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1.
Blood ; 106(5): 1749-54, 2005 Sep 01.
Article in English | MEDLINE | ID: mdl-15920008

ABSTRACT

Despite profound T-cell immunodeficiency, most patients treated with chemotherapy do not succumb to infection. The basis for residual protective immunity in lymphopenic patients is not known. We prospectively measured T-cell numbers, thymopoiesis, and T-cell memory in 73 children undergoing a 2-year chemotherapy regimen for acute lymphoblastic leukemia (ALL) and compared them to an age-matched cohort of 805 healthy children. Most patients had profound defects in CD4 and CD8 T-cell numbers at diagnosis that did not recover during the 2 years of therapy. Thymic output and the fraction of naive T cells were significantly lower than in healthy controls. However, the remaining T-cell compartment was enriched for antigen-experienced, memory T cells defined both by phenotype and by function. This relative sparing of T-cell memory may, in part, account for the maintenance of protective immunity in lymphopenic patients treated for ALL. Moreover, because the memory T-cell compartment is least affected by ALL and its treatment, strategies to induce immunity to pathogens or tumor antigens in cancer patients may be most successful if they seek to expand pre-existing memory T cells.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Immunologic Memory , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Adolescent , Age Factors , Child , Child, Preschool , Cohort Studies , Humans , Infant , Infant, Newborn , Mortality , Precursor Cell Lymphoblastic Leukemia-Lymphoma/diagnosis , Precursor Cell Lymphoblastic Leukemia-Lymphoma/immunology , Survival Analysis , Thymopoietins/immunology , Time Factors
2.
Int J Neurosci ; 115(2): 223-43, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15764003

ABSTRACT

This study was undertaken to investigate T-cell maturation in hyperplastic thymi of patients suffering from myasthenia gravis (MG). For this purpose, the expression of the major differentiational molecules (CD4, CD8, and CD3/TCRalphabeta) and that of the regulatory and activation molecules on thymocytes from MG patients and control subjects were estimated by flow cytometric analysis. In the MG patients the increase in relative proportion of immature (CD4-8- TCRalphabeta-) and the most mature (CD4+8- TCRalphabetahigh and CD4-8- TCRhigh encompassing immunoregulatory NKT) thymocytes followed by a decrease in that of CD4+8+CD3-/TCRalphabeta- cells was found. Furthermore, in these patients the relative proportion of CD4+HLA-DR+ and CD4+71+ cells was increased, whereas that of CD4+25+ cells was slightly, but significantly, decreased (reflecting, most likely, decreased contribution of T reg cells bearing this phenotype). Moreover, in MG thymi the percentage of CD45RA+ cells was reduced indicating changes in the selection processes. In keeping with this finding the reduced thymocyte apoptotic index and percentage of cells bearing apoptosing (CD4-8- TCRalphabetalow) phenotype were detected. In conclusion, the study demonstrates substantial changes in intrathymic differentiation of T cells in hyperplastic MG thymi and suggests alterations in selection events providing an increased escape of potentially autoreactive T-cell clones, on one side, and an altered maturation and/or selection of immunoregulatory cells (NKT and CD4+8-25+ T reg cells) keeping these cell clones under control, on the other side.


Subject(s)
Myasthenia Gravis/metabolism , Myasthenia Gravis/physiopathology , Thymopoietins/physiology , Adult , Antigens, Surface/immunology , Antigens, Surface/metabolism , CD4 Antigens/immunology , CD4 Antigens/metabolism , CD8 Antigens/immunology , CD8 Antigens/metabolism , Disease Progression , Female , Flow Cytometry , Fluorescence , HLA-DR Antigens/immunology , HLA-DR Antigens/metabolism , Humans , Leukocyte Common Antigens/immunology , Leukocyte Common Antigens/metabolism , Myasthenia Gravis/immunology , Receptors, Antigen, T-Cell/immunology , Receptors, Antigen, T-Cell/metabolism , Receptors, Antigen, T-Cell, alpha-beta/immunology , Receptors, Antigen, T-Cell, alpha-beta/metabolism , Thymopoietins/immunology , Thymopoietins/metabolism
3.
Inmunología (1987) ; 24(1): 44-54, ene.-mar. 2005. ilus, tab
Article in English | IBECS | ID: ibc-93538

