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M Bilej

Publications and source records attributed to M Bilej.

At least 19 recordsLinked to original sources
PubMed1998

Cellular expression of the cytolytic factor in earthworms Eisenia foetida.

Coelomic fluid of earthworms contains a 42 kDa protein designated CCF-1 (coelomic cytolytic factor 1), which accounts for approximately 40% of cytolytic activity of the entire coelomic fluid. CCF-1 was documented to be present on cells of the mesenchymal lining of the coelomic cavity as well as on free coelomocytes. Both cellular and humoral levels of CCF-1 were significantly increased after parenteral injection of endotoxin. Moreover, CCF-1 seems to be involved in cell mediated cytotoxicity, because cytotoxic activity is blocked in the presence of anti-CCF-1 monoclonal antibody (mAb).

Animals↗
PubMed1997

Lesion of leukocytes, erythrocytes, and mesothelial cells by the coelomic fluid of Eisenia foetida earthworms.

Coelomic fluid of Eisenia foetida earthworms is known to exert strong proteolytic, hemolytic, bacteriostatic, and cytolytic properties. Ultrastructural observations revealed that coelomic fluid causes multiple ruptures and defects in the erythrocyte membrane as well as in the membrane of murine peritoneal leukocytes. Incubation of peritoneal cells in coelomic fluid resulted in a disorganization of the macrophage surface microvilli, changes in the organization of cytoplasmic organelles and disruption and degranulation of mast cells. Severe mesothelial damage was observed after intraperitoneal administration of the coelomic fluid.

Animals↗
PubMed1997

[Charge interactions of immune deposits in glomeruli (experimental study)].

An i.v. injection of 8-40 mg (kg cationized and heat-aggregated rabbit or human Ig (cat-aggr RIg,-HuIg; pI 9.5) elicited a strong diffuse linear fixation in rat glomerular capillaries revealed by one-step immunofluorescence or immunoenzyme histochemistry 1 and 2 h post-injection. Preferential binding to the lamina rara externa (LRE) was documented in ultrastructure by preembedding and postembedding assays (HRP-coupled antibody and protein A-colloidal gold, respectively). After 24 and 48 h the glomeruli were negative. Polyethylenimine (PEI)-reactive polyanion of LRE was significantly reduced 1 h after cat-aggr-Ig; depletion persisted even after 48 h. Non-cationized Ig aggregates did not bind to the glomerular capillaries. A subsequent i.p. injection of swine anti-rabbit-Ig antibody (SwAR, 15 mg i.p. after 4 h) produced the same linear binding of both two antigens which, however, persisted after 10 days and assumed a granular pattern. After presensitization with RIg (1-2 mg i.p. or s.c.; 4 days before cat-aggr RIg) the early linear fixation underwent a gradual transformation into the granular pattern and deposits of mesangial, rarely of epimembranous type were found 1 week after cat-aggr RIg and later. RIg and SwIg were proved in both types of deposits. After 2 weeks both rat Ig and C 3 were present, too. Rarefaction of deposits and their concentration in the vascular poles took place during 3 months, and deposits also appeared in the media of vas afferens. The antigen load did not produce an acute glomerulonephritis or significant proteinuria; slight focal mesangial sclerosis and a discrete increase in serum creatinine were noted after 2-3 months. To sum up: The one-shot charge interaction is prompt but short-lived whereas the local binding of additional proteins, especially after a specific preimmunization, significantly prolongs the contamination of glomeruli and promotes the build-up of immune complex-type deposits which gradually retreat to the mesangial stalk and vascular pole.

Animals↗
PubMed1995

Characterization of the limited specificity of antigen recognition in earthworms.

Earthworms respond to parenteral stimulation with a protein antigen by the formation of an antigen-binding protein. Earthworms were parenterally stimulated with different proteins and the protein binding was estimated in vitro on both humoral and cellular levels. The binding was significantly higher when the same protein was used for in vivo stimulation. The degree of specificity of the antigen-binding protein after the secondary in vivo challenge increased, but even so it was considerably lower than that of vertebrate immunoglobulins.

Adjuvants, Immunologic↗
PubMed1995

Identification of a cytolytic protein in the coelomic fluid of Eisenia foetida earthworms.

Total coelomic fluid of earthworms Eisenia foetida (Oligochaeta, Annelida) is capable of lysing different mammalian tumor cell lines. This cytolytic activity is different from tumor necrosis factor (TNF)-mediated lysis and is not due to proteolysis. Total coelomic fluid was subjected to ion-exchange chromatography separation and a fraction with prominent cytolytic activity was used to elicit monoclonal antibodies that were screened for their capacity to neutralize the cytolytic effect of total coelomic fluid. One of the prepared neutralizing IgG antibodies was used for the immunoaffinity purification of a cytolytic factor from total coelomic fluid. SDS-PAGE and Western blot analyses revealed a protein band with an apparent molecular weight of 42 kDa. This cytolytic protein (termed CCF-1 or coelomic cytolytic factor 1) can be adsorbed on the surface of opsonized particles and may be involved in opsonizing and hemolytic effects of coelomic fluid.

Animals↗
PubMed1994

Antigen processing in earthworms.

The administration of protein antigens into earthworms Lumbricus terrestris and Eisenia foetida induces the formation of antigen-binding protein (ABP) with the maximum response occurring between days 4 and 8. High proteolytic activities observed both in coelomocytes and in coelomic fluids cause rapid antigen degradation; the majority of antigen is digested during the first 24 h. To analyze the role of proteolytic processing of antigen in ABP response in vitro, the intact antigen (ARS-HSA) as well as its proteolytic fragments were added to tissue explant cultures and ABP formation followed by indirect ELISA using mAb to ABP. The response to small fragments was comparable to that induced by intact ARS-HSA. Furthermore, the response to intact antigen was almost completely blocked by the non-toxic serine protease inhibitor Pefabloc, while the response to small fragments was only slightly reduced. The kinetics of response to intact antigen significantly differed from that induced by small (< 3 kDa) fragments. We suggest that proteolytic processing is involved in the stimulation of ABP formation.

