RESUMO
Coinfections are common in natural populations and the outcome of their interactions depends on the immune responses of the host elicited by the parasites. Earlier we showed that immunization with BmAFII (Sephadex G-200 eluted) fraction of human lymphatic filaria Brugia malayi inhibited progression of Leishmania donovani infection in golden hamsters. In the present study we identified cross reactive molecules of B. malayi, and investigated their effect on L. donovani infection and associated immune responses in the host. The sequence alignment and sharing of linear T- and B-cell epitopes in protein molecules of B. malayi and L. donovani counterparts were studied in silico. Hamsters were immunized with robustly cross reactive SDS-PAGE resolved fractions F6 (54.2-67.8kDa) and F9 (41.3-45.0kDa) of B. malayi and subsequently inoculated with amastigotes of L. donovani intracardially. F6 inhibited (â¼72%) L. donovani infection and upregulated Th1 cytokine expression, lymphoproliferation, IgG2, IgG2/3 levels and NO production, and downregulated Th2 cytokine expression. Sequences in HSP60 and EF-2 of F6 and L. donovani counterparts were conserved and B- and T-cell epitopes in the proteins shared antigenic regions. In conclusion, leishmania-cross reactive molecules of filarial parasite considerably inhibited leishmanial infection via Th1-mediated immune responses and NO production. Common B- and T-cell epitope regions in HSP60 and EF-2 of the parasites might have contributed to the inhibitory effect on the L. donovani infection. Thus, leishmania-cross reactive filarial parasite molecules may help in designing prophylactic(s) against L. donovani.
Assuntos
Brugia Malayi/imunologia , Leishmania donovani , Leishmaniose Visceral/prevenção & controle , Animais , Cricetinae , Citocinas/sangue , Epitopos de Linfócito T , Humanos , Imunização , Leishmania donovani/imunologia , MesocricetusRESUMO
As part of our drug discovery program for anti-filarial agents from Indian medicinal plants, leaves of Eucalyptus tereticornis were chemically investigated, which resulted in the isolation and characterization of an anti-filarial agent, ursolic acid (UA) as a major constituent. Antifilarial activity of UA against the human lymphatic filarial parasite Brugia malayi using in vitro and in vivo assays, and in silico docking search on glutathione-s-transferase (GST) parasitic enzyme were carried out. The UA was lethal to microfilariae (mf; LC100: 50; IC50: 8.84 µM) and female adult worms (LC100: 100; IC50: 35.36 µM) as observed by motility assay; it exerted 86% inhibition in MTT reduction potential of the adult parasites. The selectivity index (SI) of UA for the parasites was found safe. This was supported by the molecular docking studies, which showed adequate docking (LibDock) scores for UA (-8.6) with respect to the standard antifilarial drugs, ivermectin (IVM -8.4) and diethylcarbamazine (DEC-C -4.6) on glutathione-s-transferase enzyme. Further, in silico pharmacokinetic and drug-likeness studies showed that UA possesses drug-like properties. Furthermore, UA was evaluated in vivo in B. malayi-M. coucha model (natural infection), which showed 54% macrofilaricidal activity, 56% female worm sterility and almost unchanged microfilaraemia maintained throughout observation period with no adverse effect on the host. Thus, in conclusion in vitro, in silico and in vivo results indicate that UA is a promising, inexpensive, widely available natural lead, which can be designed and developed into a macrofilaricidal drug. To the best of our knowledge this is the first ever report on the anti-filarial potential of UA from E. tereticornis, which is in full agreement with the Thomson Reuter's 'Metadrug' tool screening predictions.
Assuntos
Brugia Malayi/efeitos dos fármacos , Filariose/tratamento farmacológico , Filaricidas/uso terapêutico , Folhas de Planta , Plantas Medicinais , Triterpenos/uso terapêutico , Animais , Simulação por Computador , Filaricidas/farmacologia , Humanos , Técnicas In Vitro , Triterpenos/farmacologia , Ácido UrsólicoRESUMO
Here we report the synthesis of novel chalcone-thiazole compounds and their antifilarial activity. The antifilarial properties of these hybrids were assessed against microfilariae as well as adult worms of Brugia malayi. Among all the synthesized compounds, only two compounds, namely 4g and 4n were identified to be promising in vitro. These active compounds were tested in B. malayi-jird (Meriones unguiculatus) and B. malayi-Mastomys coucha models. Compound 4n showed 100% embryostatic effect and 49% macrofilaricidal in jirds and M. coucha models, respectively. This study provides a new structural clue for the development of novel antifilarial lead molecules.
