Influence of heat inactivation of human serum on the opsonization of Streptococcus mutans

Michelle A. Moore, Zaid W. Hakki, Richard Gregory, Linda E. Gfell, Wan K. Kim-Park, Michael Kowolik

Research output: Contribution to journalArticle

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Abstract

Phagocytosis of bacteria, such as Streptococcus mutans, is important to host defense. One mechanism by which phagocytosis can be enhanced is by antibody or complement-mediated opsonization of bacteria. Many studies utilize opsonization of bacteria to enhance a cellular response, but little information has been found examining methodology or validity of the opsonization process following the denaturization of the serum. Human serum was inactivated by heat in order to disrupt the classical and alternative pathways of the complement cascade. S. mutans isolated from human subjects were opsonized with heat-inactivated human serum before exposing them to viable neutrophils in vitro. Luminol-dependent chemiluminescence (CL) was used to measure neutrophil activation. Human serum used to opsonize the bacteria was denatured by incubation at 57°C for intervals of 30 and 60 min to inactivate complement. The results from the opsonization data indicated that there was significantly increased CL with 60-min inactivation of the serum (34% increase in mean integration mV.min; p ≤ 0.05) over the nonopsonized control. This indicated a successful opsonization of the bacteria. In addition, the data demonstrate that the inactivation of serum requires a minimum of 60 min at 57°C to disrupt the complement cascade, while 30- and 15-min inactivations produced no significant increase in CL activity over the control. Standard sandwich ELISA assays, detecting complement binding to S. mutans, confirmed successful heat inactivation of serum showing a significant decrease (p ≤ 0.001) in complement binding to S. mutans after 30 min, but could not explain the increased CL response after 60-min heat deactivation of the serum.

Original languageEnglish
Pages (from-to)383-393
Number of pages11
JournalAnnals of the New York Academy of Sciences
Volume832
DOIs
StatePublished - Dec 15 1997

Fingerprint

Streptococcus mutans
Chemiluminescence
Bacteria
Hot Temperature
Luminescence
Serum
Luminol
Phagocytosis
Classical Complement Pathway
Assays
Alternative Complement Pathway
Neutrophil Activation
Chemical activation
Heat
Antibodies
Neutrophils
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Influence of heat inactivation of human serum on the opsonization of Streptococcus mutans. / Moore, Michelle A.; Hakki, Zaid W.; Gregory, Richard; Gfell, Linda E.; Kim-Park, Wan K.; Kowolik, Michael.

In: Annals of the New York Academy of Sciences, Vol. 832, 15.12.1997, p. 383-393.

Research output: Contribution to journalArticle

Moore, Michelle A. ; Hakki, Zaid W. ; Gregory, Richard ; Gfell, Linda E. ; Kim-Park, Wan K. ; Kowolik, Michael. / Influence of heat inactivation of human serum on the opsonization of Streptococcus mutans. In: Annals of the New York Academy of Sciences. 1997 ; Vol. 832. pp. 383-393.
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