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PubMed1976

Human thymus/leukemia-associated antigen in normal and leukemic cells.

A saline extractable human thymus/leukemia-associated antigen (HThy-L) has been detected in immunodiffusion using rabbit antisera raised to human thymus extracts. HThy-L was demonstrated in extracts of normal thymocytes, lymphocytes from E-rosetting T-cell lines and from the blast cells of patients with E-rosetting acute lymphoblastic leukemia. Extracts of leukemic cells from patients with acute myeloblastic leukemia and E-rosette-negative acute lymphoblastic leukemia contained significantly lower quantities of HThy-L. Only trace amounts of the antigen were detected in normal spleen tonsil, peripheral blood, bone marrow and PHA-induced lymphoblasts. HThy-L appears to be an antigen associated with the intrathymic differentiation of human T cells and a marker of leukemic cells in certain forms of acute leukemia.

Antigens, Neoplasm↗
PubMed1988

Evidence for the presence of ANP-precursor material in the rat thymus.

Acidic extraction of the thymus from two day old rats followed by purification on Sephadex G-50 gelfiltration revealed the presence of atrial natriuretic peptide-like material (IR-ANP) by radioimmunoassay. Verification of the obtained immunoreactivity has been achieved by the use of two different types of antisera, i.e. two antisera directed against ANP (99-126), the other antiserum recognizing the N-terminal fragment (11-37) of the precursor ANP (1-126) molecule. In addition, reverse phase high performance liquid chromatography (RP-HPLC) and high performance gel permeation chromatography (HP-GPC) monitored by the three antisera have been employed for analysis of the extracted IR-ANP. In both systems the IR-ANP corresponded to the 15 kDa-ANP molecule (1-126). Furthermore, by using immunohistochemical techniques a distinct localization of the IR-ANP material could be demonstrated. The outer cortical area of the thymus, containing mostly lymphoid cells, showed extensive immunostaining with the three different antisera. The data reported here indicate that the rat thymus is a source of ANP.

Animals↗
PubMed1996

Purification and characterization of the phosphatidylinositol-3,4,5-trisphosphate phosphatase in bovine thymus.

Using phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P3] prepared from phosphatidylinositol 4,5-bisphosphate and inositolphospholipid 3-kinase, we identified in bovine thymus extracts the enzyme activity which catalyzed dephosphorylation of PtdIns(3,4,5)P3, to produce phosphatidylinositol biphosphate. Since bovine thymus exhibited the highest level of activity among tissues screened, we tried to purify this enzyme PtdINs(3,4,5)P3 phosphatase from bovine thymus. After sequential chromatographies using S-Sepharose, heparin-Sepharose, blue Sepharose, and Toyopearl HW55, the enzyme was purified 1875-fold with a yield of 10%. SDS/PAGE analysis revealed that a 120-kDA protein band copurified with the enzyme activity. The apparent molecular mass of the active protein was 120 kDa on size-exclusion chromatography, suggesting that the 120-kDa band on SDS/PAGE is the PtdIns(3,4,5)P3 phosphatase. Since PtdIns(3,4,5)P3 phosphatase seemed to be the only activity that metabolized PtdIns(3,4,5)P3, and the enzyme did not hydrolyze phosphatidylinositol 4,5-biphosphate, the enzyme may play a critical role in the inositolphospholipid 3-kinase signalling.

Animals↗
PubMed1999

Immunopotentiating and immunotherapeutic effects of thymic hormones and factors with special emphasis on thymic humoral factor THF-gamma2.