Assuntos
Brugia Malayi/efeitos dos fármacos , Chalcona/análogos & derivados , Chalcona/farmacologia , Filariose/tratamento farmacológico , Filaricidas/síntese química , Filaricidas/farmacologia , Tiazóis/farmacologia , Animais , Chalcona/química , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Feminino , Filariose/parasitologia , Filaricidas/química , Gerbillinae , Masculino , Estrutura Molecular , Murinae , Testes de Sensibilidade Parasitária , Relação Estrutura-Atividade , Tiazóis/químicaRESUMO
We have recently identified disorganized muscle protein-1 (DIM-1) in one of the proinflammatory fractions of the human filaria Brugia malayi adult worm. The present study was undertaken to characterize B. malayi DIM-1 (DIM-1bm) and explore its vaccine potential. In this study we cloned and expressed the DIM-1bm gene, investigated its sequence homology with other nematodes, constructed in silico structural model, purified the recombinant DIM-1bm (rDIM-1bm) protein, and studied the effect of immunization with rDIM-1bm on the establishment of B. malayi infection in Mastomys coucha. DIM-1bm showed similarity with DIM-1 of Caenorhabditis elegans, Ascaris suum and Loa loa. Structural modeling revealed three immunoglobulin domains in DIM-1bm indicating that it is a member of immunoglobulin superfamily (IgSF) and 'blastn' results showed that DIM-1bm coding sequence (CDS) have almost no homology with human and mouse nucleotide sequences. Immunization with rDIM-1bm partially protected M. coucha against establishment of infection as inferred by a low recovery of microfilariae (37-64%) and parasite burden (â¼50%). The enhanced activity of macrophages, and IFN-γ and NO responses, and elevated levels of specific IgG, IgG1, IgG2a and IgG2b correlated with parasitological findings. This is the first report on cloning, expression, structural modeling and purification of rDIM-1bm and its ability to partially prevent establishment of B. malayi infection. DIM-1bm's almost complete lack of homology with the human counterpart makes it an attractive protein for exploring its vaccine potential.
Assuntos
Antígenos de Helmintos/genética , Brugia Malayi/genética , Proteínas Musculares/genética , Sequência de Aminoácidos , Animais , Antígenos de Helmintos/química , Antígenos de Helmintos/imunologia , Clonagem Molecular , Feminino , Gerbillinae , Imunoglobulina G/sangue , Interferon gama/imunologia , Macrófagos/imunologia , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Murinae , Proteínas Musculares/química , Proteínas Musculares/imunologia , Óxido Nítrico/metabolismo , FagocitoseRESUMO
We report here cloning and expression of full length mitochondrial HSP60 gene of Brugia malayi adult worm (mtHSP60bm), purification of the gene product by affinity chromatography, its in silico 3D structure and the sequence homology of the protein with Escherichia coli GroEL/ES and human HSP60. The ATP binding pocket of human HSP60 and mtHSP60bm were analyzed and compared using in silico models. The distribution of HSP60 in different life-stages of the parasite was determined using antibodies raised against recombinant mtHSP60bm (rmtHSP60bm). mtHSP60bm was present in all life-stages of the parasite except third stage infective larvae, in which it could be induced by heat-shock, and showed high degree of homology with E. coli GroEL/ES. The ATP binding pocket of HSP60 in humans, E. coli and B. malayi were also found structurally conserved. This similarity between human and mtHSP60bm might be useful in understanding the host-parasite interactions. This is the first ever report on distribution, cloning, sequence homology and ATP binding site of mtHSP60bm.
