Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1988 Dec;56(12):3015–3020. doi: 10.1128/iai.56.12.3015-3020.1988

Evidence for occurrence of passively adsorbed I antigen activity on a cultured strain of Mycoplasma pneumoniae.

K Uemura 1, L M Loomes 1, R A Childs 1, T Feizi 1
PMCID: PMC259693  PMID: 3141277

Abstract

The aim of this study was to investigate whether I antigen occurs in association with Mycoplasma pneumoniae in a form that may be immunogenic during natural infection or experimental immunization. I antigen activity was detected by radioimmunoassay in suspensions of M. pneumoniae MY11965 and in the soluble phase of mycoplasma lysates prepared with Triton X-100. There was evidence for the occurrence of I antigen in at least two macromolecular forms. The first form partitioned in the lipid phase following chloroform-methanol extraction and chromatographed on thin-layer chromatograms as a ceramide decasaccharide. The second form was associated with the residue after lipid extraction and was solubilized by treatment with sodium dodecyl sulfate or pepsin; this component was tentatively designated a glycoprotein or polysaccharide and was not investigated further. In a lipid extract from mycoplasmas that had been surface labeled by the galactose oxidase-NaB3H4 method, two 3H-labeled glycolipids were detected as minor components which chromatographed on thin-layer chromatograms in the region of an authentic I-active ceramide decasaccharide. However, no significant radioactivity was incorporated into glycolipids after metabolic labeling with [3H]glucosamine. These observations suggested that the mycoplasmas contained surface-associated glycolipids with I antigen activity that were of exogenous origin. This was supported by the observations that horse, rabbit, and fetal calf sera contained I antigen activity and that the I antigen activity in M. pneumoniae cultures reflected the levels found in the sera included in the culture media. From rabbit serum, which expressed the highest antigen activity, an I-active glycolipid was isolated that chromatographed as a ceramide decasaccharide. I-active substances passively adsorbed onto M. pneumoniae are potentially immunogenic. However, we consider these unlikely to be the main stimulus for autoantibody production in natural infection, since the autoantibodies elicited are restricted to the I carbohydrate antigen and there is a lack of antibodies to other glycolipids that may be adsorbed from serous and cellular components of the host tissues. In our view, the more likely stimulus is the specific complex formed between the mycoplasma and the sialo-oligosaccharide receptors of the Ii antigen type, as suggested previously.

