1887

Abstract

High molecular weight (Hmw) proteins 1 and 2, type IV pilin protein (PilA), outer-membrane protein P5 (OmpP5), protein D (Hpd) and adhesive protein (Hap) are surface proteins involved in the adherence of non-typeable . One hundred clinical isolates were evaluated for the presence of the genes encoding these proteins by PCR and for their adherence capacity (AC) to Detroit 562 nasopharyngeal cells (D562). The majority of isolates were from blood (77/100); other sites were also represented. Confluent D562 monolayers (1.2×10 cells per well) were inoculated with standardized minimal infective doses (m.o.i.) of 10, 10 or 10 c.f.u. per well. The AC was categorized as low (<10 %) or high (≥10 %) depending on the percentage of c.f.u. adhering per well. All the isolates evaluated showed adherence: 69/100 (69 %) demonstrated high adherence, while 31/100 (31 %) showed low adherence. Of all the genes evaluated, and/or were detected in 69/100 (69 %) of isolates. The presence of and/or was associated with increased adherence to D562 cells (≤0.001). Dot immunoblots were performed to detect protein expression using mAbs 3D6, AD6 and 10C5. Among the high-adherence isolates ( = 69), 72 % reacted with 3D6 and 21 % with 10C5. Our data indicate that the absence of Hmw1 and/or Hmw2 was associated with decreased adherence to D562 cells.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.058222-0
2013-11-01
2019-12-08
Loading full text...

Full text loading...

/deliver/fulltext/jmm/62/11/1649.html?itemId=/content/journal/jmm/10.1099/jmm.0.058222-0&mimeType=html&fmt=ahah

