1887

Journal of Medical Microbiology logo

Volume 55, Issue 3

Monoclonal antibodies binding to the cell surface of serotype 4b Free

Abstract

Serotype 4b strains of the food-borne pathogen are responsible for a large portion of sporadic listeric infections and all major food-borne listeriosis outbreaks in humans. Hybridomas were produced from three fusions with lymphocytes of ND4 mice immunized either with the insoluble antigens of serotype 4b or with formalin-killed bacterial cells and screened for monoclonal antibodies (mAbs) reactive to serotype 4b. A set of 35 mAbs was identified by ELISA as having reactivity with both the insoluble antigen fraction and the whole-cell antigens. Thirteen of these mAbs belonged to immunoglobulin subclass G1 (IgG1), fifteen were IgG2a and seven mAbs were IgM. Only 20 out of the 35 mAbs were capable of detecting protein bands of various sizes ranging from 20 to 88 kDa in Western blots. Two of these mAbs, M2365 and M2367, were capable of binding to cell-surface antigens of live serotype 4b, as demonstrated by immunofluorescence microscopy and immunogold transmission electron microscopy. Immunofluorescence microscopy showed that M2365 and M2367 failed to bind to the cell surfaces of O157 : H7, (serotype Typhimurium DT104) or . Evaluation of the cross-reactions of all 35 mAbs with whole-cell antigens of O157 : H7, . Typhimurium, and by ELISA indicated that the majority of the mAbs, including M2365 and M2367, did not cross-react with O157 : H7, . Typhimurium or and showed no or a very weak reaction with . Furthermore, M2365 and M2367 showed no reaction with whole-cell antigens derived from serotypes 1/2a, 1/2b and 3a, and from , and , in an ELISA. Collectively, these data suggest that M2365 and M2367 have potential use in the development of immunological methods of laboratory diagnosis for serotype 4b in clinical or food samples.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): H+L, heavy and light , IC, insoluble cellular and WC, whole cell
SGM
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.46305-0
2006-03-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/55/3/291.html?itemId=/content/journal/jmm/10.1099/jmm.0.46305-0&mimeType=html&fmt=ahah