The essential role played by the thymus in the development of the immune response was well documented in many publications. These findings prompted a long series of studies devised to define the factors produced and secreted by thymus cells, which are involved in the development and nature of immunological responsiveness. First experiments done with crude thymus extracts were followed by isolation of purified products and finally by chemical characterization and synthesis of immunologically active thymus-derived peptides. In this article we review the various thymic hormones and factors described, that is, thymosin fractions 5, the thymosins, prothymosin alpha, thymulin (FTS-Zn), thymopoietin, thymostimulin (TP-1), Thymic humoral factor (THF), and THF-gamma2. Studies demonstrating the activity of the various thymic factors in increasing the immunocompetence potential in both in vitro and in vivo conditions are discussed. The immunostimulatory potential of thymic factors was also investigated in experimental models where beneficial therapeutic effects were sought in a situation of immunological malfunction. The last part of the review is dedicated to clinical trials with thymic factors that revealed improvement in the immunocompetence potential in cases of immunodeficiencies, viral infections, and cancer and its correlation with therapeutic effectiveness. It seems that more research is required in order to better define conditions for the use of thymic factors in immunotherapy.

Animals↗
PubMed1991

Partial purification and characterization of the lck protein-tyrosine kinase from bovine thymus.

Soluble extracts prepared from bovine thymus contain an angiotensin-I-phosphorylating activity that is activated several-fold by high concentrations of NaCl. Fractionation of this protein-tyrosine kinase activity by chromatography on DEAE-cellulose yields a major diffuse peak of activity. The enzymes responsible for this activity are found at much higher levels in extracts from bovine thymus than from bovine spleen. The peak of activity from the DEAE-cellulose column can be further separated into two major peaks by chromatography on heparin-agarose. The second peak to elute from the heparin-agarose column was previously purified through several chromatographic steps to yield a 40 kDa protein-tyrosine kinase (p40). We have now partially purified the early-eluting peak of kinase activity by chromatography on columns of butyl-agarose, protamine-agarose and Sephacryl S200. The enzyme was identified following covalent modification with 5'-fluorosulphonylbenzoyladenosine (FSBA) by reactivity with anti-FSBA antibodies. This procedure labelled a series of 52-56 kDa proteins. These proteins were also recognized by polyclonal anti-peptide antibodies raised against the C-terminal 33 amino acids of p56lck, a major T lymphocyte protein-tyrosine kinase. Peptide maps of the partially purified enzyme were identical to maps generated from p56lck obtained from LSTRA cells. These data suggest that bovine thymus p56lck is responsible for the activity found in the early-eluting peak from heparin-agarose. Antibodies raised against a peptide corresponding to amino acids 39-64 of p56lck, a sequence found near the N-terminus, recognized the slower-migrating, but not the faster-migrating, form of the enzyme, indicating that a fraction of the protein had been proteolysed near the N-terminus during purification. The partially purified bovine enzyme exhibited a restricted substrate specificity in vitro and did not readily phosphorylate human erythrocyte band 3, casein or histone, but was able to phosphorylate acid-treated enolase. The dilute enzyme present in fractions eluting from chromatography columns was unable to catalyse an autophosphorylation reaction. Autophosphorylation could be detected in more concentrated enzyme samples and was readily observed in immune-complex assays. The phosphorylation of angiotensin I by bovine thymus p56lck was weakly activated by polyionic compounds such as heparin and polylysine, and was strongly activated by high concentrations of NaCl.

Amino Acid Sequence↗
PubMed1996

Binding of thalidomide to alpha1-acid glycoprotein may be involved in its inhibition of tumor necrosis factor alpha production.

In addition to its well known sedative and teratogenic effects, thalidomide also possesses potent immunomodulatory and antiinflammatory activities, being most effective against leprosy and chronic graft-versus-host disease. The immunomodulatory activity of thalidomide has been ascribed to the selective inhibition of tumor necrosis factor alpha from monocytes. The molecular mechanism for the immunomodulatory effect of thalidomide remains unknown. To elucidate this mechanism, we synthesized an active photoaffinity label of thalidomide as a probe to identify the molecular target of the drug. Using the probe, we specifically labeled a pair of proteins of 43-45 kDa with high acidity from bovine thymus extract. Purification of these proteins and partial peptide sequence determination revealed them to be alpha1-acid glycoprotein (AGP). We show that the binding of thalidomide photoaffinity label to authentic human AGP is competed with both thalidomide and the nonradioactive photoaffinity label at concentrations comparable to those required for inhibition of production of tumor necrosis factor alpha from human monocytes, suggesting that AGP may be involved in the immunomodulatory activity of thalidomide.