ABSTRACT

La infección VIH conduce a una depleción severa de células TCD4+, alteraciones del fenotipo de las subpoblaciones linfocitarias,disminución de la función tímica que a su vez produce un deterioro progresivo de la función inmunitaria. La introducción de la terapia antirretroviral de gran actividad (TARGA) ha disminuido la progresión clínica, suprime la carga viral (CV) e incrementa las células TCD4+ en pacientes infectados por el VIH. Sin embargo, los niños en TARGA suelen tener CV más elevada y peor respuesta virológica que en adultos. No obstante, en los niños infectados por el VIH con bajos valores de células T CD4+, la TARGA produce un aumento consistente de células T CD4+ y de los llamados T-cell rearrangement excision circles (TRECs). Estos resultados indican que la recuperación de la función tímica juega un papel clave en la reconstitución inmunitaria de los niños con VIH. Entre las citocinas identificadas como posibles reguladoras de la timopoyesis, la IL-7 puede jugar un papel esencial.La IL-7 participa en la diferenciación de los timocitos a células Tvirgen maduras en el timo que posteriormente salen a sangre periférica.Varios trabajos han demostrado la correlación inversa entre niveles plasmáticos de IL-7 y las células T CD4+. Por consiguiente, el aumento de IL-7 se ha propuesto como una respuesta homeostática a la depleción de células T CD4+. También se ha comprobado una inhibición de la timopoyesis por cepas virales X4 independientemente de la edad debido a la capacidad de las cepas X4 de infectar a los timocitos.Sin embargo se ha demostrado que la TARGA disminuye el número de aislados virales con fenotipo X4 en los niños con VIH. Enconclusión, la TARGA ha mostrado ser eficaz disminuyendo la CV y aumentando el número de células T CD4+, proceso que depende de la función tímica y estaría regulado por IL-7 (AU)


HIV-infection leads to a severe depletion of CD4+ T-cells, phenotypicalterations of T-cell subsets and a decline in thymic function,which in turn produces a progressive impairment of the immune function.The introduction of highly active antiretroviral therapy (HAART)decreased mortality rates in HIV-infection, and proved to be effective in suppressing plasma viral loads (VL) and increasing CD4+ T-cellcounts in children. However, children receiving treatment usually have higher VL and lower virological response rates than adults. But after severe depletion of T-cells, HAART results in a consistent increase in CD4+ T-cell counts and T-cell rearrangement excision circles(TREC) levels in young HIV-infected patients. These findings indicate that recovery of thymic function is a pivotal event in immunere constitution. Among the cytokines and hormones identified as possible regulators of thymopoiesis, IL-7 may play an essential role. IL-7 has been shown to take part in the differentiation of thymocytes intomature T-cells that will leave the thymus and move to the periphery.Several studies have reported higher plasma IL-7 levels in HIVpatients, which inversely correlate with CD4+ T-cell counts. The increaseof IL-7 has therefore been suggested to be a homeostatic response to T-cell depletion. Furthermore, thymopoiesis is inhibited by X4strains, independent of the age of the infected person. This could be due to the capacity of X4 strains to infect thymocytes. However,HAART produces a decrease in the amount of X4 strain isolates inHIV-infected children. In conclusion, HAART has shown to be effective in decreasing VL and increasing the immune function of CD4+ Tcells. This process is dependent on the function of the thymus as well as the homeostatic mechanisms of IL-7 (AU)


Subject(s)
Humans , Male , Female , Child , HIV Infections/immunology , Immune Reconstitution Inflammatory Syndrome/chemically induced , Anti-Retroviral Agents/adverse effects , HIV Infections/congenital , HIV Infections/drug therapy , Thymus Gland , Interleukin-7/immunology , Viral Load/immunology , Thymopoietins/immunology
4.
Clin Orthop Relat Res ; (430): 142-8, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15662316

ABSTRACT

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.


