Thymic peptides restrain the inflammatory response in mice with experimental autoimmune encephalomyelitis.

Modulation of autoimmune inflammation by the thymic peptides thymulin and thymopentin was studied in mice with acute experimental autoimmune encephalomyelitis (EAE), which resembles multiple sclerosis in humans. EAE was induced in NZW mice by a single immunisation with myelin basic protein coupled with adjuvants. Visible signs of pathology appeared on days 12-14 after the immunisation, peaked on days 20-25, were retained up to day 45, and then reverted. A biphasic cytokine response was also detected. In the "early" phase, which started at day 35, increased levels of interferon-gamma and interleukin-6 in the blood were observed; during the "delayed" phase, which started at day 48, the levels of plasma interleukin-17 and tumour necrosis factor-alpha were also raised. In addition, the phosphorylation of NF-kappaB signalling proteins and the production of heat shock protein Hsp72 were significantly increased in splenic lymphocytes from EAE-bearing mice. When applied intraperitoneally every other day for 30 days, either thymulin or thymopentin (15 µg per 100g of body weight) significantly reduced the disease severity compared to untreated EAE mice. The effect of thymulin but not thymopentin remained after its withdrawal. Thymulin reduced the cytokine response in both the early and the delayed phases, whereas thymopentin only reduced the "early phase cytokines" (IL-6 and interferon-gamma). Both peptides significantly reduced the level of phosphorylation of the NF-kappaB signalling protein IKK and the production of Hsp72 protein. The data presented here indicate the presence of time-dependent immune responses in EAE-bearing mice, which may be associated with the Th1 and Th17 subpopulations of T-cells. Thymulin and thymopentin demonstrated different patterns of activity, most likely via mechanisms involved in NF-kappa B signalling and Hsp72 expression.

[Correction of immunodeficiency status caused by herbicide 2,4-D in experiment]. / Korrektsiia immunodefitsitnogo sostoianiia, vyzvannogo gerbitsidom 2,4-D v éksperimente.

A possibility to correct the immunodeficiency state, caused by herbicide 2.4-D (20 mg/kg of weight daily during 5 days), by using immunomodulators Levamisole (perorally), Tactivinum (hypodermically) and Spleninum (hepodermically) in reactions Graft-versus-Host (GH), Antibody-Forming Cells (AFG) and phagocytic activity of peritoneal macrophages in vivo and in vitro etc, was studied in experiments with 360 mice of lines CBA and F1 (CBAxC57B1/6) weighing 18 to 22 g. It was established that Levamisole (2.5 mg/kg per body weight, for 1 to 3 days) and Tactivinum (0.2 mkg/mouse, for 1 to 3 days) had an immunocorrecting effect in poisoning by 2.4-D. Spleninum (10 mcl/mouse) corrected mainly the humoral chain in the immune response with the AFG recovery to the level observed in intact animals (controls). The data on the influence produced by the immunomodulators on the phagocytic activity in vitro correlated with such data obtained in vivo.

Lipid microparticles as a parenteral controlled release device for peptides.

To investigate the potential of physiological lipids as an alternative to synthetic polymeric materials such as poly(lactide-co-glycolide), peptide-containing glyceryl tripalmitate microparticles were prepared. A modified solvent evaporation method and a melt dispersion technique without the use of organic solvent were employed. Thymocartin (TP-4), an immunomodulating tetrapeptide and insulin were chosen as model peptides and incorporated as a solid or dissolved in 100 microl aqueous solution. The resulting microparticles were characterized with respect to particle size and morphology, biocompatibility, drug content (encapsulation efficiency) and in vitro release behavior. Electron spectroscopy for chemical analysis was used to investigate the adsorption of the model peptides to the lipid matrix material. The modified solvent evaporation as well as the melt dispersion method were suitable for the preparation of microparticles in the size range of 20-150 microm with an acceptable yield. The biocompatibility of the glyceryl tripalmitate microparticles after implantation into NMRI-mice was comparable to poly(lactide-co-glycolide) microparticles. The encapsulation efficiency for both model peptides was high (>80%) even at high theoretical loadings when the peptide was incorporated as a solution with the melt dispersion technique. The in vitro release behavior was substantially influenced by the physicochemical properties of the model peptides used in this study.

Preparation of liposome formulations containing immunomodulatory peptides.

