ABSTRACT
The effect of botulinic toxin of type A (BoNT) on urinary bladder (UB) function was studied during operation in situ in healthy rats using the method of infusion cystomanometry after a single injection of the toxin into the UB wall 3 to 30 days after the injection. Contractility of the muscular fragments in vitro was assessed in the isometric regimen in electric stimulation, depolarization of cellular membranes by ion [K+] excess, addition of noradrenalin (10(-5) M). In response to botulinic toxin injection 47-64% rats vs control animals (23%) operated on without toxin had no phasic UB contractions, detrusor pressure (Pdet) after the infusion was higher which indicated retension. In rats with intact UB contractility, contractions were stronger (Pdet 24-34 cm H2O), but rare (3-5/min) vs controls (19 cm H2O, 5/min, respectively). Contractility in vitro in response to electric stimulation regressed by 60-85% after toxin infusion. Contractility in solution [K+] regressed less (by 65, 36, 16% in 3, 10 and 30 days, respectively). No significant changes of noradrenalin-induced reactions were found. The results illustrate possible direct myogenic effect of the toxin on detrusor muscle.
Subject(s)
Botulinum Toxins, Type A/pharmacology , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Urinary Bladder/drug effects , Urodynamics/drug effects , Animals , Botulinum Toxins, Type A/administration & dosage , Female , RatsABSTRACT
A human interleukin-2 (IL-2) gene was isolated from genomic DNA library. The isolated gene with 5'- and 3'-flanking sequences of various lengths was inserted into plasmids derived from the retroviral vector pPSneo. The recombinant plasmids were transfected into myeloma X63Ag8-653 cells. The transfected cells, harbouring the IL-2 gene with the shortened (to position -165) or totally deleted 5'-flanking sequence, constitutively expressed biologically active IL-2. Deletion of 3'-flanking region on did not affect the IL-2 expression.
Subject(s)
Chromosomes, Human , Gene Expression , Interleukin-2/genetics , Animals , Base Sequence , Cell Transformation, Neoplastic , Genomic Library , Humans , Mice , Molecular Sequence Data , Multiple Myeloma/genetics , Plasmids , Recombination, Genetic , Restriction Mapping , TransfectionABSTRACT
Nine monoclonals against human Ig lambda chains were produced, 4 antibodies react with C-domain, 5--with V-domain of the lambda chain. Anti-C lambda domain antibodies recognize not less than 3 epitopes and one of them is expressed only on the isolated chain. Anti-V lambda antibodies bind both isolated lambda chain and intact IgG, IgM, IgA. Four epitopes are expressed by few lambda Bence Jones proteins of the III subgroup, the immunogen possessing the same isotype. The 4 mentioned epitopes represent private idiotypic determinants. The epitope 3E10 is characteristic of 50% Bence Jones proteins of the II and III V lambda-subgroups thus representing a common idiotypic determinant. Using anti-V lambda antibodies germ line variability of V lambda III proteins was analysed and the similarity of antigenic structure of normal and myeloma human Ig lambda chains was demonstrated.
Subject(s)
Antibodies, Monoclonal , Epitopes/analysis , Immunoglobulin lambda-Chains/immunology , Antibody Specificity , Bence Jones Protein/immunology , Humans , Immunoenzyme Techniques , Immunoglobulin Constant Regions/immunology , Immunoglobulin G/immunology , Immunoglobulin Variable RegionABSTRACT
The review is devoted to the mechanisms of generation and coding of human and murine autoantibodies.
Subject(s)
Autoantibodies/genetics , Genes, Immunoglobulin , Animals , Autoantibodies/biosynthesis , Humans , Infections/immunology , Mice , Mice, Inbred BALB C , MutationABSTRACT
We have analysed reassociated Ig molecules, containing heavy (H) or light (L) chains of Ig-B2 monoclonal antibody with human fibronectin binding activity and L of H chains of normal mouse serum immunoglobulin (Ig-NM). Examination of Ig-B2 idiotype expression in reassociated Ig indicated that 0.4% L-NM and 0.8% H-NM were able to restore Ig-B2 idiotype. The analysis of antigen binding capacity of reassociated Ig demonstrated, that only 4% H-NM created antigen binding site in complex with L-B2. We have determined the leading role of L-chain in creation of idiotype and binding site of Ig-B2. Selectivity of interaction between H and L chains is discussed. The results indicate, that not more than 4-6% of random H--L combinations produce functional Ig.
Subject(s)
Binding Sites, Antibody , Immunoglobulin G/immunology , Immunoglobulin Idiotypes/immunology , Peptides/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Fibronectins/immunology , Humans , Hybridomas , Immunoglobulin Heavy Chains/immunology , Immunoglobulin Heavy Chains/isolation & purification , Immunoglobulin Light Chains/immunology , Immunoglobulin Light Chains/isolation & purification , MiceABSTRACT
The investigation of 750 B-lymphocyte hybridoma clones obtained by fusion of mouse myeloma and newborn heterozygous Igk-la/Igk-1b rat splenocytes has revealed that 9,8% of Ig kappa-chain genes are rearranged productively. Seventeen hybridomas secrete kappa-chains of both allelic variants. The analysis of IgM molecules of nine such clones demonstrated that in six cases only one L-chain allotype is present in IgM. Thus for the first time the high frequency of selective association of H and L chains was shown. Evidently this selectively may function as one of the allelic exclusion mechanisms at the Ig assembly stage.
Subject(s)
Alleles , Immunoglobulin Allotypes/genetics , Immunoglobulin Light Chains/genetics , Animals , Fluorescent Antibody Technique , Hybridomas/immunology , Immunoglobulin Heavy Chains/analysis , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Light Chains/analysis , Peptides/analysis , Radioimmunoassay , RatsABSTRACT
Expression of RI-1a and RI-1b allelic genes controlling the production of rat lg kappa L-chains by hybridoma cells in vitro was studied. By fusing mouse myeloma cells with RI-1a/RI-1b heterozygous rat splenocytes the unique cloned hybrid cell line secreting both allelic variants has been established. This line may have appeared because cell hybridization made it possible to fix the rare case of correct rearrangement of the kappa chain gene segments on both homologous chromosomes.