Subject(s)
Bone Neoplasms/drug therapy , Bone Neoplasms/immunology , Immunotoxins/therapeutic use , Osteosarcoma/drug therapy , Osteosarcoma/immunology , Peptide Fragments/therapeutic use , Thymopoietins/therapeutic use , Animals , Disease Models, Animal , Dose-Response Relationship, Drug , Humans , Mice , Mice, SCID , Peptide Fragments/immunology , Thymopoietins/immunology , Treatment Outcome , Xenograft Model Antitumor Assays/methods
5.
Bull Exp Biol Med ; 136(2): 155-8, 2003 Aug.
Article in English | MEDLINE | ID: mdl-14631497

ABSTRACT

We studied antioxidant properties of immunofan, bursin, cyclobursin, thymopoietin II fragment, glycine, and Siberian ginseng. Experiments were performed in 2 model systems: Fe(2+)-induced oxidation of multilamellar phospholipid liposomes in a heterogeneous water-lipid system and oxidation of luminol induced by alpha,alpha'-azo-bis(isobutyramidine dihydrochloride) in a homogenous aqueous system. By the ability to entrap lipid peroxyl radicals, antioxidant activity of substances decreased in the following order: Siberian ginseng extract>bursin>cyclobursin>thymopoietin II fragment>immunofan, glycine. Siberian ginseng extract and thymopoietin II fragment interacted with Fe(2+), which contributed to elimination of catalyst of lipid peroxidation from the system. The ability of substances to interact with aqueous peroxyl radicals and luminol radicals decreased in the following order: Siberian ginseng extract>thymopoietin II fragment>immunofan>glycine, cyclobursin, bursin. Substances with high antioxidant activity improved the state of the endogenous antioxidant system and protected cells from oxidative stress. They entrapped reactive oxygen species formed in the cytoplasm, modulated free radical processes, and regulated the synthesis of bioactive molecules.


Subject(s)
Antioxidants/metabolism , Oligopeptides/immunology , Free Radicals/metabolism , Glycine/immunology , Glycine/metabolism , Iron/chemistry , Iron/metabolism , Liposomes/metabolism , Luminescent Measurements , Oligopeptides/chemistry , Oligopeptides/metabolism , Oxidation-Reduction , Panax/chemistry , Panax/immunology , Peptide Fragments/immunology , Peptide Fragments/metabolism , Plant Extracts/chemistry , Plant Extracts/immunology , Reactive Oxygen Species/metabolism , Thymopoietins/immunology , Thymopoietins/metabolism , Water/chemistry
6.
Histol Histopathol ; 16(1): 1-14, 2001 01.
Article in English | MEDLINE | ID: mdl-11193183

ABSTRACT

Papillary thyroid carcinomas (PTCs) have characteristic nuclear shape changes compared to follicular-type thyroid epithelium. We tested the hypothesis that the altered nuclear shape results from altered distribution or expression of the major structural proteins of the nuclear envelope. Lamin A, lamin B1, lamin C, lamin B receptor (LBR), lamina-associated polypeptide 2 (LAP2), emerin, and nuclear pores were examined. PTC's with typical nuclear features by H&E were compared to non-neoplastic thyroid and follicular neoplasms using confocal microscopy, and semi-quantitative immunoblotting. Lamin A/C, lamin B1, LAP2, emerin, and nuclear pores all extend throughout the grooves and intranuclear inclusions of PTC. Their distribution and fluorescent intensity is not predictably altered relative to nuclear envelope irregularities. By immunoblotting, the abundance (per cell) and electrophoretic mobilities of lamin A, lamin B1, lamin C, emerin, and LAP2 proteins do not distinguish PTC, normal thyroid, or follicular neoplasms. These results do not support previously published predictions that lamin A/C expression is related to a loss of proliferative activity. At least three LAP2 isoforms are identified in normal and neoplastic thyroid. LBR is sparse or undetectable in all the thyroid samples. The results suggest that the irregular nuclear shape of PTC is not determined by these nuclear envelope structural proteins per se. We review the structure of the nuclear envelope, the major factors that determine nuclear shape, and the possible functional consequences of its alteration in PTC.