Some series of liposomal formulations are described, loaded with three oligopeptides: TP5, TP4, and TP3 derived from the thymic hormone thymopoietin. Liposomes, both multilamellar (MLV) and stable plurilamellar vesicles (SPLV) were prepared with and without cholesterol, and contained stearylamine or dipalmitoylphosphatidylserine to induce a positive or negative charge, respectively. The encapsulation efficiency and the rate of release of the entrapped drugs, after incubation at 37 degrees C, were evaluated and results were discussed as a function of liposome type, charge and composition. In vitro evaluation of cytotoxic activity of the three peptides on K562 and IM9 human tumor lines showed in some cases (TP3 and TP5-loaded positively charged MLV; IM9 cells) a slight increase of the activity of peptides. However, a relevant intrinsic toxicity of pure phospholipids (empty liposomes) on cell cultures was observed, which altered the evaluation of drug-loaded formulations activity.

[Combined chemotherapy of toxic hepatitis in patients with pulmonary tuberculosis]. / Kombinirovannaia farmakoterapiia toksicheskikh gepatitov u bol'nykh tuberkulezom legkikh.

The follow-up involved patients with infiltrative pulmonary tuberculosis who developed toxic medicamentous hepatitis associated with antituberculosis drugs. They were treated by the method of combined pharmacotherapy which comprised sodium nucleinate (0.5 g 4 times daily), splenin (2 ml twice a day) and quercetin. The given combination of drugs rapidly improved clinical parameters and normalized immunologic tests.

[Enterosorption in the treatment of ulcerative colitis patients]. / Enterosorbtsiia v lechenii bol'nykh iazvennym kolitom.

Enterodes and splenin were used in the treatment of patients with ulcerative colitis (idiopathic proctocolitis) in association with routine drug treatment while steroids were not employed. Results indicate increase of the therapeutic effect due to improvement of regeneration of the intestine and improvement of the immunity status.

[Effect of humoral factors of the spleen after irradiation]. / Osobennosti deistviia gumoral'nykh faktorov selezenki pri obluchenii.

It is stated that therapeutic effect of humoral factors isolated from the cattle spleen is associated with its influence on alpha 2-macroglobulin (alpha 2-MG) performing a protective function. The results obtained permit recommending the isolated preparations to increase total resistivity of the organism under irradiation.

[Intra-lesional beta-interferon in combination with systemic thymopentin. Evaluation of results in 35 cases of CIN III and 2 cases of VAIN associated with HPV]. / beta-interferone intralesionale più timopentina sistemica. Valutazione su 35 CIN III e 2 VAIN III associate ad HPV.

We performed an open study on 37 patients (average age 35 years), with CIN III or VAIN III and Viral Cytopatic Effects (VCE), who underwent a new standardized bifasic therapy by means of intralesional beta-interferon, topic beta-interferon and subcutaneous timopentine injection. Each therapeutic and checking step was made by colposcopic and microcolpohysteroscopic inspection, which showed spreading necrotic zones in the dysplastic places and peripheral typical epithelium replacement. Microcolpohysteroscopy allowed us to obtain correct diagnosis of the lesion and its location, to discriminate each pathologic aspects (CIN, VAIN, VCE), to perform an adeguated biopsy and intralesional therapy and to follow-up lesion course without repeated biopsies. After two months of therapy as maximum safety limit, we performed conization (in CIN case) in order to confirm the effects of therapy by hystology and especially to evaluate the deep lesional border. The istologic examination underlined the previous microcolpohysteroscopic report of dysplastic regression until its disapperance, with lasting VCE in all the cases.

Thymopentin antagonizes stress-induced changes of GABA/benzodiazepine receptor complex.

Intraperitoneal administration of thymopentin, a thymopentin II-derived pentapeptide, had no stable and evident effect in the two anxiety models (elevated plus-maze and licking-conflict test) studied. However, in the elevated plus-maze test thymopentin antagonized the behavioral effects of DMCM, a beta-carboline derivative with anxiogenic properties. Further, it was demonstrated that the licking-conflict test procedure itself produced a significant elevation of plasma corticosterone levels, increased the number of [3H]flunitrazepam and decreased the number of [3H]muscimol binding sites in rat hippocampus. The forced-swimming stress similarly to the licking-conflict test also caused an increase in hippocampal [3H]flunitrazepam binding sites. Although ineffective behaviorally in the tests for anxiety, thymopentin pretreatment effectively reversed the changes in corticosterone levels caused by the licking-conflict test. Moreover, it normalized the changed number of benzodiazepine and GABA receptors after stressful stimuli. It is well known that not all anxiolytic drugs (i.e. buspirone) are equally active in behavioral tests for anxiety. According to our data we propose that thymopentin has stress-protective activity. As in vivo and in vitro thymopentin did not change [3H]-flunitrazepam and [3H]muscimol binding, the direct effect of this peptide on the GABA-benzodiazepine-Cl- ionophore receptor complex is unlikely. The action of this peptide on GABA release and/or metabolism can be suggested.

