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.

TP-3 immunotoxins improve antitumor activity in mice with osteosarcoma.

We measured the antitumor activity of two types of TP-3 immunotoxins that target an antigen expressed in tumors associated with osteosarcoma. Development of novel agents for treatment of patients with osteosarcoma is important. We previously described a monovalent-disulfide-stabilized recombinant immunotoxin made from the TP-3 antibody. This agent is called TP-3(dsFv)-PE38 and is cytotoxic to human osteosarcoma cells in vitro. To improve antigen binding, we designed and produced a bivalent immunotoxin, TP-3(dsFv)2-PE38. We evaluated the activity of both molecules in vitro and in vivo using tumor-bearing mice. Compared with the monovalent TP-3 immunotoxin, the bivalent TP-3 immunotoxin showed an approximately sevenfold increase in cytotoxic activity against three osteosarcoma cell lines which react with the TP-3 monoclonal antibody. The apparent affinity of the bivalent TP-3 immunotoxin was 12-fold greater than that of the monovalent TP-3 immunotoxin. The antitumor activities of both TP-3 immunotoxins were measured using severe combined immunodeficient mice bearing osteosarcoma cell line OHS-M1 tumors. The dose at which the bivalent TP-3 immunotoxin produces complete regressions of tumors is (1/2) that of the monovalent TP-3 immunotoxin. Increasing the avidity of TP-3(dsFv)-PE38 significantly improves its cytotoxic activity in vitro and results in a twofold increase in antitumor activity in vivo. Because TP-3-based immunotoxins have good antitumor activity in mice, these molecules merit additional development for possible treatment of osteosarcoma in humans.

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.

[Incidence of SLE in a patient thymectomized for myasthenia gravis]. / SLE fellépése myasthenia gravis miatt thymectomizált betegen.

The appearance of systemic lupus erythematosus (SLE) as well as thymopoietin (TP3) treatment in a patient who has undergone thymectomy because of myasthenia gravis (MG) is described. To the authors' knowledge this is the first case of this treatment upon the diagnoses above. The therapy was performed for 12 weeks by the Hungarian Thymotrinan (TP3) preparate according to a previous protocol worked out for Hodgkin's disease. At the beginning and during therapy, cellular and humoral immune parameters were monitored. Twenty months after the therapy the patient has remained in clinical remission for both diseases, MG and SLE, respectively. There is a brief survey of the biochemistry of thymic hormones as well as their clinical use, especially in the treatment of autoimmune disorders. The relative rarity of SLE after operative treatment of MG is under discussion together with the effectiveness of substitutional thymic hormone therapy.

[Immunologic and cytologic aspects of Hashimoto's thyroiditis during therapy with thymopentin]. / Aspetti immunologici e citologici della tiroidite di Hashimoto in corso di terapia con timopentina.

Hashimoto's thyroiditis is an autoimmune disease characterized by the presence of thyroid autoantibodies and frequent coexistence of other autoimmune disorders. The object of our research was to examine the peripheral blood and fine-needle cytology modifications in patients with this disease during therapy with Timopentina. Our results suggested a possible therapeutic effect of Timopentina, as an alternative to traditional cortisone treatment.

[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.

Evaluación clínica-inmunológica del tratamiento con Tp-5 en sujetos infectados con HIV / Clinical immunological evaluation of the treatment with tp-5 in subjects infected with HIV

We studied the effect of Tp-5 treatment on the immune system of 15 HIV+ males. The control groups consisted of 7 asymphtomatic HIV+ homosexual men and 33 health individuals. At the time of admission, the HIV+ subjects showed a significant decrease in the number of CD4+ T-cell and an increase in CD8+ cell when compared to normal controls. During Tp-5 treatment, 4 out of 15 of the HIV infected individuals expressed a transient increase of the CD4+ lymphocyte subpopulation. The number of CD8+ subset was not modified by the pentapepotide. The ability of pokeweed mitogen driven B lymphocytes to differentiate into plasma cell, already significantly low at admission, persisted without change at the end of the treatment. Thus, Tp-5 was not able to restore B-lynphocyte function. The T-lymphocyte PHA proliferative response of T-lymphocytes of HIV infected individuals did not differ from normal controls and no variation was observed after 3 months of the Tp-5 therapy. Nevertheless, the delayed skin reactivity to the 7 antigens tested, increased in 5 out of 9 patients, suggesting a partial restoration of T cell immunity by Tp-5. We conclude that Tp-5 in vivo can induce a transient partial variation at the T lymphocyte level in HIV infected patients

[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.

