c-Fos regulated by TMPO/ERK axis promotes 5-FU resistance via inducing NANOG transcription in colon cancer.

Acquired drug resistance is one of the most common limitations for the clinical response of colon cancer to 5-Fluorouracil (5-FU)-based chemotherapy. The relevant molecular mechanisms might be diversity, but still not be elucidated clearly. In this study, we aimed to investigate the potential mechanisms of c-Fos, a subfamily of activator protein-1, in 5-FU chemoresistance. We determined that phosphorylated c-Fos promoted colon cancer cells resistance to 5-FU by facilitating the cancer stemness. Mechanically, 5-FU treatment induced autolysosome-dependent degradation of TMPO, which subsequently triggered ERK-mediated phosphorylation of c-Fos. Additionally, c-Fos was found to bind to the promoter of NANOG and phosphorylation of c-Fos at Ser 374 was required for its regulation of NANOG expression. NANOG ablation impaired c-Fos/p-c-Fos induced 5-FU resistance and stemness. Taken together, these findings revealed that TMPO-mediated phosphorylation of c-Fos conferred 5-FU resistance by regulating NANOG expression and promoting cell stemness in colon cancer cells. c-Fos could be as a therapeutic target for colon cancer.

Antitumor activity of TP3(anti-p80)-pokeweed antiviral protein immunotoxin in hamster cheek pouch and severe combined immunodeficient mouse xenograft models of human osteosarcoma.

TP3-pokeweed antiviral protein (PAP) immunotoxin is directed against the p80 antigen on osteosarcoma cells. Previous studies have demonstrated that TP3-PAP kills clonogenic human osteosarcoma cells in vitro and shows significant antitumor activity in a murine soft tissue sarcoma model (P. M. Anderson, et al, Cancer Res., 55: 1321-1327, 1995.) In this study, we demonstrate that TP3-PAP elicits potent in vivo antitumor activity in a hamster cheek pouch model of human osteosarcoma. Furthermore, treatment with TP3-PAP at nontoxic dose levels significantly delayed the emergence and progression of leg tumors and markedly improved tumor-free survival in severe combined immunodeficient mice challenged with OHS human osteosarcoma cells. Thus, TP3-PAP may be useful in the treatment of poor risk osteosarcoma.

Thymopentin and splenopentin as immunomodulators. Current status.

Splenopentin (SP-5, Arg-Lys-Glu-Val-Tyr) and thymopentin (TP-5, Arg-Lys-Asp-Val-Tyr) are synthetic immunomodulating peptides corresponding to the region 32-34 of a splenic product called splenin (SP) and the thymic hormone thymopoietin (TP), respectively. TP was originally isolated as a 5-kDa (49-amino acids) protein from bovine thymus while studying effects of the thymic extracts on neuromuscular transmission and was subsequently observed to affect T cell differentiation and function. TP I and II are two closely related polypeptides isolated from bovine thymus. A radioimmunoassay for TP revealed a crossreaction with a product found in spleen and lymph node. This product, named splenin, differs from TP only in position 34, aspartic acid for bovine TP and glutamic acid for bovine splenin and it was called TP III as well. Synthetic pentapeptides (TP-5) and (SP-5), reproduce the biological activities of TP and SP, respectively. It is now evident that various forms of TPs were created by proteolytic cleavage of larger proteins during isolation. cDNA clones have been isolated for three alternatively spliced mRNAs that encodes three distinct human T cell TPs. The immunomodulatory properties of TP, SP, TP-5, SP-5 and some of their synthetic analogs reported in the literature have been briefly reviewed.

Use of thymic grafts or thymic factors to augment immunologic recovery after bone marrow transplantation: brief report with 2 to 12 years' follow-up.

Thymus tissue implants, thymic epithelial cells obtained from third party donors sharing one HLA-A and -B locus with the recipient, or the thymic hormones thymosin fraction 5 and thymopentin were given to recipients of HLA-identical sibling bone marrow to prevent chronic graft-versus-host disease (GVHD) and accelerate immunologic reconstitution. The clinical courses of 17 patients receiving thymus tissue and 18 patients receiving thymic hormones were reported initially 5 years ago and showed no difference in the incidence of chronic GVHD or immunologic recovery from those of concurrent or historical controls. We report here for the first time nine new patients who received thymus tissue implants with modifications of the culture method to lower the number of lymphocytes in the transplanted tissue with the intent of reducing rejection of the thymus tissue grafts. The clinical outcomes and immunologic functions of these nine patients were similar to those of the recipients of the earlier thymus tissue implants. With follow-up now ranging from 2.2 to 12.3 years (median 6.7) for the total group, 16 patients are alive. Seven never developed chronic GVHD. Nine were treated for chronic GVHD, seven of whom recovered and are leading normal lives, one has chronic pulmonary insufficiency, and one is disabled from chronic GVHD. We conclude that thymus tissue grafts or thymic epithelial cells partially HLA-matched to the recipient, thymosin fraction 5, or thymopentin used as described were not effective in reducing the incidence of chronic GVHD, improving immunologic recovery, or altering long-term survival.

