RESUMO
We have studied anti-tumor properties of bone marrow derived peptide Phe-Leu-Gly-Phe-Pro-Thr (MP-1) synthesized by classical methods. It was shown that MP-1 enhanced the effect ofcytostatic therapy of lymphatic leukemia P388 and increased latent growth period of P388 tumors implanted in irradiated mice. MP-1 also decreased metastasis of mouse breast adenocarcinoma Ca-755 after surgery.
Assuntos
Antineoplásicos , Leucemia P388 , Oligopeptídeos , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/síntese química , Linhagem Celular Tumoral , Cisplatino/administração & dosagem , Terapia Combinada , Leucemia P388/tratamento farmacológico , Leucemia P388/radioterapia , Camundongos , Oligopeptídeos/administração & dosagem , Oligopeptídeos/síntese químicaRESUMO
The bone marrow myelopeptides Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro (MP-4) and Val-Asp-Pro-Pro (MP-6) have been synthesised by a classical method and by a solid phase synthesis. The differentiating activity of MP-4 and MP-6 in human leukemia cells HL-60 and K-562 has been studied. Both peptides induce terminal differentiation of these cell lines but the mechanism of action of peptides MP-4 and MP-6 is distinguished.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Oligopeptídeos/síntese química , Oligopeptídeos/farmacologia , Células HL-60 , Humanos , Células K562 , Oligopeptídeos/químicaRESUMO
The Val-Val-Tyr-Pro-Asp bone marrow peptide (MP-5) and an analogue (MP-5-Lys) were synthesized. Fluorescent derivatives, Ftc-MP-5 and MP-5-Lys(Ftc), were prepared. The biological activity of MP-5 and MP-5-Lys was studied in vitro and in vivo. The MP-5 peptide caused 60-84% inhibition of growth of the following mouse cancers: lymphatic leukemia P-388, melanoma B-16, and cervical carcinoma CUC-5. These peptides also restored functional activity of T lymphocytes that was inhibited by metabolic products of the HL-60 leukemic cell line. MP-5-Lys(Ftc) was shown to preserve the functional properties of MP-5 toward T lymphocytes, but Ftc-MP-5 was practically inactive.
Assuntos
Antineoplásicos/síntese química , Corantes Fluorescentes/síntese química , Fatores Imunológicos/síntese química , Oligopeptídeos/síntese química , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacologia , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Camundongos , Oligopeptídeos/química , Oligopeptídeos/farmacologia , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Combined application of mumps and measles vaccine strains in equal doses results in significant decrease of immune response to the former component in humans. It is possible that this phenomenon is related with well-known immunodepressive effect of measles virus, which was demonstrated both in vivo and in vitro. It was previously shown that myelopeptide-2 (MP-2) partially neutralizes suppressive effect of measles vaccine on blast transformation of activated human lymphocytes in vitro. Partial supression of immune response to mumps vaccine by live measles vaccine was reproduced in laboratory animals. It was shown that in experiment MP-2 partially neutralized suppressive effect of measles vaccine.
Assuntos
Adjuvantes Imunológicos/farmacologia , Imunização , Vacina contra Sarampo/administração & dosagem , Sarampo/imunologia , Morbillivirus/imunologia , Vacina contra Caxumba/administração & dosagem , Vacina contra Caxumba/imunologia , Vírus da Caxumba/imunologia , Caxumba/imunologia , Oligopeptídeos/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Anticorpos Antivirais/sangue , Avaliação Pré-Clínica de Medicamentos , Feminino , Cobaias , Esquemas de Imunização , Terapia de Imunossupressão , Injeções Intramusculares , Sarampo/sangue , Caxumba/sangue , Oligopeptídeos/administração & dosagem , Ratos , Vacinas Combinadas/imunologiaRESUMO
The lifetimes of channels formed by natural gramicidin and its dimeric analog in monoglyceride lipid bilayers of various compositions were investigated. The bilayer surface tension was altered by changing the length of the monoglycerides' fatty acid chain or the chain length of hydrocarbon solvent by isomerization or saturation of the lipid, by varying the amount of solvent in the bilayer, and by changing the salt composition of the aqueous solutions. The logarithms of mean channel lifetimes were found to be proportional to the surface tension of the membrane irrespective of how the surface tension was changed. In contrast, no simple relationship between channel conductance and surface tension or bilayer thickness was found.