[Therapeutic use of splenopentin (DA SP-5) in patients with psoriasis arthropathica]. / Therapeutische Anwendung von Splenopentin (DA SP-5) bei Patienten mit Psoriasis arthropathica.

8 patients with confirmed psoriatic arthritis were treated with splenopentin for 12 months. A relief of joint pain could be detected under this treatment both after 6 weeks (improvement of the Ritchie-indices: 44%) and after 12 months (improvement: 28%). In contrast to this, X-ray investigations show a worsening of the disease after 12 months. Therefore we conclude that splenopentin alone is not a sufficient therapeutic drug in patients suffering from psoriatic arthritis.

Effect of immunomodulator thymopentin on impaired seroresponse to influenza vaccine in patients on haemodialysis.

In a double-blind placebo-controlled multicentre study, the effect of the immunomodulator thymopentin (TP5) on the antibody production to trivalent influenza vaccine was tested in 108 patients on chronic intermittent haemodialysis (HD). Antibody production was determined in pre- and postvaccination sera. Compared to a group of 35 young healthy adult control subjects, HD patients showed a clearly impaired seroresponsiveness to all three vaccine components, regardless of treatment with TP5 or placebo. We conclude that TP5 administration is not able to enhance humoral immunity in patients on chronic intermittent HD.

Thymopentin in the treatment of juvenile chronic arthritis.

An immunological imbalance is probably one of the major pathogenetic causes of rheumatoid arthritis in adults as well as in children. This aspect is the rationale for the use of immuno-modulating drugs. In our study we evaluated the effects of intravenously-administered thymopentin on systemic and local features in 10 children affected with systemic onset juvenile chronic arthritis (JCA). We also considered the effects of intra-articular thymopentin in 3 children affected with pauciarticular onset JCA.

[Levamisole and splenin in combined therapy of patients with bronchiectasis]. / Levamizol i splenin v kompleksnoi terapii bol'nykh bronkhoéktaticheskoi bolezn'iu.

Follow-up of 114 patients with bronchiectasis has demonstrated that immunomodulation agents levamisole and splenin included in combined treatment augment their therapeutic effect and normalize the immune status of patients. This is manifested by normalization of the amount and functional activity to T-lymphocytes, the content of IgG, IgM and IgA and the autoimmune shifts. A favourable effect was encountered significantly more often than that in patient who were treated by the traditional methods without the use of the immunomodulation agents and in those who were given only levamisole and splenin together with anti-bacterial and hygienic therapy.

[The effect of immunotherapy with thymopentin on the parameters of cellular immunity and the clinical course of gynecologic tumor patients]. / Einfluss einer Immuntherapie mit Thymopentin auf Parameter der zellulären Immunität und den klinischen Verlauf von gynäkologischen Tumorpatientinnen.

In 25 patients with cervical, uterine and ovarian cancer with a depletion of T-helper cells and a lack of reactivity in the LMI-Test a course of immunotherapy with thymopentin (Timunox, Cilag Biotech) at a dosage of 100 mg/die for 2 weeks and subsequently 100 mg 3 x/week for 4 weeks was performed and a comparison was made with a control group of 59 patients receiving no immunotherapy. The following parameters of cell-mediated immunity were determined: (1) leucocyte-migration-inhibition-test (LMI-test) against the tumor and varidase, and (2) number of T-helper/inducer-, T-suppressor/cytotoxic-, total T-, and natural killer cells in the peripheral blood. After a course of immunotherapy no changes in the mean value of the percentage of the cell-subpopulations could be observed, but in a quarter of all patients a reaction in the LMI-test could be shown. In the control group, under chemotherapy, the percentage of T-helper-, T-suppressor- and total T-cells decreased strongly, whereas in the immunotherapy group the percentage of these populations remained unchanged. Under the restriction of a mean observation time of only 18 (5-32) months no differences concerning the survival time between both groups could be observed; only in advanced ovarian cancer patients could a slight positive effect of immunotherapy be shown. These results support the indication of a course of immunotherapy with thymopentin in combination with chemotherapy, especially in advanced cancer patients.

