Thymus factor X (TFX-Jelfa)

Trichinella spiralis: Effect of thymus factor X on apoptosis and necrosis in mice

The aim of the study was to determine the effect of thymus factor X (TFX-Jelfa) on the percentage of apoptotic and necrotic lymphocytes in the spleen, mesenteric lymph nodes, and muscle tissue of mice infected with 200 larvae of Trichinella spiralis. TFX was administered subcutaneously at a dose of 15mg/kg. On days 7, 14, 21, 28, 35, 42, and 60 after infection, apoptotic and necrotic cells were detected by flow cytometry after staining with the Annexin V-Fluos Staining Kit. TFX increased the percentage of apoptotic lymphocytes in the spleen, mesenteric lymph nodes, and muscle tissue of mice infected with T. spiralis. The effect of TFX on the percentage of necrotic lymphocytes was weaker and less clear. Parasite load was lower in infected mice treated with TFX than in the untreated control mice. The effect of TFX on the host immune response and the survival of parasite larvae was therefore probably affected by the extent of inflammatory infiltrates, and not by the percentage of lymphocytes undergoing apoptosis.

Modulatory effects of calf thymus extract on the subset of T lymphocytes in Trichinella spiralis-infected mice

The immunotropic properties of calf thymus extract (TFX-Jelfa) is connected with the mimic action of the thymus to modulate the differentiation, maturation and function of prothymocytes and mature thymus dependent (T) cells. The studies were carried out on CFW male mice aged 3 months. The animals were infected per os with 200 larvae of Trichinella spiralis. TFX-Jelfa was administered i.p. at a dose of 10 mg/kg seven times at 24 hour intervals prior to infection. The percentage of CD4+ and CD8+ in suspension of splenocytes and mesenteric lymphonode cells by flow cytometry using monoclonal antibodies coupled with fluorescein isothiocyanate (FITC) or phycoerythrin (PE) were determined. At the same time, cryostat preparations, made from jejunum and muscle samples, were examined by the direct immunofluorescence method using FITC-labeled antibody to mouse CD4+ and CD8+. It has been found that infection with T. spiralis in mice decreases the percentage of CD8+ splenocytes, while the percentage of CD8+ mesenteric lymphonode cells does not change. However, in infected mice the percentage of CD4+ spleen cells and mesenteric lymphonode cells is increased. It has been also found that during the course of infection an increase in the number of CD8+ and CD4+ cells in the basal lamina propria of the intestines was observed. In infected mice, CD4+ lymphocytes were visible in the inflammatory infiltrates of the muscle tissue on the 14th day, whereas CD8+ lymphocytes were first observed a week later. Pretreatment with TFX does not change the inhibitory effect of infection on the percentage of CD8+ splenocytes, but potentiates the percentage of CD4+ spleen cells and mesenteric lymphonode cells increased by infection. Furthermore, administration of TFX prior to infection also potentiates the stimulatory effect of T. spiralis on the number of CD8+ and CD4+ in the basal lamina propria of the jejunum, and on the number of CD8+ cells in the inflammatory infiltrates of the muscle tissue.

[Effect of TFX preparation on the course of pregnancy in rabbits with experimental antiphospholipid syndrome]

