Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice

2018 ◽  
Vol 63 (2) ◽  
pp. 252-260 ◽  
Author(s):  
Xuliang Zhang ◽  
Lixin Xu ◽  
Xiaokai Song ◽  
Xiangrui Li ◽  
Ruofeng Yan
Keyword(s):  
Recombinant Protein ◽  
Trichinella Spiralis ◽  
Flow Cytometric Analysis ◽  
Recombinant Plasmids ◽  
Flow Cytometric ◽  
Th2 Immune Response ◽  
Wide Range ◽  
Muscle Larvae ◽  
Ifn Γ ◽  
Negative Controls

Abstract Trichinella spiralis, the main pathogen of trichinosis, infects a wide range of mammalian hosts and is one of the most widespread parasites worldwide. For parasites, glycolysis is the most important way to generate energy. Previous studies showed that some enzymes involved in the glycolytic pathway play roles in regulation the host immunity. In this paper, enolase from T. spiralis was cloned and the protective potentials were studied. One hundred and sixty ICR mice were divided into four groups and vaccinated with recombinant enolase (pET-ENO), eukaryotic recombinant plasmid encoding enolase (pVAX1-ENO) and negative controls (pVAXl and PBS), respectively. Two weeks after the second immunization, each mouse was challenged orally with 200 muscle larvae (MLs) of T. spiralis. Results showed that mice vaccinated with pET-ENO and pVAX1-ENO induced specific antibodies of IgG, IgA, IgM, but no IgE. Subclasses of IgG antibodies showed that mice immunized with recombinant protein and recombinant plasmids induced a Th1/Th2 immune response. Concentrations of serum cytokines were detected and showed significant increase of IFN-γ, IL-4 and TGFβ1, while IL-17 in each group was not significantly different. Flow cytometric analysis showed significant increase of CD4+ and CD8+ T lymphocytes in the groups immunized with recombinant protein and recombinant plasmids. Challenge infection demonstrated that immunized groups had a reduced number of worm burdens. The reductions of larvae per gram muscle (LPG) in pET-ENO and pVAX1-ENO group were 17.7% and 15.8% when compared with PBS control.

scholarly journals Analysis of Circulating Haemocytes fromBiomphalaria glabratafollowingAngiostrongylus vasorumInfection Using Flow Cytometry

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Thales A. Barçante ◽  
Joziana M. P. Barçante ◽  
Ricardo T. Fujiwara ◽  
Walter S. Lima
Keyword(s):  
Control Strategies ◽  
Flow Cytometric Analysis ◽  
Intermediate Hosts ◽  
Flow Cytometric ◽  
Wide Range ◽  
Activation Mechanisms ◽  
Labelling Procedure ◽  
Dual Colour ◽  
Cellular Defence ◽  
Cellular Components

Angiostrongylus vasorumis an emerging parasite of dogs and related to carnivores that have an indirect life cycle, with a wide range of terrestrial and aquatic gastropods as the obligatory intermediate host. Unfortunately, the relationship betweenA. vasorumand their snail hosts remains poorly understood. Circulating haemocytes are the main line of cellular defence involved in the destruction of helminths in snails. Aiming to further characterize the haemocyte subsets inBiomphalariasnails, we have performed a flow cytometric analysis of whole haemolymph cellular components using a multiparametric dual colour labelling procedure. Our findings demonstrated thatB. glabratainfected withA. vasorumhave two major circulating haemocyte subsets, referred to as small and large haemocytes. Differences in the cell proportion occurred over time. The development of better invertebrate infection control strategies would certainly result in the better control of human diseases caused by other species of the genusAngiostrongylus. Such knowledge will assist in the establishment of novel control strategies aimed at parasites that use molluscs as intermediate hosts and clarify new aspects of the parasite-host relationship regarding cell recognition and activation mechanisms, which are also found in the innate response of vertebrates.

scholarly journals False-positive flow cytometric platelet glycoprotein IIb/IIIa expression in myeloid leukemias secondary to platelet adherence to blasts

