lactoferrin and Aggressive-Periodontitis

Aggressive periodontitis is a rare but rapidly progressing form of periodontal disease that usually affects otherwise systemically healthy individuals, at a young age. It usually affects first molars and incisors, which are usually lost if treatment is not properly and early rendered. Although of low prevalence, it affects individuals of African descent at a higher prevalence, and usually multiple members within the same family. Several studies have been performed in the attempt to evaluate specific single nucleotide polymorphisms (SNPs) that could be associated with this disease. To the best of our knowledge, the present article provides the first review of the literature focusing on studies that evaluated SNPs in patients of African descent with aggressive periodontitis. Several SNPs have been evaluated in different genes according to their role in the pathogenesis of the disease, with positive and negative associations (such as IL1, FCGR3B, FPR1, LTF, CYBA, GLT6D1, TLR4) with both the localized and generalized forms of aggressive periodontitis. Given the complexity of periodontitis, the difficulty in gathering large cohorts diagnosed with this rare form of disease, and the fact that candidate gene studies may only determine part of the genetic risk of a disease, the search for specific SNPs associated with aggressive periodontitis seems to be a long one, most likely to result in the combination of multiple SNPs, in multiple genes.

Topics: Aggressive Periodontitis; Black or African American; Databases, Factual; Genetic Predisposition to Disease; GPI-Linked Proteins; Humans; Interleukin-1; Lactoferrin; NADPH Oxidases; Periodontal Diseases; Polymorphism, Genetic; Polymorphism, Single Nucleotide; Receptors, Formyl Peptide; Receptors, IgG; Risk Factors; Toll-Like Receptor 4; United States

Lactoferrin is one of a number of multifunctional proteins that are present in or on all mucosal surfaces throughout the body. Levels of lactoferrin are consistently elevated in inflammatory diseases such as arthritis, inflammatory bowel diseases, corneal disease, and periodontitis. Single-nucleotide polymorphisms (SNPs) in lactoferrin have been shown to be present in individuals susceptible to Escherichia coli-induced travelers' diarrhea and in tear fluid derived from virally associated corneal disease. Here, we review data showing a lactoferrin SNP in amino acid position 29 in the antimicrobial region of lactoferrin that acts against caries associated bacteria. This SNP was initially discovered in African American subjects with localized aggressive periodontitis (LAP) who had proximal bone loss but minimal proximal caries. Results were confirmed in a genetic association study of children from Brazil with this same SNP who showed a reduced level of caries. In vitro data indicate that lactoferrin from whole saliva derived from subjects with this SNP, recombinant human lactoferrin containing this SNP, or an 11-mer peptide designed for this SNP kills mutans streptococci associated with caries by >1 log. In contrast, the SNP has minimal effect on Gram-negative species associated with periodontitis. Moreover, periodontally healthy subjects homozygous for this lysine (K) SNP have lactoferrin in their saliva that kills mutans streptococci and have reduced proximal decay. The review summarizes data supporting the ecologic plaque hypothesis and suggests that a genetic variant in lactoferrin with K in position 29 when found in saliva and crevice fluid can influence community biofilm composition. We propose that, for caries, this SNP is ethnicity independent and protective by directly killing caries-provoking bacteria (reducing proximal decay). However, the clinical effect of this SNP in LAP is ethnicity dependent, destructive (increases LAP incidence), and complex with mechanisms still to be determined.

Topics: Aggressive Periodontitis; Anti-Infective Agents; Biofilms; Dental Caries; Dental Plaque; Humans; Lactoferrin; Lysine; Polymorphism, Single Nucleotide; Streptococcus mutans