Subject(s)
Carcinoma, Papillary, Follicular/ultrastructure , Nuclear Envelope/ultrastructure , Thyroid Neoplasms/ultrastructure , Blotting, Western , Cell Fractionation , Centrifugation, Density Gradient , Electrophoresis , Fluorescent Antibody Technique, Direct , Humans , Isomerism , Laminin/immunology , Laminin/metabolism , Laminin/ultrastructure , Membrane Proteins/immunology , Membrane Proteins/metabolism , Membrane Proteins/ultrastructure , Microscopy, Confocal , Nuclear Proteins/metabolism , Nuclear Proteins/ultrastructure , Thymopoietins/immunology , Thymopoietins/metabolism , Tissue Fixation
7.
J Neurol Sci ; 173(2): 97-102, 2000 Feb 15.
Article in English | MEDLINE | ID: mdl-10675651

ABSTRACT

To clarify the occurrence of apoptosis in skeletal muscle in pathological conditions, we studied 44 muscle biopsy specimens by immunohistochemical staining with monoclonal antibody against emerin, which is localized in muscle nuclear membrane, and by ApopTag Plus to detect DNA fragmentation. Five of six patients with myotonic dystrophy (DM) showed three to 35 myonuclei stained with anti-emerin antibody and ApopTag Plus in 1500 muscle fibers. Four of the 18 patients with polymyositis, one of those with thyroid myopathy and one with neurogenic atrophy showed a few myonuclei stained positively by these methods. Our study revealed that DNA fragmentation in myonuclei occurred in skeletal muscle fibers regardless of the type of disease, although the frequency was rather low in all of these diseases except DM. The DNA fragmentation detected in most of the patients with DM suggested a significant role of apoptosis in the pathomechanism of this disease.


Subject(s)
Apoptosis , Cell Nucleus/chemistry , DNA Fragmentation , In Situ Nick-End Labeling , Membrane Proteins/analysis , Muscle Proteins/analysis , Muscle, Skeletal/pathology , Myotonic Dystrophy/metabolism , Thymopoietins/analysis , Adolescent , Adult , Antibodies, Monoclonal/analysis , Antibodies, Monoclonal/immunology , Biomarkers , Cell Nucleus/ultrastructure , Female , Humans , Male , Membrane Proteins/immunology , Middle Aged , Muscle Proteins/immunology , Muscle, Skeletal/chemistry , Muscular Diseases/metabolism , Muscular Diseases/pathology , Myotonic Dystrophy/pathology , Nuclear Proteins , Polymyositis/metabolism , Polymyositis/pathology , Thymopoietins/immunology
8.
Differentiation ; 66(4-5): 208-17, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11269947

ABSTRACT

Emerin is an ubiquitous protein localized at the nuclear membrane of most cell types including muscle cells. The protein is absent in most patients affected by the X-linked form of Emery-Dreifuss muscular dystrophy, a disease characterized by slowly progressive muscle wasting and weakness, early contractures of the elbows, Achilles tendons, and post-cervical muscles, and cardiomyopathy. Besides the nuclear localization, emerin cytoplasmic distribution has been suggested in several cell types. We studied the expression and the subcellular distribution of emerin in mouse cultured C2C12 myoblasts and in primary cultures of human myoblasts induced to differentiate or spontaneously differentiating in the culture medium. In differentiating myoblasts transiently transfected with a cDNA encoding the complete emerin sequence, the protein localized at the nuclear rim of all transfected cells and also in the cytoplasm of some myoblasts and myotubes. Cytoplasmic emerin was also observed in detergent-treated myotubes, as determined by electron microscopy observation. Both immunofluorescence and biochemical analysis showed, that upon differentiation of C2C12 cells, emerin expression was decreased in the resting myoblasts but the protein was highly represented in the developing myotubes at the early stage of cell fusion. Labeling with specific markers of myogenesis such as troponin-T and myogenin permitted the correlation of increased emerin expression with the onset of muscle differentiation. These data suggest a role for emerin during proliferation of activated satellite cells and at the early stages of differentiation.


Subject(s)
Membrane Proteins/biosynthesis , Muscle, Skeletal/cytology , Thymopoietins/biosynthesis , Animals , Blotting, Western , Cell Differentiation/physiology , Cell Division/physiology , Cell Line , Cytoplasm/metabolism , Cytoskeleton/metabolism , DNA, Complementary/genetics , Epitopes/genetics , Flow Cytometry , Humans , Membrane Proteins/genetics , Membrane Proteins/immunology , Mice , Microscopy, Electron , Microtubules/metabolism , Muscle, Skeletal/metabolism , Nuclear Proteins , Subcellular Fractions/metabolism , Thymopoietins/genetics , Thymopoietins/immunology , Transfection
9.
Biol Chem ; 380(6): 653-60, 1999 Jun.
Article in English | MEDLINE | ID: mdl-10430029