Assuntos
Trifosfato de Adenosina/metabolismo , Brugia Malayi/metabolismo , Chaperonina 60/química , Chaperonina 60/genética , Aedes , Animais , Sítios de Ligação , Brugia Malayi/genética , Brugia Malayi/isolamento & purificação , Chaperonina 60/isolamento & purificação , Chaperonina 60/metabolismo , Cromatografia de Afinidade , Clonagem Molecular , DNA Complementar/genética , DNA de Helmintos/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Gerbillinae , Interações Hospedeiro-Parasita , Humanos , Imunização , Masculino , Conformação Molecular , Dados de Sequência Molecular , Murinae , RNA de Helmintos/genética , RNA de Helmintos/isolamento & purificação , Homologia de SequênciaRESUMO
BACKGROUND & OBJECTIVES: Earlier we demonstrated that immunization with F6, a proinflammatory molecular fraction isolated from the human filarial parasite Brugia malayi, protected the host and eliminated the infection in Mastomys coucha by a Th1/Th2 response including IgG2a antibody response. Whether F6 molecules become accessible to human host during natural course of infection and elicit similar response is not known. The present study was undertaken to determine the profile of IgG subclasses specifically reactive to F6 in different categories of bancroftian filariasis cases to infer any relationship between the levels of a particular F6-specific IgG subclass and the infection or disease status. METHODS: Serum samples of normal individuals from filariasis non-endemic regions of India like Jammu & Kashmir, Uttarakhand, and Chandigarh [(NEN-W; n=10), healthy subjects from USA (NEN-U; n=10) and three categories of bancroftian filariasis cases from endemic areas: endemic normals (EN; n=10) with no symptoms and no microfilariae, asymptomatic microfilaremics (ASM; n=10) and chronic symptomatic amicrofilaremics (CL; n=10) were assayed for F6-specific IgG1, IgG2, IgG3 and IgG4 by ELISA using SDS-PAGE-isolated F6 fraction of B. malayi adult worms. RESULTS: Significantly high levels of F6-specific IgG1, IgG2 and IgG3 were found in CL (P<0.001) and EN (P<0.01-0.001) bancroftian filariasis cases compared to NEN-U. Significant levels of F6-specific IgG1 (P<0.01) and IgG2 (P<0.01) but not IgG3 were found in ASM cases compared to NEN-U. The most abundant was IgG2 which when compared to NEN-U, was significantly high in CL (P<0.001) and EN cases (P<0.001), followed by ASM (P<0.01). F6-specific IgG4 response in EN, ASM and CL subjects was not significantly different from the levels of NEN-U. Among the non-endemic normals, the NEN-W subjects showed significant reactivity with IgG2 (P<0.001) but not with IgG1, IgG3 and IgG4 as compared to NEN-U subjects. IgG subclass levels were different in different categories. INTERPRETATION & CONCLUSIONS: The high levels of F6 reactive IgG1, IgG2 and IgG3 in endemic normals and chronic symptomatic bancroftian patients, and IgG1 and IgG2 in asymptomatic microfilaraemics, suggest that F6 molecules of parasite are accessible in these subjects for IgG subclass-specific immune response and IgG2 may be related to pathogenesis. Studies using individual F6 molecules will be done to identify the molecule(s) involved in infection and protective immunity.
Assuntos
Brugia Malayi/imunologia , Filariose/imunologia , Imunoglobulina G , Animais , Eletroforese em Gel de Poliacrilamida , Filariose/sangue , Humanos , Imunidade Ativa , Imunoglobulina G/sangue , Imunoglobulina G/classificação , Imunoglobulina G/imunologia , Índia , Inflamação/imunologiaRESUMO
A series of 3,6-epoxy [1,5]dioxocines were synthesized and evaluated for their antifilarial activity against adult parasites of human lymphatic filarial parasite Brugia malayi (sub-periodic strain) in vitro. Out of these, six compounds (4a-f) possessed improved in vitro anti-filarial activity and examples 4d and 4f were also found to be active in the in vivo experiments. These results demonstrate that 3,6-epoxy [1,5]dioxocines exhibits potent antifilarial activity and might be developed into a new class of antifilarial drug.