While Aggregatibacter actinomycetemcomitans (Aa) is highly associated with localised aggressive periodontitis (LAP) many Aa-carriers do not develop LAP. This study was designed to determine whether specific salivary factors could distinguish between subjects who have Aa initially and remain healthy (H/AA) as compared to those who develop LAP (LAP/AA).. H/AA subjects and healthy controls with no Aa (H) were enrolled in a longitudinal cohort study to investigate initiation of bone loss (LAP) over 3 years. After detection of LAP, stored saliva from 10 H, 10 H/AA, and 10 LAP/AA subjects was thawed, processed, and tested for (1) lactoferrin (Lf) concentration and iron levels; (2) agglutination of Aa; (3) killing of Gram-positive bacteria.. LAP/AA saliva levels of Lf iron were low prior to and after bone loss (3.6+1.7ngFe/μg) (LAP/AA vs. H and H/AA p≤0.01). Saliva from H/AA subjects caused Aa to agglutinate significantly more than H or LAP/AA saliva (p≤0.01). LAP/AA saliva killed Streptococcus mutans, Streptococcus sanguis and Lactobacillus in vitro by >83%. Saliva from H individuals killed these bacteria by <3.3% (LAP/AA vs. H; p≤0.01). H/AA killing was intermediate.. LAP/AA saliva showed: low levels of Lf iron, minimal Aa agglutinating activity, and high killing activity against Gram-positive bacteria. Aa-positive healthy saliva (H/AA) showed: higher levels of Lf iron, maximal Aa agglutinating activity, and moderate killing of Gram-positive bacteria. A salivary activity profile can distinguish between subjects who are Aa-positive and remain healthy from those who develop LAP.

Topics: Adolescent; Agglutination; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Alveolar Bone Loss; Anti-Bacterial Agents; Bacterial Load; Case-Control Studies; Child; Cohort Studies; Female; Follow-Up Studies; Humans; Immunoglobulin A, Secretory; Iron; Lactobacillus; Lactoferrin; Longitudinal Studies; Male; Pasteurellaceae Infections; Retrospective Studies; Saliva; Streptococcus mutans; Streptococcus sanguis

Among the innate defense mechanisms in the oral cavity, lactoferrin (LF) is a vital antimicrobial that can modify the host response against periodontopathogens. Aggregatibacter actinomycetemcomitans is the main periodontopathogen of localized aggressive periodontitis. The aim of this study is to evaluate the role of LF during A. actinomycetemcomitans-induced periodontitis.. Differences in the expression levels of cytokines, chemokines, chemokine receptors, and bone loss markers between wild-type (WT) and LF knockout mice (LFKO(-/-)) were evaluated by real time-PCR. Serum IgG and LF levels were quantified by ELISA. Alveolar bone loss among the groups was estimated by measuring the distance from cemento-enamel junction (CEJ) to the alveolar bone crest (ABC) at 20 molar sites.. Oral infection with A. actinomycetemcomitans increased LF levels in periodontal tissue (P = 0.01) and saliva (P = 0.0004) of wild-type infected (WTI) mice compared to wild-type control mice. Pro-inflammatory cytokines such as interferon-γ, tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and IL-12 were increased in the infected LF knockout (LFKO(-/-)I) mice compared to the WTI mice, whereas the anti-inflammatory cytokines IL-4 and IL-10 were decreased. Chemokines and chemokine receptors showed different expression patterns between WTI and LFKO(-/-)I mice. The LFKO(-/-)I mice developed increased bone loss (P = 0.002), in conjunction with increased expression of receptor activator of nuclear factor-κB ligand and decrease in osteoprotegerin, compared to WTI mice.. These results demonstrate that the infected LFKO(-/-) mice were more susceptible to A. actinomycetemcomitans-induced alveolar bone loss, with different patterns of immune responses compared to those of WTI mice.

Topics: Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Alveolar Bone Loss; Animals; Chemokines; Cytokines; Disease Susceptibility; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukins; Lactoferrin; Mice; Mice, Knockout; Osteoprotegerin; Pasteurellaceae Infections; Periodontium; RANK Ligand; Receptors, Chemokine; Saliva; Tooth Cervix; Tumor Necrosis Factor-alpha; Vesicular Transport Proteins