ABSTRACT

Thymopoietins (TMPOs) are a group of ubiquitously expressed nuclear proteins. They are suggested to play an important role in nuclear envelope organization and cell cycle control, as has been shown for lamina-associated polypeptides 2 alpha and beta, which are the rat homologs of human TMPOalpha and TMPObeta, respectively. The recent isolation and characterization of seven mouse TMPO mRNA transcripts named TMPO-alpha, beta, beta', gamma, epsilon delta and zeta, suggest that more than the three previously reported transcripts, alpha, beta, and gamma forms, may exist in humans. Here we report on the demonstration of putative human TMPOdelta and epsilon by immunoblotting of human cell lines using a newly prepared polyclonal antiserum against the common N-terminal region of TMPO. Furthermore, we prepared the first truly TMPO-beta-specific, affinity-purified polyclonal antiserum, using a part of the human analog of the beta-specific domain of mouse TMPO 220-259 for immunization. We showed that human TMPObeta is highly expressed in all cancerous cells tested, while hardly any cross-reactivities with other proteins could be detected. In contrast to the high expression of human TMPObeta in the cancer-derived neuroblastoma cell lines SK-N-MC and SMS-KAN, we found very low expression of human TMPObeta in low-proliferative nerve tissue. These data led us to the assumption that expression of TMPObeta may correlate with the occurrence of cancer, and therefore may serve as a new tumor marker, or even as a new target for cancer therapy.


Subject(s)
Cell Division/physiology , Thymopoietins/physiology , Amino Acid Sequence , Animals , Antibodies/immunology , Antibody Specificity , Cerebral Cortex/metabolism , Humans , Immunohistochemistry , Mice , Molecular Sequence Data , RNA, Messenger/genetics , Rats , Thymopoietins/genetics , Thymopoietins/immunology
10.
Hum Mol Genet ; 8(2): 353-9, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9949197

ABSTRACT

Emerin is a nuclear membrane protein which is missing or defective in Emery-Dreifuss muscular dystrophy (EDMD). It is one member of a family of lamina-associated proteins which includes LAP1, LAP2 and lamin B receptor (LBR). A panel of 16 monoclonal antibodies (mAbs) has been mapped to six specific sites throughout the emerin molecule using phage-displayed peptide libraries and has been used to localize emerin in human and rabbit heart. Several mAbs against different emerin epitopes did not recognize intercalated discs in the heart, though they recognized cardiomyocyte nuclei strongly, both at the rim and in intranuclear spots or channels. A polyclonal rabbit antiserum against emerin did recognize both nuclear membrane and intercalated discs but, after affinity purification against a pure-emerin band on a western blot, it stained only the nuclear membrane. These results would not be expected if immunostaining at intercalated discs were due to a product of the emerin gene and, therefore, cast some doubt upon the hypothesis that cardiac defects in EDMD are caused by absence of emerin from intercalated discs. Although emerin was abundant in the membranes of cardiomyocyte nuclei, it was absent from many non-myocyte cells in the heart. This distribution of emerin was similar to that of lamin A, a candidate gene for an autosomal form of EDMD. In contrast, lamin B1 was absent from cardiomyocyte nuclei, showing that lamin B1 is not essential for localization of emerin to the nuclear lamina. Lamin B1 is also almost completely absent from skeletal muscle nuclei. In EDMD, the additional absence of lamin B1 from heart and skeletal muscle nuclei which already lack emerin may offer an alternative explanation of why these tissues are particularly affected.


Subject(s)
Membrane Proteins/analysis , Muscular Dystrophies/metabolism , Myocardium/chemistry , Nuclear Proteins/analysis , Thymopoietins/analysis , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Humans , Immunohistochemistry , Lamin Type A , Lamin Type B , Lamins , Membrane Proteins/immunology , Mice , Mice, Inbred BALB C , Microscopy, Fluorescence , Molecular Sequence Data , Muscular Dystrophies/pathology , Myocardium/cytology , Rabbits , Rats , Sequence Homology, Amino Acid , Thymopoietins/immunology
11.
Peptides ; 19(9): 1479-87, 1998.
Article in English | MEDLINE | ID: mdl-9864053

ABSTRACT

Taking into account the sequential homology existing between thymopoietin II, the DNA-binding domain of p53 protein and FKBP (FK-506 binding protein), a series of fragments of human and bovine FKBP containing a fragment Ser39-Pro45 were synthesized. In the humoral in vitro test all the peptides act as stimulators. Whereas in the in vivo test peptides derived from bovine FKBP show an immunostimulative and those from human FKBP an immunosuppressive activity. However, after blocking the Asp residue by a Bzl group the peptide V appears to be an immunostimulator. The data obtained suggest that these peptides can influence the immune system by blocking the FKBP receptor.