Affinity Labels↗
PubMed1984

Cooperative randomized studies on the treatment of adult acute leukemia in Poland. A comparison of two remission induction regimes and two maintenance regimes for AML.

In a prospective study 100 AML cases were randomized in two groups and underwent remission induction treatment in 4 departments with "Ara-C c. inf. from the first to the eight day plus Adr. from the first to the third day" or "Ara-C + 6-TG + Adr., 6 + 6 + 2 days". The CR patients were divided into two subgroups. Subgroup A obtained maintenance therapy with 5 day course of Ara-C s.c. in addition to 6-TG, Vincristine plus Prednisone and Adriamycin sequentially, with each cycle repeated at 6 week intervals. Subgroup B obtained the calf thymus extract (TFX-Polfa) between the cycles additionally. The current results of remission induction treatment are similar to "8 + 3" and "6 + 6" regimens as measured by the CR rate (45% and 44%), survival curves (survival after 24 months - 23% and 28% respectively) and relapse rates (3.5%/pt.mo.obs. and 5.3%/pt.mo.obs.). It is still too early to evaluate the efficacy of this maintenance treatment, because the programme is still open to patient entries. At present 13 patients received maintenance regimen A and 12 patients received maintenance regimen B, with an average survival being equal to 14 and 13.5 months respectively.

Adult↗
PubMed1985

A factor from bursa of Fabricius inhibits in vitro the chorionic gonadotropin response of the chick testis.

The effect of bursa of Fabricius on the endocrine function of the chick testis was studied using an isolated testis cell preparation. Testosterone secretion, both basal and under hCG stimulation, was measured in the incubation medium. The bursal extract inhibited the response of the testis cells to hCG detected as a reduced testosterone secretion. The normal basal secretion of the testis cells was not modified by the bursal extract. To analyze some characteristics of the bursal factor, fractions of approximately known molecular weight were obtained by filtering through Amicon membranes. The active factor was found in the fraction corresponding to 1000-10000 Da. Its activity disappeared after heating of trypsin incubation, suggesting the peptidic nature of the bursal factor. Tissue extracts from gut and spleen did not modify the hCG response of the testis cells. There was inhibition of the hCG response by thymus extracts of the newly hatched chicken. These results add new evidence of a modulatory effect of the immunogenic organs on the endocrine function of the testis in the newly hatched chicken.

Animals↗
PubMed1975

The role of a thymus humoral factor in the proliferation of bone marrow CFU-S from thymectomized mice.

The colony forming capacity of bone marrow cells from thymectomized mice was shown to be reduced as compared to that of bone marrow cells from normal donors. A further indication of changes in the proliferative capacity of colony forming cells (CFU-S), following thymectomy, was given by examination of the sensitivity of these cells to chlorambucil and by 3H-thymidine 'suicide' experiments; both showed that CFU-S from thymectomized mice were not cycling at the same rate as normals. It was also found that in late pregnancy of thymectomized females, there is an elevation in the number of bone marrow CFU-S and an increase in cell cycling. Such an increase could also be achieved by implantation, into thymectomized mice, of thymus lobes in closed diffusion chambers. Finally, in vitro administration of a thymus hormone (THF) reversed the suppressive effect of thymectomy on DNA synthesis in bone marrow CFU-S. Since the action of THF was restricted to bone marrow cells of thymectomized mice it is plausible that normal bone marrow contains at least two subpopulations of CFU-S, one of which is dependent upon a humoral product of the thymus.

Animals↗
PubMedDate not supplied

Application-dependent immunomodulating and antimetastatic efficacy of thymic peptides in BALB/c-mice.

