RESUMEN
A novel actively targeted polymer carrier for anticancer drugs based on an N-(2-hydroxypropyl)methacrylamide copolymer (PHPMA) is proposed. An oligopeptide sequence GE7, attached to the polymer, is a specific ligand for the EGF receptor overexpressed on most tumor cells. Co-attachment of selected chemotherapeutics will therefore lead to formation of tumor-specific polymer therapeutics, further enhanced by the EPR effect. FACS measurements prove elevated binding activity of the fluorescently labeled PHPMA/GE7 conjugate in EGFR-rich cells (FaDu, MCF-7), compared to conjugates of scrambled peptides. Cell lines with low EGFR level (SW620, B16F10) bind the GE7 conjugate significantly less.
Asunto(s)
Antineoplásicos/química , Sistemas de Liberación de Medicamentos/métodos , Receptores ErbB/metabolismo , Ácidos Polimetacrílicos/uso terapéutico , Antineoplásicos/metabolismo , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Citometría de Flujo , Humanos , Oligopéptidos/genética , Oligopéptidos/metabolismo , Ácidos Polimetacrílicos/síntesis química , Ácidos Polimetacrílicos/química , Tomografía de Emisión de Positrones , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espectrofotometría UltravioletaRESUMEN
Herein, new biodegradable star polymer-doxorubicin conjugates designed for passive tumor targeting were investigated, and their synthesis, physico-chemical characterization, drug release, biodegradation, biodistribution and in vivo anti-tumor efficacy are described. In the conjugates, the core formed by poly(amidoamine) (PAMAM) dendrimers was grafted with semitelechelic N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers bearing doxorubicin (Dox) attached by hydrazone bonds, which enabled intracellular pH-controlled drug release. The described synthesis facilitated the preparation of biodegradable polymer conjugates in a broad range of molecular weights (200-1000g/mol) while still maintaining low polydispersity (~1.7). The polymer grafts were attached to the dendrimers through either stable amide bonds or enzymatically or reductively degradable spacers, which enabled intracellular degradation of the high-molecular-weight polymer carrier to excretable products. Biodegradability tests in suspensions of EL4 T-cell lymphoma cells showed that the rate of degradation was much faster for reductively degradable conjugates (close to completion within 24h of incubation) than for conjugates linked via an enzymatically degradable oligopeptide GFLG sequence (slow degradation taking several days). This finding was likely due to the differences in steric hindrance in terms of the accessibility of the small molecule glutathione and the bulky enzyme cathepsin B to the polymer substrate. Regarding drug release, the conjugates were fairly stable in buffer at pH 7.4 (model of blood stream) but released doxorubicin under mild acidic conditions that model the tumor cell microenvironment. The star polymer-Dox conjugates exhibited significantly prolonged blood circulation and enhanced tumor accumulation in tumor-bearing mice, indicating the important role of the EPR effect in its anti-cancer activity. The star polymer conjugates showed prominently higher in vivo anti-tumor activities than the free drug or linear polymer conjugate when tested in mice bearing EL4 T-cell lymphoma, with a significant number of long-term surviving (LTS). Based on the results, we conclude that a M(w) of HPMA copolymers of 200,000 to 600,000g/mol is optimal for polymer carriers designed for the efficient passive targeting to solid tumors. In addition, an expressive therapy-dependent stimulation of the immune system was observed.
