Serum Antibody Response to Antigens of Oral Gram-negative Bacteria by Cats with Plasma Cell Gingivitis-Pharyngitis

Sims, Tom J.; Moncla, Bernard J.; Page, Roy C.

doi:10.1177/00220345900690031001

Cited by 46 publications

(27 citation statements)

References 25 publications

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“…Enzyme‐linked immunosorbent assays (ELISA) were performed as described previously (61), but with the following modification. Briefly, ninety six well microtiter plates were preincubated with 75 μl per well of poly‐ l ‐lysine at 1 mg/ml in PBS at room temperature for 30 min to promote bacterial cell adhesion to the well, and then coated with 50 μl per well of P. gingivalis whole cells at an optical density at 660 nm of 0.3 in PBS for 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Functional characteristics of antibodies induced by Arg‐gingipain (HRgpA) and Lys‐gingipain (Kgp) from Porphyromonas gingivalis

Nakagawa

Sims

Fan

et al. 2001

Oral Microbiology and Immunology

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Arginine-specific gingipain (HRgpA) and lysine-specific gingipain (Kgp), enzymes produced by Porphyromonas gingivalis, may be candidates for an anti-P. gingivalis vaccine. The purpose of our study was to determine whether HRgpA and Kgp have opsonic target sites and whether these sites are available and accessible on intact P. gingivalis cells. Rabbits were used to generate polyclonal antibodies to both proteins. Animals were immunized and immunoglobulin G (IgG) fractions were isolated from preimmune and immune sera. Functional characteristics of the antibodies were assessed by determining antibody titers by enzyme-linked immunosorbent assay (ELISA), generating Western immunoblots, and measuring antibody enhancement of P. gingivalis opsonization, phagocytosis and killing by polymorphonuclear leukocytes (PMN) of intact cells of strains of P. gingivalis representative of the four serotypes. Strains studied included 33277 (serotype A), A7A1-28 (serotype B), W50 (serotype C) and 381 (serotype D). Both HRgpA and Kgp induced high titers of IgG antibody. Anti-HRgpA and anti-Kgp bound to both HRgpA and Kgp demonstrating a large proportion of shared antigenic epitopes. The two antibodies bound equally well to all four P. gingivalis serotypes with titers ranging from 77 to 205 ELISA units when compared to preimmune IgG set at 1 ELISA unit. The immunoblot patterns of binding of the two antibodies to HRgpA and Kgp and to sonicates of the four P. gingivalis serotypes were virtually identical. Both antibodies detected components in HRgpA at 27, 35 and 45 kDa and in Kgp at 27, 32, 35, 40 and 55 kDa. The antibodies also detected components at or near these same positions in addition to multiple high molecular mass components in the cell sonicates of P. gingivalis. Both proteins induced antibodies that significantly enhanced opsonization as assessed by chemiluminescence, with values ranging from 130 mV to 375 mV for anti-HRgpA IgG and from 240 mV to 475 mV for anti-Kgp IgG. Both antibodies significantly enhanced PMN-mediated bacterial killing of the four P. gingivalis serotypes, although the percentage of killing varied among the serotypes (24-81% for anti-HRgpA and 37-89% for anti-Kgp). Thus, both HRgpA and Kgp express opsonic target sites and induce high titers of antibodies that opsonize and enhance killing of all four serotypes of P. gingivalis. These two proteins appear to be potential candidate antigens for an anti-P. gingivalis vaccine.

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Section: Methodsmentioning

confidence: 99%

Functional characteristics of antibodies induced by Arg‐gingipain (HRgpA) and Lys‐gingipain (Kgp) from Porphyromonas gingivalis

Nakagawa

Sims

Fan

et al. 2001

Oral Microbiology and Immunology

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“…Bacterial endotoxin was purified from Actinobacillus actinomycetemcomitans (Sims et al, 1990). Human junctional epithelial cells, newborn-human aortic smooth-muscle cells, and bovine aortic endothelial cells were obtained from Drs.…”