References

  1. Ahrén I. L., Janson H., Forsgren A., Riesbeck K.. ( 2001;). Protein D expression promotes the adherence and internalization of non-typeable Haemophilus influenzae into human monocytic cells. . Microb Pathog 31:, 151–158. [CrossRef][PubMed]
    [Google Scholar]
  2. Avadhanula V., Rodriguez C. A., Ulett G. C., Bakaletz L. O., Adderson E. E.. ( 2006;). Nontypeable Haemophilus influenzae adheres to intercellular adhesion molecule 1 (ICAM-1) on respiratory epithelial cells and upregulates ICAM-1 expression. . Infect Immun 74:, 830–838. [CrossRef][PubMed]
    [Google Scholar]
  3. Barenkamp S. J., St Geme J. W. III. ( 1996;). Identification of surface-exposed B-cell epitopes on high molecular-weight adhesion proteins of nontypeable Haemophilus influenzae.. Infect Immun 64:, 3032–3037.[PubMed]
    [Google Scholar]
  4. Bookwalter J. E., Jurcisek J. A., Gray-Owen S. D., Fernandez S., McGillivary G., Bakaletz L. O.. ( 2008;). A carcinoembryonic antigen-related cell adhesion molecule 1 homologue plays a pivotal role in nontypeable Haemophilus influenzae colonization of the chinchilla nasopharynx via the outer membrane protein P5-homologous adhesin. . Infect Immun 76:, 48–55. [CrossRef][PubMed]
    [Google Scholar]
  5. Cardines R., Giufrè M., Mastrantonio P., degli Atti M. L. C., Cerquetti M.. ( 2007;). Nontypeable Haemophilus influenzae meningitis in children: phenotypic and genotypic characterization of isolates. . Pediatr Infect Dis J 26:, 577–582. [CrossRef][PubMed]
    [Google Scholar]
  6. Crook J., Tharpe J. A., Johnson S. E., Williams D. B., Stinson A. R., Facklam R. R., Ades E. W., Carlone G. M., Sampson J. S.. ( 1998;). Immunoreactivity of five monoclonal antibodies against the 37-kilodalton common cell wall protein (PsaA) of Streptococcus pneumoniae.. Clin Diagn Lab Immunol 5:, 205–210.[PubMed]
    [Google Scholar]
  7. Davis G. S., Sandstedt S. A., Patel M., Marrs C. F., Gilsdorf J. R.. ( 2011;). Use of bexB to detect the capsule locus in Haemophilus influenzae.. J Clin Microbiol 49:, 2594–2601. [CrossRef][PubMed]
    [Google Scholar]
  8. Dawid S., Barenkamp S. J., St Geme J. W. III. ( 1999;). Variation in expression of the Haemophilus influenzae HMW adhesins: a prokaryotic system reminiscent of eukaryotes. . Proc Natl Acad Sci U S A 96:, 1077–1082. [CrossRef][PubMed]
    [Google Scholar]
  9. Duim B., Bowler L. D., Eijk P. P., Jansen H. M., Dankert J., van Alphen L.. ( 1997;). Molecular variation in the major outer membrane protein P5 gene of nonencapsulated Haemophilus influenzae during chronic infections. . Infect Immun 65:, 1351–1356.[PubMed]
    [Google Scholar]
  10. Ecevit I. Z., McCrea K. W., Pettigrew M. M., Sen A., Marrs C. F., Gilsdorf J. R.. ( 2004;). Prevalence of the hifBC, hmw1A, hmw2A, hmwC, and hia genes in Haemophilus influenzae isolates. . J Clin Microbiol 42:, 3065–3072. [CrossRef][PubMed]
    [Google Scholar]
  11. Giufrè M., Carattoli A., Cardines R., Mastrantonio P., Cerquetti M.. ( 2008;). Variation in expression of HMW1 and HMW2 adhesins in invasive nontypeable Haemophilus influenzae isolates. . BMC Microbiol 8:, 83. [CrossRef][PubMed]
    [Google Scholar]
  12. Hardy G. G., Tudor S. M., St Geme J. W. III. ( 2003;). The pathogenesis of disease due to nontypeable Haemophilus influenzae.. Methods Mol Med 71:, 1–28.[PubMed]
    [Google Scholar]
  13. Hendrixson D. R., St Geme J. W. III. ( 1998;). The Haemophilus influenzae Hap serine protease promotes adherence and microcolony formation, potentiated by a soluble host protein. . Mol Cell 2:, 841–850. [CrossRef][PubMed]
    [Google Scholar]
  14. Janson H., Carlén B., Cervin A., Forsgren A., Magnusdottir A. B., Lindberg S., Runer T.. ( 1999;). Effects on the ciliated epithelium of protein D-producing and -nonproducing nontypeable Haemophilus influenzae in nasopharyngeal tissue cultures. . J Infect Dis 180:, 737–746. [CrossRef][PubMed]
    [Google Scholar]
  15. Johnson R. W., McGillivary G., Denoël P., Poolman J., Bakaletz L. O.. ( 2011;). Abrogation of nontypeable Haemophilus influenzae protein D function reduces phosphorylcholine decoration, adherence to airway epithelial cells, and fitness in a chinchilla model of otitis media. . Vaccine 29:, 1211–1221. [CrossRef][PubMed]
    [Google Scholar]
  16. Jurcisek J. A., Bookwalter J. E., Baker B. D., Fernandez S., Novotny L. A., Munson R. S. Jr, Bakaletz L. O.. ( 2007;). The PilA protein of non-typeable Haemophilus influenzae plays a role in biofilm formation, adherence to epithelial cells and colonization of the mammalian upper respiratory tract. . Mol Microbiol 65:, 1288–1299. [CrossRef][PubMed]