Asunto(s)
Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/uso terapéutico , Dendrímeros/química , Doxorrubicina/química , Doxorrubicina/uso terapéutico , Metacrilatos/química , Animales , Antibióticos Antineoplásicos/metabolismo , Antibióticos Antineoplásicos/farmacocinética , Materiales Biocompatibles/química , Materiales Biocompatibles/metabolismo , Línea Celular Tumoral , Dendrímeros/metabolismo , Doxorrubicina/metabolismo , Doxorrubicina/farmacocinética , Humanos , Linfoma/tratamiento farmacológico , Metacrilatos/metabolismo , RatonesRESUMEN
Radioactive decay of some radionuclides produces a shower of Auger electrons, potent ionizing radiation within a very short range in living tissue (typically ca. 100 nm). Therefore, they must be brought to DNA-containing cell compartments and preferentially directly to DNA to be fully biologically effective. They may be used for a triple-targeting approach (first targeting, polymer-based system targeting into tumor tissue due to EPR effect; second targeting, pH-controlled release of intercalator-bound Auger electron emitter in slightly acidic tumor tissue or endosome; third targeting, into DNA in cell nucleus by the intercalator) minimizing radiation burden of healthy tissues. We describe a first system of this type, an ellipticine derivative-bound iodine-125 attached to hydrazide moieties containing poly[N-(2-hydroxypropyl)methacrylamide]. The system is stable at pH 7.4 (0% intercalator released after 24 h incubation), while iodine-containing biologically active intercalator is released upon decrease of pH (25% intercalator released after 24 h incubation at pH 5.0-model of late endosomes). Both 2-N-(2-oxobutyl)-9-iodoellipticinium bromide and the noniodinated 2-N-(2-oxobutyl)ellipticinium bromide are potent intercalators, as proven by direct titration with DNA and ethidium displacement assay, and readily penetrate into cell nuclei, as proven by confocal microscopy. They retain chemotherapeutical antiproliferative properties of ellipticine against Raji, EL-4, and 4T1cells with IC(50) in the range 0.27-8.8 µmol/L. Polymer conjugate of 2-N-(2-oxobutyl)-9-iodoellipticinium bromide is internalized into endosomes, releases active drug, possesses cytotoxic activity, and the drug accumulates in cell nuclei.
Asunto(s)
Electrones , Elipticinas/farmacología , Orgánulos/química , Ácidos Polimetacrílicos/farmacología , Animales , Línea Celular Tumoral , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Supervivencia Celular/efectos de los fármacos , ADN/química , Relación Dosis-Respuesta a Droga , Elipticinas/química , Humanos , Hidrazinas/química , Concentración de Iones de Hidrógeno , Radioisótopos de Yodo , Ratones , Estructura Molecular , Orgánulos/efectos de los fármacos , Ácidos Polimetacrílicos/síntesis química , Ácidos Polimetacrílicos/química , Estereoisomerismo , Relación Estructura-Actividad , Distribución TisularRESUMEN
There is a wide range of techniques utilizing fluorescence of doxorubicin (Dox) commonly used for analysis of intracellular accumulation and destiny of various drug delivery systems containing this anthracycline antibiotic. Unfortunately, results of these studies can be significantly influenced by doxorubicin degradation product, 7,8-dehydro-9,10-desacetyldoxorubicinone (D*) forming spontaneously in aqueous environment, whose fluorescence strongly interfere with that of doxorubicin. Here, we define two microscopy techniques enabling to distinguish and separate Dox and D* emission based either on its spectral properties or on fluorescence lifetime analysis. To analyze influx and nuclear accumulation of Dox (free or polymer-bound) by flow cytometry, we propose using an indirect method based on its DNA intercalation competition with Hoechst 33342 rather than a direct measurement of doxorubicin fluorescence inside the cells.
Asunto(s)
Antibióticos Antineoplásicos/metabolismo , Núcleo Celular/metabolismo , ADN/metabolismo , Doxorrubicina/análogos & derivados , Doxorrubicina/metabolismo , Sistemas de Liberación de Medicamentos , Células 3T3 , Animales , Antibióticos Antineoplásicos/farmacología , Doxorrubicina/farmacología , Citometría de Flujo , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Linfoma de Células T/tratamiento farmacológico , Linfoma de Células T/metabolismo , Ratones , Polímeros/metabolismo , Espectrometría de Fluorescencia , Células Tumorales CultivadasRESUMEN
