Unopposed matrix metalloproteinase-9 expression in human tuberculous granuloma and the role of TNF-alpha-dependent monocyte networks.

Tuberculosis is characterized by granuloma formation and caseous necrosis, but the factors causing tissue destruction are poorly understood. Matrix metalloproteinase (MMP)-9 (92-kDa gelatinase) secretion from monocytes is stimulated by Mycobacterium tuberculosis (M. tb) and associated with local tissue injury in tuberculosis patients. We demonstrate strong immunohistochemical MMP-9 staining in monocytic cells at the center of granuloma and adjacent to caseous necrosis in M. tb-infected patient lymph nodes. Minimal tissue inhibitor of MMPs-1 staining indicated that MMP-9 activity is unopposed. Because granulomas characteristically contain few mycobacteria, we investigated whether monocyte-monocyte cytokine networks amplify MMP-9 secretion. Conditioned medium from M. tb-infected primary human monocytes or THP-1 cells (CoMTB) stimulated MMP-9 gene expression and a >10-fold increase in MMP-9 secretion by monocytes at 3-4 days (p < 0.009, vs controls). Although CoMTB stimulated dose-dependent MMP-9 secretion, MMP-1 (52-kDa collagenase) was not induced. Anti-TNF-alpha Ab but not IL-1R antagonist pretreatment decreased CoMTB-induced MMP-9 secretion by 50% (p = 0.0001). Anti-TNF-alpha Ab also inhibited MMP-9 secretion from monocytic cells by 50%, 24 h after direct M. tb infection (p = 0.0002). Conversely, TNF-alpha directly stimulated dose-dependent MMP-9 secretion. Pertussis toxin inhibited CoMTB-induced MMP-9 secretion and enhanced the inhibitory effect of anti-TNF-alpha Ab (p = 0.05). Although chemokines bind to G protein-linked receptors, CXCL8, CXCL10, CCL2, and CCL5 did not stimulate monocyte MMP-9 secretion. However, the response to cholera toxin confirmed that G protein signaling pathways were intact. In summary, MMP-9 within tuberculous granuloma is associated with tissue destruction, and TNF-alpha, critical for antimycobacterial granuloma formation, is a key autocrine and paracrine regulator of MMP-9 secretion.

[Adenosine deaminase in differential diagnosis of peripheral lymph node tuberculosis]. / Adenosindezaminaza v differentsial'noi diagnostike tuberkuleza perifericheskikh limfaticheskikh uzlov.

Examining the activity of serum adenosine deaminase in 112 patients with lymphadenopathy of various etiology revealed its increase (at the discriminant level equal or exceeding 20 U/l) in 86% of 50 patients with active tuberculous infection. The specificity of the parameter as a test for tuberculous peripheral lymph nodal lesion is 90.3%, the positive and negative prognostic significances are 83 and 89%, respectively, the diagnostic efficiency is 88.4%.

[Effectiveness of intratracheal administration of contrykal in combined treatment of children with intrathoracic tuberculosis complicated by endobronchial pathology]. / Effektivnost' intratrakheal'nogo vvedeniia kontrikala v kompleksnom lechenii detei s vnutrigrudnym tuberkulezom, oslozhnennym éndobronkhial'noi patologiei.

Local balance in the system of proteinases-inhibitors in bronchial secretion was studied in 97 children with intrathoracic tuberculosis involving the bronchial tree by activity of trypsin-like proteinases (TLP), free antitryptic activity (ATA) and total content of locally synthesized low molecular acid resisting proteinase inhibitors (ARI). In the children with endobronchial pathology there was observed imbalance in the system evident from increasing activity of TLP and simultaneous decreasing of ATA and the content of ARI. The maximum inhibition of ARI local secretion and increasing of the TLP activity were stated in the patients with purulent and specific endobronchitis. In the children with catarrhal endobronchitis the shifts were moderate. Local use of contrykal in the form of ultrasonic inhalations in complex therapy of tuberculosis promoted in short periods reduction of the inhibitory potential, a 2-fold decrease in the terms of clinical recovery from endobronchitis and a decrease in the frequency of marked fibrous lesions in the bronchi and lungs as compared to analogous patients in the control group treated without the use of the proteinase inhibitor.

