Protective effect of excretory-secretory proteins from Trichinella spiralis muscle larvae against myocardial injury in septic mice / 南方医科大学学报

OBJECTIVE@#To evaluate the protective effect of excretory-secretory proteins from Trichinella spiralis muscle larvae (Ts-MES) on sepsis-induced myocardial injury in mice.@*METHODS@#Eighty male BALB/C mice were randomized equally into sham-operated group, myocardial injury group, Ts-MES treatment group and dexamethasone treatment group. In the latter 3 groups, sepsis-induced myocardial injury models were established by cecal ligation and perforation; the sham operation was performed by exposure of the cecum without ligation or perforation. Forty minutes after the operation, the mice were given intraperitoneal injections 150 μL PBS, 20 μg TS-MES or 0.3 mg/kg dexamethasone as indicated. At 12 h after the operation, 6 mice were randomly selected from each group for echocardiography, and 8 mice were used for observing the survival rate within 72 h. The remaining 6 mice were examined for myocardial pathologies with HE staining and serum levels of NTPro-BNP and cTnI with ELISA; the expressions of TNF-α, IL-6, IL-10 and TGF-β in the serum and myocardial tissue were detected using ELISA and qRT-PCR.@*RESULTS@#Compared with the sham-operated mice, the septic mice showed significantly decreased cardiac function indexes (LVEF, LVFS, and E/A) with lowered survival rate within 72 h (P < 0.001) and significantly higher myocardial injury scores and serum levels of NTPro-BNP and cTnI (P < 0.01). Treatment with TS-MES significantly improved the cardiac function and 72-h survival rate (P < 0.05) and lowered the myocardial injury scores and serum levels of NTPro-BNP and cTnI (P < 0.05) in the septic mice. Compared with the sham-operated mice, the septic mice had obviously increased TNF-α and IL-6 levels in the serum and myocardial tissue (P < 0.001), which were significantly lowered by treatment with TS-MES (P < 0.05). TS-MES and dexamethasone both increased the levels of IL-10 and TGF-β in the septic mice, but the changes were significant only in TS-MES-treated mice (P < 0.05).@*CONCLUSION@#Ts-MES are capable of protecting against myocardial injury in septic mice by reducing the production of pro-inflammatory cytokines and enhancing the levels of regulatory cytokines.

Molecular cloning and characterization of a novel aspartyl aminopeptidase from Trichinella spiralis

@#Trichinellosis is an important zoonotic parasitic disease worldwide and is principally caused by ingesting animal meat containing Trichinella infective larvae. Aspartyl aminopeptidase is an intracytoplasmic metalloproteinase that specifically hydrolyzes the N-terminus of polypeptides free of acidic amino acids (aspartic acid and glutamate), and plays an important role in the metabolism, growth and development of organisms. In this study, a novel T. spiralis aspartyl aminopeptidase (TsAAP) was cloned and expressed, and its biological properties and roles in worm growth and development were investigated. The results revealed that TsAAP transcription and expression in diverse T. spiralis stages were detected by RT-PCR and Western blotting, and primarily localized at cuticle, stichosome and intrauterine embryos of this nematode by immunofluorescence test. rTsAAP has the enzymatic activity of native AAP to hydrolyze the substrate H-Glu-pNA. There was a specific binding between rTsAAP and murine erythrocyte, and the binding site was localized in erythrocyte membrane proteins. Silencing of TsAAP gene by specific dsRNA significantly reduced the TsAAP expression, enzymatic activity, intestinal worm burdens and female fecundity. The results demonstrated that TsAAP participates in the growth, development and fecundity of T. spiralis and it might be a potential target molecule for anti-Trichinella vaccines.

Specific binding of aspartic protease and enterocytes promotes Trichinella spiralis invasion of murine intestinal epithelium cells

@#Trichinella spiralis is an important foodborne zoonotic parasite and it is necessary to develop vaccine to prevent T. spiralis infection in food animals. T. spiralis aspartic protease-2 (TsASP2) has been demonstrated to play a crucial role in larval invasion of intestinal epithelium cells (IECs). The purpose of this study was to assess the interaction between TsASP2 and IECs and to investigate the immune protection elicited by vaccination with rTsASP2. The results showed that the enzymatic activity of native aspartic protease was detected in crude proteins of all T. spiralis development stages other than NBL stage, the highest activity was observed in the IIL stage. The results of Western blot showed that TsASP2 protein was expressed at ML, IIL and AW but not NBL, and the TsASP2 expression level at IIL stage was significantly higher than those of other three worm stages (P < 0.05). The specific binding between rTsASP2 and IECs was observed by immunofluorescence test (IFT) and confocal microscopy, and the binding site was localized at the IEC membrane and this binding ability was inhibited by aspartic protease specific inhibitor pepstain A. The results of ELISA showed that the binding ability was protein dose-dependent. Vaccination with rTsASP2 triggered a mixed Th1/Th2 humoral and mucosal immune responses, as demonstrated by the elevation levels of Th1/Th2 cytokines (IFN-γ and IL-4) secreted by the spleen and mesenteric lymph nodes (MLNs) of immunized mice. The mice vaccinated with rTsASP2 exhibited a 54.17% reduction in enteral adult worms and a 54.58% reduction in muscle larvae after T. spiralis challenge. The results demonstrated that TsASP2 might be a potential molecular target for anti-Trichinella vaccines.

Trichinella spiralis y Trichinella patagoniensis: estudio de la carga globular aplicando el método de Azul Alcian / Trichinella spiralis and Trichinella patagoniensis: study of globular charge applying Alcian Blue method / Trichinella spiralis e Trichinella patagoniensis: estudo da carga globular pelo método de Azul de Alcian

Resumen Los residuos de ácido siálico de las glicoproteínas de superficie son los principales responsables de la carga negativa eritrocitaria. El objetivo de este trabajo fue estudiar alteraciones de carga globular producidas por Trichinella spiralis y Trichinella patagoniensis. Se trabajó con concentrados de larvas musculares de ambas especies y con eritrocitos frescos. Se incubó el sedimento globular con igual volumen de concentrado larval (1 y 2 horas). Los controles fueron incubados con solución salina. Se aplicó el método de Azul Alcian y se determinó el coeficiente experimental de carga aniónica eritrocitaria (CexpCAE). Los resultados mostraron que la carga disminuyó con el aumento del tiempo de tratamiento para ambas especies. Los valores de CexpCAE de las suspensiones incubadas con T. spiralis fueron menores que con T. patagoniensis, indicando que T. spiralis produjo mayor disminución de carga que T. patagoniensis. Se concluye que la desialización producida por ambas especies no es la misma, lo que sugiere que la relación hospedador-parásito que se establecería in vivo sería distinta.