Some patients suffering from aggressive periodontitis (Ag-P) also display neutrophil chemotaxis dysfunction. In this study, we attempted to identify the proteins involved in Ag-P associated with neutrophil chemotaxis dysfunction using proteome analysis.. A two-dimensional fluorescence difference gel electrophoresis system was used to detect differences in protein expression between neutrophils from four patients suffering from Ag-P combined with neutrophil chemotaxis dysfunction and those from four controls. Moreover, the mRNA levels of the proteins identified by the above method were examined in neutrophils from four types of subjects using the real-time polymerase chain reaction: twenty patients suffering from Ag-P with or without the dysfunction, 15 patients with chronic periodontitis, and 15 controls.. Four proteins, lactoferrin, caldesmon, heat shock protein 70, and stac, displayed a higher protein expression level in the neutrophils from the patients suffering from Ag-P combined with the neutrophil dysfunction than in those from the control group. The caldesmon mRNA levels in the neutrophils from the patients suffering from Ag-P combined with the neutrophil dysfunction were high compared with those in the neutrophils from the patients suffering from the other two types of periodontitis and those from the control group.. Caldesmon may be a marker of Ag-P combined with neutrophil chemotaxis dysfunction.

Topics: Adult; Aggressive Periodontitis; Biomarkers; Calmodulin-Binding Proteins; Case-Control Studies; Chemotaxis, Leukocyte; Chronic Periodontitis; Electrophoresis, Gel, Two-Dimensional; Female; Gingival Hemorrhage; HSP70 Heat-Shock Proteins; Humans; Lactoferrin; Leukocyte Count; Male; Nerve Tissue Proteins; Neutrophils; Periodontal Attachment Loss; Periodontal Pocket; Periodontium; Polymerase Chain Reaction; Proteome; RNA, Messenger; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

A dramatic difference in the frequencies of the Lys/Arg single nucleotide polymorphism in the lactoferrin genotype between a small population of patients with localized juvenile periodontitis and healthy subjects has been reported. As the single nucleotide polymorphism could be associated with ethnicity, the present study aimed to investigate the association between polymorphisms of the lactoferrin gene and periodontitis.. Sixty-five patients with aggressive periodontitis, 278 with chronic periodontitis and 88 healthy controls were genotyped for the Lys/Arg polymorphism of the lactoferrin gene at position 29 [reference sequence (rs) 1126478] in the N-terminal alpha-helical region.. The frequencies of the GG genotype and the G allele were highest in the aggressive periodontitis group, followed by the chronic periodontitis group and then the healthy controls. The frequency of the G allele was significantly higher in aggressive periodontitis and chronic periodontitis groups than in healthy controls (p = 0.0037 and 0.0212). Although the difference of the GG genotype distribution between subjects with chronic periodontitis and healthy controls did not reach significance, the distribution of genotypes between aggressive periodontitis and healthy controls was significantly different. The association of the gene polymorphism and aggressive periodontitis still existed, even after adjusting for age, gender and smoking status by logistic regression analysis (GG/AG+AA: odds ratio = 2.16, 95% confidence interval = 1.09-4.35, p = 0.0287). After the study, subjects were further stratified by their smoking status; the GG genotype was still significantly associated with the risk of aggressive periodontitis in the nonsmoking group (odds ratio = 2.69, p = 0.018). However, there were no statistical differences between chronic periodontitis vs. healthy controls and aggressive periodontitis vs. healthy controls in the smoking group.. The present study revealed that the A/G polymorphism in the lactoferrin gene might be associated with aggressive periodontitis. The A allele might reduce the risk of development of aggressive periodontitis in a Taiwanese population. Our results also support the hypothesis that lactoferrin genetic polymorphisms could play a role in the risk for periodontitis separate from the smoking factor. The functionality of this gene's polymorphisms has to be further elucidated.

Topics: Adult; Aggressive Periodontitis; Arginine; Asian People; Case-Control Studies; Chronic Periodontitis; Female; Gene Frequency; Humans; Lactoferrin; Logistic Models; Lysine; Male; Middle Aged; Mutation, Missense; Polymorphism, Single Nucleotide; Smoking; Taiwan