Recurrent thrombo-embolic episodes and pregnancy loss, thrombocytopenia and the presence of antiphospholipid antibodies, first described in 1983 by Hughes and defined by Harris in 1987, are characteristic of the primary antiphospholipid syndrome (APS). APS is the cause of obstetrical problems in the form of recurrent miscarriages, intrauterine fetal death or growth retardation, and EPH gestosis. Clinical symptoms described above are probably mediated by antiphospholipid antibodies which interact with endothelial and trophoblastic cells, blood platelets, embryonic tissue cells, as well as with coagulation factors and proteins involved in the coagulation cascade or in antibody binding. Management of APS includes antiaggregation, anticoagulation, immunosuppression, and intravenous administration of gamma globulins. Successful treatment is not always the case and search for more efficient therapies continues. The importance of animal experiments led to the design at the Department of Pathology of Pregnancy and Labour of an APS model in pregnant and nonpregnant rabbits. As Turowski et al. have confirmed the immunomodulating action of TFX in rabbits, it seemed justified to examine the properties of this preparation in pregnant rabbits with experimentally induced APS. The material consisted of 30 pregnant New Zealand rabbits, divided into the following groups: I--10 pregnant rabbits (and 63 fetuses) treated intradermally twice weekly since the 10th day of pregnancy with cardiolipin together with adjuvant; I-K--5 pregnant rabbits (and 17 fetuses) treated with cardiolipin and adjuvant in the same manner as group I and additionally with intramuscular injections of 0.9% NaCl on the 20th, 21st, and 22nd day; I-T--10 pregnant rabbits (and 66 fetuses) treated with cardiolipin with adjuvant in the same manner as group I and additionally with intramuscular injections of 10 mg/day of TFX (Jelfa, Poland) on the 20th, 21st, and 22nd day of pregnancy; K--5 pregnant rabbits (and 27 fetuses) treated with injections of 0.9% NaCl twice weekly. Blood samples for laboratory analysis were collected by cardiac puncture before immunization (sample I) and on the day of caesarean section (sample II). Platelet counts and APTT tests were done. Numbers of live and dead newborns, resorbed fetuses, body mass, newborn viability and survival rates were recorded. TFX administered to pregnant rabbits with experimentally induced antiphospholipid syndrome increased the number of live newborns, reduced the incidence of fetal resorption, increased the viability and survival rate of newborn rabbits. The beneficial effect of TFX on APTT and platelet count was revealed, such that these parameters remained within the normal range despite immunization. Morphological changes observed in the placenta were not specific.

Effect of calf thymus extract and zinc supplementation on the cellular response of mice exposed to restraint stress

The studies were carried out on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to stress exposure, the mice were treated with calf thymus extract (TFX - Jelfa) i.p. at a dose of 10 mg/kg, ten times at 24 h intervals. TFX was used per se or with zinc ions interaction, by adding zinc ions (as sulfate salt) to drinking water at a dose of 72 microg/mouse per day. The results obtained show that restraint stress dramatically decreased the total number of thymocytes and splenocytes which is also accompanied by decreasing weight ratio of the thymus and spleen. The decreasing number of thymic and spleen cells corresponded to a diminishing percentage of immature, double-positive CD4+CD8+ thymocytes, mature single-positive CD4+ thymic cells and CD4+, CD8+ and CD19+ splenocytes. Changes in the number of thymic cells affect their activity, which is expressed as a decreased proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohaemagglutinin (PHA). Besides, exposure to the restraint stress decreased interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolisacharide (LPS) from E. coli. Previous treatment with TFX counteracted restraint stress-induced immunosuppression, which is expressed as partial normalisation of the total number of thymic and spleen cells, accelerated regeneration of these two lymphatic organs, shortned suppressive action of restraint stress on the percentage of immature CD4+CD8+ thymocytes and CD4+ splenocytes and in total normalisation of the CD4+ thymocytes and CD8+ splenocytes. TFX administered prior to restraint stress not only counteracted the suppresive effects of stress on the proliferative activity of thymic cells stimulated in vitro with Con A and PHA, but also augmented the proliferative response of these cells to two mitogens. The immunorestorative effect of TFX was augmented by zinc supplementation.

[Primary sclerosing cholangitis--tests with ubiquitin treatment. Case report]

Treatment of chronic liver disease, including primary sclerosing cholangitis (CSP) is a difficult and still not fully solved problem. Both monotherapy and combined pharmacological therapy have shown little effect in inhibiting the disease process and preventing complications. The objectives of ubiquitin biotherapy were restitution of the immune system and inhibition of the disease process along with the stimulation of regenerative processes of the liver. In 1994 in Gastroenterological Clinic attempts at ubiquitin biotherapy were made with the use of thymus extract (TFX-JELFA ini.), which proved to be the ubiquitin preparation that was not only active in the immune system but also played a significant role in regenerative and adjuvant processes. It has been shown that ubiquitins play an essential role in proteolysis of proteins, their intracellular elimination and in the apoptosis. The early results of the clinical observations and laboratory tests indicate a gradual improvement and inhibition of the disease process the patient with CSP.