To avoid the side effects of the anti-cancer drug doxorubicin (Dox), we conjugated this drug to a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer backbone. Dox was conjugated via an amide bond (Dox-HPMA(AM), PK1) or a hydrazone pH-sensitive bond (Dox-HPMA(HYD)). In contrast to Dox and Dox-HPMA(HYD), Dox-HPMA(AM) accumulates within the cell's intracellular membranes, including those of the Golgi complex and endoplasmic reticulum, both involved in protein glycosylation. Flow cytometry was used to determine lectin binding and cell death, immunoblot to characterize the presence of CD7, CD43, CD44, and CD45, and high-performance anion exchange chromatography with pulsed amperometric detector analysis for characterization of plasma membrane saccharide composition. Incubation of EL4 cells with Dox-HPMA(AM) conjugate, in contrast to Dox or Dox-HPMA(HYD), increased the amounts of membrane surface-associated glycoproteins, as well as saccharide moieties recognized by peanut agglutinin, Erythrina cristagalli, or galectin-1 lectins. Only Dox-HPMA(AM) increased expression of the highly glycosylated membrane glycoprotein CD43, while expression of others (CD7, CD44, and CD45) was unaffected. The binding sites for galectin-1 are present on CD43 molecule. Furthermore, we present that EL4 treated with Dox-HPMA(AM) possesses increased sensitivity to galectin-1-induced apoptosis. In this study, we demonstrate that Dox-HPMA(AM) treatment changes glycosylation of the EL4 T cell lymphoma surface and sensitizes the cells to galectin-1-induced apoptosis.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Doxorrubicina/análogos & derivados , Linfoma de Células T/tratamiento farmacológico , Ácidos Polimetacrílicos/farmacología , Amidas/química , Animales , Apoptosis , Western Blotting , Línea Celular Tumoral/efectos de los fármacos , Proliferación Celular , Doxorrubicina/farmacología , Portadores de Fármacos , Retículo Endoplásmico/metabolismo , Citometría de Flujo , Galectina 1/metabolismo , Glicosilación , Aparato de Golgi/metabolismo , Leucosialina/metabolismo , Linfoma de Células T/metabolismo , Linfoma de Células T/patología , RatonesRESUMEN
PURPOSE: In vivo efficacy and safety of HPMA-based copolymers armed with doxorubicin via a spacer containing pH-sensitive linkage that can be prepared within a broad range of attached drug contents (1) was tested in murine tumor models. METHODS: Mice bearing T cell lymphoma EL4 or B cell lymphoma 38C13 were treated with a single dose of the conjugate (15, 25, and 75 mg Dox eq./kg i.v.) in a therapeutic regime. Anti-tumor resistance of the cured animals was proved by a second challenge with a lethal dose of tumor cells without additional treatment. RESULTS: The content of drug bound to the polymer is an important parameter in relation to the conjugate therapeutic efficacy. The best anti-tumor effects were produced by conjugates with 10 - 13 wt% of bound doxorubicin. Free doxorubicin up to 4.6% relative to total drug content had no impact on the treatment efficacy and acute toxicity. The conjugates induced a complete cure of mice and regular treatment-dependent development of specific anti-tumor resistance. No myelosuppression or organ damage was observed. CONCLUSIONS: A well-defined HPMA copolymer-doxorubicin conjugate with pH-sensitive drug release is a good candidate for clinical trials as it has remarkable anti-tumor efficacy and a favorable safety profile.
Asunto(s)
Doxorrubicina/análogos & derivados , Portadores de Fármacos/síntesis química , Inmunomodulación/efectos de los fármacos , Polímeros , Ácidos Polimetacrílicos/farmacología , Ácidos Polimetacrílicos/farmacocinética , Animales , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Doxorrubicina/síntesis química , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacología , Concentración de Iones de Hidrógeno , Masculino , Ratones , Polímeros/síntesis química , Polímeros/farmacocinética , Polímeros/farmacología , Ácidos Polimetacrílicos/síntesis química , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Cancer research is focused on processes which influence in vivo tumor growth dynamics, tumor microenvironment and antitumor immune responses. Recently, it was documented that some cytostatics, including their polymeric derivatives, were able to trigger an anticancer immune response. Such interactions are studied mainly in vitro but relevant in vivo studies are necessary and were only recently started. Whole-body imaging of fluorescently labeled tumors, which enables visualization of the whole-body down to single cell-cell interactions, is therefore a promising tool in understanding these processes. MATERIALS AND METHODS: EL-4 T-cell lymphoma cells were transfected with plasmid containing either fusion construct of enhanced green fluorecent protein (EGFP) with H2B histone or pure EGFP gene under cytomegalovirus promotor and resistance to neomycin. Stability of expression was determined by flow cytometry and cellular localization of green fluorescence signal was tested using fluorescent microscopy. An in vivo whole-body imaging system was used to evaluate growth in vivo. RESULTS: EL-4 cells were successfully transfected and established stable transfectants with a proliferation rate comparable to that of wild-type EL-4. Clone 12, with very strong whole-cell expression, enables tracking of metastatic spreading, whereas clone 3, with EGFP within the cell nucleus, allows frozen section analysis and observing of interaction with immunocompetent cells. CONCLUSION: Established imageable EL-4-EGFP(+) cell lines are a magnificent tool for the study of tumor growth and the tumor microenvironment.
