scholarly journals Preparation and characterization of monoclonal antibodies against ES antigens of Trichinella spiralis

2014 ◽  
Vol 51 (4) ◽  
pp. 253-261 ◽  
Author(s):  
J. Cui ◽  
F. Jing ◽  
G. Fu ◽  
H. Ren ◽  
L. Liu ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Western Blot ◽  
Post Infection ◽  
Trichinella Spiralis ◽  
Internal Organs ◽  
Cross Reactions ◽  
Muscle Larvae ◽  
Somatic Antigens

AbstractHybridomas secreting monoclonal antibodies (MAbs) against excretory-secretory (ES) antigen of Trichinella spiralis muscle larvae were produced. The 12 monoclones (designated 3A11, 7D9, 7F6, 5A7, 6H7, 39F3, 38H2, 37E7, 35B9, 29A11, 39C9 and 39A6) secreted IgM, IgG3 and IgG2b, respectively. Western blot using T. spiralis ES antigens showed that ten of 12 MAbs recognized the bands between 119.8 − 19.3kDa (mainly 68.4 − 28.7kDa) and two MAbs (3A11, 37E7) did not recognize any protein constituents of ES antigens. Nine of 12 MAbs also recognized the larval antigens of T. nativa, T. pseudospiralis and T. nelsoni at 40 days post-infection (dpi). No cross-reactions were found with the somatic antigens of adult worms of P. westermani, C. sinensis, and T. solium cysticercus. However, three (3A11, 6H7 and 39A6) were cross-reacted with the somatic antigens of adult worms of S. japonicum. Immunofluorescent location showed that the nine MAbs reacted with the cuticle and internal organs of T. spiralis larvae. Additionally, five and eight MAbs generated in this study can recognize the early antigens of pre-encapsulated T. spiralis larvae at 11 dpi and 13 dpi, respectively. The generation and characterization of the MAbs against T. spiralis ES antigens provide foundation for the development of specific early serodiagnostic techniques for trichinellosis.

scholarly journals Characterization of a Novel Glutamine Synthetase From Trichinella spiralis and Its Participation in Larval Acid Resistance, Molting, and Development

2021 ◽  
Vol 9 ◽  
Author(s):  
Tong Xu Zhuo ◽  
Zhen Wang ◽  
Yan Yan Song ◽  
Shu Wei Yan ◽  
Ruo Dan Liu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Life Cycle ◽  
Glutamine Synthetase ◽  
Enzymatic Activity ◽  
Post Infection ◽  
Catalytic Domain ◽  
Trichinella Spiralis ◽  
Acid Resistance ◽  
Muscle Larvae

Trichinella spiralis is a major foodborne parasite worldwide. After the encapsulated muscle larvae (ML) in meat are ingested, the ML are liberated in the stomach of the host and activated into intestinal infectious larvae (IIL), which develop into adult worm after molting four times. A novel glutamine synthetase (TsGS) was identified from T. spiralis IIL at 10 h post-infection, but its biological role in T. spiralis life cycle is not clear. The aim of this study was to investigate the biological characteristics of TsGS and its functions in larval acid resistance, molting, and development. TsGS has a glutamine synthetase (GS) catalytic domain. Complete TsGS sequence was cloned and expressed in Escherichia coli BL21. rTsGS has good immunogenicity. qPCR and Western blotting showed that TsGS was highly expressed at IIL stage, and immunofluorescence revealed that TsGS was principally localized at the cuticle and intrauterine embryos of this nematode. rTsGS has enzymatic activity of natural GS to hydrolyze the substrate (Glu, ATP, and NH4+). Silencing of TsGS gene significantly reduced the IIL survival at pH 2.5, decreased the IIL burden, and impeded larval molting and development. The results demonstrated that TsGS participates in T. spiralis larval acid resistance, molting and development, and it might be a candidate vaccine target against Trichinella molting and development.

scholarly journals Production and characterization of monoclonal antibodies to the edta extract of Leptospira interrogans, serovar icterohaemorrhagiae

1996 ◽  
Vol 29 (5) ◽  
pp. 483-489
Author(s):  
Lilian Terezinha de Queiroz Leite ◽  
Mauricio Resende ◽  
Wanderley de Souza ◽  
Elizabeth R.S. Camargos ◽  
Matilde Cota Koury
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Silver Staining ◽  
Colloidal Gold ◽  
Leptospira Interrogans ◽  
Outer Envelope ◽  
Lethal Challenge ◽  
Sds Page

