ABSTRACT
Three different procedures were used to isolate lipopolysaccharides from the Salmonella enteritidis strain 477: phenol-water extraction with ethanol precipitation (LPS 1), phenol-water extraction with methanol precipitation (LPS 2) and FPLC purification (LPS 1/1). Production of interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) was observed in the supernatants of adherent spleen cells of BALB/c mice after the stimulation and cultivation of the cells. The quantity of IL-1 beta and IL-6 depended on the method of LPS isolation. The highest level of IL-1 beta was recorded at LPS 2, and of IL-6 at the stimulation of cells by means of LPS 1.
Subject(s)
Interleukin-1/biosynthesis , Interleukin-6/biosynthesis , Lipopolysaccharides/pharmacology , Spleen/drug effects , Spleen/metabolism , Animals , Cells, Cultured , Female , Lipopolysaccharides/biosynthesis , Mice , Mice, Inbred BALB C , Salmonella enteritidis/metabolism , Spleen/cytologyABSTRACT
A monoclonal antibody giving a dominant reaction with the group-specific polysaccharide of streptococcus group B in an ELISA test has been developed. The purified polysaccharide exhibited a high positivity with reference anti B streptococcal antiserum in the ELISA test. Cross-tests of antibodies with other groups of streptococci provided a minimum cross-reaction only in the case of G streptococci. Monoclonal antibodies were prepared using Streptococcus agalactiae S 589 MT strain isolated from a case of bovine mastitis which does not express Ia, Ib, II, III, IV and V type antigens, nor C, R and X protein antigens.
Subject(s)
Antibodies, Monoclonal , Polysaccharides, Bacterial/immunology , Streptococcus agalactiae/immunology , Animals , Antibody Specificity , Cattle , Cross Reactions , Enzyme-Linked Immunosorbent Assay , Female , Humans , Mastitis, Bovine/microbiology , Mice , Streptococcal Infections/diagnosis , Streptococcal Infections/microbiology , Streptococcal Infections/veterinary , Streptococcus agalactiae/isolation & purification , Streptococcus agalactiae/pathogenicityABSTRACT
The paper describes the isolation, purification and characterization of F107-fimbrial proteins, obtained by thermoelution from Escherichia coli 107/86. Isolation of the pure F107 protein was done by FPLC chromatography, employing Superose 12, Mono Q, and Phenyl-Superose columns. The highest purity of the F107 protein was achieved with Superose 12 HR 10/30. Purity checking by a HPLC system Waters 625 LC (Millipore) proved the absence of protein admixtures in a fraction from Superose 12. Analysis of the molar mass of F107 proteins by SDS PAGE revealed that F107 fimbriae consist of two proteins, one of M = 43 kDa (minor), and other of M = 18.9 kDa (major). Western blot analysis with rabbit polyclonal antiserum confirmed that the 18.9 kDa protein was the major characteristic unit of F107 fimbriae.