Asunto(s)
Modelos Animales de Enfermedad , Linfoma de Células T/patología , Animales , Procesos de Crecimiento Celular/fisiología , Línea Celular Tumoral , Colorantes Fluorescentes/química , Proteínas Fluorescentes Verdes/biosíntesis , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/genética , Histonas/genética , Linfoma de Células T/genética , Linfoma de Células T/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Microscopía Fluorescente , TransfecciónRESUMEN
The main limitation of contemporary anticancer chemotherapy remains to be the insufficient specificity of the drugs for tumor tissue, which decreases the maximum tolerated dose due to severe side effects. Micellar drug delivery systems based on amphiphilic block copolymers with a very narrow size distribution (10 to 100 nm in diameter) is a novel innovative approach. Here, we report biological and pharmacological properties of polymeric micellar conjugate containing doxorubicin (DOX) covalently bound via hydrolytically cleavable hydrazone bonds to the micelle core. The system had a very low systemic toxicity (almost 20 times lower than free DOX) and long circulation in the bloodstream (with half of the dose after 24 h). Significant accumulation of tested micelles within the tumor was confirmed by fluorescent whole body imaging. Our new micellar system showed promising therapeutic activity against established murine EL-4 T-cell lymphoma; it was found that it is able to completely cure about 75% of tumor-bearing mice (with doses of either 1 x 150 mg DOX kg(-1) or 2 x 75 mg DOX kg(-1), administered i.v.). Moreover, treatment with micelles in cured mice induced tumor-specific resistance. Up to 80% of these mice survived rechallenge with original but not with distinct tumor cells.
Asunto(s)
Antineoplásicos/administración & dosificación , Doxorrubicina/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Micelas , Polímeros/uso terapéutico , Animales , Antineoplásicos/farmacocinética , Doxorrubicina/farmacocinética , Semivida , Hidrazonas , Hidrólisis , Linfoma de Células T/tratamiento farmacológico , Ratones , Nanopartículas/química , Polímeros/química , Polímeros/farmacocinética , Inducción de RemisiónRESUMEN
Synthesis, physicochemical and biological properties and preliminary anticancer activity of new star-shaped polymer-doxorubicin (DOX) conjugates targeted with anti-CD20 monoclonal antibody were investigated. Mild reduction of antibody (Ab) with dithiothreitol (DTT) resulted in introduction of thiol groups into Ab. Polymer precursors used for the synthesis of the conjugates were based on N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers with a functional group at the polymer chain end. The copolymers were linked to the thiol groups of the reduced Ab via one-point attachment forming a star-shaped structure with central antibody surrounded by hydrophilic polymer chains. Neither reduction nor polymer modification of Ab influenced binding activity of the Ab to its specific cancer cell membrane antigen as it was confirmed in vitro by standard flow cytometry. The anticancer drug DOX was attached to the HPMA copolymer chain in an Ab-polymer system via a pH-labile hydrazone linkage or via an oligopeptide sequence degradable by lysosomal enzymes. Such Ab-polymer-DOX conjugates were fairly stable in aqueous solution at pH 7.4 and the drug was readily released in mildly acid environment at pH 5-5.5 by hydrolysis of hydrazone bond or more slowly by enzymolysis with lysosomal enzymes. The cytostatic activity of the anti-CD20 monoclonal Ab-targeted conjugates tested on several CD20-positive or negative human and mouse cancer cell lines confirmed considerable targeting capacity of the monoclonal Ab after its binding to the polymer carrier. New method of synthesis of star antibody-targeted polymer-drug conjugates with pH-controlled drug release described in this paper opens new perspectives for development of new therapeutics intended for cancer therapy.