Assuntos
Colesterol , Glicerídeos , Gramicidina , Bicamadas Lipídicas , Canais Iônicos , Potenciais da MembranaRESUMO
The bone marrow myelopeptide MP-2 (Leu-Val-Val-Tyr-Pro-Trp), exhibiting antitumor activity, and its retro-analogue (Trp-Pro-Tyr-Val-Val-Leu) were synthesized, and their properties were studied. The in vitro and in vivo activities of retro-MP-2 were comparable with those of MP-2. Both peptides equally restored the functional activity of T-lymphocytes inhibited by toxins released by HL-60 cells and inhibited by 70-82% the growth of various types of transplantable solid tumors: Ca-755 adenocarcinoma of the mammary gland, Lewis adenocarcinoma of the lung, and S180 sarcoma. The positions and intensities of the Cotton effects in CD spectra of the MP-2 peptide and its retro-analogue in various solvents are almost indistinguishable. The positions of extrema and integral intensities of the amide I and amide A bands in IR spectra of both peptides were practically identical. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.
Assuntos
Adenocarcinoma/tratamento farmacológico , Adjuvantes Imunológicos/síntese química , Adjuvantes Imunológicos/farmacologia , Carcinoma Pulmonar de Lewis/tratamento farmacológico , Neoplasias Mamárias Experimentais/tratamento farmacológico , Oligopeptídeos/síntese química , Oligopeptídeos/farmacologia , Adjuvantes Imunológicos/química , Animais , Células HL-60 , Humanos , Camundongos , Oligopeptídeos/químicaRESUMO
Along with known lymphokines involved in the regulation of hematopoiesis, a new differentiating factor (myelopeptide-4, MP-4) for myeloid cells was found. The peptide (Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro) originally isolated from the culture medium of porcine bone marrow cell culture was examined for its ability to induce differentiation in two human myeloid leukemia cell lines, HL-60 and K-562. Agents with well-known differentiation-inducing activity, such as phorbol myristate acetate, dimethylsulfoxide and the lymphokines were used as a reference. It has been shown that MP-4 significantly influences the integral characteristics of metabolism, expression of surface antigens and morphology of these cells. It decreased the level of chromosomal DNA synthesis and, in parallel, increased the total protein synthesis in both HL-60 and K-562 cells. MP-4 induced the expression of CD14 monocyte-specific surface antigen and the appearance of mature monocytes/macrophages in HL-60 cell cultures. There was a good correlation of cell metabolic/morphological changes and the CD14 marker expression for HL-60 cells. A similar phenomenon was observed in K-562 cells treated with MP-4 when the levels of hemoglobin synthesis were detected in their cytoplasm. Thus, we consider MP-4 as a new endogenous differentiating factor for myeloid cells.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Monócitos/citologia , Monócitos/efeitos dos fármacos , Oligopeptídeos/farmacologia , Sequência de Aminoácidos , Divisão Celular/efeitos dos fármacos , Tamanho Celular/efeitos dos fármacos , Meios de Cultivo Condicionados/farmacologia , DNA/biossíntese , Dimetil Sulfóxido/farmacologia , Células HL-60 , Hemoglobinas/biossíntese , Humanos , Células K562 , Receptores de Lipopolissacarídeos/metabolismo , Linfocinas/imunologia , Linfocinas/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Monócitos/metabolismo , Oligopeptídeos/química , Biossíntese de Proteínas , Linfócitos T/imunologia , Acetato de Tetradecanoilforbol/farmacologia , Regulação para Cima/efeitos dos fármacosRESUMO
