alpha-chymotrypsin and Brain-Neoplasms

In cancer treatment an attempt has been made to pharmacologically regulate the proteasome functions, thus the aim was to test whether 20S proteasome chymotrypsin-like (ChT-L) activity has a role in glial brain tumors. Furthermore, we analyzed the correlation between proteasome activity and IL-8, CCL2, NF-κB1 and NF-κB2 concentrations, which impact on brain tumors has already been indicated.. Plasma 20S proteasome ChT-L activity was assayed using the fluorogenic peptide substrate Suc-Leu-Leu-Val-Tyr-AMC in the presence of SDS. IL-8, CCL2, NF-κB1 and NF-κB2 concentration was analyzed with the use of ELISA method. Immunohistochemistry for IDH1-R132H was done on 5-microns-thick formalin-fixed, paraffin-embedded tumor sections with the use of antibody specific for the mutant IDH1-R132H protein. Labelled streptavidin biotin kit was used as a detection system.. Brain tumor patients had statistically higher 20S proteasome ChT-L activity (0.649 U/mg) compared to non-tumoral individuals (0.430 U/mg). IDH1 wild-type patients had statistically higher 20S proteasome ChT-L activity (1.025 U/mg) compared to IDH1 mutants (0.549 U/mg). 20S proteasome ChT-L activity in brain tumor patients who died as the consequence of a tumor (0.649) in the following 2 years was statistically higher compared to brain tumor patients who lived (0.430 U/mg). In brain tumor patients the 20S proteasome ChT-L activity positively correlated with IL-8 concentration.. Elevated 20S proteasome ChT-L activity was related to the increased risk of death in glial brain tumor patients. A positive correlation between 20S proteasome ChT-L activity and IL-8 concentration may indicate the molecular mechanisms regulating glial tumor biology. Thus research on proteasomes may be important and should be carried out to verify if this protein complexes may represent a potential therapeutic target to limit brain tumor invasion.

Topics: Adult; Aged; Biometry; Brain Neoplasms; Chemokine CCL2; Chymotrypsin; Cysteine Endopeptidases; Female; Glioma; Humans; Interleukin-8; Male; Middle Aged; Neuroglia; NF-kappa B; Peptides; Proteasome Endopeptidase Complex; Proteolysis; Serine Endopeptidases

The expression of glial fibrillary acidic protein, fibronectin (FN), factor VIII-related antigen (FVIII/RAG), and of three monohistiocytic markers, lysozyme, alpha-1-antitrypsin and alpha-1-antichymotrypsin was examined in five gliosarcomas (GS) by peroxidase-antiperoxidase immunostaining of formalin-fixed and paraffin-embedded specimens, and compared with vascular changes in 16 glioblastomas (GB). In contrast to GB, endothelial proliferations of GS were sheathed by sarcomatous tissue (perivascular sarcoma), which was contiguous with fibrosarcomatous areas. Cells with conspicuous intracytoplasmic FN content (FN+ cells) were seen in the vascular stroma of GB and dominated in the sarcomatous parts of GS. Most FN+ cells of GS were of varying size and shape and clearly neoplastic. Monohistiocytic markers were demonstrable in small infiltrating mononuclear cells as well as in many sarcomatous cells including FN+ cells. FVIII/RAG was restricted to lumen-lining endothelium and was not found in sarcomatous cells. These results suggest that a major part of sarcoma in GS is less likely to develop from proliferated endothelial cells than from histiocytic cells in the perivascular spaces of GB. By FN mediation, histiocytic cells might also guide and promote sarcomatous proliferations of other mesenchymal cells, leading to fibrosarcomatous development. Prominent monstrous giant cells of one GS seemed to be degenerating glioma cells.

Topics: Adult; Antigens; Brain Neoplasms; Cell Division; Chymotrypsin; Factor VIII; Fibronectins; Glial Fibrillary Acidic Protein; Glioma; Histiocytes; Humans; Male; Middle Aged; Muramidase; Neoplasm Proteins; Trypsin; von Willebrand Factor

The phenomenon of glioma killing by lymphokine activated killer cells (LAK) was studied. We demonstrate that LAK cells generated by culturing the lymphokine interleukin-2 (IL-2) with peripheral blood lymphocytes from brain tumour patients destroys autologous glioma. The rat 9L glioma model was used to show that LAK killing was tumour-selective as glioma but not syngeneic normal brain tissue was destroyed. The susceptibility of both human and 9L rat glioma to LAK cell killing was markedly diminished by pretreating glioma cells with trypsin or chymotrypsin, but was unaffected by pretreatment with neuraminidase, glycosidases, sodium periodate or hydrocortisone. These results suggest that the cell surface determinant on glioma cells responsible for its tumour selective lysis by LAK is a protein sensitive to trypsin and chymotrypsin. The tumour-selective killing of glioma by LAK in vitro prompted the initiation of a Phase I study in which ten patients with malignant glioma have been treated with direct intracerebral injection of IL-2 or LAK without evidence of systemic or brain toxicity.

Topics: Adult; Aged; Animals; Borohydrides; Brain Neoplasms; Cell Line; Chymotrypsin; Cytotoxicity, Immunologic; Disease Models, Animal; Female; Glioma; Humans; Interleukin-2; Killer Cells, Natural; Lymphocyte Activation; Male; Middle Aged; Periodic Acid; Rats; Rats, Inbred F344; Trypsin

Normal and diseased human central nervous system (CNS) tissues were studied immunohistochemically by a monoclonal antibody to human macrophages (EBM/11), antisera to glial fibrillary acidic protein (anti-GFAP), and alpha-1-antichymotrypsin (alpha 1-ACT). EBM/11 reacted with brain macrophages located mainly around blood vessels in normal brain; it also reacted with resting microglia in normal brain and with numerous reactive microglia and macrophages in brain tumours and inflammatory lesions. Microglia did not react with anti-GFAP or alpha 1-ACT. An EBM/11 positive phenotype, therefore, is shared by microglia and macrophages and suggests that microglial cells form a specialised part of the mononuclear phagocyte system.

Topics: alpha 1-Antichymotrypsin; Antibodies, Monoclonal; Astrocytes; Astrocytoma; Brain; Brain Neoplasms; Chymotrypsin; Glial Fibrillary Acidic Protein; Glioma; Humans; Macrophages; Neuroglia; Trypsin Inhibitors