Treatment of marrow graft recipients with thymopentin.

Four adult patients with acute non-lymphocytic leukemia were given marrow grafts from HLA-identical siblings following 120 mg/kg cyclophosphamide and 10-12 Gy total body irradiation. All received intermittent intravenous methotrexate as prophylaxis against graft-versus-disease (GVHD). In an attempt to accelerate immune recovery and prevent GVHD, each patient received thymopentin (TP5) for 100 days after grafting. No adverse effects were seen with TP5 administration. All four patients developed acute GVHD (one grade I, one grade II, and two grade III). Two patients died of late infections: one at 6 months from Pneumocystis carinii pneumonia and one at 11 months from disseminate Pseudomonas, Candida and cytomegalovirus infection. Two patients survive more than 3.9 years after transplantation with Karnofsky scores of 100%. One required treatment for chronic GVHD and recovered. Delayed-type hypersensitivity, antibody production to specific antigen in vivo, and results of in vitro immunologic studies were not altered by TP5 treatment. We conclude that while the administration of TP5 in these patients as described was not harmful, it did not prevent opportunistic infection, improve immunologic reconstitution or lower the incidences of acute or chronic GVHD from that of our previous experiences without thymopentin.

Immunomodulators in the treatment of peritonitis in burned and malnourished animals.

Deficiencies in the immune system that lead to increased morbidity and mortality from infectious complications have been well documented in patients suffering from trauma, malnutrition, sepsis, and thermal injuries. We investigated the potential benefit of immune stimulation for preventing infection in such conditions in an animal model by evaluating three drugs: Corynebacterium parvum, thymopentin (TP-5), and CP-46,665. One-hundred eighty female guinea pigs were rendered immunodeficient by first inflicting a 30% total body surface burn and then placing the animals on diets with calories inadequate to maintain body weight. One half of the animals were then given one of the three immunomodulators on the first, third, and fifth days after burn injury, to try to reverse immunodeficiency. The remaining animals received saline solution injections. Animal responses were evaluated by inserting a clot containing Escherichia coli and Bacillus fragilis into their peritoneal cavity 6 days after burn injury. The animals were followed for 21 days after burn injury. Autopsies on those that died revealed peritonitis and/or pneumonia; autopsies on these that survived showed no pneumonia and there was consistent resolution of peritonitis. TP-5 and CP-46,665, but not C. parvum, significantly improved survival rates and mean survival time in those animals receiving 100 kcal/kg/day. TP-5 and CP-46,665 may be of benefit to the severely stressed, malnourished surgical patient who is at risk of bacterial infection.

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.

Immunostimulation by TP-5 in immunocompromised patients and animals--current status of investigation.

TP-5 is a pentapeptide containing active site of the thymic hormone, thymopoietin. It has been shown to exert effects on multiple components of human and animal immune system both in vivo and in vitro. Initial studies demonstrated it to be necessary for T lymphocyte maturation. Its effects on other parts of the immune system are primarily immunostimulatory, and the drug can be used to correct immunodeficiencies resulting from several causes. TP-5 has been shown to improve survival rates in multiple animal studies where the animal has been rendered immunodeficient prior to septic challenge. It has also been successful in preventing and treating infections in a number of human studies. Evaluation of the exact mechanism by which the drug improves survival has demonstrated that it is of complex nature, involving interactions between various types of white blood cells.

Age-dependent antileukemic action and suppression of immune response by 1,4-benzoquinone-guanylhydrazone-thiosemicarbazone (ambazone) in mice.

The antineoplastic activity of 1,4-benzoquinone-guanylhydrazone-thiosemicarbazone (ambazone) against murine leukemia P388 was found to be markedly reduced in 12- and 18-month-old mice as compared to young animals. The immune response against sheep red blood cells (SRBC), a T cell-dependent antigen, was also strongly diminished in tumor-free old mice and was further suppressed after ambazone treatment. Since the antileukemic effect of ambazone disappeared more or less in congenitally athymic nude mice, in neonatally thymectomized or silica-pretreated animals, it has been concluded that the action of the compound seems to be limited to young adult immunocompetent tumor-bearing hosts. Therefore immunosenescence, primarily of T cell functions of old tumor-bearers, may represent a decisive factor influencing the antileukemic, especially curative effect of ambazone in aged animals. A combined treatment with ambazone and immunomodulators (thymalin or a splenopentin derivative) failed to improve the antileukemic effect in young and old leukemia P388-bearing mice.

Therapeutic possibilities of thymopoietin fragments (TP3 and TP4) based on experimental animal models.