Abstract The sialic acid residues of the surface glycoproteins are mainly responsible for the erythrocyte negative charge. The objective of this work was to study alterations of globular charge produced by Trichinella spiralis and Trichinella patagoniensis. Work was carried out on muscle larvae concentrates of both species and fresh erythrocytes. The treatment was performed by incubating the globular pellet with equal volume of larval concentrate (1 and 2 hours). Controls were incubated with saline solution. The Alcian Blue method was applied and the experimental coefficient of erythrocyte anion charge (expCEAC) was determined. The results showed that the globular charge decreased with increasing treatment time for both species. The expCEAC values of the suspensions incubated with T. spiralis were lower than with T. patagoniensis, indicating that T. spiralis produced a greater decrease in charge than T. patagoniensis. It is concluded that the desialization produced by both species is not the same, suggesting that the host-parasite relationship that would be established in vivo would be different.

Resumo Os resíduos de ácido siálico das glicoproteínas de superfície são os principais responsáveis pela carga negativa dos eritrócitos. O objetivo desse trabalho foi estudar as alterações da carga globular produzidas por Trichinella spiralis e Trichinella patagoniensis. Trabalhamos com concentrados de larvas musculares de ambas as espécies e com eritrócitos frescos. O tratamento foi realizado incubando o sedimento globular com igual volume de concentrado larval (1 e 2 horas), Os controles foram incubados com solução salina. Foi aplicado o método de Azul de Alcian e se determinou o coeficiente experimental de carga aniônica de eritrócitos (CexpCAE). Os resultados mostraram que a carga diminuiu com o aumento do tempo de tratamento para ambas as espécies. Os valores de CexpCAE das suspensões incubadas com T. spiralis foram menores que com T. patagoniensis, indicando que T. spiralis produziu uma diminuição maior na carga que T. patagoniensis. Conclui-se que a dessalinização produzida por ambas as espécies não é a mesma, sugerindo que a relação hospedeiro parasita que seria estabelecida in vivo é diferente.

Trichinella spiralis y Trichinella patagoniensis: cinética de desialización eritrocitaria / Trichinella spiralis and Trichinella patagoniensis: kinetics of erythrocyte desialization / Trichinella spiralis e Trichinella patagoniensis: cinética de dessialização eritrocitária

El objetivo del trabajo fue estudiar la cinética de desialización eritrocitaria producida por larvas infectantes de Trichinella spiralis y Trichinella patagoniensis. Se trabajó con 7 suspensiones eritrocitarias incubadas con 1.000±200 larvas musculares/mL, durante 2 horas, tomando muestra al tiempo inicial y cada 15 minutos. Los respectivos eritrocitos controles se incubaron de la misma manera con solución salina. Se aplicaron el método de titulación por Polibrene calculando el CexpST y un análisis de varianza (ANOVA) con las comparaciones múltiples según Tukey. Los resultados mostraron que el valor promedio de CexpST disminuyó con el aumento del tiempo de incubación, para ambas especies. En el tratamiento con T. spiralis no hubo diferencias significativas entre el valor medio del coeficiente a tiempo 60 y 75 minutos, mientras que con T. patagoniensis, a 45 y 60 minutos. Todas las restantes diferencias fueron significativas. La comparación entre los tratamientos, para cada uno de los tiempos, mostró que al tiempo inicial el coeficiente promedio no difirió entre las especies, pero que a todos los otros tiempos fue significativamente menor en la incubación de los eritrocitos con T. spiralis. Se concluye que la relación hospedador-parásito que se establece en ambos casos es distinta y probablemente también la capacidad de adaptación y de daño al hombre.

The objective of this work was to study the kinetics of erythrocyte desialization produced by infective larvae of Trichinella spiralis and Trichinella patagoniensis. It was performed on 7 erythrocyte suspensions incubated with 1,000±200 muscle larvae/ mL for 120 minutes, taking samples at the initial time and every 15 minutes. The respective control erythrocytes were incubated in the same way with saline solution. The Polybrene Titration method calculating the CexpST and variance analysis (ANOVA) with the multiple comparisons according to Tukey were applied. The results showed that the average value of CexpST decreased with the increase in incubation time, for both species. There were no significant differences between the mean value of the coefficient at 60 and 75 minutes in the treatment with T. spiralis, while neither were there any differences between 45 and 60 minutes in the incubation with T. patagoniensis. All other differences were significant. The comparison between the two treatments, for each of the times, showed that at the initial time the average coefficient did not differ between the species, but at all other times it was significantly lower in the incubation of the erythrocytes with T. spiralis. It is concluded that the parasite host relationship that is established in both cases is different and probably also is the ability to adapt and cause harm to man.

O objetivo do trabalho foi estudar a cinética de dessialização eritrocitária. produzida por larvas infectantes de Trichinella spiralis e Trichinella patagoniensis. O trabalho foi feito com 7 suspensões eritrocitárias incubadas com 1.000±200 larvas musculares/mL por 2 horas, colhendo amostras no tempo inicial e a cada 15 minutos. Os respectivos eritrócitos-controle foram incubados da mesma forma com solução salina. Foi aplicado o método de titulação por Polibreno calculando o CexpST e também uma análise da variância (ANOVA) com as comparações múltiplas de acordo com Tukey. Os resultados mostraram que o valor médio de CexpST diminuiu com o aumento do tempo de incubação para ambas as espécies. No tratamento com T. spiralis não houve diferenças significativas entre o valor médio do coeficiente no tempo 60 e 75 minutos, ao passo que com T. patagoniensis, aos 45 e 60 minutos. Todas as diferenças restantes foram significativas. A comparação entre os tratamentos, para cada um dos tempos, mostrou que no tempo inicial o coeficiente médio não diferiu entre as espécies, mas que em todos os outros tempos foi significativamente menor na incubação dos eritrócitos com T. spiralis. A conclusão é que a relação hospedeiro-parasita, estabelecida em ambos os casos, é diferente e provavelmente também a capacidade de adaptação e dano ao homem.

Protective effect of recombinant adult serine protease inhibitor from Trichinella spiralis on sepsis-associated acute kidney injury in mice / 中国血吸虫病防治杂志

