Abstract
IgG subclass antibodies to Pseudomonas aemginosa outer membrane proteins (OMP) were investigated in serum from cystic fibrosis (CF) patients by immunoblotting and enzyme-linked immunosorbent assay (ELISA). Fifteen patients (eight in good and seven in poor clinical condition) have been followed for an average of 13 years with multiple serum samples covering the preinfection, and early and late stages of chronic infection. Laser-scanning densitometry of photographs taken from immunoblots was used to quantify antibody level and compare with ELISA titres. The earliest anti-OMP antibodies to appear were of the IgG1 subclass. There was no significant difference in IgG subclass antibody levels to OMPs between patients in relatively good and poor clinical condition. Data presented indicate a high positive correlation among measurements of IgG subclass antibodies to P. aeruginosa OMPs using ELISA and immunoblotting.
Similar content being viewed by others
References
Albus A, Saalmann M, Tesch H, Pedersen SS, Döring G (1989) Increased levels of IgG subclasses specific for Pseudomonas aeruginosa exoenzyme and polysaccharide antigens in chronically infected patients with cystic fibrosis. APMIS 97:1146–1148
Alexander MD, Andrews JA, Leslie RGQ, Wood NJ (1978) The binding of human and guinea-pig IgG sublcasses to homologous macrophage and monocyte Fc receptors. Immunology 35:115–123
Aronoff SC, Stern RC (1988) Serum IgG antibody to outer membrane antigens of Pseudomonas cepacia and Pseudomonas aeruginosa in cystic fibrosis. J Infect Dis 157:934–940
Fernandes PB, Kim C, Cundy KR, Huang NN (1988) Antibodies to cell envelope proteins of Pseudomonas aeruginosa in cystic fibrosis patients. Infect Immun 33:527–532
Ferrante A, Beard LJ, Feldman RG (1990) IgG subclass distribution of antibodies to bacterial and viral antigens. Pediatr Infect Dis J 9 (Supplement 8): 16–24
Fick RB, Olchowski J, Squier SU, Merrill WW, Reynolds HY (1986) Immunoglobulin-G subclasses in cystic fibrosis; IgG2 response to Pseudomonas aeruginosa lipopolysaccharide. Am Rev Respir Dis 133:418–422
Filip C, Fletcher G, Wulff JL, Earhart CF (1973) Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium lauryl sarcosinate. J Bacteriol 115:717–722
Fomsgaard A, Dinesen B, Shand GH, Pressler T, Høiby N (1989) Antilipopolysaccharide antibodies and differential diagnosis of chronic Pseudomonas aeruginosa lung infection in cystic fibrosis. J Clin Microbiol 27:1222–1229
Hammarström L, Smith ECI (1986) IgG subclasses in bacterial infections. Monogr Allergy 19:122–133
Hancock REW, Carey AM (1979) Outer membrane of P. aeruginosa: Heat- and 2-mercaptoethanol-modifiable proteins. J Bact 140:902–910
Hancock REW, Mouat ECA, Speert DP (1984) Quantitation and identification of antibodies to outer-membrane proteins of Pseudomonas aeruginosa in sera of patients with cystic fibrosis. J Infect Dis 149:220–226
Høiby N (1974) Pseudomonas aeruginosa infection in cystic fibrosis. Relationship between mucoid strains of P. aeruginosa and the humoral immune response. Acta Pathol Microbiol Immunol Scand [B] 82:551–558
Høiby N (1977) Pseudomonas aeruginosa infection in cystic fibrosis. Diagnostic and prognostic significance of Pseudomonas aeruginosa precipitins determination by means of crossed immunoelectrophoresis. Acta Pathol Microbiol Immunol Scand [C] [Suppl] 262:1–96
Kubesch P, von Specht BU, Tümmler B (1988) Immune response in cystic fibrosis to outer membrane proteins of Pseudomonas aeruginosa. Zentralbl Bacteriol Microbiol Hyg 269:395–410
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the folin phenol reagent. J Biol Chem 193:265–275
Ojeniyi B, Høiby N, Rosdahl VT (1991) Prevalence and persistence of polyagglutinable Pseudomonas aeruginosa in isolates from cystic fibrosis patients. APMIS 99:187–195
Perelmuter L (1983) IgG4 and the immune system. Clin Rev Allergy 1:267–287
Pressler T, Mansa B, Jensen T, Pedersen SS, Høiby N, Koch C (1988) Increased IgG2 and IgG3 concentration is associated with advanced Pseudomonas aeruginosa infection and poor pulmonary function in cystic fibrosis. Acta Paediatr Scand 77:576–582
Pressler T, Pedersen SS, Espersen F, Høiby N, Koch C (1990) IgG subclass antibodies to Pseudomonas aeruginosa in sera from patients with chronic P. aeruginosa infection investigated by ELISA. Clin Exp Immunol 81:428–434
Schumaker VN, Calcott M, Siegelberg HL, Muller-Eberhard HJ (1976) Ultracentrifuge studies of the binding of IgG of different subclasses to the C1q subunit of the first component of complement. Biochemistry 15:5175–5181
Shand GH, Anvar H, Kadurugamuwa J, Brown MRW, Siverman SH, Melling J (1985) In vivo evidence that bacteria in urinary tract infection grow under iron-restricted conditions. Infect Immun 48:35–39
Shand GH, Pedersen SS, Tilling R, Brown MRW, Høiby N (1988) Use of immunoblot detection of serum antibodies in the diagnosis of chronic Pseudomonas aeruginosa lung infection in cystic fibrosis. J Med Microbiol 27:169–177
Shand GH, Pedersen SS Brown MRW, Høiby N (1991) Serum antibodies to Pseudomonas aeruginosa outher membrane proteins and iron-regulated membrane proteins at different stages of chronic cystic fibrosis lung infection. J Med Microbiol 34:203–212
Shwachman H, Kulczycki LL (1958) Long-term study of one hundred and five patients with cystic fibrosis; studies made over a five- to-fourteen year period. Am J Dis Child 96:9–15
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Wood RE, Boat TF, Doershuk CF (1976) Cystic fibrosis. Am Rev Respir Dis 113:833–878
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pressler, T., Kronborg, G., Shand, G.H. et al. Determination of IgG subclass antibodies to Pseudomonas aeruginosa outer membrane proteins in cystic fibrosis lung infection using immunoblotting and enzyme-linked immunosorbent assay. Med Microbiol Immunol 181, 339–349 (1992). https://doi.org/10.1007/BF00191546
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00191546