Myelopeptide-2 (MP-2) Leu-Val-Val-Tyr-Pro-Trp originally isolated from the supernatant of porcine bone marrow cell culture was examined for its capacity to restore the mitogen responsiveness of human T lymphocytes inhibited by conditioned media from HL-60 leukemia cells (HL-60 CM). MP-2 added to phytohemagglutinin (PHA)-stimulated T lymphocytes together with HL-60 CM abolished the suppression of T-lymphocyte proliferative response in a dose-dependent manner. Another bone marrow hexapeptide Phe-Leu-Gly-Phe-Pro-Thr, MP-1, did not display this action in that experimental system. MP-2 was also effective being added after T-lymphocyte exposure to HL-60 CM which suggests its recovery but not protective effect on T-lymphocytes treated with tumor cell products. Flow cytometry analysis revealed HL-60 CM influence on the expression of CD3 and CD4 T-cell surface antigens. It decreased the content of CD3- and CD4-positive cells and induced the appearance of T lymphocytes with reduced density of CD3 and CD4 antigens. MP-2 was able to restore the T-cell phenotype altered by HL-60 CM. MP-2 seems to be promising in anti-tumor therapy.
Assuntos
Medula Óssea/imunologia , Complexo CD3/metabolismo , Antígenos CD4/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Oligopeptídeos , Peptídeos/farmacologia , Linfócitos T/imunologia , Células da Medula Óssea , Antígenos CD8/fisiologia , Citometria de Fluxo , Células HL-60 , Humanos , Imunidade Celular , Mitógenos/farmacologia , Fito-Hemaglutininas/farmacologia , Regulação para CimaRESUMO
Myelopeptide 1 (MP-1) is hexapeptide originally isolated from porcine bone marrow cell culture. It was synthesized and its immunoregulatory properties were studied. MP-1 caused a 1.5-2-fold dose-dependent increase of antibody production in the culture of mouse immune lymph node cells. It abolished Con A induction of T suppressors in the suspension of mouse spleen cells and counteracted the inhibitory effect of T suppressors on antibody production. The inoculation of MP-1 (1 x 10(-9) g/mouse) to mice two weeks after their gamma-irradiation (2 Gy) resulted in an increase of antibody production up to 80.2 +/- 15.5% as compared to that in the irradiated control 37.6 +/- 12.0%. Immunofluorescent analysis revealed the specific binding of MP-1 with receptors on the target cells in the suspension of mouse spleen cells. It is supposed that MP-1 participates in the immunoregulatory processes in the living organism.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Medula Óssea/fisiologia , Oligopeptídeos/farmacologia , Linfócitos T Reguladores/imunologia , Animais , Formação de Anticorpos/efeitos da radiação , Células Cultivadas , Concanavalina A , Feminino , Imunofluorescência , Raios gama , Fatores Imunológicos/farmacologia , Linfonodos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Oligopeptídeos/isolamento & purificação , Oligopeptídeos/metabolismo , Baço/imunologia , Suínos , Linfócitos T , Linfócitos T Reguladores/efeitos dos fármacosRESUMO
Bone marrow cells of various animal species and men produce a group of bioregulatory peptides called myelopeptides (MPs). A highly purified MP fraction and some individual molecules have been isolated from the supernatant of porcine bone marrow cell cultures by reverse phase chromatography. MPs have a wide spectrum of functional activities: immunoregulatory, differentiating and opiate-like. They evoke 2.5-fold stimulation of antibody production to various antigens. They correct some immune defects in MRL/lpr mice with spontaneous autoimmune disorders that results in 2-fold prolongation of the life span of these mice. MPs influence the differentiation of bone marrow and peripheral blood cells derived from healthy and leukemic donors. They induce terminal differentiation in the leukemic human HL-60 cell line. MPs also show an effect on pain sensitivity. A new immunocorrective drug Myelopid has been developed on the basis of MP mixtures. This drug is effectively used in Russia both in medicine and veterinary practice for prophylaxis and treatment of diseases accompanied by immunodeficiency. Two individual MPs were isolated and identified: Phe-Leu-Gly-Phe-Pro-Thr (MP-1) and Leu-Val-Val-Tyr-Pro-Trp (MP-2). MP-1 displays immunoregulatory activity; MP-2 abolishes the inhibitory effect of leukemic cells on T-lymphocyte functional activity. MPs seem to provide not only immunoregulation but also to participate in complex interactions between different systems in the organism.