Assuntos
Brugia Malayi/efeitos dos fármacos , Compostos de Epóxi/síntese química , Filaricidas/farmacologia , Oxocinas/síntese química , Animais , Compostos de Epóxi/química , Compostos de Epóxi/farmacologia , Feminino , Filaricidas/síntese química , Filaricidas/química , Concentração Inibidora 50 , Estrutura Molecular , Oxocinas/química , Oxocinas/farmacologiaRESUMO
Filarial parasites survive by inducing tolerance in host but the antigens and mechanisms involved are not clear. Recently we found that BmAFI, a Sephadex G-200 eluted fraction of Brugia malayi adult worm extract, stimulates IL-10 release from THP-1 cells. In the present study, we determined the SDS-PAGE profile of BmAFI and infective 3rd stage larva (L3), investigated the effect of pre-sensitization of host with BmAFI on the survival and development of L3 in the non-permissive peritoneal cavity (p.c.) of the permissive host Mastomys coucha and in the p.c. of non-permissive Swiss mice, and studied immunological correlates for the observed effects. The parasite development and burden in p.c., was determined in sensitized infected M. coucha and Swiss mice and the release of TGF-ß, IL-4, IL-10, IL-13, IFN-γ and NO, cellular proliferative response to Con A and BmAFI and levels of IgG subclasses and IgE were determined in sensitized infected M. coucha. Cellular proliferative response to Con A and BmAFI, mRNA expression of GATA-3, CTLA-4 and T-bet were determined in sensitized Swiss mice. In addition, the parasitological parameter was also studied in BmAFI-sensitized M. coucha exposed to the infection by standard subcutaneous (s.c.) route to assess whether sensitization enhances the intensity of infection. BmAFI-sensitization permitted survival of L3 and their development to adult stage by day 60 p.i. in the p.c. of M. coucha; in non-sensitized animals L3 could molt to L4 only and no parasite could be recovered beyond day 30 p.i. In M. coucha that received infection by s.c. route, pre-sensitization with BmAFI enhanced the microfilaraemia and adult worm recovery. In sensitized Swiss mice L3 could successfully molt to L4 in p.c. with improved recovery of parasite. BmAFI sensitization upregulated TGF-ß and IL-10 release, IgG1 and IgG2b levels, GATA-3 and CTLA-4 mRNA expression, suppressed the cellular proliferative response and downregulated Con A stimulated response, IgE, IL-13, IFN-γ and NO responses. Immunoblot analysis showed that the BmAFI antiserum also strongly reacts with some L3 molecules. The results show, for the first time, that sensitization with the anti-inflammatory BmAFI which shares some of its molecules with those in L3, facilitates parasite survival in the non-permissive p.c. of the permissive host M. coucha, render a non-permissive Swiss mouse partially permissive to infection and enhances parasite load in M. coucha receiving the infection through permissive s.c. route by evoking a modified Th2 type of response and anti-inflammatory milieu. In conclusion, the findings suggest that the anti-inflammatory BmAFI fraction facilitates survival of B. malayi infection even in non-permissive environment.
Assuntos
Antígenos de Helmintos/metabolismo , Brugia Malayi/patogenicidade , Filariose/parasitologia , Evasão da Resposta Imune , Murinae/parasitologia , Cavidade Peritoneal/parasitologia , Fatores de Virulência/metabolismo , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Brugia Malayi/crescimento & desenvolvimento , Brugia Malayi/imunologia , Proliferação de Células , Citocinas/metabolismo , Filariose/imunologia , Filariose/patologia , Tolerância Imunológica , Imunoglobulina G/sangue , Leucócitos Mononucleares/imunologia , Masculino , Camundongos , Murinae/imunologia , Óxido Nítrico/metabolismo , Fatores de Virulência/imunologiaRESUMO
Presently available marketed alum adsorbed hepatitis B vaccine used for prophylactic immunization, can effectively elicit humoral immunity but is poor inducer of cell-mediated immunity (CMI). Besides, conventional alum-adjuvant vaccines require multiple injections to achieve long-lasting protective immune responses. Therefore, as a result of insufficient immunization, infections are still the leading killer among diseases. The present investigation was therefore, aimed at developing "single-shot" HBsAg adsorbed microspheres of poly (DL)-lactide-co-glycolide (PLGA) (L/G 50:50 and 75:25) and their capability to stimulate the cell mediated immune response against hepatitis B surface antigen. These microspheres were characterized in vitro for their size, shape polydispersity index, percentage HBsAg adsorption efficiency and in vitro release profile. The immune-stimulating activities were also studied following subcutaneous injection of HBsAg adsorbed PLGA microspheres (single-dose on day 0) and compared with alum adsorbed vaccines (two-doses on 0 and 28 days) in Balb/c mice. Specific cell-mediated immune responses such as lymphocyte transformation assay (stimulation-index) including release of interferon-gamma (IFN-γ), interleukin-2 (IL-2) and nitric-oxide were determined. Cellular responses in case of alum adsorb HBsAg vaccine was very low. These studies demonstrate the potential of cationic polymeric microspheres based vaccine in stimulating cell mediated immune response along with humoral response against hepatitis B.
