ABSTRACT
ABSTRACT: Advances in the fields of glycobiology and immunology have provided many insights into the role of carbohydrate-protein interactions in the immune system. Jacalin of Artocarpus integrifolia (JCA) and structural mannoprotein of Saccharomyces uvarum (MPS) are molecules with immunomodulatory properties. JCA is an IgA human lectin binding molecule that causes the mitogenic stimulation of immune cells, production of cytokines, chemotaxis, and activation of leukocytes. Studies on the immunomodulatory properties of JCA and MPS in mammals and fish suggest that they have an action on antibody production. The aim of this study was to investigate the possible action of JCA and MPS on the production of specific antibodies in laying hens. For this, laying hens were inoculated with an intra abdominal injection of sheep red blood cells (SRBC) with either JCA (0.075 µg, 0.75 µg, and 7.5 µg) or MPS (20 µg and 100 µg). Levels of anti-SRBC antibodies of the IgY, IgM, and IgA classes were evaluated by ELISA. Results showed that JCA and MPS have immunomodulatory effects on levels of anti-SRBC IgM, IgA, and IgY. An immunostimulatory effect of JCA was observed in primary immune response on anti-SRBC IgY, while an inhibitory effect of JCA and MPS was observed in secondary immune response on the production of IgM and IgA anti-SRBC. These results suggested that MPS and JCA have immunomodulatory effects on antibody production and could be used in future studies on humoral immune response in poultry.
RESUMO: Avanços nos campos glicobiologia e imunologia forneceram muitas informações sobre o papel das interações da proteína-carboidrato na modulação do sistema imunológico. A jacalina extraída de Artocarpus integrifolia (JCA) e a manoproteína da parede celular de Saccharomyces uvarum (MPS) são moléculas com propriedades imunomoduladoras. JCA é uma lectina com afinidade pela IgA humana e tem ação mitogênica sobre células do sistema imunológico estimulando a produção de citocinas, a quimiotaxia e a ativação de leucócitos. Estudos sobre as propriedades imunomoduladoras de JCA e MPS em mamíferos e peixes sugerem que essas moléculas podem ter um efeito sobre a produção de anticorpos. O objetivo deste estudo foi investigar a ação da JCA e MPS sobre a produção de anticorpos específicos em galinhas poedeiras. Para isso, galinhas poedeiras foram inoculadas por via intra-abdominal com eritrócitos de carneiro (SRBC) em associação com o JCA (0,075 µg, 0,75 µg, e 7,5 µg) ou MPS (20 µg e 100 µg). Os níveis de anticorpos anti-SRBC das classes IgY, IgM, e IgA foram avaliados por ELISA. Os resultados mostraram que a JCA e a MPS têm um efeito imunomodulador sobre a produção IgY, IgM, ou IgA anti-SRBC. Um efeito imunoestimulador da JCA foi observado sobre a produção de anticorpos IgY na resposta imune primária, enquanto um efeito imuno inibitório da JCA e da MPS sobre a produção de IgM e IgA anti-SRBC na resposta imune secundária. Estes resultados sugerem que o MPS e JCA tem efeito modulador sobre a produção de anticorpos e podem ser utilizados em estudos futuros sobre a imunidade humoral em aves comerciais.
ABSTRACT
The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.
Subject(s)
Animals , Humans , Antibodies, Helminth/blood , Antigens, Helminth , Taenia solium/immunology , Antigens, Helminth/immunology , Case-Control Studies , Chromatography, Affinity , Enzyme-Linked Immunosorbent Assay , Immunoblotting , Immunologic Tests/methods , Neurocysticercosis/diagnosis , Neurocysticercosis/immunology , Sensitivity and SpecificityABSTRACT
Total antigen from Leishmania (Leishmania) amazonensis and isolates from the Leishmania braziliensis complex, along with their respective antigenic fractions obtained by affinity chromatography on concanavalin-A-Sepharose and jacalin-agarose columns evaluated using immunoenzymatic ELISA assay. For this, serum samples from 229 patients were used, grouped as American tegmental leishmaniasis (nº=58), visceral leishmaniasis (nº=28), Chagas disease (nº=49), malaria (nº=32), tuberculosis (nº=13) and healthy volunteers (nº=49). Samples from American tegmentary leishmaniasis showed higher reactivity with antigens isolated from the Leishmania braziliensis complex than with antigens from Leishmania amazonensis (p<0.001). ELISA assays showed a sensitivity range from 60 percent to 95 percent with antigens isolated from the Leishmania braziliensis complex. There was marked nonspecific reactivity among serum samples with the use of antigenic fractions binding with concanavalin-A and jacalin from both Leishmania complexes, in comparison with other antigens (p<0.001). The results presented in this study suggest that the use of homologous antigens increases the efficiency of anti-Leishmania immunoglobulin detection, which may be very valuable for diagnostic purposes.