Objective To investigate the protective effect of recombinant adult serine protease inhibitor from Trichinella spiralis (TsadSPI) on sepsis-associated acute kidney injury in mice. Methods A total of 18 male BALB/c mice were randomly divided into the sham-operation group, the model group, and the TsadSPI treatment group, of 6 mice in each group. Sepsis-associated acute kidney injury was modeled in the model group and TsadSPI treatment group by cecal ligation puncture (CLP), while mice in the sham-operation group were only given exploratory laparotomy without ligation or perforation of the cecum. After 30 min of CLP, mice in the sham-operation group and the model group were intraperitoneally injected with PBS (100 μL), and mice in the TsadSPI treatment group were intraperitoneally injected with PBS (100 μL) containing TsadSPI (2 μg). At 12 h following modeling, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (Cr) and urea nitrogen (BUN) were measured to assess the liver and kidney functions, and the changes of the mouse kidney structure were observed using HE staining. In addition, the serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and transforming growth factor (TGF)-β were measured using an enzyme-linked immunosorbent assay (ELISA), and the myeloid differentiation factor 88 (MyD88) and nuclear factor kappa-B (NF-κB) p65 expression was determined in kidney tissues using immunohistochemical staining. Results At 12 h following CLP, there were significant differences in the serum levels of ALT (F = 41.031, P < 0.001), AST (F = 54.757, P < 0.001), Cr (F = 24.142, P < 0.001) and BUN (F = 214.849, P < 0.001) among the three groups, and higher levels of ALT, AST, Cr and BUN were measured in model group than in the sham-operation group (P < 0.001), while lower ALT, AST, Cr and BUN levels were found in the TsadSPI treatment group than in the model group (P < 0.001). HE staining showed severe mouse kidney injuries following CLP, and TsadSPI treatment resulted in remarkable alleviation of the injury. ELISA measured significant differences in the TNF-α (F = 47.502, P < 0.001) and IL-6 levels (F = 222.061, P < 0.001) among the three groups, and showed a remarkable reduction in the TNF-α and IL-6 levels in the TsadSPI treatment group as compared to those in the model group (P < 0.001). In addition, there were significant differences in serum IL-10 (F = 16.227, P < 0.001) and TGF-β levels (F = 52.092, P < 0.001) among the three groups, and higher IL-10 and TGF-β levels were seen in the TsadSPI treatment group than in the model group (P < 0.001). Immunohistochemical staining showed greater MyD88 expression and a higher nuclear positive rate of NF-κB p65 in kidney tissues in the model group than in the TsadSPI treatment group. Conclusions TsadSPI may reduce the MyD88 expression and nuclear positive rate of NF-κB p65 in mouse kidney tissues to up-regulate the expression of immunomodulatory factors and down-regulate the expression of pro-inflammatory cytokines, thereby protecting sepsis-associated acute kidney injury.

Polarization of bone marrow-derived macrophages induced by recombinant Trichinella spiralis cysteine protease inhibitors in vitro / 中国血吸虫病防治杂志

Objective To investigate the regulatory role of recombinant Trichinella spiralis cysteine protease inhibitors (rTs-Cys) in induction of polarization of bone marrow-derived macrophages (BMDMs) in vitro. Methods BMDMs were captured and cultured in conditioned medium for 7 days. Then, mature BMDMs were harvested and assigned into four groups. Cells in Group A (negative control) were given 10 ng/mL IFN-γ combined with 100 ng/mL LPS, cells in Group B (positive control) were treated with IL-4 and IL-10 (at 10 ng/mL both), and cells in Group C (recombinant protein alone) were stimulated with 1 μg/mL rTs-Cys, while cells in Group D (protein co-culture) were simultaneously treated with 1 μg/mL rTs-Cys, 10 ng/mL IFN-γ and 100 ng/mL LPS. Cells and culture supernatant were collected 24 hour post-treatment, and the proportions of F4/80+, CD11b+, CD206+ and CD11c+ cells were detected by flow cytometry. The levels of interleukin IL-6 (IL-6), tumor necrosis factor-α (TNF-α), IL-10 and transforming growth factor-β (TGF-β) in the cell culture supernatant were measured by ELISA and the CD86+ and CD206+ phenotypes were identified by immunofluorescent staining. Results Flow cytometry detected no significant difference in the proportion of F4/80+ CD11b+ CD11c+ cells among the four groups (F = 46.184, P < 0.001), and a lower proportion of F4/80+ CD11b+ CD11c+ cells was seen in groups C and D than in group A (all P values < 0.001). There was a significant difference in the proportion of F4/80+ CD11b+ CD206+ cells among the four groups (F = 11.032, P < 0.001), and a greater proportion of F4/80+ CD11b+ CD206+ cells was seen in groups C and D than in group A (all P values < 0.01). Immunofluorescent staining showed higher CD206+ expression and lower CD86+ expression in groups C and D than in Group A. There were significant differences in the IL-6 and (F = 3.950, P < 0.001) and TNF-α (F = 205.827, P < 0.001) levels in the cell culture supernatants among the four groups, and significantly lower IL-6 and TNF-α levels were measured in groups C and D than in Group A (both P < 0.05). There were significant differences in the IL-10 and (F = 8.274, P < 0.001) and TGF-β (F = 13.559, P < 0.01) levels in the cell culture supernatants among the four groups, and greater IL-10 and TGF-β levels were measured in Group C than in Group A (both P values < 0.01). In addition, the TGF-β level was significantly higher in Group D than in Group A (P < 0.05); however, there was no significant difference in the IL-10 level between groups D and A (P > 0.05). Conclusion rTs-Cys may induce the polarization of BMDMs to antiinflammatory M2 macrophages in vitro and inhibit the activation of M1 macrophages.

Compromiso neurológicoy cardiovascularen pacientes con triquinosis / Neurological and cardiovascular involventin patients with trichinosis

Introducción: La triquinosis es una infección parasitaria causada por nematodos del género Trichinella. El compromiso cardíaco no es habitual, pero representa la causa más frecuente de muerte por triquinosis, mientras que la afectación neurológica ocurre principalmente en pacientes severamente enfermos. Se sabe que a mayor cantidad de larvas ingeridas menor es el tiempo de incubación y mayor es la severidad de la enfermedad. Se presentan dos pacientes con compromiso del sistema nervioso central, uno de ellos cardiovascular, ambos pertenecientes a un brote de triquinosis ocurrido en la ciudad de Bahía Blanca.

Background: Trichinosis is a parasitic infection caused by nematodes of the genus Trichinella. Cardiac involvement is not usual but represents the most frequent cause of death by trichinosis, while neurological affectation occurs mainly in severely ill patients. A greater number of larvae ingested, the shorter the incubation time and the greater the severity of the disease. We present two patients with compromise of the central nervous system and one of them cardiovascular, both belonging to an outbreak of trichinosis in the city of Bahía Blanca

Larvas recién nacidas de Trichinella spiralis: cinética de agregación eritrocitaria / Newborn larvae of Trichinella spiralis: erythrocyte aggregation kinetics

El objetivo de este trabajo fue estudiar la cinética de agregación eritrocitaria producida por dos concentraciones de larvas recién nacidas (LRN) de Trichinella spiralis. Se trabajó con 5 suspensiones eritrocitarias incubadas con 500 y 1.000±100 LRN/mL durante 120 minutos, con tomas de muestra al tiempo inicial y cada 15 minutos. Los respectivos controles se incubaron de la misma manera con solución salina. Se aplicaron el método de titulación por Polibrene calculando el CexpST y la técnica de análisis de la variancia con las comparaciones múltiples según Tukey. Los resultados mostraron para ambas concentraciones de LRN, que el coeficiente promedio disminuyó con el aumento del tiempo de incubación. En el tratamiento con 1.000 LRN/mL, el coeficiente promedio no presentó diferencias significativas a tiempo 0 y 15 minutos, ni entre 60 y 75 minutos, mientras que con 500 LRN/mL no hubo diferencias entre los tiempos 0, 15 y 30 minutos. Todas las restantes diferencias fueron significativas para ambas concentraciones larvales. El valor promedio de CexpST no difirió significativamente entre los dos tratamientos a tiempo 0 y 15 minutos, pero a todos los otros tiempos fue menor a mayor concentración de larvas. La experiencia realizada indicaría que in vivo, la cantidad de LRN y el tiempo que permanecen en circulación determinan el grado de desializacion eritrocitaria, y por lo tanto el riesgo de activación T y de alteraciones hemorreológicas en el hospedador.