Assuntos
Portadores de Fármacos/química , Antígenos da Hepatite B/imunologia , Vacinas contra Hepatite B/administração & dosagem , Imunidade Celular/imunologia , Imunidade Humoral/imunologia , Ácido Láctico/química , Ácido Poliglicólico/química , Animais , Cátions , Estabilidade de Medicamentos , Eletroforese em Gel de Poliacrilamida , Antígenos da Hepatite B/administração & dosagem , Antígenos da Hepatite B/química , Injeções Subcutâneas , Interferon gama/sangue , Interferon gama/imunologia , Interleucina-2/sangue , Interleucina-2/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura , Microesferas , Peso Molecular , Óxido Nítrico/sangue , Óxido Nítrico/imunologia , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Propriedades de Superfície , Tecnologia FarmacêuticaRESUMO
There is increasing interest in phytoestrogens as potential alternatives to synthetic selective estrogen receptor modulators (SERMs) in the prevention and therapy of breast cancer. The present study is aimed at determining whether dietary glycine soya (Glycine max seeds; GS), which is rich in phytoestrogens, can enhance the anti breast cancer efficacy of the SERM tamoxifen (TAM) and the effect of TAM and GS, either alone or in combination, on DMBA-initiated hepatocarcinogenesis in rat. For determination of enhancing effect, rats bearing palpable 7, 12-dimethylbenz[α] anthracene (DMBA)-induced mammary tumors were treated with TAM (10 mg kg(-1)/day) while being fed AIN-93G diet with or without added GS (3×10(4) mg kg(-1)), and the tumor growth was monitored up to 5 weeks of treatment. For determining the effect on hepatocarcinogenesis, DMBA-initiated rats were exposed to TAM and dietary GS as above for 6 weeks during promotion stage in a medium-term bioassay, and the development of placental form of glutathione-S-transferase (GST-P)-expressing preneoplastic liver lesions was quantified. Exposure to both TAM and dietary GS enhanced the anti tumor efficacy of TAM via a combination of tumor cell apoptosis (determined by TUNEL) and inhibition of tumor cell proliferation (determined by PCNA immunostaining) and suppressed the growth of GST-P-positive liver lesions. The findings show that dietary GS enhances the therapeutic efficacy of TAM against mammary tumors and minimizes TAM's hepatocarcinogenesis promotion potential.
Assuntos
Antineoplásicos/uso terapêutico , Dieta , Glycine max , Neoplasias Mamárias Experimentais , Tamoxifeno/toxicidade , 9,10-Dimetil-1,2-benzantraceno/toxicidade , Animais , Antineoplásicos/administração & dosagem , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Glutationa S-Transferase pi/genética , Glutationa S-Transferase pi/metabolismo , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/patologia , Tamanho do Órgão , Ratos , Ratos Sprague-DawleyRESUMO
New chemical entities are desperately needed that overcome the limitations of existing drugs for neglected diseases. Screening a diverse library of 10,000 drug-like compounds against 7 neglected disease pathogens resulted in an integrated dataset of 744 hits. We discuss the prioritization of these hits for each pathogen and the strong correlation observed between compounds active against more than two pathogens and mammalian cell toxicity. Our work suggests that the efficiency of early drug discovery for neglected diseases can be enhanced through a collaborative, multi-pathogen approach.
