Four influenza A-H3N2 viruses isolated in pigs from different herds in Central Italy in the period 1981/82 have been antigenically and biochemically analysed. Three of them A/Sw/Italy/2/81, A/Sw/Italy/7/81, A/Sw/Italy/8/82 were found to be serologically related to A/Bangkok/1/79 (H3N2). These three viruses were shown to have an identical electrophoretic pattern, as regards virus induced polypeptides and were clearly distinguishable from the virus A/Sw/Italy/6/81 which was antigenically related to A/England/42/72 (H3N2) and A/Sw/Taiwan/7310/70 as shown by specific monoclonal and polyclonal antisera. The observed biochemical variations underline the importance of the changes occurring by genetic reassortments or mutations in human influenza viruses, during their maintenance in pigs.
The sensitivities of haemagglutination-inhibition (HI) and single radial haemolysis (SRH) techniques in detecting antibodies against influenza A/Bangkok/1/79, A/Brazil/11/78, B/Singapore/222/79, B/Hong Kong/5/72 strains, in human sera were compared. For antibodies to influenza B viruses the HI tests employing ether-treated antigens were also evaluated. The SRH technique appears to be more sensitive for detecting protective titres of antibodies against influenza B strains and influenza A/Brazil/11/78.
Thyroid hormone entry into the thymocyte, a thyroid hormone target, was investigated by incubating the cells with tracer amounts of [125I]L-T3. At 37 C T3 uptake was linear with time up to 2 min, and then approached a plateau. The specific T3 uptake, obtained by subtracting the uptake in the presence of excess unlabeled T3, represented 48 +/- 6% of the total at equilibrium. Unlabeled L-T4, D-T3, and triiodothyroacetic acid were less effective than L-T3 in reducing [125I]T3 uptake. Kinetic studies on the initial rate of T3 uptake indicated, for the saturable process, a maximum velocity of approximately 1 pmol/10(6) cells.min and a Km of approximately 0.8 nM. Lowering incubation temperature to 4 C resulted in a two thirds reduction of the total T3 uptake. Washout experiments indicated a different hormone release, being more rapid for cells incubated at 4 C than at 37 C; at 30 min 70% of labeled T3 was released when incubation was carried out at 4 C compared to only 35% after incubation at 37 C, indicating the major intracellular location of the hormone at the latter temperature. An energy requirement of T3 uptake in thymocytes was shown by sensitivity to oligomycin; the effect was dose dependent, showing a maximal decrease in specific uptake of 85%. The involvement of cation movement in the entry process of T3 was indicated by the sensitivity to ouabain. These results indicate the existence of a stereospecific, energy-dependent, saturable process for T3 entry in thymocytes.
Summary.We have investigated the kinetics of the amplification of the progenitor cell compartments (CFC) in haemopoietic organs during murine ontogenesis and compared the growth requirements of fetal and adult CFC. Two haemopoietic phases were recognized in the fetal liver (FL): an exponential growth phase, from 11·5 to 15·5 d post conception (p.c.), during which the mean number of nucleated cells and of CFC in the FL increased from 4·9 × 10 5 to 7·0 × 10 7 and from 4·5 × 10 3 to 2·7 × 10 5 , respectively, and a recessive phase after 15·5 d p.c., during which the CFC number in the FL gradually decreased, although some CFC were still detectable in the liver after birth. In serum-deprived cultures, FL and adult marrow (AM) CFC had similar responses to GM-CSF, and did not respond to G-CSF or IL-3. In contrast, FL, but not AM, erythroid colonies grew Epo-independently whereas SCF alone induced formation of maximal numbers of erythroid bursts from FL, but not from AM cells. The proliferative and differentiative effect of SCF alone on fetal cells was confirmed in serum-deprived cultures of purified early progenitor cells isolated by cell sorting on the basis of multiple parameters from FL and AM light-density cells. In culture of purified FL cells, SCF alone induced a similar amplification of total cells (maximal amplification at day 12: 800-300-fold) and total CFC (11-38-fold of maximal amplification at day 6) to the combination of SCF plus IL-3 (1300-800-fold amplification of total cells and 31-88-fold amplification of CFC). In contrast, SCF alone allowed only survival of purified AM early progenitor cells. Therefore FL early progenitor cells have an intrinsic higher potential than their adult counterpart to respond to SCF, confirming the potent role of this growth factor in the development of the murine haemopoietic system.
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