The aim of this work was to study the kinetics of erythrocyte aggregation produced by two concentrations of Trichinella spiralis newborn larvae (NL). Work was performed with 5 erythrocyte suspensions incubated with 500 and 1000 ± 100 NL/mL for 120 minutes, taking samples at the initial time and every 15 minutes. The respective controls were incubated in the same way with saline solution. The Polybrene titration method calculating the CexpST and the variance analysis technique with the multiple comparisons according to Tukey were applied. The results showed that the average coefficient decreased with the rise in incubation time for both NL concentrations. The average coefficient did not present significant differences between the initial time and 15 minutes, nor between 60 and 75 minutes in the treatment with 1000 NL/mL, while there were no differences between 0,15 and 30 minutes in the treatment with 500 NL/mL. All other differences were significant for both larval concentrations. The average value of CexpST did not differ significantly between the two time treatments at zero time and 15 minutes, but at all other times it was less at a higher concentration of larvae. The experience carried out would indicate that in vivo, the amount of NL and the time that they remain in circulation determines the degree of erythrocyte desialylation, and therefore, the risk of T activation and hemorrheological alterations in the host.

O objetivo desse trabalho foi estudar a cinética de agregação eritrocitária produzida por duas concentrações de larvas recém nascidas (LRN) de Trichinella spiralis. O trabalho foi feito com 5 suspensões eritrocitárias incubadas com 500 e 1.000 ± 100 LRN/mL por 120 minutos, colhendo amostras no tempo inicial e a cada 15 minutos. Os respectivos controles foram incubados da mesma forma com solução salina. Foi aplicado o método de titulação por Polibreno e se calculou CexpST. e a técnica de análise da variância com as comparações múltiplas de acordo com Tukey. Os resultados mostraram, para ambas as concentrações de LRN, que o o coeficiente médio diminuiu com o aumento do tempo de incubação. No tratamento com 1.000 LRN/mL o coeficiente médio não mostrou diferenças significativas no tempo 0 e 15 minutos ou entre 60 e 75 minutos, ao passo que não houve diferenças com 500 LRN/mL entre tempos 0, 15 e 30 minutos. Todas as restantes diferenças foram significativas para ambas as concentrações de larvas. O valor médio de CexpST não diferiu significativamente entre os dois tratamentos no tempo de 0 e 15 minutos, mas em todos os outros tempos foi menor em maior concentração de larvas. A experiência realizada indicaria que in vivo a quantidade de LRN e o tempo que permanecem em circulação determina o grau de dessialização dos eritrócitos e, portanto, o risco de ativação T e de alterações hemorreológicas no hospedeiro.

Efecto de Trichinella spiralis y de Trichinella patagoniensis n.sp. sobre la desialización / Effect of Trichinella spiralis and Trichinella patagoniensis n.sp. on desialization

El ácido siálico tiene importantes funciones biológicas, muchas de las cuales determinan su participación en la respuesta inmune. El objetivo del trabajo fue comparar el efecto de Trichinella spiralis y Trichinella patagoniensis n.sp. sobre la desialización eritrocitaria. Se trabajó con 10 concentrados de larvas musculares de T. spiralis y 10 de T. patagoniensis de la misma concentración larval. Se realizó el tratamiento incubando el sedimento de eritrocitos frescos con igual volumen de concentrado larval (37 ºC), tomando muestra a los 30, 60 y 90 minutos. Los controles fueron incubados de la misma forma con solución salina. Se aplicó el método de Titulación de la Agregación por Polibrene y se determinó el CexpST. Los resultados mostraron que el valor medio del CexpST en los eritrocitos tratados con T. spiralis fue significativamente menor que en los glóbulos tratados con T. patagoniensis, para todos los tiempos estudiados. El aumento del tiempo de tratamiento también disminuyó significativamente el valor medio del CexpST para las dos especies. Éste fue significativamente menor a los 90 minutos de incubación que a los 60 minutos y éstos a su vez menores que a los 30 minutos. Se concluye que T. spiralis provocó mayor desialización eritrocitaria que T. patagoniensis en las condiciones experimentales estudiadas.

Sialic acid has important biological functions, many of which determine its participation in the immune response. The objective of this paper was to compare the effect of Trichinella spiralis and Trichinella patagoniensis n.sp. on erythrocyte desialization. Work was performed on 10 larval concentrates of muscle larvae of T. spiralis and 10 of T. patagoniensis of the same larval concentration. The treatment was carried out incubating the sediment of fresh erythrocytes with an equal volume of larval concentrate (37 °C), taking samples at 30, 60 and 90 minutes. The controls were incubated in the same way treated with saline solution. Titration of aggregation by Polybrene Method was applied and the CexpST was determined. The results showed that the mean value of CexpST in erythrocytes with T. spiralis was significantly lower than in the globules treated with T. patagoniensis, for all the studied times. The increase in treatment time also significantly decreased the mean value of CexpST for the two species, being significantly lower at 90 minutes of incubation than at 60 minutes and these in turn lower than at 30 minutes. It is concluded that T. spiralis caused greater erythrocyte desialization than T. patagoniensis in the experimental conditions studied.

O ácido siálico tem importantes funções biológicas, muitas das quais determinam sua participação na resposta imune. O objetivo foi comparar o efeito de Trichinella spiralis e Trichinella patagoniensis n.sp. sobre a dessialização eritrocitária. Trabalhou-se com 10 concentrados de larvas musculares de T. spiralis e 10 de T. patagoniensis da mesma concentração larval. Realizou-se o tratamento incubando o sedimento de eritrócitos frescos com igual volume de concentrado larval (37 ºC), tomando amostra aos 30, 60 e 90 minutos. Os controles foram incubados da mesma forma com solução salina. Foi aplicado o método de Titulação da Agregação por Polibrene e se determinouo CexpST. Os resultados mostraram que o valor médio do CexpST nos eritrócitos Tratados com T. spiralis foi significativamente menor que nos glóbulos tratados com T. patagoniensis, para todos os tempos estudados. O aumento do tempo de tratamento também diminuiu significativamente o valor médio do CexpST para as duas espécies, sendo significativamente menor aos 90 minutos de incubação que aos 60 minutos e eles por sua vez menores que aos 30 minutos. Conclui-se que T. spiralis provocou maior dessialização eritrocitária que T. patagoniensis nas condições experimentais estudadas.

