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.

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.

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.

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.

First observations on high-dosed and long-term thymopentin treatment in active rheumatoid arthritis.

In this pilot study carried out in two centres, six male and two female patients with severe active rheumatoid arthritis (RA) (average duration over 10 years) were treated with thymopentin 50 mg in the form of prolonged i.v. injection (over 10 min), 3 times weekly for 3 to 20 weeks. Two of these patients were subsequently treated with different s.c. doses of thymopentin in a crossover fashion for more than two years, including periods without any treatment or treatment with placebo. The overall clinical efficacy was judged by assessing pain patterns and joint status and the functional stage of the patients according to Steinbrocker; in addition, the sedimentation rate was measured before and after the therapy. Seven out of eight patients showed definite improvement in their clinical status as assessed by the Steinbrocker scale. Most of the symptoms, particularly pain, capsular swelling, tenderness and morning stiffness, were remarkably reduced within 3 weeks of thymopentin treatment. Sedimentation rate decreased in five out of eight patients. Prolonged i.v. injections seemed to have somewhat better effects than s.c. administration; in the latter group the highest dose (3 X 100 mg/week or higher) produced the best results. During placebo treatment and during the medication-free intervals both groups of patients got worse. No side-effects occurred during the study.

Interim results on the clinical effects of i.v. administered thymopentin in active rheumatoid arthritis.

Forty-one patients with active rheumatoid arthritis entered a controlled, double-blind, randomized study; 21 received prolonged i.v. injections (10 min) of thymopentin 50 mg 3 times a week for 3 consecutive weeks; the other 20 received placebo under the same conditions. The groups were comparable at the start of the study. Statistical tests of changes within the treatment groups after 3 weeks showed that the improvement achieved in the thymopentin group was significant (p less than 0.05 or p less than 0.01) for each clinical parameter, except for left-hand grip strength. On the other hand, no significant improvement was observed for any parameter except morning stiffness in the patients on placebo. The intergroup comparison showed significant differences, favouring thymopentin over placebo treatment, in the Ritchie index, scores for swollen joints, assessment of severity of pain and scores for changes in the activity of the disease. Only minor side-effects were experienced in the two treatment groups. The present placebo-controlled double-blind study confirms the previous positive results achieved in open studies, i.e., the beneficial therapeutic effect of prolonged i.v. injections of thymopentin in patients with severe rheumatoid arthritis observed after 3 weeks of therapy. The drug appears to be safe at the dose regimen used.

Immunomodulation with thymopentin in humans.

The effect of thymopentin administered i.v. or s.c. on the levels of circulating specific IgM and IgG KLH (key-hole limpet haemocyanine) antibodies and non-specific immunoglobulins were measured at weekly intervals in elderly volunteers for three subsequent weeks after vaccination with 500 micrograms KLH. As compared with the placebo group, specific IgM and IgG antibody responses significantly increased in the s.c. treated group, but remained at significantly lower levels in the i.v. treated groups. Increases in non-specific immunoglobulin levels were observed after vaccination in the placebo group; no such increases appeared in the groups treated with thymopentin. The results demonstrate the immunomodulatory effect of thymopentin in humans. It is assumed that, depending on the route of application (which indirectly represents different doses), thymopentin can either stimulate or inhibit immune processes. As an immunomodulator it may represent a new therapeutic tool for immunostimulation as well as for specific immunosuppression.

Preliminary results on clinical and immunological effects of thymopentin in AIDS.

In an open study, 10 African patients with AIDS were treated with thymopentin 50 mg i.v. 3 times a week for 2 consecutive months: 1 month by i.v. direct injections and 1 month by 30-min i.v. infusions. In group A there were 6 patients with AIDS-lymphadenopathy characterized by weight loss, chronic fever, generalized lymphadenopathy and OKT-4 to OKT-8 ratio below 0.2. Group B consisted of 4 patients with AIDS and opportunistic infections. Immunological studies performed before, during and after therapy included lymphocyte count, T-cell subsets assessment, study of blastogenic response of lymphocytes to PHA and delayed hypersensitivity skin testing to 5 antigens. In group A, results after thymopentin i.v. direct injections showed a significant increase in OKT-3 and OKT-8 cells. After thymopentin i.v. infusion, blastogenic response to PHA increased significantly as compared with pretherapy values and to the values after i.v. direct injections. At the end of infusion therapy, skin tests became positive for 3 antigens (range 2-4). Furthermore, all 6 patients noted subjective improvement associated with significant weight gain and disappearance of fever. In contrast, in group B the clinical and immunological status worsened during therapy and two patients died from opportunistic infections. This preliminary study suggests that i.v. infusion with thymopentin may be useful in the early phase of the acquired immune deficiency syndrome as it produces symptomatic and immunological improvement.

The effect of thymopentin treatment on the relapse rate in frequently relapsing herpes simplex virus infections.

Twenty-four patients suffering from longstanding severe recurrent herpes simplex, who had not responded to prior therapy, were treated with s.c. thymopentin injections 50 mg, three times weekly, over a period of six weeks. They were followed up at weekly intervals over this period and then six weeks later. Moreover, the longest relapse-free period observed in the year after the treatment was recorded in the investigator's documentation. Thirteen of the 14 patients with labial herpes simplex and 10 of the 13 patients with genital herpes simplex improved markedly as shown by a decrease in the relapse rate of at least 50%, shorter episodes of relapse and improvement of symptoms such as pain and itching. Fourteen of these 27 patients experienced no relapse for a period longer than four months after cessation of the therapy. No serious side-effects were observed. Laboratory examinations before, during and after thymopentin did not reveal significant alterations except for an increase in the T-helper/T-suppressor ratio. The effect of thymopentin is assumed to be due to T-helper cell activation resulting in enhanced interleukin-2 production with subsequent proliferation of cytotoxic T lymphocytes and natural killer cells which are capable of producing immune interferon.

[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 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.

[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.

[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.

[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.

[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.

[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.

[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.

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.