Antígeno total de Leishmania (Leishmania) amazonensis e isolado do complexo Leishmania brazilienis, assim como suas respectivas frações antigênicas obtidas por cromatografia de afinidade em coluna de concanavalina-A ligada a sepharose e Jacalina ligada a agarose foram avaliadas por ensaio imunoenzimático ELISA. Para tanto, foram utilizadas amostras de soros de 229 pacientes agrupadas em leishmaniose tegumentar americana (nº=58), leishmaniose visceral (nº=28), doença de Chagas (nº=49), malaria (nº=32), tuberculose (nº=13) e voluntários saudáveis (nº=49). Houve maior reatividade das amostras de leishmaniose tegumentar americana com a utilização dos antígenos obtidos do isolado do complexo Leishmania braziliensis quando comparado com antígenos de Leishmania amazonensis (p<0,001). Observou-se ainda que a sensibilidade do teste ELISA variou de 60 a 95 por cento entre os antígenos obtidos do isolado do complexo Leishmania braziliensis. Houve acentuada reatividade inespecífica das amostras de soros com a utilização das frações antigênicas ligantes de Concanavalina-A e Jacalina de ambos os complexos Leishmania em comparação aos demais antígenos (p<0,001). Os resultados apresentados no presente trabalho sugerem que a utilização de antígenos homólogos aumentam a eficiência de detecção de imunoglobulina anti-Leishmania o que pode ser de grande valia para o propósito de diagnóstico.
Subject(s)
Animals , Humans , Antigens, Helminth , Leishmania braziliensis/immunology , Leishmania mexicana/immunology , Leishmaniasis, Mucocutaneous/diagnosis , Leishmaniasis, Visceral/diagnosis , Antigens, Helminth/isolation & purification , Case-Control Studies , Chromatography, Affinity , Cross Reactions , Chagas Disease/immunology , Enzyme-Linked Immunosorbent Assay , Leishmaniasis, Mucocutaneous/immunology , Leishmaniasis, Visceral/immunology , Malaria/immunology , Plant Lectins , Sensitivity and Specificity , Sepharose/analogs & derivatives , Sepharose , Tuberculosis/immunologyABSTRACT
Naturally occurring serum IgG against terminal α-galactoside epitopes (anti- Gal), present exclusively in man, apes and old world monkeys, was used as probe for these epitopes in human brain. Human brain grey matter soluble glycoproteins enriched in α galactosyl groups by affinity chromatography on jacalin-sepharose, specifically binds to human anti-Gal in immuno dot blots. Anti-Gal recognized exclusively the terminal α galactoside epitope in human brain glycoproteins since binding was abolished by the presence of 1-0-methyl α-D-galactopyranoside as well as by pretreatment of glycoproteins with coffee bean α-galactosidase. Anti-Gal-peroxidase staining of jacalin-binding human brain glycoproteins in western immuno blots revealed mainly five anti-Gal-binding polypeptides with Mr (in kDa) of 94, 108, 180, 210 and 230 respectively. Since the presence of anti-Gal in higher animals accompanies suppression of the corresponding epitope in most tissues, apparently to maintain immunological balance, possible implications of the above observation for autoimmunity, tumor metastasis and infection are discussed.
ABSTRACT
Jacalin has been found to agglutinate Ehrlich ascites cells. The agglutination was inhibited by a-glycosides of D-Gal and ß -D-Gal(1 → 3)-D-GalNAc suggesting that the lectin-ascites interaction was carbohydrate-specific. There was 21·8% inhibition of tumour (ascites) cell growth in vivo in mice administered 50μg of jacalin by injection for 6 days following intraperitoneal injection of ascites cells. Administration of 100, 150 and 200 μg jacalin resulted in 40·2, 57·5 and 83% inhibition respectively. The in vivo inhibition of tumour cells growth by jacalin was due to its preferential binding with D-Gal-a -( 1→6) present as terminal residues in the glycoprotein on tumour cell surface.