Immunohistochemical observation of intracytoplasmic lysozyme in proliferative and neoplastic fibrohistiocytic lesions.

Distribution of intracytoplasmic lysozyme in proliferative and neoplastic fibrohistiocytic lesions and non fibrohistiocytic tumors was studied by immunoperoxidase technique on formalin fixed, paraffin-embedded sections. The cases examined were 161 fibrohistiocytic lesions and 86 non-fibrohistiocytic tumors. Contrary to our expectation, the lysozyme positive cells were found only in the minority of cases with fibrohistiocytic lesions. Cells positive for lysozyme were found only in 13 out of 100 cases of dermatofibroma, one out of 4 cases of xanthogranuloma and 8 out of 33 cases of malignant fibrous histiocytoma. Dermatofibrosarcoma protuberans and non-fibrohistiocytic tumors were negative for lysozyme. It is suggested that in proliferative fibrohistiocytic lesions, induction of lysozyme synthesis is weak or absent. Some malignant fibrous histiocytomas showed scattered lysozyme positive neoplastic cells, indicating their probable histiocytic origin or differentiation. On the other hand, evidence of histiocytic differentiation of dermatofibrosarcoma protuberans was not obtained using lysozyme immunohistiochemistry.

Angiotensin-converting enzyme. I. Activity and correlation with serum lysozyme in sarcoidosis, other chest or lymph node diseases and healthy persons.

Serum angiotensin-converting enzyme (ACE) activity was studied in healthy controls, in 57 untreated sarcoidosis patients, and in 164 patients with other chest or lymph node diseases. The serum ACE activity of healthy persons was independent of sex, intake of meals, and smoking habits. There were no diurnal variations. Healthy children had a significantly higher ACE mean value than adults, whose ACE activity was not affected by age. The sarcoidosis patients had the highest ACE mean values, but those of patients with silicosis and asbestosis were also significantly elevated. Pulmonary cancer patients had decreased serum ACE activity, which was probably due to antimitotic treatment. Serum lysozyme (LZM) concentrations did not correlate with normal ACE activity, but the correlation between elevated ACE and LZM was significant in sarcoidosis and silicosis, and the trend was clearly the same for asbestosis. This indicates separate sources for these enzymes when ACE activity is normal, and a common source, i.e. macrophages, when ACE activity is increased. ACE production in certain diseases involving macrophages may be due to the bradykinin inhibiting effect of this enzyme.

Histochemistry of normal and diseased human lymph nodes.

Gomori's metal precipitate technique was used to demonstrate the phosphatase activity of the human cervical lymph node in health and disease, using four different phosphate esters (sodium beta-glycerophosphate and adenosine triphosphate at pH 9, riboflavin 5'-phosphate at pH 9.2 and 5'-monophosphoric acid at pH 8.3). In fetal lymph nodes, using 5'-monophosphoric acid, an outstanding positive activity was noticed in the lymphatic follicles. With the other three substrates there was either no nodular reaction or just a narrow rim of positive activity around the follicles, the internodular tissue being negative with all four substrates used. With chronic non-specific lymphadenitis the enzyme hydrolysing the three substrates (beta-glycerophosphate, riboflavin 5'-phosphate and adenosine triphosphate) began to make their appearance. It seems that with lymphadenitis, a qualitative change of the phosphatase activity takes place. A special characteristic pattern of phosphatase activity has been described in both 'early' and 'caseating' tuberculous lymphadenitis. In malignant lymphomas it was noticed that no activity was encountered with any of the four substrates in reticulum cell sarcoma. However, in lymphosarcoma a positive activity was obtained when either beta-glycerophosphate or adenosine triphosphate substrates was used, to the extent that one can depend upon this characteristic phosphatase activity in differentiating between reticulum cell sarcoma and lymphosarcoma. However, no enzymatic activity was obtained when the other two phosphate esters were used.