Subject(s)
Adjuvants, Immunologic , Immunophilins/immunology , Amino Acid Sequence , Animals , Biological Assay , Cattle , Humans , Interleukin-1/analysis , Interleukin-2/analysis , Interleukin-6/analysis , Mice , Mice, Inbred BALB C , Mice, Inbred CBA , Molecular Sequence Data , Oligopeptides/immunology , Tacrolimus Binding Proteins , Thymopentin/immunology , Thymopoietins/immunology , Tumor Necrosis Factor-alpha/analysis , Tumor Suppressor Protein p53/immunology
12.
Immunol Res ; 17(3): 345-68, 1998.
Article in English | MEDLINE | ID: mdl-9638477

ABSTRACT

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.


Subject(s)
Adjuvants, Immunologic/pharmacology , Peptide Fragments/pharmacology , Thymopentin/pharmacology , Thymopoietins/pharmacology , Animals , Autoimmune Diseases/drug therapy , Child , Dermatitis/drug therapy , Humans , Immunologic Deficiency Syndromes/drug therapy , Infections/drug therapy , Myasthenia Gravis/immunology , Neoplasms/drug therapy , Neuromuscular Agents/pharmacology , Peptide Fragments/immunology , Peptide Fragments/therapeutic use , Thymopentin/immunology , Thymopentin/therapeutic use , Thymopoietins/immunology , Thymopoietins/isolation & purification , Thymopoietins/therapeutic use
13.
J Cell Sci ; 111 ( Pt 6): 781-92, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9472006

ABSTRACT

The product of the X-linked Emery-Dreifuss muscular dystrophy gene is a protein called emerin, which is localized to the nuclear membrane. We have expressed full-length recombinant human emerin in an in vitro coupled reticulocyte system; it has a molecular mass of 34 kDa, inserts into microsomes in a type II orientation, and does not exhibit any N-linked glycosylation or cleavage event. Affinity-purified human emerin antiserum cross-reacts with the in vitro-expressed emerin and with a 34 kDa band present in a wide range of human tissue samples. Expression and subcellular distribution of emerin were studied in lymphoblastoid cell lines established from four patients with Emery-Dreifuss muscular dystrophy containing different mutations in the emerin gene. Emerin protein was detected in two of these patients by immunoblotting. In striking contrast to wild-type emerin, which was localized to the nuclear fraction and was insoluble in non-ionic detergents and high salt, emerin from these two patients exhibited a more random subcellular localization and increased solubility. On the basis of the mutations present in these patients, it would appear that emerin possesses two non-overlapping nuclear envelope targeting sequences. We have also demonstrated that emerin can occur in four different phosphorylated forms, three of which appear to be associated with the cell cycle. The mutant forms of emerin taken from the two patients exhibited aberrant cell cycle-dependent phosphorylated forms. This data suggests that for emerin to function normally it must be correctly localized, retained at the nuclear membrane and phosphorylated by cell cycle-mediated events.


Subject(s)
Cell Cycle , Intracellular Fluid/metabolism , Membrane Proteins/metabolism , Muscular Dystrophies/metabolism , Muscular Dystrophies/pathology , Thymopoietins/metabolism , Adult , Alkaline Phosphatase/pharmacology , Amino Acid Sequence , Animals , Cell Cycle/genetics , Cell Line, Transformed , Cloning, Molecular , DNA, Complementary/isolation & purification , Humans , Immune Sera/isolation & purification , Male , Membrane Proteins/genetics , Membrane Proteins/immunology , Molecular Sequence Data , Muscular Dystrophies/genetics , Muscular Dystrophy, Emery-Dreifuss , Nuclear Proteins , Octoxynol/pharmacology , Phenotype , Phosphorylation , Rats , Recombinant Proteins/biosynthesis , Sodium Chloride/pharmacology , Solubility , Subcellular Fractions/metabolism , Thymopoietins/genetics , Thymopoietins/immunology
14.
Exp Cell Res ; 230(1): 133-44, 1997 Jan 10.
Article in English | MEDLINE | ID: mdl-9013715