Monoclonal antibodies (MABs) ivere produced against an etbylenediaminetetraacetate (EDTA) extract of Leptospira interrogans serovar icterohaemorrhagiae being characterized by gel precipitation as IgM and IgG (IgGl and IgG2b). The EDTA extract was detected as several bands by silver staining in SDS-PAGE. In the Western blot the bands around 20 KDa reacted with a monoclonal antibody, 47B4D6, and was oxidized by periodate and was not digested by pronase, suggesting that the determinant is of carbohydrate nature, lmmunocytochemistry, using colloidal gold labeling, showed that an EDTA extract determinant recognized by monoclonal antibody 47B4D6, is localized under the outer envelope of serovar icterohaemorrhagiae. Hoe AIAB raised against the EDTA extract was not able to protect hamsters from lethal challenge with virulent homologous leptospires.

scholarly journals Detection of circulating antigens in serum samples of mice experimentally infected with Trichinella spiralis by a sandwich ELISA based on IgY

2014 ◽  
Vol 51 (3) ◽  
pp. 181-189 ◽  
Author(s):  
F. Jing ◽  
J. Cui ◽  
R. Liu ◽  
L. Liu ◽  
P. Jiang ◽  
...  
Keyword(s):  
Post Infection ◽  
Trichinella Spiralis ◽  
Sandwich Elisa ◽  
Egg Yolk ◽  
Mouse Serum ◽  
Serum Samples ◽  
Egg Yolk Immunoglobulin ◽  
Muscle Larvae ◽  
Sandwich Elisa Assay ◽  
Circulating Antigens

AbstractIn the present study, a sandwich ELISA based on IgY (egg yolk immunoglobulin) was developed for detection of circulating antigens (CAg) in sere of mice experimentally infected with Trichinella spiralis. The IgY-sandwich ELISA assay involved the use of chicken antibody IgY against excretory-secretory (ES) antigens of Trichinella spiralis muscle larvae as a capture antibody and mouse polyclonal antibody IgG to ES antigens as a detecting antibody. This method was able to detect as little as 3 ng/ml of ES antigens added to normal mouse serum. A group of sixteen mice was orally inoculated with 500 T. spiralis muscle larvae per animal. The serum samples from the infected mice were taken during 1–35 days post-infection (dpi). The CAg was detectable as early as 8 dpi in the sera of infected mice. The level of CAg increased dramatically during 13–15 dpi and reached a peak at 22 dpi and remained a plateau for 3 days, then declined gradually. Another peak of CAg occurred at 31 dpi. The anti-Trichinella antibodies was first detected in 14.3 % of the infected mice at 2 weeks post-infection (wpi), and reached a peak positive rate of 100 % at 5 wpi. Moreover, the infected mice were treated with abendazole at 5 wpi and the serum CAg levels increased significantly during 2–6 days posttreatment (dpt) and then declined rapidly during 8–14 dpt. By 42 dpt, the CAg levels decreased to the undetected level, but the detection rate of antibodies was still 100 %. The IgY-sandwich ELISA appears to be a sensitive for detection of antigenemia of T. spiralis and valuable to judge the efficacy of chemotherapy in trichinellosis.

scholarly journals Cellular and molecular changes and immune response in the intestinal mucosa during Trichinella spiralis early infection in rats

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
María Priscila Saracino ◽  
Cecilia Celeste Vila ◽  
Melina Cohen ◽  
María Virginia Gentilini ◽  
Guido Hernán Falduto ◽  
...  
Keyword(s):  
Immune Response ◽  
Lamina Propria ◽  
Trichinella Spiralis ◽  
Mucosal Immune Response ◽  
Mesenteric Lymph Nodes ◽  
Muscle Larvae ◽  
Gut Immunity ◽  
Gut Mucosa

Abstract Background: The main targets of the host’s immune system in Trichinella spiralis infection are the adult worms (AW), at the gut level, and the migrant or newborn larvae (NBL), at systemic and pulmonary levels. Most of the studies carried out in the gut mucosa have been performed on the Payer’s patches and/or the mesenteric lymph nodes but not on the lamina propria, therefore, knowledge on the gut immune response against T. spiralis remains incomplete. Methods This study aimed at characterizing the early mucosal immune response against T. spiralis, particularly, the events taking place between 1 and 13 dpi. For this purpose, Wistar rats were orally infected with muscle larvae of T. spiralis and the humoral and cellular parameters of the gut immunity were analysed, including the evaluation of the ADCC mechanism exerted by lamina propria cells. Results A marked inflammation and structural alteration of the mucosa was found. The changes involved an increase in goblet cells, eosinophils and mast cells, and B and T lymphocytes, initially displaying a Th1 profile, characterised by the secretion of IFN-γ and IL-12, followed by a polarization towards a Th2 profile, with a marked increase in IgE, IgG1, IL-4, IL-5 and IL-13 levels, which occurred once the infection was established. In addition, the helminthotoxic activity of lamina propria cells demonstrated the role of the intestine as a place of migrant larvae destruction, indicating that not all the NBLs released in the gut will be able to reach the muscles. Conclusions The characterization of the immune response triggered in the gut mucosa during T. spiralis infection showed that not only an effector mechanism is directed toward the AW but also towards the NBL as a cytotoxic activity was observed against NBL exerted by lamina propria cells.