Assuntos
Ativinas , Medula Óssea/metabolismo , Fatores Imunológicos/fisiologia , Oligopeptídeos , Peptídeos/fisiologia , Sequência de Aminoácidos , Analgésicos Opioides , Animais , Formação de Anticorpos , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/uso terapêutico , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/isolamento & purificação , Peptídeos/uso terapêutico , Linfócitos T/imunologiaRESUMO
Two hexapeptides affecting pain sensitivity were isolated from the bone marrow cells supernatant, shown to have primary structure Leu-Val-Val-Tyr-Pro-Trp and Phe-Leu-Gly-Phe-Pro-Thr, and synthesized by the solid phase method. In physico-chemical and biological properties the synthetic peptides were identical to the peptides isolated.
Assuntos
Medula Óssea/química , Oligopeptídeos/isolamento & purificação , Sequência de Aminoácidos , Animais , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Dados de Sequência Molecular , Oligopeptídeos/química , Oligopeptídeos/fisiologia , Limiar da Dor , SuínosRESUMO
Previously unknown bioregulators from bone marrow, myelopeptides, were isolated, identified, and synthesized, and their biological properties and mechanism of action were studied in detail. Phe-Leu-Gly-Phe-Pro-Thr (MP-1) manifests an immunocorrecting effect by restoring the level of antibody production in animals suffering from immunodeficiencies of various etiologies; Leu-Val-Val-Tyr-Pro-Trp (MP-2) manifests an antitumor effect by abolishing the inhibitory action of tumor cells on the functional activity of T-lymphocytes; Leu-Val-Cys-Tyr-Pro-Gln (MP-3) stimulates phagocytosis by macrophages and, in this way, protects animals from infections caused by pathogenic microorganisms; and Phe-Pro-Arg-Ile-Met-Thr-Pro (MP-4) is a new factor of cell differentiation inducing terminal cell differentiation in the HL-60 and K-562 leukemic cell lines.
Assuntos
Adjuvantes Imunológicos/química , Oligopeptídeos/química , Oligopeptídeos/farmacologia , Adjuvantes Imunológicos/farmacologia , Sequência de Aminoácidos , Animais , Diferenciação Celular/efeitos dos fármacos , Humanos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Fagocitose/efeitos dos fármacos , Relação Estrutura-Atividade , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Células Tumorais CultivadasRESUMO
Peptide Leu-Val-Cys-Tyr-Pro-Gln, identical to the bone marrow peptide MP-3, and its Val3 and Ser3 analogs, lacking SH-group, were synthesized by conventional methods of peptide chemistry in solution and, along with the MP-3 S-S-dimerization product, were studied with respect to their effect on the macrophage phagocytic activity. It was shown that the activity was only enhanced by peptide MP-3, which demonstrated the essential role of the SH-group in this function. The dimer analog of MP-3, unlike dimer analogs of other monocysteine-containing peptides, glutathione and HP5b, did not exhibit the inhibitory effect.