Protective effect of excretory-secretory protein from adult Trichinella spiralis on ovalbumin-induced allergic rhinitis in mice / 中国血吸虫病防治杂志

Objective To investigate the protective effect of excretory-secretory protein (AES) from adult Trichinella spiralis on ovalbumin (OVA)-induced allergic rhinitis in mice. Methods Eighteen female BALB/c mice were randomly divided into three groups, including the blank control group (Group A), OVA-induced rhinitis group (Group B) and AES treatment group (Group C). Mice in Group A were given PBS. Mice in Group B were intraperitoneally injected with antigen adjuvant suspension for systemic sensitization, once every other day for seven times; then, local excitation was intranasally induced with 5% OVA solution once a day for seven times to establish a mouse model of allergic rhinitis. In addition to induction of allergic rhinitis, mice in Group C were given 25 μg AES at baseline sensitization and local excitation. Following the final challenge, mice were observed for 30 min in each group, and the behavioral score was evaluated. The serum levels of IFN-γ, IL-4, IL-5, IL-10 and TGF-β were determined using an enzyme-linked immunosorbent assay in mice, and the pathological changes of mouse nasal mucosa were observed under a microscope. Results There was a significant difference in the mouse behavioral scores among the three groups (F = 110.12, P < 0.01). The mouse behavioral score was significantly higher in Group B than in Group A (7.17 ± 0.75 vs. 1.33 ± 0.52, P < 0.01), and more remarkable pathological damages of mouse nasal mucosa were seen in Group B than in Group A, while the mouse behavioral score was significantly decreased in Group C than in Group B (P < 0.01), and the pathological damages of mouse nasal mucosa remarkably alleviated in Group C relative to Group B. There was a significant difference in serum IFN-γ level among the three groups (F = 7.50, P < 0.01) and the serum IFN-γ level in Group B was significantly lower than in group A and C (both P < 0.05). There were significant differences in serum IL-4 (F = 470.81, P < 0.01) and IL-5 levels (F =68.20, P < 0.01) among the three groups, and significantly greater serum IL-4 and IL-5 levels were detected in Group B than in Group A (P < 0.01), while significantly lower serum IL-4 and IL-5 levels were detected in Group C than in Group B (P < 0.01). There were significant differences in serum IL-10 (F = 174.91, P < 0.01) and TGF-β levels (F = 9.39, P < 0.01) among the three groups, and significantly greater serum IL-10 and TGF-β levels were seen in Group C than in Group B (both P < 0.05). Conclusion T. spiralis AES has a remarkable protective activity against OVA-induced allergic rhinitis in mice.

Protective effect of excretory-secretory protein from adult Trichinella spiralis on ovalbumin-induced allergic rhinitis in mice / 中国血吸虫病防治杂志

Objective To investigate the protective effect of excretory-secretory protein (AES) from adult Trichinella spiralis on ovalbumin (OVA)-induced allergic rhinitis in mice. Methods Eighteen female BALB/c mice were randomly divided into three groups, including the blank control group (Group A), OVA-induced rhinitis group (Group B) and AES treatment group (Group C). Mice in Group A were given PBS. Mice in Group B were intraperitoneally injected with antigen adjuvant suspension for systemic sensitization, once every other day for seven times; then, local excitation was intranasally induced with 5% OVA solution once a day for seven times to establish a mouse model of allergic rhinitis. In addition to induction of allergic rhinitis, mice in Group C were given 25 μg AES at baseline sensitization and local excitation. Following the final challenge, mice were observed for 30 min in each group, and the behavioral score was evaluated. The serum levels of IFN-γ, IL-4, IL-5, IL-10 and TGF-β were determined using an enzyme-linked immunosorbent assay in mice, and the pathological changes of mouse nasal mucosa were observed under a microscope. Results There was a significant difference in the mouse behavioral scores among the three groups (F = 110.12, P < 0.01). The mouse behavioral score was significantly higher in Group B than in Group A (7.17 ± 0.75 vs. 1.33 ± 0.52, P < 0.01), and more remarkable pathological damages of mouse nasal mucosa were seen in Group B than in Group A, while the mouse behavioral score was significantly decreased in Group C than in Group B (P < 0.01), and the pathological damages of mouse nasal mucosa remarkably alleviated in Group C relative to Group B. There was a significant difference in serum IFN-γ level among the three groups (F = 7.50, P < 0.01) and the serum IFN-γ level in Group B was significantly lower than in group A and C (both P < 0.05). There were significant differences in serum IL-4 (F = 470.81, P < 0.01) and IL-5 levels (F =68.20, P < 0.01) among the three groups, and significantly greater serum IL-4 and IL-5 levels were detected in Group B than in Group A (P < 0.01), while significantly lower serum IL-4 and IL-5 levels were detected in Group C than in Group B (P < 0.01). There were significant differences in serum IL-10 (F = 174.91, P < 0.01) and TGF-β levels (F = 9.39, P < 0.01) among the three groups, and significantly greater serum IL-10 and TGF-β levels were seen in Group C than in Group B (both P < 0.05). Conclusion T. spiralis AES has a remarkable protective activity against OVA-induced allergic rhinitis in mice.

Source analysis of lymphocytes secreting interleukin-22 from spleen of mice infected with Trichinella spiralis at the early encapsulated stage / 中华地方病学杂志

Objective To investigate the cell origin of interleukin (IL)-22-secreting cell of mice infected with Trichinella spiralis (T.spiralis) at the early encapsulated stage.Methods Twelve Balb/c mice were divided into the infected group and the control group according to body weight by random number table.The infected mice were intragastrically administrated with 300 muscle larvae of T.spiralis,and the control mice were given the same amount of normal saline.The IL-22-secreting cell subsets in mouse splenic lymphocytes were detected by flow cytometry at the fourth week after infection.Results The proportion of IL-22-secreting cells in splenic lymphocytes of T.spiralis infected mice was increased when compared with control group [(0.88 ± 0.25)％ vs (0.28 ±0.17)％,t =-4.899,P ＜ 0.05].There was no significant difference between the proportion of CD3+IL-22+ cells and CD3-IL-22+ cells in the splenic lymphocytes of the infected group [(0.29 ± 0.17)％ vs (0.51 ± 0.17)％,t =-2.195,P ＞ 0.05],and the percentage of CD3-IL-22+ cells were similar between the infected group and the control group [(0.51 ± 0.17)％ vs (0.44 ± 0.22)％,t =-0.600,P ＞ 0.05].The proportion of CD3+IL-22+ cells in the infected group was significantly higher than that in the control group [(0.29 ± 0.17)％ vs (0.07 ± 0.06)％,t =-3.068,P ＜ 0.05],and the percentage of CD4+IL-22+ T cells and γδTCR+IL-22+ T cells were obviously increased in CD3+ lymphocytes [(1.28 ± 0.54)％ vs (0.16 ± 0.07)％,(0.33 ± 0.22)％ vs (0.02 ± 0.00)％,t =-4.997,-3.342,P ＜ 0.05].Conclusions The proportion of IL-22-secreting splenic lymphocytes is increased in mice infected with T.spiralis at the early encapsulated stage.The rise is caused by increased numbers of IL-22-secreting CD3 + lymphocytes,especially CD4+ T cells and γδT cells.