    [Google Scholar]
  17. Kimaro Mlacha S. Z., Peret T. C., Kumar N., Romero-Steiner S., Dunning Hotopp J. C., Ishmael N., Grinblat-Huse V., Riley D. R., Erdman D. D.. & other authors ( 2013;). Transcriptional adaptation of pneumococci and human pharyngeal cells in the presence of a virus infection. . BMC Genomics 14:, 378. [CrossRef][PubMed]
    [Google Scholar]
  18. MacNeil J. R., Cohn A. C., Farley M., Mair R., Baumbach J., Bennett N., Gershman K., Harrison L. H., Lynfield R.. & other authors ( 2011;). Current epidemiology and trends in invasive Haemophilus influenzae disease–United States, 1989-2008. . Clin Infect Dis 53:, 1230–1236. [CrossRef][PubMed]
    [Google Scholar]
  19. Mantel N., Haenszel W.. ( 1959;). Statistical aspects of the analysis of data from retrospective studies of disease. . J Natl Cancer Inst 22:, 719–748.[PubMed]
    [Google Scholar]
  20. Meats E., Feil E. J., Stringer S., Cody A. J., Goldstein R., Kroll J. S., Popovic T., Spratt B. G.. ( 2003;). Characterization of encapsulated and noncapsulated Haemophilus influenzae and determination of phylogenetic relationships by multilocus sequence typing. . J Clin Microbiol 41:, 1623–1636. [CrossRef][PubMed]
    [Google Scholar]
  21. Murphy T. F., Faden H., Bakaletz L. O., Kyd J. M., Forsgren A., Campos J., Virji M., Pelton S. I.. ( 2009;). Nontypeable Haemophilus influenzae as a pathogen in children. . Pediatr Infect Dis J 28:, 43–48. [CrossRef][PubMed]
    [Google Scholar]
  22. Pittman M.. ( 1931;). Variation and type specificity in the bacterial species Haemophilus influenzae.. J Exp Med 53:, 471–492. [CrossRef][PubMed]
    [Google Scholar]
  23. Reddy M. S., Bernstein J. M., Murphy T. F., Faden H. S.. ( 1996;). Binding between outer membrane proteins of nontypeable Haemophilus influenzae and human nasopharyngeal mucin. . Infect Immun 64:, 1477–1479.[PubMed]
    [Google Scholar]
  24. Rodriguez C. A., Avadhanula V., Buscher A., Smith A. L., St Geme J. W. III, Adderson E. E.. ( 2003;). Prevalence and distribution of adhesins in invasive non-type b encapsulated Haemophilus influenzae.. Infect Immun 71:, 1635–1642. [CrossRef][PubMed]
    [Google Scholar]
  25. Romero-Steiner S., Pilishvili T., Sampson J. S., Johnson S. E., Stinson A., Carlone G. M., Ades E. W.. ( 2003;). Inhibition of pneumococcal adherence to human nasopharyngeal epithelial cells by anti-PsaA antibodies. . Clin Diagn Lab Immunol 10:, 246–251.[PubMed]
    [Google Scholar]
  26. Rubach M. P., Bender J. M., Mottice S., Hanson K., Weng H. Y. C., Korgenski K., Daly J. A., Pavia A. T.. ( 2011;). Increasing incidence of invasive Haemophilus influenzae disease in adults, Utah, USA. . Emerg Infect Dis 17:, 1645–1650. [CrossRef][PubMed]
    [Google Scholar]
  27. St Geme J. W. III, Falkow S., Barenkamp S. J.. ( 1993;). High-molecular-weight proteins of nontypable Haemophilus influenzae mediate attachment to human epithelial cells. . Proc Natl Acad Sci U S A 90:, 2875–2879. [CrossRef][PubMed]
    [Google Scholar]
  28. St Geme J. W. III, Kumar V. V., Cutter D., Barenkamp S. J.. ( 1998;). Prevalence and distribution of the hmw and hia genes and the HMW and Hia adhesins among genetically diverse strains of nontypeable Haemophilus influenzae.. Infect Immun 66:, 364–368.[PubMed]
    [Google Scholar]
  29. Tondella M. L., Popovic T., Rosenstein N. E., Lake D. B., Carlone G. M., Mayer L. W., Perkins B. A..the Active Bacterial Core Surveillance Team ( 2000;). Distribution of Neisseria meningitidis serogroup B serosubtypes and serotypes circulating in the United States. . J Clin Microbiol 38:, 3323–3328.[PubMed]
    [Google Scholar]
  30. Wang X., Mair R., Hatcher C., Theodore M. J., Edmond K., Wu H. M., Harcourt B. H., Carvalho M. G., Pimenta F.. & other authors ( 2011;). Detection of bacterial pathogens in Mongolia meningitis surveillance with a new real-time PCR assay to detect Haemophilus influenzae.. Int J Med Microbiol 301:, 303–309. [CrossRef][PubMed]
    [Google Scholar]
  31. Winter L. E., Barenkamp S. J.. ( 2006;). Antibodies specific for the high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae are opsonophagocytic for both homologous and heterologous strains. . Clin Vaccine Immunol 13:, 1333–1342. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.058222-0
Loading
/content/journal/jmm/10.1099/jmm.0.058222-0
Loading

Data & Media loading...

Supplements

Supplementary material 

PDF
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