[Nonspecific active stimulation of the immune system in the combined treatment of rectal cancer]. / Nespetsificheskaia aktivnaia stimuliatsiia immunnoi sistemy v kompleksnom lechenii raka priamoi kishki.

Results of investigations on the efficacy of splenin as an immunomodulator in the complex treatment of 70 patients with cancer of the rectum revealed its favourable effect on the T- and B-systems of immunity. It was found that immunotherapy effected favourably the immunological properties of the body and this was also supported by clinical data. This makes it possible to recommend splenin as in immunomodulator in the complex treatment of patients with cancer of the rectum.

[Thymopentin in a case of autoimmune thrombocytopenia secondary to chronic lymphatic leukemia]. / Timopentina in un caso di piastrinopenia autoimmune secondaria a leucemia linfatica cronica.

The Authors describe a case of chronic lymphocytic leukemia complicated by immune thrombocytopenic purpura. They consider the various causes of thrombocytopenia during lympho-proliferative disorders and underline the different prognostic significance. They evaluate the efficacy of treatment with timopentina.

[Thymopentin in the treatment of cutaneous melanoma]. / La timopentina nel trattmento del melanoma cutaneo.

Thirty-three patients were treated with thymopentin (synthetic thymic-hormone) after complete surgical removal of cutaneous malignant melanoma without clinical evidence of metastases and with evidence of immunocellular deficiency, with the aim of obtaining a normalization of the immunological parameters. The cell-mediated immunity was evaluated by utilizing monoclonal antibodies CD3, CD4, CD8, CD21, NK and the Multitest C.M.I. (Institut Merieux-Lyon-France), before treatment and then every three months. A follow-up at 29 months showed an improvement of the immune parameters: respectively 58%, 67% and 42% of the patients undergoing therapy had an increase of 40% of the CD3, CD4 and NK lymphocytes. Seven patients, with pre-existing hypergy following tests of skin-reactions, present a normalization of this parameter during the treatment. In the follow-up three patients had metastases. The results show the improvement of the considered immunological parameters and low percentage of relapses. It may be considered as a preventive measure for immunological control of relapses.

[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.

[Disorders of immune system function and their correction in melanomas with regional metastases]. / Narusheniia funktsii immunnoi sistemy i ikh korrektsiia pri melanomakh s regionarnymi metastazami.

Dysfunction of the thymus and the immune system in patients with melanoma was shown to be in direct correlation with tumor extent. The use of thymosin-like immunomodulator was followed by the most apparent recovery of the parameters studied in patients with localized melanoma resulting in improved clinical course of the disease. In cases of tumor spread into regional lymph nodes, immunologic disorders observed were identical to those registered prior to treatment. Tumor did not disseminate in patients retaining normal immune status.

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.

Phenotypical and functional heterogeneity of the large granular lymphocytes increased after various treatments in a patient with combined immunodeficiency.

A boy with combined immunodeficiency having low natural killer (NK)-cell activity received thymopoietin pentapeptide (TP-5) treatment, transplanted with T cell-depleted HLA-haploidentical bone marrow (BMT) cells from his father and with thymus tissue from an infant at different times during the first year of life. He showed a marked increase in large granular lymphocytes (LGL) both during the treatment with TP-5 and after BMT. The LGL generated following TP-5 injection had a T3+Leu11- surface phenotype and low NK activity. In contrast, the LGL appearing after BMT showed T3-, Leu7+, and/or Leu11+ surface phenotypes, had high NK- and K-cell activities, and were lymphokine-activated killer (LAK)-cell precursors. These killer activities were assigned to the Leu7-Leu11+ subset and proved to be of recipient origin. LGL proliferation following BMT was accompanied by neutropenia, which was improved in association with a reduction in the number of LGL and the appearance of T cells of BMT donor origin following thymus transplantation. This suggested the inhibition of granulopoiesis by the LGL and an in vitro study revealed that the Leu7+Leu11- subset of LGL suppressed the growth of granulocyte/macrophage colony-forming units. These results indicated that phenotypically different LGL could be generated by different treatments and that the LGL showing NK activity were distinct from those regulating granulopoiesis. It was also suggested that the generation of LGL was controlled by T cells.