ABSTRACT

The nuclear envelope contains three distinct membrane domains, the outer membrane, the inner membrane, and the pore membrane, that reversibly vesiculate in mitosis. We previously suggested from single-labeling immunofluorescence microscopy analysis of mitotic cells in culture that mitotic vesiculation of the nuclear membranes may proceed in a domain-specific manner (Chaudhary and Courvalin, J. Cell. Biol. 122, 295-306, 1993). In the present study, we add biochemical support to this hypothesis by sorting domain-specific mitotic vesicles. Antibodies directed against the lamin B receptor, a marker of the inner membrane, and glycoprotein gp210, a marker of the pore membrane, were used to isolate by affinity two populations of mitotic vesicles that were selectively enriched in each of these markers. These two vesicle populations were of different size distribution; the pore membrane-derived vesicles were smaller (80% being < or = 200 nm) than the inner membrane-derived vesicles (80% > or = 200 nm). Double-labeling immunofluorescence microscopy analysis of mitotic cells in culture showed that the time course and topology of disassembly and reassembly of inner and pore membrane domains were different, confirming that domain-specific vesicles are generated during mitosis. In these studies, protein LAP2/thymopoietin beta, another marker of the inner nuclear membrane, was segregating as lamin B receptor, suggesting that both proteins were contained in the same mitotic vesicles.


Subject(s)
DNA-Binding Proteins , Mitosis/physiology , Nuclear Envelope/metabolism , Animals , Antibodies , Biomarkers , Cell Fractionation , Cell Line , HeLa Cells , Humans , Membrane Glycoproteins/immunology , Membrane Glycoproteins/metabolism , Membrane Proteins/immunology , Membrane Proteins/metabolism , Microscopy, Electron , Microscopy, Fluorescence , Nuclear Envelope/ultrastructure , Nuclear Pore Complex Proteins , Nuclear Proteins/immunology , Nuclear Proteins/metabolism , Rabbits , Receptors, Cytoplasmic and Nuclear/immunology , Receptors, Cytoplasmic and Nuclear/metabolism , Thymopoietins/immunology , Thymopoietins/metabolism , Lamin B Receptor
15.
Biochem Biophys Res Commun ; 226(2): 431-8, 1996 Sep 13.
Article in English | MEDLINE | ID: mdl-8806652

ABSTRACT

An expression vector was constructed to produce a common region of human thymopoietin family proteins. The recombinant protein was expressed in Escherichia coli as a fusion protein with a biotinylated tag region and purified by affinity chromatography on a monomeric avidin resin. The thymopoietin family-specific antibody was induced in rabbits by immunization with the recombinant fusion protein. Western blotting analysis using the antibody revealed that the expression of thymopoietin family proteins was remarkably tissue specific. Among those, thymopoietin beta/lamina-associated polypeptide 2 appears to be specifically expressed in tissues with high proliferative activity.


Subject(s)
DNA-Binding Proteins , Membrane Proteins/genetics , Nuclear Proteins/genetics , Thymopoietins/genetics , Amino Acid Sequence , Animals , Antibodies/chemistry , Antibodies/immunology , Antibody Specificity , Base Sequence , Blotting, Western , Cells, Cultured , Escherichia coli/genetics , Factor Xa/metabolism , Humans , Immunochemistry , Membrane Proteins/immunology , Membrane Proteins/isolation & purification , Molecular Sequence Data , Nuclear Proteins/immunology , Nuclear Proteins/isolation & purification , Rabbits , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/isolation & purification , Thymopoietins/immunology , Thymopoietins/isolation & purification
16.
Acta Virol ; 37(2-3): 181-3, 1993.
Article in English | MEDLINE | ID: mdl-8105663

ABSTRACT

The influence of the peptide diacetylsplenopentin (SP5) on an early protection of guinea pigs against foot-and-mouth disease (FMD) was investigated. 80% protection was achieved if SP5 was applied in a dose of 2 mg one day before challenge with FMD virus (FMDV) type O1 Lausanne strain. In comparison with this a conventional commercial adsorbate vaccine protected guinea pigs about 7 days after vaccination. An earlier protection can be obtained in general by vaccination with a higher content of the immunogen. A tenfold increase of the 146 S particle dose in a conventional oil adjuvanted FMDV vaccine protected pigs about 2 days earlier as observed after inoculating a "normal" vaccine (about 11 days).


