cows, but increases were not affected by quantity of dietary iodine.

Animals↗

Tumor-associated antigens of rat moloney sarcoma cells. II. Cytosol antigens.

Rat Moloney sarcoma cells (MST) were pulsed with 35S-L-methionine for 10 and 60 min and lysed by vortexing in 0.5% deoxycholate, 0.5% NP40, 0.02 M Tris, 0.05 M NaCl, pH 7.5, for 30 sec. The lysate was centrifuged at 16,300 X G for 10 min and the supernatant was co-precipitated with Ig fractions of normal BN serum, normal Lewis serum, BN antiserum to Moloney sarcoma cells (BNaMST), BN antiserum to tumor-associated antigens (BNaTAA), BN antiserum to murine leukemia virus (BNaMuLV), BN antiserum to p30 (BNap30), BN antiserum to gp70 (BNagp70), Lewis antiserum to BN (LeaBN), and BN antiserum to BC5 tumor (BNaBC5). With BNaTAA and BNaMST, a cytoplasmic TAA with m.w. 85,000 was detected. In addition, BNaTAA detected three other species of cytoplasmic TAA with m.w. 220,000, 170,000 and 39,000.

Animals↗

Tumor-associated antigens of rat moloney sarcoma cells. I. Cell-surface antigens.

The dissociated cell surface membranes of a rat Moloney sarcoma (MST), derived from a BN rat, were extracted with 2 M KI, with 6 M guanidine thiocyanate, or by papain digestion. Extracts obtained with these three reagents were fractionated on columns of controlled-pore glass, 170 A pore size. A fraction was eluted from each preparation that contained tumor-associated antigens (TAA), viral, fetal, and histocompatibility antigens. With an antibody specific for TAA, the TAA, devoid of detectable viral, fetal, and histocompatibility antigens, were co-precipitated by addition of goat antibody to rat immunoglobulin. After electrophoresis, on slab gels, three bands were detected with estimated m.w. of 185,000, 150,000, and 70,000. No such bands were detected on slab gel electrophoresis with extracts of BC5, a chemically induced tumor, of normal BN lymphoid cells, of M-MuLV, or of fetuses, after incubation with anti-TAA antibody and goat antibody to rat immunoglobulin. TAA extracts prepared with 2 M KI, 6 M guanidine thiocyanate, or papain digestion showed immunologic reactivity. Cold TAA inhibited the co-precipitation of labeled TAA by rat antibody specific for TAA; they elicited antibody in guinea pigs but not in rats; and antibodies specific for TAA were cytotoxic to MST in the presence of C.

Animals↗

Deoxycytidine transport in the presence of a cytidine deaminase inhibitor and the transport of uracil in Escherichia coli B.

Tetrahydrouridine, a cytidine deaminase inhibitor, prevents periplasmic degradation of deoxycytidine by Escherichia coli B. It does not inhibit deoxycytidine transport and therefore allows an accurate determination of deoxycytidine transport. Data obtained using tetrahydrouridine show that deoxycytidine is transported in E. coli B as the intact nucleoside by an active transport process, with a K-m of 6 times 10-minus 6 M. Cytidine and deoxyadenosine inhibit transport competitively, whereas guanosine has no effect on transport. Arsenate or KCN greatly reduces transport. In a mutant resistant to the nucleoside antibiotic, showdomycin, the active transport of deoxycytidine is lost, and residual slow uptake occurs by passive diffusion. Uracil is accumulated in E. coli B by an active transport process with a K-m of 5 times 10-minus 7 M.

Adenine Phosphoribosyltransferase↗

Long-term incubation of rat thymus cells: Cytolytic actions of glucocorticoids in vitro.