Asunto(s)
Citostáticos/metabolismo , Portadores de Fármacos/síntesis química , Sistemas de Liberación de Medicamentos , Metacrilatos/química , Polímeros/química , Acrilamidas/química , Animales , Antibióticos Antineoplásicos/química , Anticuerpos Monoclonales/inmunología , Linfoma de Burkitt/tratamiento farmacológico , Linfoma de Burkitt/metabolismo , Línea Celular Tumoral , Doxorrubicina/química , Portadores de Fármacos/química , Evaluación Preclínica de Medicamentos , Humanos , Concentración de Iones de Hidrógeno , Linfoma de Células T/tratamiento farmacológico , Linfoma de Células T/metabolismo , Masculino , Ratones , Modelos Químicos , Peso Molecular , Solubilidad , Agua/químicaRESUMEN
The successful treatment of cancer with conventional drugs is frequently complicated by the resistance of tumor cells to such a non-specific therapy. Over the last few years, immunotherapy has gained attention as a tumor-specific approach. Recent findings demonstrated that some conventional cytostatics stimulate local anticancer responses. New anticancer drugs, including their polymeric derivatives, are currently being developed with the aim of destroying tumors more effectively and more specifically. Among these, the water-soluble conjugates of doxorubicin with N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer carrier have emerged as efficient therapeutics because they are able to not only directly destroy cancer cells but also elicit systemic tumor-specific anticancer responses. Here, we discuss new insights into their mechanisms of immune surveillance, which could suggest novel approaches to cancer therapy.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Citotoxinas/administración & dosificación , Factores Inmunológicos/administración & dosificación , Modelos Biológicos , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , Polímeros/administración & dosificación , Animales , Antineoplásicos/administración & dosificación , HumanosRESUMEN
BALB/c mice bearing syngeneic BCL1 leukemia, a mouse model of human chronic lymphocytic leukemia, were treated with polymer-bound doxorubicin conjugate targeted with BCL1-specific monoclonal antibody. Such treatment can cure up to 100% of mice and the cured mice show long-lasting resistance to BCL1 leukemia. We show that both CD4+ and CD8+ T cells are required for establishment of the resistance, but only CD8+ T cells are necessary for its maintenance. BCL1 cells express MHC class I and II and also costimulatory molecules CD80 and CD86, which can aid eliciting of antitumor response. On the other hand, BCL1 cells also use several immunoescape mechanisms, such as expression of PD-L1, PD-L2, and interleukin-10. BCL1 cells thus can be recognized by BCL1-specific T cells, but instead of effective priming, such T cells are anergized or deleted by apoptosis. Moreover, BCL1 leukemia progression is accompanied by robust expansion of CD4+CD25+Foxp3+ regulatory T (Treg) cells. Although it has been shown that depletion of Treg cells in tumor-bearing mice can retard tumor growth, direct evidence that expansion of Treg cells can promote tumor growth was lacking. In this study, we provide first direct evidence that expanded Treg cells can indeed promote tumor progression by using mice with selectively expanded Treg cells before inoculation of BCL1 leukemia. Finally, we have also shown that elimination of some immunoescape mechanism (e.g., deletion of Treg) can significantly improve the therapeutic outcome of chemotherapy.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Doxorrubicina/análogos & derivados , Leucemia de Células B/tratamiento farmacológico , Leucemia de Células B/inmunología , Ácidos Polimetacrílicos/farmacología , Animales , Antibióticos Antineoplásicos/farmacología , Anticuerpos Monoclonales/inmunología , Doxorrubicina/farmacología , Femenino , Inmunoconjugados/inmunología , Inmunoconjugados/farmacología , Ratones , Ratones Endogámicos BALB C , Linfocitos T Reguladores/inmunología , Escape del Tumor/efectos de los fármacos , Escape del Tumor/inmunologíaRESUMEN
Doxorubicin is one of the most potent anti-tumor drugs with a broad spectrum of use. To reduce its toxic effect and improve its pharmacokinetics, we conjugated it to an HPMA copolymer carrier that enhances its passive accumulation within solid tumors via the EPR effect and decreases its cytotoxicity to normal, noncancer cells. In this study, we compared the antiproliferative, pro-survival, and death signals triggered in EL-4 cancer cells exposed to free doxorubicin and doxorubicin conjugated to a HPMA copolymer carrier via either enzymatically (PK1) or hydrolytically (HYD) degradable bonds. We have previously shown that the intracellular distribution of free doxorubicin, HYD, and PK1 is markedly different. Here, we demonstrated that these three agents greatly differ also in the antiproliferative effect and cell death signals they trigger. JNK phosphorylation sharply increased in cells treated with HYD, while treatment with free doxorubicin moderately decreased and treatment with PK1 even strongly decreased it. On the other hand, treatment with free doxorubicin greatly increased p38 phosphorylation, while PK1 and HYD increased it slightly. PK1 also significantly increased ERK phosphorylation, while both the free doxorubicin and HYD conjugate slightly decreased it. Long-term inhibition of JNK significantly increased both proliferation and viability of EL-4 cells treated with free doxorubicin, showing that the JNK signaling pathway could be critical for mediating cell death in EL-4 cells exposed to free doxorubicin. Both activation of caspase 3 and decreased binding activity of the p50 subunit of NFkappaB were observed in cells treated with free doxorubicin and HYD, while no such effects were seen in cells incubated with PK1. Analysis of the expression of genes involved in apoptosis and regulation of the cell cycle demonstrated that free doxorubicin and HYD have very similar mechanisms of action, while PK1 has very different characteristics.