Animals↗
PubMed1993

Hemolytic function of opsonizing proteins of earthworm's coelomic fluid.

Synthetic 2-hydroxyethylmethacrylate copolymer particles (HEMA) can be opsonized in the coelomic fluid of Eisenia foetida earthworms. The incomplete coelomic fluid (i.e., the coelomic fluid after incubation with HEMA particles) exerts a lower level of hemolytic activity compared to complete coelomic fluid. The decreased hemolysis can be compensated by the addition of isolated opsonins. On the other hand, isolated opsonins do not possess direct hemolytic capacity. It can be suggested that at least one of the isolated opsonins is involved in the hemolytic process. These results support the hypothesized cooperation of humoral and cellular mechanisms in earthworm defence.

Animals↗
PubMed1993

The fate of protein antigen in earthworms: study in vivo.

Parenteral administration of antigen induces in the earthworms Lumbricus terrestris and Eisenia foetida the formation of antigen-binding protein. Labelled antigen was efficiently digested in the coelomic fluids of both species since the total amount of radioactivity in the coelomic fluids decreased. Antigen internalized in the free coelomocytes of both species was present in a TCA-precipitable form, although the decline of total radioactivity was higher than in the coelomic fluid, indicating that the TCA-nonprecipitable fragments are rapidly released from coelomocytes into the coelomic fluid. Within the first 24 h after administration, labelled antigen was detected throughout all tissues except for the epidermis and cuticle, and after 4 days it was found in the chloragogenic tissue around the gut, dorsal vein and typhlosole.

Animals↗
PubMed1993

The fate of protein antigen in earthworms: study in vitro.

Earthworm coelomocytes digest protein antigen in vitro. Proteolytic activity was detected both in cell-free coelomic fluid and in cell cultures of free coelomocytes which are effectors of earthworm immunodefense mechanisms. Antigen is cleaved either intracellularly or by proteolytic enzymes released by coelomocytes into the medium. Proteolysis was observed both in non-stimulated and antigen-stimulated cultures. Since significantly higher proteolysis was shown in supernatants from cultures of antigen-stimulated coelomocytes, we can assume that the release of proteolytic enzymes was inducible.

Animals↗
PubMed1992

The effect of an antigenic stimulation on proliferative response in athymic mice.

The proliferation activity of the main cellular categories of bone marrow after infusion of 3H-thymidine was studied in nu/nu and +/+ 1-month- and 3-month-old BALB/c mice in comparison with lymphoid cells in the spleen and mesenteric lymph nodes. The stem cell defect in nu/nu mouse bone marrow is compensated by an increased proliferation in myeloid series and in agranulocytes. The increase of proliferation activity among lymphoid cells in peripheral lymphoid organs was observed only in the 3-month-old mice with a delay in the nudes.

Animals↗
PubMed1992

Repeated antigenic challenge induces earthworm coelomocyte proliferation.

Annelids are able to protect themselves against foreign materials by natural and acquired cellular immunity. The aim of this study was to characterize the kinetic of antigen induced proliferation of earthworm coelomocytes. The proliferative activity of free coelomocytes of Eisenia foetida decreased after a second contact with the same antigen. Precursor cells in the mesenchymal lining of the coelomic cavity responded to stimulation immediately. These results indicate that for a successful triggering of the proliferative response of the free coelomocytes repeated contact with stimulating agents is needed.

Animals↗
PubMed1992

New trypsin inhibitors are present in the coelomic fluid of the earthworm Lumbricus terrestris.

Proteinase-inhibiting components of the coelomic fluid of the earthworm Lumbricus terrestris were examined. Inhibition of proteinases of serine, aspartate and thiol families was tested. Very strong inhibition was observed only in the case of trypsin. Additional data suggest that the inhibition is related to proteins of molar mass of 42 kDa and 20 kDa, respectively. These two proteins are present in the coelomic fluid in several forms which differ in their isoelectric points.

Animals↗
PubMed1991

The immunosuppressive effects of bilirubin.

The strong effects of bilirubin on various levels of the immune system are multifactorial. Concerning the mechanisms of these effects, we hypothesize that the primary causes of the described actions of bilirubin are the direct interaction of bilirubin molecules with cell membranes.

Bilirubin↗
PubMed1991

Characterization of antigen-binding protein in earthworms Lumbricus terrestris and Eisenia foetida.

Injection of antigen into the annelid worms Lumbricus terrestris (LT) and Eisenia foetida (EF) results in a marked increase of coelomic fluid protein concentration and the formation of a protein which binds the stimulating antigen (3). In this report we show that the increases in total protein concentration after first and second doses of antigen were higher and were achieved earlier in LT than in EF, while the accumulation of antigen-binding protein in coelomic fluid was similar in both species. Antigen-binding protein isolated by affinity chromatography retained its original binding activity. Its molecular weight in coelomic fluid as well as after isolation was 56 kD when analyzed by SDS-PAGE and immunoblotting. Under reducing conditions, two bands with mol./wt. 31 and 33 kD appeared which did not reveal detectable binding activity. This suggests that the 56 kD binding protein of annelids is composed of two disulphide-linked polypeptide chains both of which participate in antigen-binding site formation.

Animals↗