The immunomodulating and antimetastatic activity of clinically approved, low molecular weight, standardized thymic peptide (TP) preparations was evaluated in BALB/c-mice. Daily applications (subcutaneously, s.c.; intraperitoneally, i.p.; intramusculary, i.m.) of two commercially available TP preparations (7 consecutive days, 10, 50 and 100 micrograms per mouse and injection) up-regulated the thymus weight and thymocyte counts as well as peripheral blood leukocyte and lymphocyte counts in liver metastases-bearing mice. The immunomodulating activity of TP application was most pronounced and statistically significant for thymus weight and counts of thymocytes, leukocytes and lymphocytes after s.c. administration of both TP preparations and concentrations. I.p. and i.m. TP-injections were less effective at reaching statistical significance, however, for defined dosages and parameters, only. To evaluate the influence of TP on experimental liver metastases, RAW 117 lymphosarcoma cells were intravenously inoculated into BALB/c-mice. TP (10, 50, 100 micrograms/mouse) were s.c., i.p. and i.m. administered daily for 7 consecutive days starting 24 hours after tumor cell challenge. Liver colonization was investigated on day 14 after tumor cell inoculation and demonstrated a statistically significant (p < 0.05) reduction of experimental liver metastases for s.c. (both preparations and concentrations) as well as i.p. and i.m. (dose-dependent) TP-treated mice.

Adjuvants, Immunologic↗
PubMed1979

[Action of thymic extracts on the growth of different cell populations in vitro].

Thymic, splenic and renal cellular extracts were obtained by tissular grinding and ultra centrifugation, from animals having undergone diverse hormonal treatments. These extracts were added to different cellular populations in culture (carcinomatous HeLa strain, established MRCs strain, primo-culture of human foetal lung strain). The thymic extracts of non treated animals and the splenic extracts rich in FK substance from intact guinea pigs treated by oestrogenes or FSH, provoque an inhibition of tumoral cell growth. The most important effects are obtained with thymus extracts rich in FK substance. The FK substance seems capable of concentrating certain antitumoral thymic fractions.

Animals↗
PubMed1985

Single stranded DNA binding proteins derive from hnRNP proteins by proteolysis in mammalian cells.

As we have previously demonstrated, mammalian single stranded DNA binding proteins (ssDBP) and heterogeneous nuclear RNA binding proteins (hnRNP proteins) are antigenically and structurally related. In this paper we show that ssDBP are specific proteolytic products of hnRNP core proteins. Proteolysis can be observed in crude extract, both total and nuclear and is not inhibited by the most commonly used protease inhibitors. Such phenomenon can be observed in HeLa cells, human fibroblasts and calf thymus extracts. A trypsin-like protease that cleaves purified hnRNP proteins to give ssDBP of Mr = 24-28 Kd can be purified from HeLa cells. A precursor-product relationship can be established between hnRNP core proteins type A and an ssDBP of 24 Kd (UP1).

Animals↗
PubMed1984

Characterization of the autoimmune antigenic determinant for ribonucleoprotein (RNP) antibody.

Small nuclear ribonucleoprotein complexes are antigens in various autoimmune diseases. The serological pattern of high titers of circulating antibody to nuclear ribonucleoprotein (RNP) antigen is a diagnostic marker for mixed connective tissue disease (MCTD); whereas antibody to Sm is prevalent in systemic lupus erythematosus (SLE). Both calf thymus and rabbit thymus are commonly used, excellent sources for preparation of the corresponding antigens RNP and Sm in clinical and research laboratories (A. M. Boak et al., accompanying paper). Thus, biochemical and structural characterization of the minimal antigenic determinant in these preparations is important for its use in the laboratory, as well as significant for understanding MCTD, SLE, and other examples of autoimmunity. Purification and biochemical analyses of immunologically active RNP from many different preparations of calf thymus extract has revealed that the majority of antibody in monospecific MCTD patient sera recognizes an antigen composed of the 165 nucleotide RNA, U1 RNA, and five peptides. Calf thymus U1 RNA was found to be identical in sequence to that of man. A sequence of 55 nucleotides within the 165 nucleotide RNA was the minimal RNA fragment found in RNP particles that were still immunologically active. Two of the RNP peptides react with patient sera monospecific for RNP and thus, are presumably the antigenic peptides complexed with the 55 nucleotide RNA sequence.

Animals↗