ABSTRACT
Interleukin (IL) -12 is a heterodimeric cytokine mainly produced by monocytes, macrophages, and dendritic cells in mammals. IL-12p70 composed of IL-12p35 and IL-12p40, is known to play a crucial role in promoting cell-mediated immunity (CMI) through Th1 differentiation and IFN-γ production. Although two types of IL-12p35 (p35a, p35b) and three types of IL-12p40 (p40a, p40b and p40c) have been identified in several fish species, the knowledge on functional characteristics of teleost IL-12 is still limited. In the present study, we cloned two types of IL-12p35 and three types of IL-12p40 genes in amberjack and yellowtail, and analyzed their expressions in response to stimulation with Nocardia seriolae in amberjack. As a result, four types of IL-12 (IL-12p35a, p35b, p40a and p40b) and IFN-γ mRNA were increased by live-N. seriolae stimulation but not by formalin-killed N. seriolae, suggesting that four types of IL-12 (p35, p35b, p40a and p40c) participate in promoting CMI. Subsequently, we produced six types of recombinant IL-12p70 (rIL12p70) protein in insect cells. Head kidney leukocytes were cultured with formalin-killed N. seriolae and six types of rIL-12p70 to elucidate the role of amberjack IL-12p70 in induction of CMI. After stimulation, IFN-γ expression was elevated whereas IL-10 expression was suppressed in Head kidney leukocytes stimulated with four types of rIL-12 (p40a/p35a, p40c/p35a, p40a/p35b, p40a/p35b). On the other hand, two types of rIL-12 (p40b/p35a, p40b/p35b) only elicited down regulation of IL-10 expression. These results indicate that all amberjack IL-12p70 isoforms are involved in Th1 -differentiation and promotion of CMI with different manners. Fish IL-12 has a potential for the promising vaccine adjuvant.
Subject(s)
Actinomycetales Infections/veterinary , Bacterial Vaccines/immunology , Fish Diseases/therapy , Fish Proteins/genetics , Interleukin-12/genetics , Nocardiaceae/immunology , Perciformes , Actinomycetales Infections/microbiology , Actinomycetales Infections/therapy , Amino Acid Sequence , Animals , Fish Diseases/microbiology , Fish Proteins/chemistry , Fish Proteins/metabolism , Interleukin-12/chemistry , Interleukin-12/metabolism , Interleukin-12 Subunit p35/genetics , Interleukin-12 Subunit p35/metabolism , Interleukin-12 Subunit p40/genetics , Interleukin-12 Subunit p40/metabolism , Phylogeny , Sequence Alignment/veterinary , Vaccines, Attenuated/immunology , Vaccines, Inactivated/immunology , Vaccines, Synthetic/immunologyABSTRACT
Tsukamurella spp. es un bacilo gram positivo, aeróbico, catalasa positivo, no móvil, no esporulado, que pertenece al orden de los actinomicetales. Los géneros incluidos en este orden son Nocardia, Gordonia, dietza, Skermania, Williamsia, Turicella, Streptomyces y Rhodococcus. Otros géneros relacionados son Corynebacterium y Mycobacterium. Las infecciones por esos microorganismos se han asociado con neumopatías crónicas, inmunodepresión (leucemia, tumores, infección por el HIV) e infecciones postoperatorias de heridas. Se notificó la presencia de tsukamurella en hemocultivos asociada al uso de sondas o catéteres, otros dispositivos médicos y en casos individuales de tenosinovitis necrosante con abscesos subcutáneos, infecciones óseas y cutáneas, meningitis, peritonitis y conjuntivitis y también como germen colonizante. Se presenta un caso de otomastoiditis en un paciente HIV positivo causado por este germen.
Subject(s)
Humans , Male , Adult , Antiretroviral Therapy, Highly Active , Diplopia/pathology , HIV , Mastoiditis/therapy , Nocardiaceae/immunologyABSTRACT
Antigenic extracts were prepared from culture filtrates of the principal agents known to cause actinomycetoma, namely Actinomadura madurae, Actinomadura pelletieri, Nocardia asteroides, Nocardia brasiliensis, Nocardia otitidis-caviarum, and Streptomyces somaliensis. These antigenic preparations were compared by immunodiffusion (ID), counterimmunoelectrophoresis (CIE), line immunoelectrophoresis (LIE) and rocket line immunoelectrophoresis (RLIE), with rabbit antisera prepared against each of the extracts. Cross-reactivity between antigenic extracts from the different actinomycetes, measured by determining the number of precipitin lines in homologous and heterologous systems, was common. Reactions were always stronger and more precipitin lines were present when antigenic extracts were tested against homologous antisera. Similarities between A. madurae and A. pelletieri antigens were of a low order and cross-reactivity did not exceed 33%. A. pelletieri resembled N. asteroides more closely than A. madurae, with 44% of detectable antigenic components cross-reacting. The three species of Nocardia had common antigenic epitopes, but the overall degree of similarity was of a low order (between 12 and 27% by LIE and RLIE). Antigenic extracts of S. somaliensis had few components in common with the other species tested and only one of the 34 lines present in the RLIE system for N. asteroides showed any reaction of identity with an antigenic component in the S. somaliensis extract. Single cross-reacting lines were also present in the CIE and LIE systems.
Subject(s)
Actinomycetales Infections/microbiology , Antigens, Bacterial/analysis , Nocardia/immunology , Nocardiaceae/immunology , Streptomyces/immunology , Counterimmunoelectrophoresis , Cross Reactions , Humans , Immunodiffusion , ImmunoelectrophoresisSubject(s)
Actinomycetales/classification , Nocardia/classification , Nocardiaceae/classification , Actinomycetales/analysis , Actinomycetales/immunology , Antigens, Bacterial , Carbohydrates/analysis , Cell Wall/analysis , DNA, Bacterial/analysis , Fatty Acids/analysis , Lipids/analysis , Nocardia/analysis , Nocardia/immunology , Nocardiaceae/analysis , Nocardiaceae/immunology , Quinones/analysisABSTRACT
Counterimmunoelectrophoresis used for the diagnosis of mycetoma was found superior to immunodiffusion (ID) especially when using neat serum and 1:2 dilution (P=less than 0.0001 with neat serum; and less than 0.01 with 1:2 dilution). It is recommended for routine use in mycology laboratories for following up patients on treatment and for confirming the diagnosis of sera that are weakly positive by ID.
