The effect of hydatid cyst protoscolex somatic antigens on full-thickness skin wound healing in mouse.

BACKGROUND: Wound healing has evolved in recent years, resulting in diverse therapeutic options. OBJECTIVE: This study evaluated the effects of the somatic antigen of the hydatid cyst protoscolex on wound healing in mice with full-thickness skin wounds. METHODS: Fifty-four adult mice, weighing 25 ± 5 g and approximately 60 days old, were divided into three groups (A, B, and C), each further divided into three subgroups. Subgroups A1, A2, and A3 were assigned negative controls. B1, B2, and B3 received hydatid cyst somatic antigen tests at 10 µg/SC, whereas C1, C2, and C3 received somatic antigen tests at 20 µg/SC. Under general anesthesia, a wound biopsy puncture of 9.8 mm in diameter was performed on the mice's back and spine. In the experimental group, antigen and alum adjuvant were administered subcutaneously around the wound, while the control group received Phosphate-Buffered Saline (PBS). Using digital images, a geometric assessment was conducted on days 0, 1, 3, 6, 9, 12, 15, 18, and 21 post-wounding. The obtained images were analyzed by Image J software and after analyzing the data by SPSS software. RESULTS: A significant difference in terms of epithelization was observed in the antigen treatment group with a dose of 20 µg on days 3 and 6 (P < 0.05). Furthermore, the 20 µg antigen group was significantly higher than the 10 µg antigen group in terms of this factor on day 3 (P < 0.05). Skin samples were taken from all wounds on days 3, 10 and 21 for microscopic evaluation. Regarding epithelization, on day 10, a significant difference was observed in the treatment group with a concentration of 10 µg with the control group and the treatment group with a concentration of 20 µg (P < 0.05). CONCLUSION: Based on the results of the present study, it can be concluded that somatic antigens of protoscolex hydatid cyst are dose-dependent and antigens with a dose of 20 µg by subcutaneous injection accelerate wound healing and epithelialization.

Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells.

BACKGROUND: Angiogenesis is defined as the formation of new vessels by sprouting of endothelial cells from pre-existing vessels in response to stimuli, such as hypoxia or inflammation. Subcutaneous dirofilariasis, caused by Dirofilaria repens, is a zoonotic disease characterized by the formation of subcutaneous nodules with the presence of at least one encapsulated worm, showing perivascular vascularization around it. The aim of this study is to analyze whether the somatic antigen of adult D. repens worms interacts with and modulates the angiogenic mechanism, cell proliferation and migration, and formation of pseudo-capillaries. METHODS: The expression of VEGF-A, VEGFR-1/sFlt, VEGFR-2, mEnd and sEnd in cultures of human vascular endothelial cells stimulated with somatic antigen of adult worms of D. repens (DrSA), vascular endothelial growth factor (VEGF) and DrSA + VEGF were evaluated by using ELISA commercial kits. Cellular viability was analyzed by live cell count, cytotoxicity assays by using a commercial kit, cell proliferation by MTT-based assay, cell migration by wound-healing assay carried out by scratching wounds and capacity of formation of pseudo-capillaries analyzing cell connections and cell groups in Matrigel cell cultures. In all cases unstimulated cultures were used as controls. RESULTS: DrSA + VEGF significantly increased the expression of VEGF-A, VEGFR-2 and mEndoglin compared to other groups and unstimulated cultures. Moreover, DrSA + VEGF produced cell proliferation and migration and increased the formation of pseudo-capillaries. CONCLUSIONS: Somatic antigen of adult D. repens worms activated the proangiogenic mechanism, cell proliferation and cell migration as well as formation of pseudo-capillaries in this in vitro human endothelial cell model. These processes could be related to the survival of adult D. repens in subcutaneous nodules in infected hosts.

Dicrocoelium dendriticum induces autophagic vacuoles accumulation in human hepatocarcinoma cells.

The relationship between Dicrocoelium dendriticum and cancer has been poorly investigated so far, but a large amount of findings suggest that other trematodes can favour cancer in both animals and humans. In this study, the effects of D. dendriticum on cell proliferation, cell death mechanisms and oxidative stress induction were evaluated in hepatocellular carcinoma (HCC) cell lines (HepG2 and HuH7). Results showed that short time exposure to low concentrations of somatic antigens from D. dendriticum caused slight proliferation in both HepG2 and HuH7 cells while high concentrations and long exposure time to extracts from D. dendriticum caused a significant growth inhibition. This effect was, however, not paralleled by apoptosis but it occurred with an about 40% increase of the formation of autophagic vacuoles. In the same experimental conditions, a strong oxidative stress was recorded with an about 100% increase of the intracellular O(2-). These data suggest the occurrence of an escape anti-apoptotic mechanism in HCC cells. In conclusion, these results suggest a role for D. dendriticum in the chronic oxidative stress and in the regulation of transformation processes in HCC warranting additional investigations in this specific area of research.

Lymphocyte Migration Inhibition Response in Trichuris muris Infected and Vaccinated Mice.

