RESUMEN
Menopause and oophorectomy without estrogen therapy (ED) have been associated with increased production of bone-active cytokines by peripheral blood mononuclear cells. The current study extended evaluation to gingival crevicular fluid (GCF) levels of interleukin (IL)-1 beta and IL-6 in such subjects compared to premenopausal and postmenopausal estrogen-treated females (ES). 13 ED and 13 ES Caucasians with a history of moderate-severe adult periodontitis provided GCF from 1-3 clinically identical sites each (5-6 mm probing depth, 5-7 mm clinical attachment loss, bleeding on probing). 30 s GCF samples were obtained and evaluated for IL-1 beta and IL-6 levels using two-site enzyme-linked immunosorbent assays (ELISAs). The frequency of GCF IL-1 beta-positive subjects was elevated in ED versus ES (92% versus 23%; p < 0.0004, chi 2 analysis). IL-6 was detected more frequently in ED subjects (23% versus 8%; not significant); however, the frequency of IL-6 detection was low in both groups due to short sampling times. These data support the concept that clinical conditions causing low estrogen environments allow increased local production of the bone-active cytokine IL-1 beta, and perhaps IL-6.
Asunto(s)
Líquido del Surco Gingival/química , Interleucina-1/análisis , Interleucina-6/análisis , Menopausia/metabolismo , Periodontitis/metabolismo , Premenopausia/metabolismo , Adulto , Factores de Edad , Resorción Ósea/metabolismo , Terapia de Reemplazo de Estrógeno , Femenino , Líquido del Surco Gingival/inmunología , Hemorragia Gingival/metabolismo , Humanos , Menopausia/inmunología , Persona de Mediana Edad , Ovariectomía , Pérdida de la Inserción Periodontal/metabolismo , Bolsa Periodontal/metabolismo , Periodontitis/inmunología , Premenopausia/inmunologíaRESUMEN
Gingival crevicular fluid (GCF) IL-8 and IL-1 beta levels were determined by sandwich enzyme-linked immunosorbent assays. Associations between IL-8 and IL-1 beta GCF levels, and between these cytokines and patient estrogen status were evaluated. IL-8 and IL-1 beta were detected more frequently and in higher amounts/30 s GCF sample in estrogen-deficient patients than in estrogen-sufficient patients. IL-8 and IL-1 beta GCF levels were significantly correlated. These findings suggest that GCF IL-8 levels are associated with patient estrogen status and local IL-1 beta concentrations.
Asunto(s)
Estrógenos/deficiencia , Líquido del Surco Gingival/metabolismo , Interleucina-1/metabolismo , Interleucina-8/metabolismo , Periodontitis/metabolismo , Adulto , Factores de Edad , Distribución de Chi-Cuadrado , Ensayo de Inmunoadsorción Enzimática , Estrógenos/metabolismo , Femenino , Humanos , Modelos Lineales , Menopausia/fisiología , Persona de Mediana Edad , Pérdida de la Inserción Periodontal/metabolismoRESUMEN
Interleukin (IL)-1 alpha and beta are cytokines which can mediate inflammatory, bone resorbing, and reparative effects in the periodontium, but few longitudinal data exist exploring their role following periodontal therapy. This study examined gingival crevicular fluid (GCF) concentrations of IL-1 alpha and IL-1 beta at sites with shallow sulci (SS) or inflamed moderate/advanced pockets (M/AP) before and 6 months after treatment with closed scaling/root planing (SC/RP) or papillary flap debridement (PFD), all in the same subject (n = 14 patients). No significant differences were noted in IL-1 alpha or beta concentrations (determined with two-site enzyme-linked immunosorbent assays) between SS and M/AP sites at baseline. While both therapies improved clinical parameters of periodontal disease, IL-1 alpha concentration increased significantly (p < 0.05) in M/AP-PFD sites 6 months after treatment, but were unchanged in other groups. IL-1 beta concentrations were numerically lower after therapy, except for a significant increase (p < 0.05) in M/AP-PFD sites. These data suggest that surgical wound healing in an inflamed, plaque-infected site (M/AP-PFD) results in prolonged production of IL-1, which may be a reflection of the extent of tissue trauma and delayed wound healing. In spite of increased IL-1 levels, these sites demonstrated significant short-term improvement in clinical attachment level (+ 1.8 mm, p < or = 0.001) postoperatively.
Asunto(s)
Líquido del Surco Gingival/química , Interleucina-1/análisis , Periodontitis/cirugía , Periodontitis/terapia , Aplanamiento de la Raíz , Colgajos Quirúrgicos , Adulto , Anciano , Índice de Placa Dental , Hemorragia Gingival/terapia , Humanos , Estudios Longitudinales , Persona de Mediana Edad , Bolsa Periodontal/metabolismo , Bolsa Periodontal/patología , Bolsa Periodontal/cirugía , Bolsa Periodontal/terapia , Periodontitis/metabolismo , Periodontitis/patologíaRESUMEN
Selected gingival bacteria and cytokine profiles associated with patients who did not respond to conventional periodontal therapy (refractory) were evaluated. 10 subjects with a high incidence of post-active treatment clinical attachment loss (> 2% sites/year lost > or = 3 mm) were compared to 10 age-, race-, and supragingival plaque-matched patients with low post-treatment clinical attachment loss (< 0.5% sites/year) relative to the following parameters at 2 sites/patient with the deepest probing depths: (1) presence of 3 selected periodontal pathogens (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens) in subgingival plaque as determined by selective culturing, and (2) gingival crevicular fluid (GCF) levels of 3 cytokines associated with bone resorption (IL-1 alpha, IL-1 beta, IL-6) as determined by two-site ELISA. Results indicated no significant differences in any clinical measurement (except incidence of clinical attachment loss), in the presence of any bacterial pathogen, or in GCF cytokine levels between refractory subject sites versus stable subject sites. However, when sites producing the greatest total GCF cytokine/patient were compared, sites from refractory patient produced significantly more IL-6 (30.1 +/- 4.0 versus 15.4 +/- 2.8 nM, p < 0.01). The subgingival presence of each of the 3 bacterial pathogens was associated with elevated GCF IL-1 concentrations. These data suggest that gingival IL-1 and IL-6 production is different in response to local and systemic factors associated with periodontitis, and that IL-6 may play a role in the identification and mechanisms of refractory periodontitis.
Asunto(s)
Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Eikenella corrodens/aislamiento & purificación , Líquido del Surco Gingival/inmunología , Líquido del Surco Gingival/microbiología , Interleucina-1/análisis , Interleucina-6/análisis , Periodontitis/inmunología , Periodontitis/microbiología , Porphyromonas gingivalis/aislamiento & purificación , Adulto , Aggregatibacter actinomycetemcomitans/inmunología , Estudios de Casos y Controles , Recuento de Colonia Microbiana , Placa Dental/inmunología , Placa Dental/microbiología , Eikenella corrodens/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Bolsa Periodontal/patología , Periodontitis/patología , Porphyromonas gingivalis/inmunologíaRESUMEN
Samples of gingival crevicular fluid (GCF) were harvested from sites manifesting features characteristic of active disease including inflammation, periodontal attachment loss, and radiographic signs of alveolar bone destruction in untreated patients with advanced periodontitis. The presence and concentrations of interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) were measured using ELISAs specific for these cytokine molecules. IL-1 alpha and/or IL-1 beta were identified in the GCF of 15 of 15 patients having untreated periodontitis. Ninety percent (71 of 79) of the sites tested contained measureable amounts of IL-1, with IL-1 beta as the more frequently occurring form. IL-1 alpha levels ranged from 0.23 nM to 13.9 nM in the GCFs. IL-1 beta levels were between 0.04 nM and 5.28 nM. Marked reductions of total IL-1 levels were observed following effective treatment. Both forms of IL-1 messenger RNA (mRNA) were detected in 17 of 17 gingival tissue samples from 6 patients. These results demonstrate that IL-1 is produced and released locally in periodontal disease at concentrations sufficient to mediate tissue inflammation and bone resorption. IL-1 may serve as a marker of periodontal tissue destruction.
Asunto(s)
Líquido del Surco Gingival/metabolismo , Gingivitis/metabolismo , Interleucina-1/análisis , Enfermedades Periodontales/etiología , Adulto , Anciano , Anticuerpos Monoclonales , Biomarcadores , Resorción Ósea/inmunología , Dinoprostona/biosíntesis , Ensayo de Inmunoadsorción Enzimática , Humanos , Persona de Mediana Edad , Enfermedades Periodontales/genética , Enfermedades Periodontales/metabolismo , ARN Mensajero/análisisRESUMEN
To determine if the release of IL-1 alpha and IL-1 beta by cultured PBMC could be independently modulated by different exogenous stimuli, we examined the effect of LPS, IFN gamma, latex beads, and indomethacin on the release of IL-1 alpha and IL-1 beta. PBMC culture supernatants were fractionated by Sephacryl-S-200 column chromatography or HPLC (TSK G3000SW), and each fraction was tested for thymocyte mitogenic activity in the presence or absence of preincubation with anti-IL-1 alpha or anti IL-1 beta monoclonal antibody (mAb) and for the presence of IL-1 alpha or IL-1 beta protein by ELISA. In all experiments, thymocyte mitogenic activity not neutralizable by anti-IL-1 alpha or anti-IL-1 beta mAb was detected in the 25K Mr range, which ranged from 12 to 50% of the total thymocyte mitogenic activity released, depending on the stimuli. Cultured PBMC from 95% of individuals release thymocyte mitogenic activity in the absence of exogenous stimuli, which was increased 1.3-to 7-fold by lopopolysaccharide (LPS) (25-50 micrograms/ml). All of this increased activity was due to increased release of IL-1 beta and non-IL-1 thymocyte mitogenic activity, with no change in the total amount of IL-1 alpha released. Indomethacin (0.1 microgram/ml) induced release of increased thymocyte mitogenic activity of 1.3- to 1.4-fold over unstimulated cultures. All of this increased activity was due to increased release of IL-1 alpha and non-IL-1 activity with a concomitant decrease in IL-1 beta release. Interferon gamma (40-100 U/ml) increased the amount of IL-1 alpha and decreased IL-1 beta and non-IL-1 activity released, resulting in no overall change in the total amount of thymocyte mitogenic activity. Molecular weight fractionation of the PBMC culture supernatants revealed that thymocyte mitogenic activity eluting in the 25K Mr range was not due to IL-1 alpha or IL-1 beta. With certain culture conditions, thymocyte mitogenic activity was detected in the 30-40K Mr range. PBMC cultured with LPS and latex beads in the absence of serum released 30-40K Mr IL-1 alpha, as well as 17K Mr IL-1 alpha and 17K Mr IL-1 beta. PBMC cultured in 2% fetal calf serum (FCS) alone from some donors released only 30-40K Mr thymocyte mitogenic activity. Both IL-1 alpha and IL-1 beta protein was detected by ELISA in this Mr range but only the IL-1 alpha was bioactive.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Interleucina-1/metabolismo , Leucocitos Mononucleares/metabolismo , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Humanos , Indometacina/farmacología , Interferón gamma/farmacología , Interleucina-1/análisis , Interleucina-1/inmunología , Lipopolisacáridos/farmacología , Activación de Linfocitos , Peso MolecularRESUMEN
Five adjuvants were compared to Freund's adjuvant for production of mouse polyclonal antibodies and monoclonal antibodies (McAbs) to human serum albumin (HSA) and interleukin-1 alpha (IL-1 alpha). Parameters examined were titer, affinity, concentration, isotype, epitope specificity and neutralizing activity of sera and hybridoma supernatants. Freund's adjuvant, while producing high titers and concentrations of antibodies in sera, was inferior to other adjuvants for eliciting antibodies with particular qualities. The adjuvants Quil A and A1(OH)3/[Thr1]muramyldipeptide elicited the highest affinity antibodies to HSA. Syntex adjuvant formulation-1 (SAF-1) elicited the highest percentage of 'protective' IgG2a antibodies to HSA. All adjuvants, particularly Quil A and Ribi adjuvant system, where superior to Freund's adjuvant in eliciting antibodies which bound native versus denatured HSA. In a comparison of SAF-1 and Freund's adjuvant, SAF-1 was superior to Freund's adjuvant in eliciting polyclonal and hybridoma antibodies which neutralized the biological activity of IL-1 alpha. These results show that adjuvants selectively and independently enhance different qualities of the antibody response. Furthermore, immunization with the appropriate adjuvant can optimize production of McAbs with desired qualities.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Afinidad de Anticuerpos , Formación de Anticuerpos , Epítopos , Isotipos de Inmunoglobulinas/análisis , Animales , Ensayo de Inmunoadsorción Enzimática , Femenino , Hibridomas , Ratones , Ratones Endogámicos BALB C , Albúmina Sérica/inmunologíaRESUMEN
The clinical and roentgenologic data from 31 excised components from 19 revision arthroplasty cases were correlated with the histology and biochemistry of the membrane at the bone-cement or bone-prosthesis interface. Twenty-seven components were cemented and four were uncemented. Twenty-four implants were clinically and roentgenologically loose, one was possibly loose, and six were well fixed. Loose components, whether cemented or not, demonstrated statistically higher prostaglandin E2 levels in the surrounding membrane compared to the nonloose group. Collagenase and M-collagenase levels were absent or insignificantly low in all specimens; no detectable interleukin 1 beta was found. This suggests that prostaglandin E2 may be associated with the bone lysis associated with prosthesis loosening.
Asunto(s)
Prótesis de Cadera , Cementos para Huesos , Dinoprostona/metabolismo , Humanos , Técnicas In Vitro , Interleucina-1/análisis , Colagenasa Microbiana/biosíntesis , Falla de Prótesis , Reoperación , Membrana Sinovial/metabolismo , Membrana Sinovial/patologíaRESUMEN
Monoclonal antibodies (McAb) were developed to the Mr 17,500 form of human recombinant interleukin 1, IL 1 beta. Four McAb have been identified that inhibit the biological activity of IL 1 beta. McAb H34 and H67, at 1 microgram/ml (6 X 10(-9) M), completely inhibit the capacity of 1 ng/ml (6 X 10(-11) M) recombinant IL 1 beta to stimulate the proliferation of murine thymocytes or human fibroblasts in vitro. McAb H6 and H21 are approximately 10-fold less potent, and completely inhibit IL 1 beta activity at 10 micrograms/ml (6 X 10(-8) M) in both assays. The McAb do not have a significant effect on the biological activity of human recombinant IL 1 alpha in either assay. These McAb block the binding of recombinant [125I]IL 1 beta to IL 1 receptors on mouse 3T3 fibroblasts and have affinity constants for IL 1 beta in the range of 10(9) to 10(10) liters/mol. Competition studies suggest that two nonoverlapping epitopes on the IL 1 beta molecule are recognized by the McAb. H6 and H34 recognize one epitope, and H21 and H67 another. McAb H6 and H67 have been used together in a two-site ELISA to detect IL 1 beta. The sensitivity of the ELISA, which is 15 pg/ml (0.86 pM), approaches the limit of sensitivity of the thymocyte proliferation assay. The ELISA and thymocyte proliferation assay were used to quantitate IL 1 beta in E. coli LPS-stimulated human monocyte culture supernatants (HMCS). The level of IL 1 beta detected by ELISA in culture supernatants from eight donors ranged from 1.7 to 5.6 ng/ml, with a mean value of approximately 3 ng/ml. By comparison, the thymocyte proliferation assay gave levels of IL 1 in HMCS that were eight fold higher when quantitated by using recombinant IL 1 beta as a standard. This discrepancy with the bioassay used was reflected by the three fold higher maximum stimulation of thymocyte proliferation by HMCS as compared with recombinant IL 1 alpha or IL 1 beta, and only 45% inhibition of HMCS IL 1 activity by McAb. Thus, factors other than IL 1 beta account for the IL 1-like activity in monocyte culture supernatant as measured by the bioassay. The ILB1 McAb and ELISA allow for the first time-sensitive, accurate, and convenient quantitation of IL 1 beta levels in biological fluids or specimens.