Blood ◽  
1992 ◽  
Vol 79 (9) ◽  
pp. 2399-2403
Author(s):  
SA Betz ◽  
K Foucar ◽  
DR Head ◽  
IM Chen ◽  
CL Willman
Keyword(s):  
De Novo ◽  
Flow Cytometric Analysis ◽  
Platelet Glycoprotein ◽  
Treatment Protocols ◽  
Megakaryoblastic Leukemia ◽  
Flow Cytometric ◽  
Wide Range ◽  
Specific Antigens ◽  
De Novo Aml ◽  
Immunologic Assays

Because neither morphologic nor routine cytochemical features are pathognomonic, the diagnosis of acute megakaryoblastic leukemia (AML- M7) is difficult, requiring either specialized ultrastructural or immunologic techniques. Because the ultrastructural techniques are cumbersome, immunologic assays for expression of several platelet- specific antigens, such as CD41a (platelet glycoprotein IIb/IIIa), have become the primary method used to detect megakaryoblastic differentiation. In our flow cytometric analysis of the immunophenotypes of over 1,000 cases of AML from patients registered to Southwest Oncology Group (SWOG) Treatment Protocols, we found that 38% of cases demonstrated CD41a reactivity. Because this frequency of CD41a expression by flow cytometry greatly exceeded the number of morphologically defined cases of AML-M7, we postulated that the reaction may be caused by platelets adherent to leukemic blasts. To investigate this hypothesis, we performed a side-by-side comparison of flow cytometric and cytospin immunofluorescence studies on 37 cases of adult de novo AML that demonstrated a wide range of CD41a expression by flow cytometric analyses. We found that the expression of CD41a detected by flow cytometric techniques was secondary to adherent platelets or platelet fragments in 85% of cases. Many of these cases also expressed the lineage-specific carbohydrate, LNF III (CD15), which may mediate platelet adhesion to mature monocytes and neutrophils. Only 15% of the CD41a flow cytometrically positive cases demonstrated true diffuse membrane and cytoplasmic positivity on cytospin slides indicative of megakaryoblastic differentiation. Cytospin immunofluorescence for CD41a should be performed on all cases of suspected AML-M7. If only flow cytometric techniques are used, adherent platelets may result in the erroneous diagnosis of this AML subtype.

scholarly journals Short-Term Relocation Stress-Induced Hematological and Immunological Changes in Saimiri boliviensis boliviensis

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Pramod N. Nehete ◽  
Bharti P. Nehete ◽  
Akash G. Patel ◽  
Sriram Chitta ◽  
Henrieta Scholtzova ◽  
...  
Keyword(s):  
Stress Responses ◽  
Functional Activity ◽  
Flow Cytometric Analysis ◽  
Squirrel Monkeys ◽  
Blood Analysis ◽  
Immunological Parameters ◽  
Social Stressors ◽  
Flow Cytometric ◽  
Abrupt Changes ◽  
Ifn Γ

Nonhuman primates are frequently transported to a new location or temporarily relocated within their colony. Both transportation and relocation expose animals to new environments, causing them to undergo a stress response (before adapting). In our NHP colony, the mentioned situations are not infrequent for many reasons, including maintenance. The objective of this study was to determine whether abrupt changes consisting of relocation, housing, separation, and grouping could influence hematological and immunological parameters and thereby functional activity. The current study used squirrel monkeys as a model to investigate the stress-inducing effects of relocation within a facility, while animals acclimated to new situations (physical, housing). A detailed blood analysis revealed significant Changes in lymphocytes, triglycerides, total protein, creatinine, and ALT. Flow cytometric analysis of peripheral blood showed reduction in CD3+, CD4+, and CD8+ T cells and monocytes, while B cells and natural killer (NK) cells changed with relocation. Simultaneously, changes in functional activity of immune cells altered proliferative responses and as shown by ELISpot (IFN γ). Though the parameters studied are not affected as severely as those in animals transported by road or air, stress responses induced by intrafacility relocation are significant and worth consideration. Our findings indicate that squirrel monkeys mimic the features seen in humans exposed to social stressors and may serve an important model for understanding the mechanisms of stress-induced immune dysfunction in humans.

scholarly journals False-positive flow cytometric platelet glycoprotein IIb/IIIa expression in myeloid leukemias secondary to platelet adherence to blasts

