Studies of the mechanism of bacterial resistance to complement-mediated killing. V. IgG and F(ab')2 mediate killing of E. coli 0111B4 by the alternative complement pathway without increasing C5b-9 deposition

Keith A Joiner, R. C. Goldman, C. H. Hammer, L. Leive, M. M. Frank

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Abstract

The mechanism of antibody-dependent complement-(c) mediated killing of Escherichia coli 0111B4, strain 12015 (12015), was examined. 12015 was resistant to serum killing when incubated in hypogammaglobulinemic serum (HγS) or pooled normal human serum (NHS) that had been previously adsorbed to remove specific antibody (Abs NHS). Presensitization with immune rabbit serum or purified immune rabbit IgG resulted in 1 to 3 log killing when 5 x 108 colony forming units (CFU)/ml were incubated in 10 to 40% Abs NHS. Binding of 125I-C3 and the nonpresensitized strain was quantitated when these organisms were incubated in 10, 20, and 40% Abs NHS. Stable binding of up to 3.0 x 106 molecules of C3 and 8.0 x 104 molecules of C9 to presensitized and nonpresensitized isolates occurred in the highest concentration of serum, but there was no killing without presensitizaton. Similar results were found when Abs NHS was chelated with ethylene bis glycoltetraacetic acid containing 2 mM MgCl2 (Mg EGTA) to block classical pathway activation, indicating that antibody mediated the bactericidal reaction through the alternative pathway. Deposition of C3 and C9 and killing of 12015 in 10% Abs NHS or 10% HγS was measured after presensitization with increasing amounts of IgG, F(ab')2, or Fab'. There was a dose-dependent increase in C3 deposition and killing, but only minimal change in C9 binding when 1.0 x 103 to 3.2 x 104 IgG or F(ab')2/CFU were bound to the bacterial surface. In contrast, there was no increase in C3 or C9 binding and no bacterial killing when 1 x 103 to 3.4 x 104 molecules Fab'/CFU were bound to the bacterial surface. These experiments show that immune IgG and F(ab')2 can mediate killing of E. Coli 0111B4 by the alternative pathway without changing the extent of terminal C component attachment to the bacterial surface.

Original languageEnglish (US)
Pages (from-to)2563-2569
Number of pages7
JournalJournal of Immunology
Volume131
Issue number5
StatePublished - 1983
Externally publishedYes

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Alternative Complement Pathway
Complement Membrane Attack Complex
Immunoglobulin G
Escherichia coli
Serum
Stem Cells
Antibodies
Rabbits
Magnesium Chloride
Egtazic Acid
Immune Sera

ASJC Scopus subject areas

  • Immunology

Cite this

@article{6221811faeea4def8df1278f6e05ed75,
title = "Studies of the mechanism of bacterial resistance to complement-mediated killing. V. IgG and F(ab')2 mediate killing of E. coli 0111B4 by the alternative complement pathway without increasing C5b-9 deposition",
abstract = "The mechanism of antibody-dependent complement-(c) mediated killing of Escherichia coli 0111B4, strain 12015 (12015), was examined. 12015 was resistant to serum killing when incubated in hypogammaglobulinemic serum (HγS) or pooled normal human serum (NHS) that had been previously adsorbed to remove specific antibody (Abs NHS). Presensitization with immune rabbit serum or purified immune rabbit IgG resulted in 1 to 3 log killing when 5 x 108 colony forming units (CFU)/ml were incubated in 10 to 40{\%} Abs NHS. Binding of 125I-C3 and the nonpresensitized strain was quantitated when these organisms were incubated in 10, 20, and 40{\%} Abs NHS. Stable binding of up to 3.0 x 106 molecules of C3 and 8.0 x 104 molecules of C9 to presensitized and nonpresensitized isolates occurred in the highest concentration of serum, but there was no killing without presensitizaton. Similar results were found when Abs NHS was chelated with ethylene bis glycoltetraacetic acid containing 2 mM MgCl2 (Mg EGTA) to block classical pathway activation, indicating that antibody mediated the bactericidal reaction through the alternative pathway. Deposition of C3 and C9 and killing of 12015 in 10{\%} Abs NHS or 10{\%} HγS was measured after presensitization with increasing amounts of IgG, F(ab')2, or Fab'. There was a dose-dependent increase in C3 deposition and killing, but only minimal change in C9 binding when 1.0 x 103 to 3.2 x 104 IgG or F(ab')2/CFU were bound to the bacterial surface. In contrast, there was no increase in C3 or C9 binding and no bacterial killing when 1 x 103 to 3.4 x 104 molecules Fab'/CFU were bound to the bacterial surface. These experiments show that immune IgG and F(ab')2 can mediate killing of E. Coli 0111B4 by the alternative pathway without changing the extent of terminal C component attachment to the bacterial surface.",
author = "Joiner, {Keith A} and Goldman, {R. C.} and Hammer, {C. H.} and L. Leive and Frank, {M. M.}",
year = "1983",
language = "English (US)",
volume = "131",
pages = "2563--2569",
journal = "Journal of Immunology",
issn = "0022-1767",
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T1 - Studies of the mechanism of bacterial resistance to complement-mediated killing. V. IgG and F(ab')2 mediate killing of E. coli 0111B4 by the alternative complement pathway without increasing C5b-9 deposition