Alteración del ácido siálico eritrocitario por efecto de Trichinella spiralis aplicando el método de Azul Alcian / Alteration of erythrocyte sialic acid by the effect of Trichinella spiralis applying the Alcian Blue method / Alteração do ácido siálico eritrocitário por efeito da Trichinella spiralis aplicando o método de Alcian Blue

Actualmente se reconoce que los ácidos siálicos están involucrados en múltiples funciones biológicas y que tienen un papel importante en la interacción parásito-hospedador. El objetivo del trabajo fue estudiar la alteración del ácido siálico eritrocitario por efecto de T. spiralis aplicando el método de Azul Alcian. Se trabajó con 11 concentrados de larvas musculares de T. spiralis (LM) y 9 de larvas recién nacidas de T. Spiralis (LRN) (total: 20) y con suspensiones de eritrocitos frescos. Se realizó el tratamiento incubando el sedimento globular con igual volumen de concentrado larval (37 ºC durante 1 hora). Los controles fueron incubados de la misma forma con solución salina. Se aplicó el método de Azul Alcian y se determinó CAE% en el control y en los glóbulos tratados. Se calculó CexpCAE. Los resultados mostraron que 5/9 de los concentrados de LRN y 9/11 de LM modificaron la carga globular. La media y la desviación estándar de los CexpCAE de las suspensiones en que la carga disminuyó por contacto con LRN y LM, fueron 0,614±0,1946 y 0,656±0,1865 respectivamente, mientras que en las que no se modificó resultó 0,955±0,0289 en el tratamiento con LRN y 0,93±0,0141 con LM. Se concluye que el Método de Azul Alcian es útil para estudiar las variaciones en la carga eritrocitaria por efecto de LRN y LM de T. spiralis.

It is now recognized that sialic acids are involved in multiple biological functions and that they have an important role in the parasite-host interaction. The objective of this work was to study the alteration of erythrocyte sialic acid by the effect of T. spiralis applying the Alcian Blue method. Work was carried out with 11 larval concentrates of ML and 9 of NL (total 20) and with fresh erythrocyte suspensions. The treatment was performed incubating the globular sediment with equal volume of larval concentrate (37 °C for 1 hour). The controls were incubated in the same way with saline solution. The Alcian Blue method was applied and CAE% was determined in the control and in the treated globules. CexpCAE was calculated. The results showed that 5/9 of the NL concentrates and 9/11 of ML modified the globular charge. The mean and standard deviation of the CexpCAE of the suspensions in which the charge decreased by contact with NL and ML were 0.614±0.1946 and 0.656±0.1865 respectively, whereas in those that did not change, it was 0.955±0.0289 in the NL treatment and 0.93±0.0141 in the ML. It is concluded that the Alcian Blue Method is useful to study the variations in erythrocyte charge due to NL and ML of T. spiralis.

Atualmente se reconhece que os ácidos siálicos estão envolvidos em múltiplas funções biológicas e que têm um papel importante na interação parasita-hospedeiro. O objetivo do trabalho foi estudar a alteração do ácido siálico eritrocitário por efeito de T. spiralis aplicando o método do Alcian Blue. Trabalhou-se com 11 concentrados de larvas musculares de T. spiralis (LM) e 9 de larvas recém-nascidas de T. Spiralis (LRN) (total: 20) e com suspensões de eritrócitos frescos. Realizou-se o tratamento incubando o sedimento globular com igual volume de concentrado larval (37 ºC durante 1 hora). Os controles foram incubados da mesma forma com solução salina. Aplicou-se o método de Azul Alcian e determinou-se CAE% no controle e nos glóbulos tratados. Calculou-se CexpCAE. Os resultados mostraram que 5/9 dos concentrados de LRN e 9/11 de LM modificaram o carga globular. A média e o desvio padrão dos CexpCAE das suspensões em que a carga diminuiu por contato com LRN e LM, foram 0,614±0,1946 e 0,656±0,1865 respectivamente, enquanto que naquelas onde não se modificou, resultou 0,955±0,0289 no tratamento com LRN e 0,93±0,0141 com LM. Conclui-se que o método de Alcian Blue é útil para estudar as variações na carga eritrocitária por LRN e LM de T. spiralis.

Desialización eritrocitaria con y sin exposición del antígeno T producida por Trichinella spiralis / Erythrocyte desialylation with and without T antigen exposure produced by Trichinella spiralis / Desialylation eritrocitaria com e sem exposição antigénio T produzida por Trichinella spiralis

El objetivo del trabajo fue estudiar la desialización de eritrocitos que desenmascaran el antígeno T y de los glóbulos que no lo exponen, por contacto con larvas recién nacidas (LRN) de T. spiralis. Se trabajó con 15 suspensiones eritrocitarias en medio enzimático, incubadas con LRN. Los GR control se incubaron con solución salina. Para analizar el efecto del parásito sobre la exposición del antígeno T, se aplicó el Método de Aglutinación anti-T con antígeno T y se estudió la agregación por los métodos de Polibrene y de Análisis Digital de Imágenes, determinando CexpST, CCA y Distribución de los agregados eritrocitarios (DAE). La media de los coeficientes en los eritrocitos que expusieron el antígeno T fueron CexpST=0,22±0,179 y coeficiente de células aisladas (CCA)=0,73±0,108 y en los que no 0,86±0,125 y 0,205±0,163 respectivamente. La DAE mostró que los GR que lo expusieron, presentaron marcada disminución de células aisladas y pronunciado aumento de grandes agregados en relación a los controles, mientras que los GR en los que no hubo exposición, la disminución de células aisladas con respecto a los controles fue menor y el aumento de agregados estuvo homogéneamente distribuido entre todas las categorías. La experiencia realizada concluye que los valores obtenidos de los coeficientes difieren significativamente en los GR que exponen el antígeno T y en los que no, por lo que estas técnicas podrían ser predictivas para detectar desenmascaramiento T.

The aim of this work was to study the desialylation of erythrocytes that expose the T antigen by contact with T. spiralis newborn larvae (NL) and of those that do not. Work was carried out with 15 red cell suspensions in enzymatic medium, which were incubated with NL. The respective Control RBC were incubated with saline solution. To analyze the effect of the parasite on the exposure of the T antigen, the Anti T- antigen T Agglutination Method was applied and the aggregation was studied by Polybrene and Digital Image Analysis Methods determining CexpTS, ICC and Distribution of Erythrocyte Aggregates (DEA). The mean of coefficients in the erythrocytes that exposed the T antigen were CexpTS= 0.22 ± 0.179 and ICC= 0.73 ± 0.108 and of those that did not it was 0.86±0.125 y 0.205±0,163 respectively. DEA showed that the RBCs that exposed it showed a marked decrease of isolated cells and a pronounced increase of large aggregates in relation to the Controls, while the RBCs in which there was no exposure, the decrease of isolated cells with respect to Controls was lower and the increase of aggregates was homogeneously distributed among all categories. The experience concludes that the obtained values of the coefficients differ significantly in the RBCs that expose the T antigen from those that do not, so these techniques could be predictive to detect T unmasking.