Blood ◽  
1992 ◽  
Vol 79 (9) ◽  
pp. 2399-2403 ◽  
Author(s):  
SA Betz ◽  
K Foucar ◽  
DR Head ◽  
IM Chen ◽  
CL Willman
Keyword(s):  
De Novo ◽  
Flow Cytometric Analysis ◽  
Platelet Glycoprotein ◽  
Treatment Protocols ◽  
Megakaryoblastic Leukemia ◽  
Flow Cytometric ◽  
Wide Range ◽  
Specific Antigens ◽  
De Novo Aml ◽  
Immunologic Assays

Abstract Because neither morphologic nor routine cytochemical features are pathognomonic, the diagnosis of acute megakaryoblastic leukemia (AML- M7) is difficult, requiring either specialized ultrastructural or immunologic techniques. Because the ultrastructural techniques are cumbersome, immunologic assays for expression of several platelet- specific antigens, such as CD41a (platelet glycoprotein IIb/IIIa), have become the primary method used to detect megakaryoblastic differentiation. In our flow cytometric analysis of the immunophenotypes of over 1,000 cases of AML from patients registered to Southwest Oncology Group (SWOG) Treatment Protocols, we found that 38% of cases demonstrated CD41a reactivity. Because this frequency of CD41a expression by flow cytometry greatly exceeded the number of morphologically defined cases of AML-M7, we postulated that the reaction may be caused by platelets adherent to leukemic blasts. To investigate this hypothesis, we performed a side-by-side comparison of flow cytometric and cytospin immunofluorescence studies on 37 cases of adult de novo AML that demonstrated a wide range of CD41a expression by flow cytometric analyses. We found that the expression of CD41a detected by flow cytometric techniques was secondary to adherent platelets or platelet fragments in 85% of cases. Many of these cases also expressed the lineage-specific carbohydrate, LNF III (CD15), which may mediate platelet adhesion to mature monocytes and neutrophils. Only 15% of the CD41a flow cytometrically positive cases demonstrated true diffuse membrane and cytoplasmic positivity on cytospin slides indicative of megakaryoblastic differentiation. Cytospin immunofluorescence for CD41a should be performed on all cases of suspected AML-M7. If only flow cytometric techniques are used, adherent platelets may result in the erroneous diagnosis of this AML subtype.

Interferon gamma delays apoptosis of mature erythroid progenitor cells in the absence of erythropoietin

Blood ◽  
2000 ◽  
Vol 95 (12) ◽  
pp. 3742-3749 ◽  
Author(s):  
Ilseung Choi ◽  
Koichiro Muta ◽  
Amittha Wickrema ◽  
Sanford B. Krantz ◽  
Junji Nishimura ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Interferon Gamma ◽  
Human Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Annexin V ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cells ◽  
Flow Cytometric ◽  
Ifn Γ ◽  
Protease Assay

Based on the hypothesis that interferon gamma (IFN-γ) may have stimulating effects on survival of hematopoietic progenitor cells, we examined the effect of IFN-γ on apoptosis of mature erythroid colony-forming cells (ECFCs) derived from human peripheral blood obtained from normal, healthy volunteers. When the cells were cultured in the presence of IFN-γ, even without erythropoietin (EPO), the viability of the cells was maintained for at least 36 hours. When apoptosis of ECFCs was assessed by flow cytometric analysis', using annexin V, IFN-γ reduced the extent of apoptosis of the cells, as well as EPO. DNA fragmentation of ECFCs was also reduced by IFN-γ. In cells cultured with IFN-γ alone, expression of Bcl-x was detected but the level of expression decreased gradually during incubation for 36 hours, and the expression level was lower than incubation with EPO. Fas expression and activation of downstream caspases were assessed by flow cytometric analysis or fluorometric protease assay. IFN-γ induced Fas expression of the cells without the activation of caspase8 or caspase3 during 16 hours of incubation, while deprivation of EPO induced expression of Fas and the activation of both caspase8 and caspase3. We propose that IFN-γ produces a stimulating signal for the survival of mature erythroid progenitor cells by reducing apoptosis through a mechanism other than modulating Fas and one related to the expression of Bcl-x.