AU - Joiner, Keith A

AU - Goldman, R. C.

AU - Hammer, C. H.

AU - Leive, L.

AU - Frank, M. M.

PY - 1983

Y1 - 1983

N2 - The mechanism of antibody-dependent complement-(c) mediated killing of Escherichia coli 0111B4, strain 12015 (12015), was examined. 12015 was resistant to serum killing when incubated in hypogammaglobulinemic serum (HγS) or pooled normal human serum (NHS) that had been previously adsorbed to remove specific antibody (Abs NHS). Presensitization with immune rabbit serum or purified immune rabbit IgG resulted in 1 to 3 log killing when 5 x 108 colony forming units (CFU)/ml were incubated in 10 to 40% Abs NHS. Binding of 125I-C3 and the nonpresensitized strain was quantitated when these organisms were incubated in 10, 20, and 40% Abs NHS. Stable binding of up to 3.0 x 106 molecules of C3 and 8.0 x 104 molecules of C9 to presensitized and nonpresensitized isolates occurred in the highest concentration of serum, but there was no killing without presensitizaton. Similar results were found when Abs NHS was chelated with ethylene bis glycoltetraacetic acid containing 2 mM MgCl2 (Mg EGTA) to block classical pathway activation, indicating that antibody mediated the bactericidal reaction through the alternative pathway. Deposition of C3 and C9 and killing of 12015 in 10% Abs NHS or 10% HγS was measured after presensitization with increasing amounts of IgG, F(ab')2, or Fab'. There was a dose-dependent increase in C3 deposition and killing, but only minimal change in C9 binding when 1.0 x 103 to 3.2 x 104 IgG or F(ab')2/CFU were bound to the bacterial surface. In contrast, there was no increase in C3 or C9 binding and no bacterial killing when 1 x 103 to 3.4 x 104 molecules Fab'/CFU were bound to the bacterial surface. These experiments show that immune IgG and F(ab')2 can mediate killing of E. Coli 0111B4 by the alternative pathway without changing the extent of terminal C component attachment to the bacterial surface.

AB - The mechanism of antibody-dependent complement-(c) mediated killing of Escherichia coli 0111B4, strain 12015 (12015), was examined. 12015 was resistant to serum killing when incubated in hypogammaglobulinemic serum (HγS) or pooled normal human serum (NHS) that had been previously adsorbed to remove specific antibody (Abs NHS). Presensitization with immune rabbit serum or purified immune rabbit IgG resulted in 1 to 3 log killing when 5 x 108 colony forming units (CFU)/ml were incubated in 10 to 40% Abs NHS. Binding of 125I-C3 and the nonpresensitized strain was quantitated when these organisms were incubated in 10, 20, and 40% Abs NHS. Stable binding of up to 3.0 x 106 molecules of C3 and 8.0 x 104 molecules of C9 to presensitized and nonpresensitized isolates occurred in the highest concentration of serum, but there was no killing without presensitizaton. Similar results were found when Abs NHS was chelated with ethylene bis glycoltetraacetic acid containing 2 mM MgCl2 (Mg EGTA) to block classical pathway activation, indicating that antibody mediated the bactericidal reaction through the alternative pathway. Deposition of C3 and C9 and killing of 12015 in 10% Abs NHS or 10% HγS was measured after presensitization with increasing amounts of IgG, F(ab')2, or Fab'. There was a dose-dependent increase in C3 deposition and killing, but only minimal change in C9 binding when 1.0 x 103 to 3.2 x 104 IgG or F(ab')2/CFU were bound to the bacterial surface. In contrast, there was no increase in C3 or C9 binding and no bacterial killing when 1 x 103 to 3.4 x 104 molecules Fab'/CFU were bound to the bacterial surface. These experiments show that immune IgG and F(ab')2 can mediate killing of E. Coli 0111B4 by the alternative pathway without changing the extent of terminal C component attachment to the bacterial surface.

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