scholarly journals Production and characterization of monoclonal antibodies raised against recombinant human granzymes A and B and showing cross reactions with the natural proteins

1993 ◽  
Vol 163 (1) ◽  
pp. 77-83 ◽  
Author(s):  
J.Alain Kummer ◽  
Angela M. Kamp ◽  
Marcel van Katwijk ◽  
Just P.J. Brakenhoff ◽  
Katarina Radošević ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cross Reactions

A panel of antigens of muscle larvae of Trichinella spiralis and T. pseudospiralis as revealed by two-dimensional Western blot and immunoelectron microscopy

Parasitology ◽  
1999 ◽  
Vol 118 (6) ◽  
pp. 615-622 ◽  
Author(s):  
Z. WU ◽  
I. NAGANO ◽  
Y. TAKAHASHI
Keyword(s):  
Western Blot ◽  
Trichinella Spiralis ◽  
Cross Reactivity ◽  
Microscopical Study ◽  
Detectable Level ◽  
Antigenic Peptides ◽  
Two Dimensional ◽  
Western Blots ◽  
Muscle Larvae ◽  
Isoelectric Points

This study characterized antigens of Trichinella spiralis and T. pseudospiralis muscle larvae recognized by mice infected with the worms. Two-dimensional (2-D) Western blot analysis revealed some profile of antigenic peptides including: (1) molecular weight (MW); (2) isoelectric points (pI), (3) reactivity to well-defined monoclonal antibodies (mAb) and (4) cross-reactivity between the 2 species. Antigenic peptides of T. spiralis consisted of about 100 spots. The MW ranged from 22 to 80 kDa, and pI ranged from 4 to 7. The mAb against TSL-1 stained most of the T. spiralis excretory–secretory (E–S) peptides migrating at 40, 45 and 50 kDa, and the mAb against TSL-4 stained non-E–S peptides. Antigenic peptides of T. pseudospiralis consisted of about 20 to 30 peptide spots. The MW ranged from 25 to 80 kDa, and pI ranged from 4 to 7. The mAb against TSL-1 stained most of the T. pseudospiralis E–S peptides migrating at 35 and 45 kDa, and the mAb against TSL-4 stained non-E–S peptides. Two-dimensional Western blots showed that the E–S products of T. spiralis and T. pseudospiralis were highly cross-reactive with each other. The non-E–S peptides were, however, not recognized by T. pseudospiralis-infected sera but were recognized by T. spiralis-infected sera. An immunoelectron microscopical study showed the similar result that stichocyte granules and cuticle surface (known to contain E–S antigen) had cross-reactive antigens between the two species. T. pseudospiralis-infected sera stained very weakly the cuticle inner layers and haemolymph (known to contain non-E–S antigen). This evidence implies that mice infected with T. pseudospiralis do not evoke antibodies against non-E–S antigen at the detectable level.

scholarly journals A comparison of antigenic peptides in muscle larvae of severalTrichinellaspecies by two-dimensional western-blot analysis with monoclonal antibodies

Parasite ◽  
2001 ◽  
Vol 8 ◽  
pp. S117-S119 ◽  
Author(s):  
M.A. Dea-Ayuela ◽  
F.M. Ubeira ◽  
A. Pitarch ◽  
C. Gil ◽  
A.R. Martínez-Fernández ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Antigenic Peptides ◽  
Two Dimensional ◽  
Muscle Larvae

scholarly journals Production and characterization of polyclonal and monoclonal antibodies against human thromboxane synthase

Blood ◽  
1990 ◽  
Vol 76 (1) ◽  
pp. 80-85 ◽  
Author(s):  
R Nusing ◽  
MP Wernet ◽  
V Ullrich
Keyword(s):  
Enzyme Activity ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Human Lung ◽  
Human Platelet ◽  
Human Lung Tissue ◽  
Western Blots ◽  
Conformational Epitopes ◽  
Platelet Thromboxane

Abstract Polyclonal and monoclonal antibodies (MoAbs) were raised against human platelet thromboxane (Tx) synthase. Neither the antiserum nor the MoAbs inhibited the enzyme activity significantly. Three MoAbs, Tu 300, Kon 6, and Kon 7, were purified and further characterized. They are monospecific as shown by activity precipitation or Western blot analysis, and recognized different epitopes on Tx-synthase. Tu 300 could precipitate the enzyme and recognized conformational epitopes, whereas Kon 6 and Kon 7 only reacted in Western blots. Antibody Tu 300 can be used in immunohistology but shows no crossreactivity with Tx- synthase from other species. In human lung tissue staining with peroxidase, coupled Tu 300 was only found in alveolar macrophages.

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