Flow cytometric analysis of intracellular IFN-γ, IL-4 and IL-10 in CD3+4+ T-cells from rat spleen

2000 ◽  
Vol 244 (1-2) ◽  
pp. 29-40 ◽  
Author(s):  
Emma M Caraher ◽  
Monique Parenteau ◽  
Heidi Gruber ◽  
Fraser W Scott
Keyword(s):  
T Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Ifn Γ

scholarly journals Adjuvanticity of β -Glucan for Vaccine Against Trichinella spiralis

2021 ◽  
Vol 9 ◽  
Author(s):  
Yi Liu ◽  
Xiaolei Liu ◽  
Li Yang ◽  
Yangyuan Qiu ◽  
Jianda Pang ◽  
...  
Keyword(s):  
T Cells ◽  
Trichinella Spiralis ◽  
Serine Protease Inhibitor ◽  
Vaccine Adjuvants ◽  
Vaccine Candidates ◽  
The Past ◽  
Th2 Immune Response ◽  
Ifn Γ

In the past 30 years, few researches focus on the efficacy of adjuvant against Trichinella spiralis infection. Identifying new, improved vaccine adjuvants for T. spiralis infection are required. β-glucan are effective and safe as adjuvant for infectious diseases. In this paper, we first observed the adjuvanticity of β-glucan as adjuvant for defensing helminth T. spiralis in vivo. We showed that IgG and IgE were elevated in the mice immunized with β-glucan combined with recombinant T. spiralis serine protease inhibitor (rTs-Serpin), which is one of the vaccine candidates. Furthermore, in vitro, the combination of β-glucan and rTs-Serpin enhanced the maturation of bone marrow dendritic cells (BMDCs) compared to rTs-Serpin alone. We showed that β-glucan + rTs-Serpin –treated BMDCs secreted higher production of IL-12 and IL-10. Moreover, β-glucan + rTs-Serpin –treated BMDCs not only promoted the population of CD4+ IFN-γ+ T cells, but also enhanced the population of CD4+ IL-4+ T cells. These findings suggested that β-glucan, as an adjuvant, have the capacity to protect against T. spiralis infection via activating both Th1 and Th2 immune response.

Interferon gamma delays apoptosis of mature erythroid progenitor cells in the absence of erythropoietin

Blood ◽  
2000 ◽  
Vol 95 (12) ◽  
pp. 3742-3749 ◽  
Author(s):  
Ilseung Choi ◽  
Koichiro Muta ◽  
Amittha Wickrema ◽  
Sanford B. Krantz ◽  
Junji Nishimura ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Interferon Gamma ◽  
Human Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Annexin V ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cells ◽  
Flow Cytometric ◽  
Ifn Γ ◽  
Protease Assay

Abstract Based on the hypothesis that interferon gamma (IFN-γ) may have stimulating effects on survival of hematopoietic progenitor cells, we examined the effect of IFN-γ on apoptosis of mature erythroid colony-forming cells (ECFCs) derived from human peripheral blood obtained from normal, healthy volunteers. When the cells were cultured in the presence of IFN-γ, even without erythropoietin (EPO), the viability of the cells was maintained for at least 36 hours. When apoptosis of ECFCs was assessed by flow cytometric analysis', using annexin V, IFN-γ reduced the extent of apoptosis of the cells, as well as EPO. DNA fragmentation of ECFCs was also reduced by IFN-γ. In cells cultured with IFN-γ alone, expression of Bcl-x was detected but the level of expression decreased gradually during incubation for 36 hours, and the expression level was lower than incubation with EPO. Fas expression and activation of downstream caspases were assessed by flow cytometric analysis or fluorometric protease assay. IFN-γ induced Fas expression of the cells without the activation of caspase8 or caspase3 during 16 hours of incubation, while deprivation of EPO induced expression of Fas and the activation of both caspase8 and caspase3. We propose that IFN-γ produces a stimulating signal for the survival of mature erythroid progenitor cells by reducing apoptosis through a mechanism other than modulating Fas and one related to the expression of Bcl-x.

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