Full text

PDF
3015

Images in this article

3018Image
on p.3018

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Costea N., Yakulis V. J., Heller P. Inhibition of cold agglutinins (anti-I) by M. pneumoniae antigens. Proc Soc Exp Biol Med. 1972 Feb;139(2):476–479. doi: 10.3181/00379727-139-36167. [DOI] [PubMed] [Google Scholar]
  3. Costea N., Yakulis V. J., Heller P. The mechanism of indction of cold agglutinins by mycoplasma pneumoniae. J Immunol. 1971 Mar;106(3):598–604. [PubMed] [Google Scholar]
  4. Egge H., Kordowicz M., Peter-Katalinić J., Hanfland P. Immunochemistry of I/i-active oligo- and polyglycosylceramides from rabbit erythrocyte membranes. Characterization of linear, di-, and triantennary neolactoglycosphingolipids. J Biol Chem. 1985 Apr 25;260(8):4927–4935. [PubMed] [Google Scholar]
  5. Feizi T. Significance of carbohydrate components of cell surfaces. Ciba Found Symp. 1987;129:43–58. doi: 10.1002/9780470513484.ch4. [DOI] [PubMed] [Google Scholar]
  6. Feizi T., Taylor-Robinson D. Cold agglutinin anti-I and Mycoplasma pneumoniae. Immunology. 1967 Oct;13(4):405–409. [PMC free article] [PubMed] [Google Scholar]
  7. Feizi T., Taylor-Robinson D., Shields M. D., Carter R. A. Production of cold agglutinins in rabbits immunized with human erythrocytes treated with Mycoplasma pneumoniae. Nature. 1969 Jun 28;222(5200):1253–1256. doi: 10.1038/2221253a0. [DOI] [PubMed] [Google Scholar]
  8. Feizi T. The blood group Ii system: a carbohydrate antigen system defined by naturally monoclonal or oligoclonal autoantibodies of man. Immunol Commun. 1981;10(2):127–156. doi: 10.3109/08820138109050693. [DOI] [PubMed] [Google Scholar]
  9. Gahmberg C. G., Hakomori S. I. External labeling of cell surface galactose and galactosamine in glycolipid and glycoprotein of human erythrocytes. J Biol Chem. 1973 Jun 25;248(12):4311–4317. [PubMed] [Google Scholar]
  10. Gooi H. C., Veyrières A., Alais J., Scudder P., Hounsell E. F., Feizi T. Further studies of the specificities of monoclonal anti-i and anti-I antibodies using chemically synthesized, linear oligosaccharides of the poly-N-acetyllactosamine series. Mol Immunol. 1984 Nov;21(11):1099–1104. doi: 10.1016/0161-5890(84)90120-2. [DOI] [PubMed] [Google Scholar]
  11. Hanfland P., Egge H., Dabrowski U., Kuhn S., Roelcke D., Dabrowski J. Isolation and characterization of an I-active ceramide decasaccharide from rabbit erythrocyte membranes. Biochemistry. 1981 Sep 1;20(18):5310–5319. doi: 10.1021/bi00521a034. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lind K. Production of cold agglutinins in rabbits induced by Mycoplasma pneumoniae, Listeria monocytogenes or Streptococcus MG. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Aug;81(4):487–496. doi: 10.1111/j.1699-0463.1973.tb02234.x. [DOI] [PubMed] [Google Scholar]
  14. Loomes L. M., Uemura K., Childs R. A., Paulson J. C., Rogers G. N., Scudder P. R., Michalski J. C., Hounsell E. F., Taylor-Robinson D., Feizi T. Erythrocyte receptors for Mycoplasma pneumoniae are sialylated oligosaccharides of Ii antigen type. Nature. 1984 Feb 9;307(5951):560–563. doi: 10.1038/307560a0. [DOI] [PubMed] [Google Scholar]
  15. Loomes L. M., Uemura K., Feizi T. Interaction of Mycoplasma pneumoniae with erythrocyte glycolipids of I and i antigen types. Infect Immun. 1985 Jan;47(1):15–20. doi: 10.1128/iai.47.1.15-20.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Magnani J. L., Brockhaus M., Smith D. F., Ginsburg V., Blaszczyk M., Mitchell K. F., Steplewski Z., Koprowski H. A monosialoganglioside is a monoclonal antibody-defined antigen of colon carcinoma. Science. 1981 Apr 3;212(4490):55–56. doi: 10.1126/science.7209516. [DOI] [PubMed] [Google Scholar]
  17. Manchee R. J., Taylor-Robinson D. Haemadsorption and haemagglutination by mycoplasmas. J Gen Microbiol. 1968 Mar;50(3):465–478. doi: 10.1099/00221287-50-3-465. [DOI] [PubMed] [Google Scholar]
  18. Plackett P., Marmion B. P., Shaw E. J., Lemcke R. M. Immunochemical analysis of Mycoplasma pneumoniae. 3. Separation and chemical identification of serologically active lipids. Aust J Exp Biol Med Sci. 1969 Apr;47(2):171–195. doi: 10.1038/icb.1969.19. [DOI] [PubMed] [Google Scholar]
  19. Razin S., Kutner S., Efrati H., Rottem S. Phospholipid and cholesterol uptake by Mycoplasma cells and membranes. Biochim Biophys Acta. 1980 Jun 6;598(3):628–640. doi: 10.1016/0005-2736(80)90042-5. [DOI] [PubMed] [Google Scholar]
  20. Razin S., Tully J. G. Cholesterol requirement of mycoplasmas. J Bacteriol. 1970 May;102(2):306–310. doi: 10.1128/jb.102.2.306-310.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saito T., Hakomori S. I. Quantitative isolation of total glycosphingolipids from animal cells. J Lipid Res. 1971 Mar;12(2):257–259. [PubMed] [Google Scholar]
  22. Taylor-Robinson D., Purcell R. H., Wong D. C., Chanock R. M. A colour test for the measurement of antibody to certain mycoplasma species based upon the inhibition of acid production. J Hyg (Lond) 1966 Mar;64(1):91–104. doi: 10.1017/s0022172400040377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Uemura K., Childs R. A., Hanfland P., Feizi T. A multiplicity of erythrocyte glycolipids of the neolacto series revealed by immuno-thin-layer chromatography with monoclonal anti-I and anti-i antibodies. Biosci Rep. 1983 Jun;3(6):577–588. doi: 10.1007/BF01120703. [DOI] [PubMed] [Google Scholar]
  24. Watkins W. M. Biochemistry and Genetics of the ABO, Lewis, and P blood group systems. Adv Hum Genet. 1980;10:1-136, 379-85. doi: 10.1007/978-1-4615-8288-5_1. [DOI] [PubMed] [Google Scholar]
  25. Wood E., Lecomte J., Childs R. A., Feizi T. A radioimmunoassay for the measurement of blood group Ii activities: its application to glycoconjugates, oligosaccharides and intact cells. Mol Immunol. 1979 Oct;16(10):813–819. doi: 10.1016/0161-5890(79)90160-3. [DOI] [PubMed] [Google Scholar]
  26. Wuthier R. E. Purification of lipids from nonlipid contaminants on Sephadex bead columns. J Lipid Res. 1966 Jul;7(4):558–561. [PubMed] [Google Scholar]
  27. Yu R. K., Ledeen R. W. Gangliosides of human, bovine, and rabbit plasma. J Lipid Res. 1972 Sep;13(5):680–686. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES