Gelatinase B [matrix metalloproteinase (MMP)-9] and collagenases (MMP-8/-13) are upregulated in cerebrospinal fluid during aseptic and bacterial meningitis in children.

We investigated the protein expression of gelatinases [matrix metalloproteinase (MMP)-2 and -9] and collagenases (MMP-8 and -13) in cerebrospinal fluid (CSF) from patients with bacterial (BM, n = 17) and aseptic (AM, n = 14) meningitis. In both, MMP-8 and -9 were increased in 100% of patients, whereas MMP-13 was detectable in 53% and 82% respectively. Three patients with clinical signs of meningitis, without CSF pleocytosis, scored positive for all three MMPs. MMP-8 appeared in two isoforms, granulocyte-type [polymorphonuclear cell (PMN)] and fibroblast/macrophage (F/M) MMP-8. Analysis of kinetic changes from serial lumbar punctures showed that these MMPs are independently regulated, and correlate only partly with CSF cytosis or levels of the endogenous inhibitor, tissue inhibitor of matrix metalloproteinase-1. In vitro, T cells, peripheral blood mononuclear cells (PBMCs) and granulocytes (PMN) release MMP-8 and -9, whereas MMP-13 could be found only in the former two cell types. Using models of exogenous (n-formyl-Met-Leu-Phe, T cell receptor cross-linking) and host-derived stimuli (interleukin-2), the kinetics and the release of the MMP-8, -9 and -13 showed strong variation between these immune cells and suggest release from preformed stocks. In addition, MMP-9 is also synthesized de novo in PBMCs and T cells. In conclusion, invading immune cells contribute only partially to MMPs in CSF during meningitis, and parenchymal cells are an equally relevant source. In this context, in patients with clinical signs of meningitis, but without CSF pleocytosis, MMPs seem to be a highly sensitive marker for intrathecal inflammation. The present data support the concept that broad-spectrum enzyme inhibition targeting gelatinases and collagenases is a potential strategy for adjunctive therapy in infectious meningitis.

Salivary albumin, total protein, IgA, IgG and IgM concentrations and occurrence of some periodontopathogens in HIV-infected patients: a 2-year follow-up study.

HIV infection reduces oral defensive mechanisms and may affect mucosal integrity. Differences in salivary protein concentrations and periodontopathogenic bacteria were studied in 56 HIV-infected patients with respect to their disease phase. Thirty-three patients were followed up for 2 years. Fifty-three healthy subjects of corresponding age and sex were studied as controls. At baseline, salivary albumin, total protein, IgA, and IgM levels were significantly higher (P<0.05-0.0001) in all phases of HIV infection, except the asymptomatic (ASX) phase, when compared with the control group. IgG levels were significantly increased in all phases except the ASX phase (P<0.05). After 2 years, salivary total protein, IgG, and IgM levels were still higher (P<0.05-0.005) in all HIV phases when compared with the control group (P<0.05-0.005). The albumin level was significantly higher in the ASX phase (P<0.005) and in the AIDS-related complex phase (ARC) (P<0.05), while the increase in IgA level was significant only in the ARC phase (P<0.005). Periodontopathogenic bacteria analyzed by PCR were detected both in the patients and the non-infected, but a statistically significant difference in the carriage percentage between the follow-up lymphadenopathy syndrome phase (LAS) and the control group was found only in Porphyromonas gingivalis (P<0.05) and Bacteroides forsythus (P< 0.0001). Thus, HIV infection appeared to cause a significant increase in the studied salivary proteins, suggesting leakage of serum components into the mouth.

72-kDa and 92-kDa gelatinases in saliva of patients with human immunodeficiency virus infection.

Human immunodeficiency virus (HIV) infection has been associated with periodontal diseases in HIV-seropositive patients. In periodontal diseases, matrix metalloproteinases (MMPs) may play key roles in the extracellular matrix, basement membrane, serpin degradation, and modification of cytokine action. We characterized the 72 kDa type IV collagenase (gelatinase A, MMP-2) and 92 kDa type IV collagenase (gelatinase B, MMP-9) in the saliva of HIV-seropositive patients and seronegative healthy controls by activity measurements and quantitative immunoblotting. Immunoblot analysis with specific antibodies against MMP-2 and MMP-9 and their tissue inhibitors (TIMP-1, TIMP-2) disclosed that, independent of the phase of the patients' HIV infection, their salivary samples contained higher amounts of MMP-2 and MMP-9 immunoreactivities in pro- and active forms and the TIMP-1 and TIMP-2 inhibitors than did the control samples. Healthy control saliva contained only slight immunoreactivities for gelatinases and TIMPs. However, as judged by the studied clinical and microbiologic indicators, HIV-seropositive patients showed only a slight tendency to develop periodontitis. Overall, an increased amount of gelatinases in saliva may reflect increased host response and defense activities in HIV infection.

Matrix metalloproteinases-1, -3 and -8 and myeloperoxidase in saliva of patients with human immunodeficiency virus infection.

OBJECTIVE: Human immunodeficiency virus (HIV)-seropositive patients have frequently severe gingival inflammation and/or attachment loss. In addition many infectious diseases affect their periodontium with varying clinical manifestations. Matrix metalloproteinases seem to play a key role in physiological periodontal remodelling and pathological tissue destruction. The aim of the present study was to characterize the presence, molecular forms, cellular sources, activities, and relative amounts of fibroblast-type (matrix metalloproteinase [MMP]-1) and neutrophil (MMP-8) collagenases, as well as their potential activator stromelysin-I (MMP-3) and myeloperoxidase in saliva of HIV-seropositive patients at different phases of HIV-infection. HIV-seronegative, healthy, age-matched patients served as controls. PATIENTS AND METHODS: Saliva samples were characterized by Western blotting using antibodies specific for MMP-1, MMP-3 and MMP-8. Interstitial collagenase activities were measured using quantitative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis/laser densitometry assay. Myeloperoxidase was analysed using quantitative dot blotting. RESULTS: Clinical and microbiological evaluation of HIV-seropositive patients' periodontium showed the presence of putative periodontopathogens ie Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Peptostreptococcus micros (Psm) and Campylobacter rectus (Cr) in their periodontal pockets. The amount of Candida increased with the severity of HIV-infection. Clinical and microbiological findings of HIV-seropositive patients suggested that they have a tendency to develop periodontal disease. Interstitial collagenase activities were found to be increased in saliva of different phases of HIV-infected patients compared to the controls. Independent of the phase of HIV-infection saliva samples contained pro- and active forms of MMP-1, -3 and -8 using Western blotting. Saliva samples from healthy controls were found to contain hardly any immunoreactivities for MMP-1 or MMP-8, but considerable amounts of MMP-3 were detected. Quantitative dot blotting demonstrated increased amounts of myeloperoxidase in HIV-patients' saliva relative to controls. CONCLUSION: The present results showed increased amounts of MMP-1, -3, -8 and myeloperoxidase in HIV-patients' saliva. MMP-1 and -8 may have been activated by MMP-3 and/or oxidants generated by myeloperoxidase. The increased amounts of MMPs and myeloperoxidase may reflect and directly participate in HIV-infection associated periodontitis.