Assuntos
Ativação de Macrófagos/efeitos dos fármacos , Oligopeptídeos/farmacologia , Compostos de Sulfidrila/farmacologia , Sequência de Aminoácidos , Medula Óssea/química , Cromatografia Líquida de Alta Pressão , Oligopeptídeos/síntese química , Oligopeptídeos/química , Compostos de Sulfidrila/síntese química , Compostos de Sulfidrila/químicaRESUMO
A new method was developed for the isolation of biologically active peptides present in small quantities in supernatants of bone marrow cell cultures. Four new peptides, which exhibited immunostimulating and differentiating activities, were isolated from the supernatant using solid phase extraction and reversed-phase HPLC. For these peptides, the following primary structures were elucidated: Leu-Val-Cys-Tyr-Pro-Gln, Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro, Val-Val-Tyr-Pro-Asp, and Val-Asp-Pro-Pro.
Assuntos
Adjuvantes Imunológicos/isolamento & purificação , Células da Medula Óssea/metabolismo , Oligopeptídeos/isolamento & purificação , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Aminoácidos/análise , Animais , Células Produtoras de Anticorpos/efeitos dos fármacos , Células Produtoras de Anticorpos/imunologia , Células da Medula Óssea/química , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Células Cultivadas , Fracionamento Químico , Cromatografia Líquida de Alta Pressão , Células HL-60 , Humanos , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Camundongos , Oligopeptídeos/química , Oligopeptídeos/farmacologia , SuínosRESUMO
A model is suggested to study the effects the myelopeptide 2 (MP2) immunomodulator on the results of live measles rash immunized by vaccine. Guinea pigs were found to respond to live measles vaccine L-16 the way man does, i.e. the antibodies' level in their sera was increasing until days 28-56 after a single intramuscular vaccine injection. An intramuscular injection of the MP2 immunomodular, made alongside with vaccination, was demonstrated to enhance the immune response to the measles vaccine and to induce its action. Finally, it was for the first time, that the possibility was proven to enhance the action of the live measles vaccine by an immunomodulator.
Assuntos
Fatores Imunológicos/administração & dosagem , Vacina contra Sarampo/imunologia , Sarampo/imunologia , Oligopeptídeos/administração & dosagem , Vacinação , Animais , Anticorpos Antivirais/sangue , Avaliação Pré-Clínica de Medicamentos , Cobaias , Injeções Intramusculares , Sarampo/sangue , Morbillivirus/imunologia , Fatores de TempoAssuntos
Fatores Imunológicos/fisiologia , Leucemia Mieloide Aguda/patologia , Oligopeptídeos , Peptídeos/fisiologia , Linfócitos T/imunologia , Adjuvantes Imunológicos , Sequência de Aminoácidos , Humanos , Fatores Imunológicos/química , Dados de Sequência Molecular , Peptídeos/química , Células Tumorais CultivadasAssuntos
Adjuvantes Imunológicos/fisiologia , Oligopeptídeos , Peptídeos/fisiologia , Sequência de Aminoácidos , Animais , Células Cultivadas , Feminino , Linfonodos/citologia , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Dados de Sequência Molecular , Baço/citologia , Baço/imunologia , Linfócitos T Reguladores/imunologiaAssuntos
Medula Óssea/química , Macrófagos Peritoneais/imunologia , Oligopeptídeos , Peptídeos/fisiologia , Fagocitose/fisiologia , Animais , Relação Dose-Resposta Imunológica , Macrófagos Peritoneais/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Peptídeos/química , Peptídeos/isolamento & purificação , Salmonella typhimurium/imunologiaRESUMO
We studied the capacity of myelopeptide-4 (regulatory peptide of the bone marrow origin) to induce terminal differentiation of HL-60 and K-562 leukemic cells. Myelopeptide-4 increased the expression of CD14 and CD38 differentiation antigens on the surface of HL-60 cells and of CD44 antigen on K-562 cells, induced the appearance of mature monocyte/macrophages in HL-60 culture and hemoglobin-producing cells in K-562 cell culture, and stimulated phagocytic activity of THP-1 leukemic cells. Myelopeptide-4 is an endogenous factor of cell differentiation, a prospective agent for antileukemic therapy.