Subject(s)
Aphthovirus/immunology , Foot-and-Mouth Disease/prevention & control , Peptide Fragments/immunology , Thymopoietins/immunology , Viral Vaccines , Adjuvants, Immunologic , Animals , Antibodies, Viral/biosynthesis , Guinea Pigs , Viral Vaccines/immunology
17.
Thymus ; 18(3): 169-84, 1991 Nov.
Article in English | MEDLINE | ID: mdl-1723818

ABSTRACT

Murine monoclonal antibodies (mAbs) were developed to discriminate thymopoietin, a human thymic hormone, and thysplenin, a closely related molecule found in spleen. Three of these recognized both native and synthetic thymopoietin as well as thysplenin. Together they define two non-overlapping epitopes which withstand sodium dodecyl sulfate denaturation and can be detected by western blotting. We used these three mAbs to demonstrate the production of thymopoietin by cultured thymic epithelial cells for up to several weeks. Three additional mAbs were selective for thysplenin. Highly specific mAbs will be useful for characterizing further these physiologically distinct polypeptides.


Subject(s)
Antibodies, Monoclonal/immunology , Epitopes/immunology , Thymopoietins/analysis , Thymus Gland/chemistry , Cells, Cultured/chemistry , Epithelial Cells , Fluorescent Antibody Technique , Humans , Sensitivity and Specificity , Spleen/chemistry , Thymopoietins/immunology , Thymus Gland/cytology
18.
Chem Pharm Bull (Tokyo) ; 37(2): 391-6, 1989 Feb.
Article in English | MEDLINE | ID: mdl-2787213

ABSTRACT

[Glu34]human splenin (hSP) was synthesized in a conventional manner by assembling ten peptide fragments followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole (molar ratio, 1:1) in trifluoroacetic acid in the presence of m-cresol and dimethylselenide. Finally, the deprotected peptide was incubated with dithiothreitol to reduce sulfoxide on the methionine side chain. Incubation of peripheral lymphocytes isolated from uremic patients with the synthetic [Glu34]hSP showed an enhancing effect on the reduced B-lymphocytes, but synthetic human thymopoietin (hTP) had no effect under the same conditions.


Subject(s)
B-Lymphocytes/drug effects , Thymopoietins/chemical synthesis , Thymus Hormones/chemical synthesis , Uremia/blood , Amino Acid Sequence , Amino Acids/analysis , Humans , In Vitro Techniques , Molecular Sequence Data , Thymopoietins/immunology , Thymopoietins/pharmacology
20.
Thymus ; 11(3): 173-83, 1988.
Article in English | MEDLINE | ID: mdl-3363629

ABSTRACT

Anti-thymosin alpha 1 monoclonal antibodies recognized, on immunoblots, 1 to 2 bands corresponding to molecules of 34 and 35 Kd when using aqueous extracts of thymus, spleen, kidney, liver, brain, pituitary and adrenal glands from rats or mice. Anti-bovine thymopoietin polyclonal antibodies, in the same conditions, labelled analogous 34, 35 and 35.5 Kd molecules in brain and thymus extracts but also a 40 Kd molecules in thymus and a 90 Kd in brain extracts. Anti-synthetic thymulin monoclonal antibodies recognized irregularly and poorly a 52 Kd molecule from thymus and brain extracts. These results suggest that thymopoietin, thymulin and specially Thymosin alpha 1 are first synthesized in large precursors. Finally, other organs seem capable of synthesizing thymosin alpha 1 and probably thymopoietin, but for thymulin, the results are too irregular to conclude.


Subject(s)
Antibodies, Monoclonal/analysis , Thymic Factor, Circulating/immunology , Thymopoietins/immunology , Thymosin/analogs & derivatives , Thymus Hormones/immunology , Tissue Extracts/immunology , Animals , Collodion , Electrophoresis, Polyacrylamide Gel , Mice , Mice, Inbred Strains , Rats , Rats, Inbred Strains , Thymalfasin , Thymosin/immunology , Thymus Gland/metabolism
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