Salivary proteomics technology can be used to evaluate the disease progression of periodontitis and the systemic screening of proteomes of saliva from subjects with aggressive periodontitis has not been available. The objective of this preliminary study was to compare the proteomic profile of whole unstimulated saliva of subjects with generalized aggressive periodontitis (GAgP) with that of healthy volunteers to identify proteins, the levels of which were significantly altered between the two groups.. Whole unstimulated saliva was obtained from five subjects with GAgP and five healthy subjects, and proteins were separated using two-dimensional gel electrophoresis. Proteins, the levels of which were significantly different between the two groups, were identified by computer image analyses and subsequent electrospray ionization tandem mass spectrometry.. Eleven proteins that exhibited a different level in the GAgP group vs. the control group were identified. Compared with whole saliva of healthy control subjects, the levels of serum albumin, immunoglobulin (Ig) gamma2 chain C region, Ig alpha2 chain C region, vitamin D-binding protein, salivary alpha-amylase and zinc-alpha2 glycoprotein were increased in whole unstimulated saliva of GAgP subjects, while those of lactotransferrin, elongation factor 2, 14-3-3 sigma, short palate, lung and nasal epithelium carcinoma-associated protein 2 precursor and carbonic anhydrase 6 were decreased.. Comparison of the proteomic profile of whole unstimulated saliva of GAgP subjects with that of healthy control subjects revealed at least 11 differential proteins. The approach applied herein might be helpful to aid understanding of the etiology of GAgP.

Topics: 14-3-3 Proteins; Adipokines; Adult; Aggressive Periodontitis; Biomarkers; Biomarkers, Tumor; Carbonic Anhydrases; Carrier Proteins; Electrophoresis, Gel, Two-Dimensional; Exonucleases; Exoribonucleases; Glycoproteins; Humans; Image Processing, Computer-Assisted; Immunoglobulin alpha-Chains; Immunoglobulin gamma-Chains; Lactoferrin; Leucine Zippers; Neoplasm Proteins; Peptide Elongation Factor 2; Phosphoproteins; Protein Precursors; Proteome; Saliva; Salivary alpha-Amylases; Salivary Proteins and Peptides; Serum Albumin; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Vitamin D-Binding Protein

The iron-binding protein lactoferrin is a ubiquitous and abundant constituent of human exocrine secretions. Lactoferrin inhibits bacterial growth by sequestering essential iron and also exhibits non-iron-dependent antibacterial, antifungal, antiviral, antitumor, anti-inflammatory, and immunoregulatory activities. All of these non-iron-dependent activities are mediated by the highly charged N terminus of lactoferrin. In this study we characterized a Lys/Arg polymorphism at position 29 in the N-terminal region of human lactoferrin that results from a single nucleotide polymorphism in exon 1 of the human lactoferrin gene. We expressed cDNAs encoding both lactoferrin variants in insect cells and purified the two proteins by ion exchange chromatography. The two lactoferrin variants exhibited nearly identical iron-binding and iron-releasing activities and equivalent bactericidal activities against a strain of the gram-negative bacterium Actinobacillus actinomycetemcomitans. When tested against the gram-positive species Streptococcus mutans and Streptococcus mitis, however, lactoferrin containing Lys at position 29 exhibited significantly greater bactericidal activity than did lactoferrin containing Arg. In addition, the Lys-containing lactoferrin stimulated bovine tracheal epithelial cells to synthesize much higher levels of tracheal antimicrobial peptide mRNA than did the Arg-containing variant. A genotyping assay that distinguished between the two alleles based on a polymorphic EarI restriction site showed that the Lys and Arg alleles had frequencies of 24% and 76%, respectively, among 17 healthy human subjects, and 72% and 28%, respectively, among nine patients with localized juvenile periodontitis. Our findings suggest that these two lactoferrin variants are functionally different and that these differences may contribute to the pathogenesis of localized juvenile periodontitis.