O objetivo do trabalho foi estudar a desialização de eritrócitos que desmarcaram o antígeno T e dos glóbulos que não o expõem, por contato com LRN de T. spiralis. Trabalhou-se com 15 suspensões de eritrócitos em meio enzimático, incubadas com LRN. Os GV controle foram incubados com solução salina. Para analisar o efeito do parasita sobre a exposição do antígeno T, foi aplicado o Método de Aglutinação anti T com antígeno T e se estudou a agregação pelos Métodos de Polibrene e de Análise Digital de Imagens, determinando CexpST, CCA e Distribuição dos agregados eritrocitários (DAE). A média dos coeficientes nos eritrócitos que expuseram o antígeno T foram CexpST= 0.22±0.179 e CCA= 0.73±0.108 e naqueles que não o expuseram 0.86±0.125 y 0.205±0,163 respectivamente. Foi demonstrado que os GV que o expuseram, apresentaram marcada diminuição de células isoladas e pronunciado aumento de grandes agregados em relação aos controles, enquanto que os GV nos quais não houve exposição, a diminuição de células isoladas com respeito aos controles foi menor e o aumento de agregados esteve homogeneamente distribuído entre todas as categorias. A experiência realizada conclui que os valores obtidos dos coeficientes diferem significativamente nos GV que expõem o antígeno T e nos que não, portanto estas técnicas poderiam ser preditivas para detectar desmascaramento T.

Effect of the somatic proteins from Trichinella spiralis muscle larvae on apoptosis of H446 small cell lung cancer cells / 中国人兽共患病学报

To probe the effects of Trichinella spiralis muscle larval somatic proteins on small cell lung cancer H446 cells and the possible mechanism of anti-tumor,H446 cells were culture with 0.2 mg/mL,0.4 mg/mL,0.6 mg/mL,0.8 mg/mL,1.0 mg/mL,and 1.2 mg/mL somatic proteins respectively.The experimental group was set and no dosing as control group.MTT colorimetric assay was used to test the effects of T.spiralis muscle larval somatic proteins on the proliferative activity of H446 cells.We used flow cytometry (FCM) to test the influence of T.spiralis muscle larval somatic proteins induced H446 cells apoptosis.The real-time PCR and Western blot methods were used to detect the expression of Cyt-C and apoptotic protease activating factor 1(Apaf-1) mRNA and protein.The MTT colorimetric assay showed that T.spiralis muscle larval somatic proteins could inhibit the proliferation of H446 cells;the flow cytometry showed that polypide proteins acted on H446 cells after 24 h appeared an obvious effect on promoting apoptosis.Results of real-time PCR and Western blot analysis indicated that compared with the control group,Cyt-C and Apaf-1 showed up-regulated expression.T.spiralis muscle larval somatic proteins could inhibit proliferation activity and induce the apoptosis of H446 cells,and its effects may be related to up-regulated expression of Cyt-C and Apaf-1.

Efficacy of Albizia julibrissin total saponins against Trichinella spiralis in mice / 中国人兽共患病学报

We investigated the therapeutic effect of Albizia julibrissin total saponins on mice infected with Trichinella spiralis.Thirty-six ICR mice infected with Trichinella spiralis were randomly divided into 6 groups (each mouse infected with 300 T.spiralis),6 mice in each.Group Ⅰ:infected non-treated group (intestinal phase);group Ⅱ..received Albizia julibrissin total saponins group (intestinal phase);group Ⅲ:received albendazole group (intestinal phase);group Ⅳ:infected nontreated group (muscular phase);group Ⅴ:received Albizia julibrissin total saponins group (muscular phase);group Ⅵ:received albendazole group (muscular phase).Mice of Ⅰ,Ⅱ,Ⅲ group were administered on the second days post-infection(dpi) and continued for 3 days.Mice in these groups were sacrificed 7th dpi and adult worms recovered from the small intestine were counted.Mice of Ⅳ,Ⅴ,Ⅵ group were administered on the 7th dpi and continued for 14 d.The mice were sacrificed on 40th dpi,and the muscle larvae were counted.HE staining counts muscle larvae and the expression of IL-1β,IL-6,TNF-α and COX-2 in the diaphragm were detected by immunohistochemistry.Results showed that the number of adult worms and larva in groups received Albizia julibrissin total saponins and albendazole were significantly lower than that of infected non-treated group (P＜0.01).The worms reduction rate was 70.34％ and 80.02％ respectively,and the larva were 65.60％ and 90.66％ respectively.Results of HE staining showed the number of encysted larval and the expression of inflammatory cell were significantly reduced.The expression of IL-1β,IL-6,TNF-α and COX-2 was decreased in drug-treated groups.In conclusion,the total saponins of Albizia julibrissin showed adequate efficacy on Trichinella spiralis adults and encapsulated larva.Although the effect is slightly inferior to albendazole,as traditional Chinese medicine extract,it is less toxic.

Levels of Th1/Th2 in spleen lymphocyte of colitic mice without or with Trichinella spiralis infection / 中国免疫学杂志

Objective:To study the levels of Th1 and Th2 in spleen lymphocyte of TNBS and OXZ colitis without or with Trichinella spiralis(T.spiralis)infection.Methods:Female BALB/c mice were randomly divided into 3 groups:50％ Ethanol,TNBS or OXZ,T.spiralis+TNBS or OXZ(at least 6 in each group when mice were killed).The ratio of Th1/Th2 lymphocyte in spleen was detected by FCM with three color intracellular cytokine staining.Results:In spleen lymphocyte of TNBS colitic mice with prior nematode infection,the ratio of Th1/Th2 were not significantly different compared with that seen in colitic mice without prior nematode infection on day 3 post-induction of colitis(P＞0.05),the ratio of Th1/Th2 was significantly lower than which on day 7 post-induction of colitis (P＜0.05).In spleen lymphocyte of OXZ colitic mice with prior nematode infection on days 3 and 7 post-induction of colitis,the ratio of Th1/Th2 was significantly higher than that seen in colitic mice without prior nematode infection(P＜0.05).Conclusion:Infection of mice with T.spiralis distracts the mucosal immune system response from a Th1 response toward a protective Th2 response and up-regulate Tr1-cytokines with an attendant reduction in the severity of colonic damage and balance of Th1/Th2.Prior T.spiralis infection doesn't reduce the severity of OXZ-induced colitis,but without aggravating colitis.