Asunto(s)
Acrilamidas/farmacología , Doxorrubicina/farmacología , Linfoma de Células T/metabolismo , Acrilamidas/análisis , Acrilamidas/química , Animales , Apoptosis/genética , Caspasa 3/metabolismo , Ciclo Celular/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular , Doxorrubicina/análisis , Doxorrubicina/química , Citometría de Flujo , Expresión Génica/efectos de los fármacos , Concentración 50 Inhibidora , Linfoma de Células T/química , Ratones , Quinasas de Proteína Quinasa Activadas por Mitógenos/antagonistas & inhibidores , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , FosforilaciónRESUMEN
In our previous work, the salivary gland extract (SGE) from Ixodes ricinus ticks impaired T-lymphocyte proliferation and clearly modulated the immune response towards the Th2 pattern in human peripheral blood mononuclear cell culture. In the present work, the results obtained on mouse splenocytes are compared with those on human leukocytes. ELISA (protein level) and RNAse protection assay (mRNA level) showed that SGE enhanced interleukin (IL)-1alpha, IL-1beta, IL-1Ra, IL-6, and IL-12p40 cytokines, whereas production of IL-2, IL-5, IL-10, and IL-13 was decreased. The minute levels of IL-9, IL-15 and IL-12p70 were not changed after the addition of tick saliva. IL-4 was upregulated, whereas the production of gamma interferon and migratory inhibition factor was downregulated after the addition of SGE. Tick saliva decreased concanavalin A-stimulated spleen cell proliferation and the percentage of activated T-cells. We conclude that the Th2 polarization did not involve all of the cytokines tested. However, the Th2 subset-augmenting effect of tick saliva was confirmed.
Asunto(s)
Citocinas/biosíntesis , Ixodes/química , Glándulas Salivales/química , Células Th2/inmunología , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos B/metabolismo , Complejo CD3/metabolismo , Extractos Celulares/farmacología , Células Cultivadas , Citocinas/análisis , Citocinas/clasificación , Citometría de Flujo , Interacciones Huésped-Parásitos , Humanos , Interleucina-1/biosíntesis , Interleucina-1/clasificación , Ixodes/citología , Ixodes/fisiología , Factores Inhibidores de la Migración de Macrófagos/análisis , Factores Inhibidores de la Migración de Macrófagos/biosíntesis , Ratones , Mitógenos/farmacología , Receptores de Transferrina , Glándulas Salivales/citología , Bazo/citología , Bazo/crecimiento & desarrollo , Células TH1/inmunologíaRESUMEN
Saliva-activated transmission of Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 was demonstrated using salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks and C3H mice. Injection of Borrelia spirochaetes together with SGE increased the level of bacteraemia and accelerated the appearance of bacteria in the urinary bladder, compared with the injection of spirochaetes alone. More I. ricinus nymphs became infected when feeding on mice inoculated with B. afzelii plus SGE. Analysis of cytokines produced by cells of draining lymph nodes from SGE-treated mice showed a suppression of proinflammatory cytokines IFN-gamma, IL-6 and GM-CSF following a transient upregulation in comparison with the control mice infected without SGE.