[Stimulating effect of biologically active substances from the spleen on the functional activity of hepatocytes]. / Stimuliruiushchii éffekt biologicheski aktivnykh veshchestv selezenki na funktsional'nuiu aktivnost' gepatotsitov.

Studies in the effect of both splenin and spleen "protein-free extract" on the monolayer culture of chick hepatic embryos have revealed that small doses exert a stimulating effect on hepatocytes, while the large ones induce degenerative changes. Hepatotrophic characters of the "protein-free extract" are determined, while utilizing lower doses than those of splenin.

[Telethermographic evaluation of the intra-articular administration of thymopentin in rheumatoid arthritis of the knee]. / Valutazione teletermografica dell'impiego della timopentina per via intraarticolare nell'artrite reumatoide del ginocchio.

Fifteen subjects with rheumatoid arthritis defined as "classical" or "definite" according to A.R.A. criteria and classified according to Steinbrocker as stages I to III were submitted to knee joint infiltration (bilateral where both knees were involved) with 50 mg thymopentin in 1 ml, once a week for five weeks. Telethermographic examination was performed in all cases in order to evaluate the thermal inflammatory component before the first and 48 hours after the last infiltration. In none of the patients with the exception of only one case, did the drug lead to a significant change of the thermal index. This evaluation does not take into account any other objective and subjective parameters the study of which was beyond the scope of the above research.

[Thymopentin in the treatment of recurrent condylomata acuminata]. / La timopentina nel trattamento dei condilomi acuminati recidivanti.

Twelve patients who had been suffering from genital and/or perianal recurrent condyloma acuminatum for a minimum of 5 to a maximum of 24 months, in spite of treatment, were studied from the immunological viewpoint and treated with 50 mg s.c. Thymopentin three times a week for 4 or 6 weeks. Six of the patients were cured at the end of treatment, five after 5 months, and one was not cured. Analysis of the clinico-laboratory data shows a significant agreement between the course of clinical signs and the immunological picture. The various cure stages are probably attributable to the basic immune arrangement which was more impaired in the 5 patients who were cured more slowly and in the non-cured case. In the latter too, however, Thymopentin permitted correcting the balance of the relationship between the various lymphocyte subpopulations.

[A case of juvenile rheumatoid arthritis treated with thymopentin]. / Un caso di artrite reumatoide infantile (ARI) trattato con timopentina.

The case of a ten year old girl with JRA presenting a systemic onset and resistant to non-steroid therapy is described. After one course of thymopentin the patient responded satisfactorily. After ten months from the beginning of treatment and a third course she is still doing well. The authors comment the modalities of therapy.

The immune-enhancing effect of perioperative thymopentin administration in elderly patients undergoing major surgery.

The effects of perioperative administration of thymopentin (TP-5) on in vivo and in vitro measurements of cell-mediated immunity in elderly patients undergoing major surgery were investigated. A placebo-controlled study was conducted in 25 patients (mean age, 67 years) with congenital or acquired heart disease undergoing surgery with cardiopulmonary bypass. Patients were divided into three groups: Group 1 patients were given 50 mg of TP-5 subcutaneously two hours preoperatively. Group 2 patients were given 50 mg of TP-5 subcutaneously two hours preoperatively and 48 hours postoperatively. Group 3 patients were given placebo at corresponding times. Cell-mediated immunity measurements were the in vivo delayed-type hypersensitivity (DTH) response on day 0 and on day 7 to an antigen skin test battery. The in vitro studies included antigen cocktail-induced lymphocyte proliferation of peripheral blood mononuclear cells. The DTH response on day 7 after surgery was significantly suppressed in group 3 patients compared with the preoperative baseline value, while it remained unaltered in group 1 and 2 patients. There was a considerable difference of DTH measurements (number of positive antigen responses and sum of their mean diameters) between group 2 and 3 patients. Antigen cocktail-induced lymphocyte proliferation, following initial suppression in the majority of patients, was significantly different between the placebo group and patients in group 2 on day 7 after surgery. The data indicate that perioperative administration of TP-5 might be of considerable clinical utility in preventing a defective cellular immune response.