Exposición de criptoantígeno eritrocitario por larvas recién nacidas de Trichinella spiralis / Exposure of erythrocyte cryptic antigen by Trichinella spiralis newborn larvae / Exposição do cripto antígeno eritrocitário por larvas recém-nascidas de Trichinella spiralis

La desialización del eritrocito puede exponer determinantes antigénicas crípticas de la membrana, entre las cuales se encuentra el antígeno T. Se ha comunicado que las larvas recién nacidas de Trichinella spiralis (LRN), las cuales circulan por el torrente sanguíneo del hospedador, captan el ácido siálico eritrocitario. El objetivo de este trabajo fue estudiar in vitro la exposición del criptoantígeno T por la desialización producida por distintas concentraciones de LRN. Se trabajó con 30 suspensiones eritrocitarias en medio enzimático, que fueron incubadas en partes iguales con concentrados de LRN (GR Tratados) durante 2 horas a 37 ºC con agitación continua. Los respectivos GR Controles se pusieron en contacto con solución salina. El tratamiento de 10/30 suspensiones globulares se realizó con 500 LRN/mL, 10 con 300 LRN/mL y las últimas 10 con 150 LRN/mL. Se aplicó el Método de Aglutinación anti-T con antígeno T (en Placa y Tubo) enfrentando las suspensiones a suero de adulto y suero de cordón. El tratamiento de 9/10 de las suspensiones con 500 LRN/mL, el de 7/10 con 300 LRN/mL y el de 2/10 con 150 LRN/mL las expuso al criptoantígeno. Se concluye que en pacientes diabéticos, hipertensos o con otra patología que produzca disminución de ácido siálico eritrocitario y que cursen simultáneamente una infección por T. spiralis, en la etapa de circulación de larvas por el torrente sanguíneo podría producirse la activación T con la consecuente hemólisis, trombocitopenia y trombosis.

Erythrocyte desialylation can expose cryptic antigenic determinants of the membrane, among which is the T antigen. It has been reported that newborn larvae (NL), which circulate in the bloodstream of the host, capture erythrocyte sialic acid. The aim of this study was to study in vitro exposure of cryptic T antigen by the desialylation produced by different concentrations of NL. Work was carried out on 30 red cell suspensions in enzymatic medium, which were incubated in equal parts with NL concentrates (Treated RBC) for 2 hours at 37 ºC with continued agitation. The respective Control RBC was incubated with saline solution. Treatment of 10/30 globular suspensions was performed with 500 NL/mL, 10 with 300 NL/mL and the last 10 with 150 NL/mL. Anti T- antigen T Agglutination Tests (Plate and Tube) were made, facing the globular suspensions against adult and cord human sera. Treatment of 9/10 suspensions with 500 NL/mL, 7/10 with 300 NL/mL and 2/10 with 150 NL/mL exposed T antigen. It is concluded that in patients with diabetes, hypertension or other diseases with lower content of erythrocyte sialic acid and who simultaneously have a T. spiralis infection, T activation may occur in the stage of larvae circulating through the bloodstream, with consequent haemolysis, thrombocytopenia and thrombosis.

A dessialização do eritrócito pode expor determinantes antigênicos crípticos da membrana, entre os quais se encontra o antígeno T. Foi comunicado que as larvas recém-nascidas de Trichinella spiralis (LRN), as quais circulam na corrente sanguínea do hospedeiro, capturam o ácido siálico eritrocitário. O objetivo foi estudar in vitro a exposição do cripto antígeno T pela dessialização produzida por diferentes concentrações de LRN. Trabalhou-se com 30 suspensões eritrocitárias em meio enzimático, incubadas em partes iguais com concentrados de LRN (GV Tratados) durante 2 horas a 37 °C com agitação contínua. Os respectivos GV Controles entraram em contato com solução salina. Tratamento de 10/30 suspensões globulares foi realizado com 500 LRN/mL, 10 com 300 LRN/mL e as últimas 10 com 150 LRN/mL. Foi aplicado o Método de aglutinação anti T-antígeno T (em Placa e Tubo) enfrentando as suspensões a soro de adulto e soro de cordão. O tratamento de 9/10 suspensões com 500 NRL / mL, o de 7/10 com 300 LRN/mL e o de 2/10 com 150 LRN/mL expuseram o cripto antígeno. Conclui-se que em pacientes diabéticos, hipertensos ou com outra patologia que produza diminuição do ácido siálico eritrocitário, e que padeçam simultaneamente uma infecção por T. spiralis, na fase de circulação de larvas pela corrente sanguínea, poderia ocorrer a ativação T com a consequente hemólise, trombocitopenia e trombose.

Análisis proteómico de los productos de excreción-secreción de cuatro aislados de Trichinella spiralis obtenidos de hospederos accidentales / Proteomic analysis of the excretion-secretion products of four Trichinella spiralis isolates obtained from accidental hosts

Resumen: Introducción: Trichinella spiralis es un nemátodo tisular que se aloja en el músculo esquelético de humanos y otros mamíferos y causa una serie de alteraciones fisiológicas. Las proteínas de los productos de excreción-secreción de T. spiralis juegan un papel importante en la aparición y regulación de estas alteraciones. Sin embargo, aún no se conoce el efecto de estos productos en la infección e invasión del parásito al hospedero. Métodos: Mediante un análisis electroforético en una dimensión, Western blot y espectrometría de masas, se evaluaron las diferencias y similitudes entre proteínas antigénicas y de superficie de cuatro aislados de T. spiralis obtenidos de hospederos accidentales (perros) y la cepa de referencia aislada de cerdos (MSUS/MEX/91/CM). Resultados: Utilizando ontología de genes, se encontraron cinco proteínas exclusivas de los hospederos accidentales. Después del análisis, se encontró que estas proteínas forman parte de la matriz extracelular del parásito, cuentan con actividad catalítica y están implicadas en la adhesión a las células del hospedero. La actividad antigénica de las cuatro cepas aisladas de hospederos accidentales es idéntica a la reportada para T. spiralis, visualizándose el triplete antigénico característico de 43, 45 y 47 kDa. Conclusiones: Las proteínas exclusivas de los hospederos accidentales proveen información para entender el mecanismo de acción de este parásito para penetrar el músculo y evadir la respuesta inmune en el hospedero.

Abstract: Background: Trichinella spiralis is an intestinal and tissue nematode specific for mammalian skeletal muscle, causing a series of physiological alterations. T. spiralis excretory-secretion products play an important role in the appearance and regulation of these alterations. However, the effect of these products on the infection and invasion of the parasite to the host is unknown. Methods: Differences and similarities between antigenic proteins and surface proteins of four accidental hosts isolates (dogs) of T. spiralis and the reference strain isolated from pigs (MSUS/MEX/91/CM) were assessed by electrophoresis, western blot and mass spectrometry. Results: Using gene ontology, five proteins exclusive to the accidental hosts were analyzed. The results showed that these proteins are part of the extracellular matrix of the parasite, present catalytic activity, and bind to host cells. The antigenic activity the four strains showed the antigenic triplet characteristic of T. spiralis of 43, 45 and 47 kDa. Conclusions: Five proteins exclusive to dog isolates provided information to understand the mechanism of action of this parasite to penetrate the muscle and evade the immune response in the host.