Assuntos
Antiparasitários/isolamento & purificação , Avaliação Pré-Clínica de Medicamentos/métodos , Doenças Negligenciadas/tratamento farmacológico , Doenças Parasitárias/tratamento farmacológico , Descoberta de Drogas/tendências , HumanosRESUMO
We evaluated the antifilarial activity of 6 flavonoids against the human lymphatic filarial parasite Brugia malayi using an in vitro motility assay with adult worms and microfilariae, a biochemical test for viability (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT)-reduction assay), and two animal models, Meriones unguiculatus (implanted adult worms) and Mastomys coucha (natural infections). In vitro, naringenin and hesperetin killed the adult worms and inhibited (>60%) MTT-reduction at 7.8 and 31.2 µg/ml concentration, respectively. Microfilariae (mf) were killed at 250-500 µg/ml. The half maximal inhibitory concentration (IC(50)) of naringenin for motility of adult females was 2.5 µg/ml. Flavone immobilized female adult worms at 31.2 µg/ml (MTT>80%) and microfilariae at 62.5 µg/ml. Rutin killed microfilariae at 125 µg/ml and inhibited MTT-reduction in female worms for >65% at 500 µg/ml. Naringin had adulticidal effects at 125 µg/ml while chrysin killed microfilariae at 250 µg/ml. In vivo, 50 mg/kg of naringenin elimiated 73% of transplanted adult worms in the Meriones model, but had no effect on the microfilariae in their peritoneal cavity. In Mastomys, the same drug was less effective, killing only 31% of the naturally acquired adult worms, but 51%, when the dose was doubled. Still, effects on the microfilariae in the blood were hardly detectable, even at the highest dose. In summary, all 6 flavonoids showed antifilarial activity in vitro, which can be classed, in a decreasing order: naringenin>flavone=hesperetin>rutin>naringin>chrysin. In jirds, naringenin and flavone killed or sterilized adult worms at 50mg/kg dose, but in Mastomys, where the parasite produces a patent infection, only naringenin was filaricidal. Thus naringenin and flavone may provide a lead for design and development of new antifilarial agent(s). This is the first report on antifilarial efficacy of flavonoids.
Assuntos
Brugia Malayi/efeitos dos fármacos , Filaricidas/farmacologia , Flavanonas/farmacologia , Flavonoides/farmacologia , Animais , Brugia Malayi/isolamento & purificação , Corantes , Modelos Animais de Doenças , Filariose Linfática/tratamento farmacológico , Feminino , Flavanonas/normas , Flavonoides/normas , Gerbillinae/parasitologia , Hesperidina/farmacologia , Humanos , Masculino , Murinae/parasitologia , Rutina/farmacologia , Análise de Sobrevida , Sais de Tetrazólio , TiazóisRESUMO
Centchroman is a non-steroidal oral contraceptive and has been found to be a candidate drug for breast cancer exhibiting partial to complete remission of lesions in 40.5% of breast cancer patients. The therapeutic efficacy of centchroman was monitored alone and together with glycine soya on growth of 7,12-dimethylbenz[alpha]anthracene-induced breast tumor in rat. The tumor regression was monitored at different doses of centchroman alone ranging from 0 to 10 mg kg(-1) and with glycine soya from 1x10(4) to 5x10(4) mg kg(-1) per day until 5weeks treatment. An optimum tumor treatment opus was established with varying treatment parameters including doses of therapeutic agents and treatment period. The tumors were found to be static with a strong anti-estrogenic effect. Overall our study shows that both centchroman and glycine soya alone and jointly combat with breast cancer.
Assuntos
9,10-Dimetil-1,2-benzantraceno/toxicidade , Carcinógenos/toxicidade , Centocromano/uso terapêutico , Antagonistas de Estrogênios/uso terapêutico , Glycine max , Neoplasias Mamárias Experimentais/tratamento farmacológico , Animais , Apoptose , Proliferação de Células , Centocromano/administração & dosagem , Antagonistas de Estrogênios/administração & dosagem , Feminino , Neoplasias Mamárias Experimentais/induzido quimicamente , Neoplasias Mamárias Experimentais/patologia , Ratos , Ratos Sprague-Dawley , Receptores de Estrogênio/metabolismoRESUMO
The present investigations were aimed to compare the humoral and cell-mediated immune responses between recombinant hepatitis B surface antigens (HBsAg) adsorbed L-PLA microspheres (Ms) vaccine (single-shot) and marketed alum-HBsAg vaccine (two-doses). The blank cationic (cetyltrimethyammoniumbromide) microspheres were prepared by the double emulsion (w/o/w) solvent evaporation technique. The HBsAg was adsorbed onto the surface of blank cationic microspheres. These microspheres were characterized in vitro for their size, shape, adsorption-efficiency, in-process stability, and HBsAg release studies. Specific humoral immune responses (IgM and IgG) and cell-mediated immune responses (cellular-proliferation) assay including release of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), and nitric oxide (NO) from host's cells stimulated with HBsAg or lipopolysaccharide (LPS)/ concanavalin A (con A) in-vitro were determined. Based on these findings, it was concluded that the single injection (using subcutaneous-route) of the polymeric microspheres produced better immune response (both humoral and cell-mediated) than two injections of a conventional alum-HBsAg vaccine. These data demonstrate high potential of polymeric microspheres for their use as a carrier adjuvant for hepatitis B vaccine.
