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1.
Recept Channels ; 4(3): 197-203, 1996.
Article in English | MEDLINE | ID: mdl-9014242

ABSTRACT

Using a recombinant yeast strain expressing human beta 2 adrenergic receptors under a galactose-inducible promoter, we established conditions for receptor production in 1-15 liter fermenter culture. Crucial factors contributing to consistent high-level expression included the use of selective glucose-free medium, the maintenance of the pH of the culture at 7.2-7.5 and the presence of an antagonist. The expression strategy and production conditions used with the beta 2 adrenergic receptor were then employed to express the human alpha 2-C2 adrenergic receptor in Saccharomyces cerevisiae. Galactose-induced yeast cells displayed specific, high-affinity [3H]rauwolscine binding and contained a 50-kDa species recognized by an alpha 2-C2 receptor specific antiserum. In fermenter culture, up to 10(5) high-affinity [3H]rauwolscine binding sites per cell (corresponding to 30-60 pmol/mg of protein) were obtained. The high expression level combined with relative ease and low cost of scaling-up make yeast a promising alternative to mammalian cells for the production of adrenergic and other G-protein-coupled receptors for structural studies.


Subject(s)
Receptors, Adrenergic/biosynthesis , Receptors, Adrenergic/genetics , Saccharomyces cerevisiae/genetics , Adrenergic alpha-Antagonists/pharmacology , DNA, Complementary/genetics , Fermentation , Genetic Vectors , Humans , Phentolamine/pharmacology , Protein Engineering , Receptors, Adrenergic, alpha-2/biosynthesis , Receptors, Adrenergic, alpha-2/drug effects , Receptors, Adrenergic, alpha-2/genetics , Receptors, Adrenergic, beta-2/biosynthesis , Receptors, Adrenergic, beta-2/genetics , Saccharomyces cerevisiae/growth & development
2.
Acta Paediatr Scand ; 72(1): 37-40, 1983 Jan.
Article in English | MEDLINE | ID: mdl-6344550

ABSTRACT

Urabe Am 9, a new strain of mumps vaccine, originally developed in Japan, was evaluated in children 14 to 20 months of age in a comparative trial with the Jeryl Lynn strain. Both vaccines performed well. The antibody responses were measured using a neutralization test and a haemolysis-in-gel test. The seroconversion rates at six weeks, as detected with either one or both tests, were 55/58 (94.8%) after the Urabe Am 9 and 58/60 (96.7%) after the Jeryl Lynn vaccine. Only mild infrequent adverse reactions were observed. It is concluded that both strains of live attenuated mumps vaccine are immunogenic and well-tolerated in this age group.


Subject(s)
Mumps Vaccine/immunology , Vaccines, Attenuated/immunology , Antibodies, Viral/analysis , Clinical Trials as Topic , Humans , Infant , Mumps Vaccine/adverse effects , Vaccines, Attenuated/adverse effects
3.
Acta Paediatr Scand ; 72(1): 41-6, 1983 Jan.
Article in English | MEDLINE | ID: mdl-6344551

ABSTRACT

Two mumps-measles vaccine combinations were evaluated for their reactogenicity and immunogenicity in children aged 14 to 20 months. The Urabe Am 9-Schwarz combination vaccine was given to 108 double seronegative children. The seroconversion rate at six weeks after vaccination was 99.1% for measles (haemagglutination-inhibition test) and 92.6% for mumps (neutralization and haemolysis-in-gel tests). The Jeryl Lynn-Moraten vaccine was administered to 85 double seronegative children; the seroconversion rates were 95.3% for measles and 83.5% for mumps. The reported post-vaccination signs and symptoms resembled those seen after monovalent measles vaccine but were more accentuated. Fever over 37.5 C degrees was reported in 66.7% and unusual restlessness and irritability in 68.5% of the Urabe Am 9-Schwarz double seronegative vaccines compared to 55.3% (p less than 0.05) and 54.1% (p less than 0.05), respectively, in the recipients of the Jeryl Lynn-Moraten vaccine. These relatively high reaction rates probably reflect the close observation of the children by their parents during the study. Nevertheless, the tendency towards increased reaction rate and, possibly, reduced immunogenicity of bivalent mumps-measles vaccines as compared to the corresponding single vaccines should be taken into account in the planning of large scale vaccination of young children.


Subject(s)
Measles Vaccine/immunology , Mumps Vaccine/immunology , Vaccines, Attenuated/immunology , Antibodies, Viral/analysis , Clinical Trials as Topic , Humans , Infant , Measles Vaccine/adverse effects , Mumps Vaccine/adverse effects , Vaccines, Attenuated/adverse effects
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