Section: Methodsmentioning

confidence: 99%

Cell Attachment Activity of Cementum Proteins and Mechanism of Endotoxin Inhibition

et al. 1991

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Cementum occupies a unique anatomical location where soft connective tissues of the periodontium are attached to root surfaces. Cell attachment properties of proteins present in cementum were studied. Human and bovine cementum were extracted with 0.5 mol/L CH3COOH followed by 4 mol/L guanidine, and proteins were separated by ion-exchange chromatography and SDS-polyacrylamide gel electrophoresis. Cells were labeled with radioactive amino acids and added to tissue-culture plastic plates incubated with cementum proteins, and attachment was measured. Results showed that cementum proteins promoted the attachment of smooth muscle cells, endothelial cells, and fibroblasts, but not epithelial cells. Fibroblasts attached more efficiently than other cell types, and they manifested spreading with re-organization of actin filaments. No attachment occurred to plates incubated with endotoxin from A. actinomycetemcomitans. Fewer fibroblasts attached to plates treated with cementum proteins in the presence of endotoxin, but cells pre-treated with endotoxin attached normally. Attachment was not inhibited when plates were incubated first with attachment proteins and then with endotoxin; however, it was decreased when endotoxin or bovine serum albumin preceded cementum proteins. Cementum proteins with Mr 68,000, 61,000, 55,000, and 36,000 (p68, p61, p55, and p36, respectively) manifested attachment activity, while protein(s) with Mr 23,000-24,000 did not. Western blots revealed that guanidine extracts contained three bands cross-reacting with anti-bovine sialoprotein-II antibody, but the p61, p55, and p36 were negative. We conclude that cementum contains bovine sialoprotein-II and at least four other fibroblast attachment proteins, and that they do not support epithelial cell attachment.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…Purified Pg protein fraction was prepared by the SDS-TCA method. 39 Briefly, lyophilized cells were suspended in 1% sodium dodecylsulfate and 1% 2-mercaptoethanol, boiled, and solubilized ultrasonically as described above. Insoluble peptidoglycan and debris were removed by centrifugation at 100,000 g for 1 hour.…”

Section: Antigen Preparationsmentioning

confidence: 99%

Humoral Immune Responses to Porphyromonas gingivalis Before and Following Therapy in Rapidly Progressive Periodontitis Patients

Bd²,

et al. 1991

Journal of Periodontology

102

105

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We have performed studies aimed at elucidating the nature of the humoral immune response in rapidly progressive periodontitis (RPP). We analyzed the sera of 36 periodontally normal subjects and 36 RPP patients for titers and avidities of IgG antibodies reactive with the antigens of Porphyromonas gingivalis using ELISA, prior to and following treatment. We used whole‐cell sonicate, purified lipopolysaccharide (LPS), and total extractable protein as plate antigens. Twelve of the patients had antibody titers at least 2‐fold greater than the median of the controls and were designated as seropositive. The remaining 24 patients had titers that did not exceed twice the median titer of the controls and were designated as seronegative. For both patient groups, antibody titers were highest when whole‐cell antigen was used, intermediate for LPS, and lowest for the protein fraction. Following treatment, median titer for seropositive patients decreased from pretreatment values of 241.7 to 76.5, while median titer for seronegative patients increased from 39.5 to 80.1. Avidities of pretreatment sera from both patient groups for all 3 antigen preparations were lower than the median avidities of the control sera. Avidity significantly increased following treatment to levels greater than those for control sera in both patient groups. Thus, some young adults with severe periodontitis mount a humoral immune response and produce high levels of serum IgG antibodies reactive with antigens of P. gingivalis, while others do not. The antibodies produced are of relatively low avidity, and may therefore be relatively ineffective biologically. Therapy, which greatly reduces antigen load, appears to stimulate production of higher avidity IgG antibodies in both patient groups; in the seropositive group, low avidity antibodies appear to be replaced by antibodies of higher avidity. Both the purified LPS and protein fractions contain reactive antigen(s), although LPS binds more antibody. Our data are consistent with the idea that many RPP patients do not produce protective levels of biologically functional antibody during the course of their natural infection, but they may be stimulated to do so by treatment. J Periodontol 1991; 62:781–791.

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Serum Antibody Response to Antigens of Oral Gram-negative Bacteria by Cats with Plasma Cell Gingivitis-Pharyngitis

Cited by 46 publications

References 25 publications

Functional characteristics of antibodies induced by Arg‐gingipain (HRgpA) and Lys‐gingipain (Kgp) from Porphyromonas gingivalis

Functional characteristics of antibodies induced by Arg‐gingipain (HRgpA) and Lys‐gingipain (Kgp) from Porphyromonas gingivalis

Cell Attachment Activity of Cementum Proteins and Mechanism of Endotoxin Inhibition

Humoral Immune Responses to Porphyromonas gingivalis Before and Following Therapy in Rapidly Progressive Periodontitis Patients

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