A system has been developed for long-term incubation of rat thymus cells in sus pension in a chemically defined medium consisting of medium 199 and methylcellulsoe. With this system it is possible to study quantitatively the full time-course of glucocorticoid actions, from the early inhibitory effects on glucose uptake to the slow cytolytic effects. Cells incubated under these conditions preserve their sensitivity to the inhibitory effects of cortisol on glucose uptake for al least 12 h. Glucocorticoids at physiological concentrations lead to cytolysis (measured by reduction in viable cell counts determined by a modification of the pronase-cetrimide method) by 12 h incubation time. The cytolytic effect exhibits well-defined dose-response relationships and specificity for glucocorticoids. By 24 h,cortisol at 10(-6)M consistently reduces the total cell count by about 15%, and the viable cell count by about 40%. Cortisol is active down to 10(-7) M. Dexamethasone is roughly 10 times as active as cortisol, and cortisone completely inactive. Cortexolone is slightly active, but also shows antiglucocorticoid activity by its ability to block the action of cortisol. Under anerobic conditions in Medium 199, no inhibitory effect of cortisol is observed on glucose uptake.

Animals↗

Induction of NF-kappa B during monocyte differentiation is associated with activation of HIV-gene expression.

Cells of the monocyte-macrophage lineage are important targets of HIV infection. We report here that the phenotypic differentiation of monocyte cell lines induced by phorbol esters or tumour necrosis factor alpha (TNF alpha) is associated with expression of nuclear factor kappa B (NF-kappa B). In parallel with such differentiation, HIV transcription, monitored using an HIV long terminal repeat reporter gene construct, is activated in such cells under the influence of enhanced NF-kappa B expression. Also, in a promonocyte cell line chronically infected with HIV, NF-kappa B expression and HIV transcription were enhanced on stimulation with phorbol ester or TNF alpha. Thus, stimulation of monocyte cell lines by phorbol esters or TNF alpha induces cell differentiation and activates HIV transcription. Such a process may have fundamental implications in AIDS pathogenesis in vivo and may be important in disease progression induced by opportunistic infections directly or indirectly involving macrophages.

Cell Differentiation↗

Daunorubicin coupled to monoclonal antibodies via a cis-aconitic anhydride linker: biochemical and cytotoxic properties revisited.

Daunorubicin (DNR) was coupled to monoclonal antibodies (Mab) reactive to breast tumor cells using the acid-labile linking agents cis-asonitic anhydride and two other non acid-labile analogs, glutaric anhydride and citraconic anhydride. The acid derivatives of DNR formed by reaction with the anhydrides were converted to their N-hydroxysuccinimide (NHS) active esters for coupling to MAb. The molar input of drug NHS ester to MAb ranged from 1:1 to 100:1. The resulting MAb-DNR conjugates were purified by gel filtration and analyzed by high performance liquid chromatography. Monomeric conjugates contained 0.2 to 11.0 moles of DNR/mole of MAb. No evidence of cell killing was observed up to a concentration of 10 micrograms/ml DNR bound to MAb, while DNR exhibited 50% killing of the breast tumor cell line MCF-7 at a concentration of 1 microgram/ml.

Aconitic Acid↗

Fetal cardiorespiratory and neurobehavioral response to zidovudine (AZT) in the baboon.

OBJECTIVE: To evaluate the effects of intravenous of zidovudine (AZT) at a dose and duration of infusion comparable to that used clinically on parameters reflective of fetal well-being. METHODS: Thirteen chronically instrumented noninfected baboons were monitored during intravenous infusions of AZT. Fetal cardiorespiratory activity and neurobehavioral function were assessed with 4-48-hour infusion of AZT to ten mothers (0.5-2.1 mg/kg per hour) and three fetuses (2-6 mg/h), which resulted in fetal plasma concentration of AZT of 194-3100 ng/ml. RESULTS: No significant differences were found in the mean values in control periods, before and after infusion with values during infusion for parameters of fetal heart rate and rate variability (n = 7), breathing activity (n = 8), electroencephalographic activity (n = 8), and behavioral state (N = 7). No correlations were found with drug level. CONCLUSIONS: The absence of associations between exposure of the fetal baboon to AZT and changes in parameters reflective of fetal condition suggests that comparable exposure of the human fetus during intravenous infusion of drug would not confound clinical monitoring used to assess fetal well-being. These findings supplement conclusions from clinical research in support of U.S. Public Health Service recommendations that intrapartum fetal monitoring be performed as clinically indicated, not specifically because pregnant patients are treated with intravenous AZT.

Animals↗