Assuntos
Portadores de Fármacos/química , Antígenos de Superfície da Hepatite B/imunologia , Vacinas contra Hepatite B/imunologia , Poliésteres/química , Animais , Cátions , Química Farmacêutica/métodos , Emulsões , Antígenos de Superfície da Hepatite B/administração & dosagem , Vacinas contra Hepatite B/administração & dosagem , Imunidade Celular , Imunidade Humoral , Injeções Subcutâneas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microesferas , Tamanho da PartículaRESUMO
Blank polymeric lamellar substrate particles (PLSP) of poly (l-lactide) were prepared and recombinant hepatitis B surface antigen (rHBsAg) was adsorbed onto these particles. The physical characteristics of blank PLSPs or PLSP-rHBsAg in vitro and its immunological responses in Balb/c mice were investigated. The average size of the particles was less than 10microm. Antigen adsorption efficiency was found to be 62.66+/-1.26%. Immunization with PLSP-rHBsAg resulted in upregulation of specific cellular (lymphoproliferation, IFN-gamma and NO release) as well as IgG response in animals. These responses were higher than those produced by two-dose schedule of alum-adsorbed antigen (alum-rHBsAg). Thus in conclusion, in terms of convenience and efficacy PLSP-rHBsAg is superior to alum-rHBsAg.
Assuntos
Adjuvantes Imunológicos/fisiologia , Portadores de Fármacos/administração & dosagem , Antígenos de Superfície da Hepatite B/imunologia , Vacinas contra Hepatite B/imunologia , Hepatite B Crônica/imunologia , Microesferas , Adjuvantes Imunológicos/administração & dosagem , Animais , Formação de Anticorpos , Esquema de Medicação , Antígenos de Superfície da Hepatite B/administração & dosagem , Vacinas contra Hepatite B/administração & dosagem , Hepatite B Crônica/prevenção & controle , Humanos , Interferon gama/biossíntese , Ativação Linfocitária , Linfócitos/imunologia , Linfócitos/metabolismo , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Camundongos , Óxido Nítrico/biossíntese , Polímeros/química , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologiaRESUMO
BACKGROUND & OBJECTIVE: Lymphatic filariasis is a disabling disease that continues to cripple population in tropical countries. Currently available antifilarial drugs are not able to control the disease. Therefore, a better antifilarial is urgently required for proper management of the disease. We undertook this study to assess the antifilarial activity of Caesalpinia bonducella-seed kernel against rodent filarial parasite in experimental model. METHODS: Microfilaraemic cotton rats and Mastomys coucha harbouring Litomosoides sigmodontis and Brugia malayi respectively, were treated with crude extract or fractions of the seed kernel C. bonducella through oral route for 5 consecutive days. Microfilaricidal, macrofilaricidal and female worm sterilizing efficacy was assessed. RESULTS: Crude extract showed gradual fall in microfilariae (mf) count in L. sigmodontis-cotton rat model from day 8 post-treatment attaining more than 95 per cent fall by the end of observation period. It also exhibited 96 per cent macrofilaricidal and 100 per cent female sterilizing efficacy. The butanol fraction F018 caused 73.7 per cent reduction in mf count and 82.5 per cent mortality in adult worms with 100 per cent female sterilization. The aqueous fraction F019 exerted more than 90 per cent microfilaricidal activity and 100 per cent worm sterilization. Two chromatographic fractions, F024 and F025 of hexane soluble fraction exhibited 64 and 95 per cent macrofilaricidal activity, respectively. Both the fractions caused gradual fall in microfilaraemia and 100 per cent worm sterilization. In B. malayi-M. coucha model F025 showed gradual reduction in microfilaraemia and caused 80 per cent sterilization of female parasites INTERPRETATION & CONCLUSION: In conclusion, C. bonducella- seed kernel extract and fractions showed microfilaricidal, macrofilaricidal and female-sterilizing efficacy against L. sigmodontis and microfilaricidal and female-sterilizing efficacy against B. malayi in animal models, indicating the potential of this plant in providing a lead for new antifilarial drug development.