Topics: Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Animals; Anti-Bacterial Agents; Cloning, Molecular; Genotype; Humans; Iron; Lactoferrin; Polymerase Chain Reaction; Polymorphism, Genetic; Recombinant Proteins; Spodoptera; Transcriptional Activation

Actinobacillus actinomycetemcomitans (Aa) is associated with localized aggressive periodontal disease in juveniles (LAgP). Lactoferrin (LF) is an iron-binding salivary protein that has been shown to kill Aa in its iron-free form (apo) and reduce binding to host cells in its iron-saturated form (halo). However, recent in vitro studies show that LF does not kill clinical isolates of Aa, and LF with reduced levels of bound iron does not interfere with its attachment. These findings suggest that colonization of Aa may occur more readily in an environment containing LF with low iron levels. The purpose of this study was to examine the relationship of LF iron levels in saliva of LAgP patients as compared to their age-, gender-, and race-matched controls.. Whole and parotid saliva was collected from LAgP patients and matched controls. Micrograms of LF/mg of protein as well as nanograms of iron/micrograms of LF were determined. Iron binding was determined in parotid saliva by addition of nonlabeled and 59Fe labeled iron.. LAgP patients' whole saliva had higher LF levels than controls, but their LF contained less iron (P < or =0.005). No iron was found in LF from parotid saliva in either group. When iron was added to parotid saliva, the LAgP saliva bound 20 to 30 times less iron than controls (P< or =0.001). Finally, LF was identified as the major iron-binding protein in parotid saliva by 59Fe autoradiography and Western blotting.. This study shows that the level of bound iron in LF is significantly reduced in LAgP patients compared to controls. These data suggest that LF from LAgP patients has a reduced capacity to bind iron and that LF iron levels may play an important role in Aa-induced LAgP.

Topics: Adolescent; Adult; Aggressive Periodontitis; Autoradiography; Blotting, Western; Case-Control Studies; Child; Female; Humans; Immunoelectrophoresis; Iron; Iron Radioisotopes; Lactoferrin; Male; Saliva; Salivary Proteins and Peptides; Statistics, Nonparametric

Prior reports have suggested that the iron-binding protein lactoferrin (LF) may either kill Actinobacillus actinomycetemcomitans (Aa) or interfere with its binding to host cells. Other studies have indicated that the degree of iron saturation of LF might play a role in these interactions. However, these studies utilized strains that had lost critical attachment characteristics found in well-preserved clinical isolates of Aa. The purpose of this work was to study the effect of LF iron levels on survival and attachment of well-preserved clinical isolates of Aa.. LF containing 0%, 30%, and 100% iron saturation was tested for its ability to kill clinical isolates of Aa and to inhibit their binding to buccal epithelial cells (BECs).. Neither iron-free LF (apo-LF) nor iron-saturated LF killed Aa clinical isolates. Increasing the iron saturation of LF resulted in an increased inhibition of Aa binding to BECs (P < or =0.005). This effect was consistent for the 3 clinical isolates tested. Pretreatment of Aa with iron-saturated LF reduced binding to BECs by 58%, 61.8%, and 64.2%, respectively, for each of the 3 clinical strains tested (P < or =0.005). Pretreatment of Aa strains with apo-LF, iron alone, or bovine serum albumin had no effect on binding. Pretreatment of BECs with LF (either apo-LF or iron-containing LF) had no influence on Aa binding.. These results indicate that reduction in binding of Aa to epithelial cells is maximized by pretreatment of Aa cells with iron-saturated lactoferrin. These in vitro results suggest that patients with lactoferrin containing lowered levels of iron would be more susceptible to Aa colonization.

Topics: Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Analysis of Variance; Animals; Apoproteins; Bacterial Adhesion; Cattle; Cells, Cultured; Child; Colony Count, Microbial; Epithelial Cells; Female; Humans; Iron; Lactoferrin; Male; Mouth Mucosa

The aim of this study was to examine the longitudinal association of selected non-immune anti-microbial host factors (peroxidases, lysozyme and lactoferrin) to the localized juvenile periodontitis (LJP) disease status.. Peroxidases, lysozyme and lactoferrin were quantitated from seven patients with LJP before and after periodontal therapy. Analyses were performed from simultaneously collected samples of peripheral blood polymorphonuclear leukocytes (PMNs), gingival crevicular fluid (GCF from diseased sites) and paraffin-stimulated whole saliva. Similar assays were done also from seven periodontally healthy controls.. During untreated phase of LJP myeloperoxidase, lysozyme and lactoferrin concentrations were remarkably elevated in peripheral blood PMNs, also reflected in their high concentrations in GCF. All these values normalised with respect to healthy controls during the periodontal therapy. No similar longitudinal changes were seen in whole saliva but during therapy salivary peroxidase concentrations declined below the control values, in accordance with our previous observations in parotid saliva samples of LJP patients.. In LJP the concentrations of lysozyme, lactoferrin and myeloperoxidase are significantly elevated in peripheral blood PMNs, also reflected in GCF. During periodontal therapy these values decline and approach those observed in healthy controls. No similar changes are seen in stimulated whole saliva.