The effects of thymic hormones are focused on the induction of T-cell subpopulations and restoration of the reactivity of an impaired immune system. TP3 and TP4 (corresponding to thymopoietin 32-34 and 32-35) exert a thymic hormone substitution effect. These peptides elicit dissimilar quantitative and qualitative effects. The aim of the present experiments was to investigate: (a) the effect of thymopoietin fragments in mice with unbalanced immune systems caused by experimental manipulation; and (b) the ratio of target cells after treatment. The distribution of Thy1, Lyt1, Lyt2 positivity was determined in a direct complement mediated cytotoxicity test. Autoantibody production was measured by Coombs' test. A count of Lewis Lung Tumour (LLT) metastases was made after two weeks of inoculation. Groups of mice were thymectomized and/or injected with cyclophosphamide (CY) (240 mg/kg) 96 h before tumour cell inoculation. The number of LLT metastases was decreased by treatment with peptides (TP3 = 72, TP4 = 97, TP5[thymopoietin 32-36] = 83.1 in %) and immunosuppression produced by CY was partly restored. After thymectomy, however, only TP3 treatment caused a decreasing effect (97.4%) on CY immunotoxicity independently of thymectomy. Inhibition of autoantibody production was detected with TP3 (5-6 weeks earlier than in mice treated with TP5). The ratio of Thy1+ and Lyt2+ cells was increased by treatment with TP3 and TP4, but the ratio of Lyt1+ cells was decreased by application of TP5. After TP3 treatment of nude mice the Lyt1+/Lyt2+ ratio increased both in bone marrow and spleen. No effect of TP4 was observed on Lyt 1+ cells, but the number of Lyt2+ increased in bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Influence of thymopentin on postsurgical immune deficiency: a clinical pilot study.

The effect of a single dose (50 mg) of thymopentin, administered subcutaneously, on antigen-induced proliferation of peripheral blood mononuclear cells (PBMCs) was investigated in 25 volunteers (all over 50 years of age) and in nine patients undergoing major surgery. Blood samples from another nine patients having the same type of surgery were used as control in the second trial. A statistically significant increase in the proliferative response was observed in the group of volunteers two hours after thymopentin administration. This increment was not demonstrable as significant 24 h later. In contrast to this observation, the surgical procedure-induced significant decrement in the proliferation of PBMCs observed on the first and third postoperative days did not occur in the thymopentin-treated patients on the first postoperative day. This preventive effect of the single dose of thymopentin administered two hours before surgery was no longer demonstrable on the third postoperative day. Further studies are ongoing in order to establish a treatment regimen with thymopentin for the therapy of trauma-induced immune deficiencies.

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.

[Thymopentin as adjuvant therapy in the hepatitis B vaccination of non- or hyporesponsive subjects]. / La timopentina come terapia adiuvante nella vaccinazione anti-epatite B in soggetti non-O iporesponsivi.

The efficacy of thymopentin as adjuvant therapy was assessed in 13 people who did not respond to standard anti-hepatitis B vaccination with Pasteur HEVAC or Merck HV-VAX. Thymopentin (Sindtomodulina, Italfarmaco)--was given in doses of 50 mg 3 times a week for 3 consecutive weeks, a booster dose of the vaccine (40 mcg HB VAX injected into the deltoid muscle, or 10 mcg HEVAC subcutaneous) being given at the start of the second week. In 69.23% of the patients whose anamnesis revealed no immune deficiency, the Merrieux Multitest showed defective cell-mediated immunity. The adjuvant treatment produced an adequate immune response to the vaccine (anti ABc antibody titre 10 mU/ml) in 76.9% of cases and normalised cell-mediated immunity in 66.6% of those found to be hypoanergic at basal screening.

[Behçet's disease. II. Etiopathogenetic and therapeutic aspects. Personal observations]. / La malattia di Behcet. Nota II. Aspetti etiopatogenetici e terapeutici. Osservazioni personali.

The aetiopathogenesis of Behcet's disease is still not known. In the light of current knowledge, the most likely hypothesis seems to be that of an autoimmune reaction set off by viral, bacterial or other antigens. The reaction may manifest itself in the vascular system, and cause vasculitic conditions in genetically susceptible subjects. The existence of a constitutional susceptibility factor would seem to be confirmed by the high and significant incidence of determinant histocompatibility antigens: HLA-B5 in Japan and in Mediterranean countries, HLA-A2 and HLA-A28 in Great Britain and the United States. Corticosteroids are most commonly used to treat the disease; immunosuppressants are also recommended--possibly in association with corticosteroids. Three personally observed cases of Behcet's disease are reported. The first was an example of the disease in its complete form (oral and genital aphthae, ocular lesions); the second case incomplete (no genital aphthae); and the third characterised by severe neurological involvement (neuro-Behcet).

[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.

[Thymopentin and immune response in patients with cancer]. / Timopentina e risposta immunitaria nel paziente neoplastico.

The results of an experimental study using thymopentin (Timunox) on cancer patients are reported. The drug was administered to 25 patients with cancers of the digestive tract in the form of subcutaneous injections of a vial of the product on alternate days for 36 days. All patients treated revealed a good immune response as indicated by assays of total lymphocytes, T4 lymphocytes the T4/T8 ratio and skin tests, apart from their excellent postoperative recovery. Timunox is therefore considered effective in improving the immune competence of cancer patients.