BACKGROUND: Immunological response of host and parasite play a key role in developing vaccination and immunization. The present study deals with the immune response and effecter mechanism, which was confirmed by migration inhibition factor (MIF). METHODS: The present work was conducted in Parasitological Lab of Postgraduate Department of Zoology, Government Holkar Science College, Indore (M.P.) during 2006-2007. For MIF assay, lymphocytes were separated from heparinized blood of experimental and control mice. Aliquots of cell suspension were placed in four wells cut in a preparation of agarose in a Petri dish. Two wells were filled with soluble test antigen, while rest two wells were filled with medium (control wells). Petri dish was incubated overnight at 37 °C in a humidified environment at 5% CO2 in air. Cells migrated under the agarose in a circle were fixed and stained. Diameters of the migration areas were measured with ocular micrometer. RESULTS: MIF reaction was maximum (44.2%) in the group IVEgESAg5 and minimum (10.8%) in the group IVASoAg1. The maximum MIF reaction was shown by eggs ES antigen and least by adult worm somatic antigen. The interesting observation was that migration inhibition increases as dose increased or we could say the reaction was dose dependent. CONCLUSION: Increased value of MIF response in vaccinated mice suggested the involvement of lymphocytes in cell-mediated immunity. This study also proves that excretory-secretory (ES) antigen of eggs from Trichuris muris was more effective in imparting immunity in mice.

Antígenos de larvas pulmonares de Ascaris suum reconocidos por anticuerpos producidos en Oryctolagus cuniculus / Ascaris suum lung larvae antigens recognized by antibodies produced in Oryctolagus cuniculus

El presente trabajo tuvo como objetivo determinar mediante la técnica de Western Blot los antígenos de larvas pulmonares de Ascaris suum que son detectados por anticuerpos producidos en Oryctolagus cuniculus inmunizado experimentalmente. Las larvas pulmonares (L3 y L4) fueron obtenidas en 120 ejemplares de Mus musculus cepa BALB/c ratón infectados experimentalmente por vía oral con huevos infectivos de A. suum. Parte de estas larvas fueron cultivadas en el medio Eagle (MEM) para la obtención de antígenos de excreción/secreción y la otra fue sonificada para la obtención de antígenos somáticos, los cuales sirvieron para inmunizar dos ejemplares de O. cuniculus, utilizando Adyuvante Completo e Incompleto de Freund. A las 5 semanas de inmunización se obtuvo sangre de los conejos por punción cardiaca a fin de recuperar el suero, parte del cual fue purificado parcialmente por precipitación salina y diálisis. Mediante la técnica de electroinmuno-transferencia (Western Blot) y usando sueros de los conejos inmunizados se detectaron en los antígenos de excreción/secreción de 20 horas de cultivo reducidos con dithiothreitol, 12 bandas antigénicas de 100, 72.4, 56.2, 42.7, 39.8, 34.6, 31.6, 30.2, 19.5, 16.9, 15.5 y 14.9 KDa, siendo las más reactivas las de 100, 72.4, 16.9, 15.5 y 14.9 KDa. En los antígenos somáticos bajo condiciones de reducción, se detectaron solamente seis bandas de 42.7, 39.8, 34.6, 30.2, 28 y 25.2 KDa de poca reactividad. Estos resultados permiten afirmar que los antígenos de excreción/secreción de A. suum de 20 horas de incubación en el medio MEM inducen la producción de un mayor número de anticuerpos de tipo IgG en conejos inmunizados experimen-talmente.

Excretory/secretory antigens (E/SAg) and somatic antigens (SAg) of Ascaris suum lung larvae that induce the immunoglobulin G antibodies production in Oryctolagus cuniculus experimentally immunized was determined. For this purposes, specimens of Mus musculus BALB/c were inoculated orally with infective eggs of A. suum obtained from pigs naturally parasitized in order to obtain the lung larvae. Part of these larvae was cultured in Minimum Essential Medium (MEM) to obtain E/SAg and another part was sonicated to obtain the SAg too. Both, E/SAg and SAg mixed with complete and incomplete Freund's adjuvant were used for rabbits immunization. Five weeks after the immunization, the rabbits were bled by cardiac puncture obtaining the immunosera by centrifugation, which was purified partially by saline precipitation and dialysis. By using an Western blot technique with purified immunoserum and E/SAg obtained to 20 hours and reduced with dithiothreitol, fourteen antigens bands of 100, 72.4, 56.2, 42.7, 39.8, 34.6, 31.6, 30.2, 19.5, 16.9, 15.5 y 14.9 KD, were detected. The bands of 100, 72.4, 16.9, 15.5 and 14.9 KDa were been the most reactives. Thereby the SAg, also reduced with dithiothreitol, seven bands of 42.7, 39.8, 34.6, 30.2, 28.0 and 25.2 KDa were detected. They were a bit clear. In conclusion, the E/SAg induce the highest production of immunoglobulin G antibodies in rabbits experimentally immunized.

M. tuberculosis Somatic Antigen Specific CD8+T cell Responses in BCG-Vaccinated Subjects / 결핵및호흡기질환

BACKGROUND: The immune responses mediated by CD8+T cells are known to be significant in controlling M. tuberculosis infections. In order to determine the role of cytotoxic CD8+T cells in the protective immune mechanism in latently infected subjects, this study examined whether or not the cytotoxic immune responses of CD8+T cells specific to the M. tuberculosis somatic antigens are induced in BCG vaccinated healthy subjects. METHODS: Cytotoxicity and IFN-gamma elispot assays were used to investigate the activities of CD8+T cells specific for the thyA30-38 peptide epitope in circulating peripheral blood mononuclear cells (PBMC) from BCG-vaccinated HLA-A*0201 and A*0206 subjects. RESULTS: The results indicate the cytotoxic and IFN-gamma immune responses of CD8+T cells specific for thyA30-38 were induced in BCG vaccinated healthy subjects. CONCLUSION: The cytotoxic and IFN-gamma responses by CD8+T cells specific for the M. tuberculosis somatic antigens are induced in BCG-vaccinated subjects, and appear to be involved in the protective immune mechanism in latently infected people against a M. tuberculosis infection.