References

  1. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A. Struhl K. (editors) 2005 Current Protocols in Molecular Biology New York: Wiley;
     [Google Scholar]
  2. Bhunia A. K., Johnson M. G. 1992; Monoclonal antibody specific for Listeria monocytogenes associated with a 66-kilodalton cell surface antigen. Appl Environ Microbiol 58:1924–1929
     [Google Scholar]
  3. Bhunia A. K., Ball P. H., Fuad A. T., Kurz B. W., Emerson J. W., Johnson M. G. 1991; Development and characterization of a monoclonal antibody specific for Listeria monocytogenes and Listeria innocua . Infect Immun 59:3176–3184
     [Google Scholar]
  4. Donnelly C. W. 2001; Listeria monocytogenes : a continuing challenge. Nutr Rev 59:183–194
     [Google Scholar]
  5. Farber J. M., Speirs J. I. 1987; Potential use of continuous cell lines to distinguish between pathogenic and nonpathogenic Listeria spp. J Clin Microbiol 25:1463–1466
     [Google Scholar]
  6. Kathariou S., Mizumoto C., Allen R. D., Fok A. K., Benedict A. A. 1994; Monoclonal antibodies with a high degree of specificity for Listeria monocytogenes serotype 4b. Appl Environ Microbiol 60:3548–3552
     [Google Scholar]
  7. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
     [Google Scholar]
  8. Lin M., Trottier E., Mallory M. 2005; Enzyme-linked immunosorbent assay based on a chimeric antigen bearing the antigenic regions of structural proteins Erns and E2 for the serodiagnosis of classical swine fever virus infection. Clin Diagn Lab Immunol 12:877–881
     [Google Scholar]
  9. Loiseau O., Cottin J., Robert R., Tronchin G., Mahaza C., Senet J. M. 1995; Development and characterization of monoclonal antibodies specific for the genus Listeria . FEMS Immunol Med Microbiol 11:219–230 [CrossRef]
     [Google Scholar]
  10. Lorber B. 1997; Listeriosis. Clin Infect Dis 24:1–9 [CrossRef]
     [Google Scholar]
  11. McLauchlin J., Taylor A. G. 1989; The use of monoclonal antibodies in the characterization and purification of cell surface antigens of Listeria monocytogenes serogroup 4. Acta Microbiol Hung 36:459–465
     [Google Scholar]
  12. McLauchlin J., Black A., Green H. T., Nash J. Q., Taylor A. G. 1988; Monoclonal antibodies show Listeria monocytogenes in necropsy tissue samples. J Clin Pathol 41:983–988 [CrossRef]
     [Google Scholar]
  13. McLauchlin J., Samuel D., Taylor A. G. 1989; The use of monoclonal antibodies to demonstrate Listeria monocytogenes in post mortem tissue using a direct immunofluorescence technique, and to detect a soluble antigen in CSF supernatants using an ELISA. Acta Microbiol Hung 36:303–308
     [Google Scholar]
  14. Mead P. S., Slutsker L., Dietz V., McCaig L. F., Bresee J. S., Shapiro C., Griffin P. M., Tauxe R. V. 1999; Food-related illness and death in the United States. Emerg Infect Dis 5:607–625 [CrossRef]
     [Google Scholar]
  15. Norton D. M. 2002; Polymerase chain reaction-based methods for detection of Listeria monocytogenes : toward real-time screening for food and environmental samples. J AOAC Int 85:505–515
     [Google Scholar]
  16. Pandiripally V. K., Westbrook D. G., Sunki G. R., Bhunia A. K. 1999; Surface protein p104 is involved in adhesion of Listeria monocytogenes to human intestinal cell line, Caco-2. J Med Microbiol 48:117–124 [CrossRef]
     [Google Scholar]
  17. Ralovich B. 1993; Detection and epidemiological typing of Listeria strains – diagnostic methods for Listeria infections (a review). Acta Microbiol Hung 40:3–38
     [Google Scholar]
  18. Rocourt J., Bille J. 1997; Foodborne listeriosis. World Health Stat Q 50:67–73
     [Google Scholar]
  19. Schlech W. F. III 1993; An animal model of foodborne Listeria monocytogenes virulence: effect of alterations in local and systemic immunity on invasive infection. Clin Invest Med 16:219–225
     [Google Scholar]
  20. Schlech W. F. III 2000; Foodborne listeriosis. Clin Infect Dis 31:770–775 [CrossRef]
     [Google Scholar]
  21. Shulman M., Wilde C. D., Köhler G. 1978; A better cell line for making hybridomas secreting specific antibodies. Nature 276:269–270 [CrossRef]
     [Google Scholar]
  22. Siragusa G. R., Johnson M. G. 1990; Monoclonal antibody specific for Listeria monocytogenes , Listeria innocua , and Listeria welshimeri . Appl Environ Microbiol 56:1897–1904
     [Google Scholar]
  23. Torensma R., Visser M. J., Aarsman C. J., Poppelier M. J., Fluit A. C., Verhoef J. 1993; Monoclonal antibodies that react with live Listeria spp. Appl Environ Microbiol 59:2713–2716
     [Google Scholar]
  24. Yu W. 2004; The gene products of Listeria monocytogenes induced specifically during rabbit infection . MSc thesis University of Ottawa; Ottawa, Canada:
  25. Ziegler H. K., Orlin C. A. 1984; Analysis of Listeria monocytogenes antigens with monoclonal antibodies. Clin Invest Med 7:239–242
     [Google Scholar]
  26. Zola H. 1987 Monoclonal Antibodies: a Manual of Techniques Boca Raton, FL: CRC Press;
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.46305-0
Loading
Monoclonal antibodies binding to the cell surface of Listeria monocytogenes serotype 4b
J. Med. Microbiol. 55, 291 (2006); https://doi.org/10.1099/jmm.0.46305-0
/content/journal/jmm/10.1099/jmm.0.46305-0
/content/journal/jmm/10.1099/jmm.0.46305-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error