Topics: Adolescent; Adult; Aggressive Periodontitis; Case-Control Studies; Female; Follow-Up Studies; Gingival Crevicular Fluid; Humans; Lactoferrin; Male; Muramidase; Neutrophils; Periodontal Index; Periodontal Pocket; Peroxidases; Saliva; Time Factors

The local, saliva-associated defense mechanisms of 28 juvenile periodontitis (JP) patients and their age- and sex-matched controls were studied. Lysozyme, lactoferrin, salivary peroxidase, myeloperoxidase, and thiocyanate concentrations were determined from both whole saliva and parotid saliva. The total concentrations of salivary IgA, IgG, and IgM were assayed. The periodontal condition and the salivary flow rates were registered. Among the JP patients, a significantly elevated concentration of IgG was found in parotid saliva but not in whole saliva. Salivary peroxidase activities were significantly low both in the whole and in the parotid saliva samples of the JP patients, and leukocyte-derived myeloperoxidase was present in significantly low amounts in whole saliva of these patients. Because both glandular (salivary peroxidase) and polymorphonuclear-cell-derived (myeloperoxidase) enzyme activities were low among the JP patients, suppressed peroxidase-mediated host defense mechanisms could be characteristic of JP.

Topics: Adolescent; Adult; Aggressive Periodontitis; Amylases; Female; Gingival Hemorrhage; Humans; Immunoglobulin A, Secretory; Immunoglobulin G; Immunoglobulin M; Lactoferrin; Male; Muramidase; Periodontal Pocket; Peroxidase; Peroxidases; Saliva

Fourteen subjects were examined for lactoferrin content in PMNs of venous blood. Eight of the subjects were diagnosed localized juvenile periodontitis (LJP) and four adult periodontitis (AP), all having subgingival occurrence of Actinobacillus actinomycetemcomitans (A.a.). Two subjects had healthy gingival conditions and no detectable A.a. Deficiency or low PMN lactoferrin amounts were found in six of the eight subjects with LJP and in two of the subjects with AP. The reduced lactoferrin content in the PMNs was suggested to be depending on a cytotoxic factor produced by A.a. adding to an intrinsic PMN defect.

Topics: Actinobacillus; Adolescent; Adult; Aggressive Periodontitis; Chemotactic Factors; Humans; Lactoferrin; Middle Aged; Neutrophils; Periodontitis

Patients with localized juvenile periodontitis (LJP) exhibit defective neutrophil functions to a variety of environmental and host stimuli. It is not clear, however, how many of the measurable functions are defective and whether individual patients exhibit single or multiple dysfunctions. The purpose of this study was to evaluate chemotaxis, phagocytosis, specific granule release and superoxide production in a group of 23 previously unreported LJP patients. Our results indicate that all 23 of these LJP patients exhibited chemotaxis depression to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) and endotoxin-activated serum (EAS). Smaller groups from the 23 chemotactically defective LJP group were used to test other function due to inability to obtain sufficient quantities of blood. Fourteen of 14 LJP patients tested exhibited defective phagocytosis. Ten LJP patients were evaluated for specific granule release, and 14 LJP patients were evaluated for superoxide production. Both granule release and superoxide production were found to be normal in chemotactically defective LJP patients. Since both defective and normal responses noted in the same neutrophil populations are mediated by the same receptor, it is hypothesized that the cellular defect lies in a post receptor pathway.

Topics: Aggressive Periodontitis; Chemotaxis, Leukocyte; Cytoplasmic Granules; Humans; Kinetics; Lactoferrin; Lactoglobulins; Neutrophils; Periodontal Diseases; Phagocytosis; Receptors, Complement; Superoxides