• Title/Summary/Keyword: immunized larvae

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Partial cross-resistance between Strongyloides venezuelensis and Nippostrongylus brasiliensis in rats

  • Baek, Byeong-Kirl;Islam, M.-Khyrul;Kim, Jin-Ho;Lee, John-Wha;Hur, Jin
    • Parasites, Hosts and Diseases
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    • v.37 no.2
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    • pp.101-107
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    • 1999
  • Rats were immunized through an initial infection with 1,000 filariform larvae (L3) of Nippostrongylus brasiliensis and after complete expulsion of worms they were challenged with 1,000 L3 of Strongyloides venezuelensis to investigate whether cross-resistance developed against a heterologous parasite. Nippostrongylus brasiliensis immunized rats developed a partial cross-resistance against S.venezuelensis migrating larvae (MSL3) in the lungs and adult worms in the small intestine. The population of MSL3 in the lungs were significantly lower (p<0.05) in immunized rats($22.0{\;}{\pm}{\;}7.4$) compared with controls ($105.0{\;}{\pm}{\;}27.6$). The populations of adult worms, egg output and fecundity were initially decreased but from day 14 post-challenge they did not show any significant difference between immunized and control rats. However, the length of worm in immunized rat was revealed as retardation. Peripheral blood eosinophilia was significantly decreased (P<0.05) on day 7 post-challenge and then gradually increased which peaked on da 42 post-challenge when most of the worms were expelled. these results suggest that peripheral blood eosinophilia is strongly involved in the worm establishment and expulsion mechanisms.

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Identification of a novel immune-related gene in the immunized black soldier fly, Hermetia illucens (L.)

  • Jung, Seong-Tae;Goo, Tae-Won;Kim, Seong Ryul;Choi, Gwang-Ho;Kim, Sung-Wan;Nga, Pham Thi;Park, Seung-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.2
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    • pp.25-30
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    • 2018
  • The larvae of Hermetia. illucens have a high probability of coming into contact with microorganisms such as bacteria and fungi. Therefore, the survival of H. illucens is primarily the protection of their own against microbial infection. This effect depends on the development of the innate immune system. Antimicrobial Peptides (AMPs) exhibit antimicrobial activity against other bacterial strains and can provide important data to understand the basis of the innate immunity of H. illucens. In this study, we injected larvae with Enterococcus. faecalis (gram-positive bacteria) and Serratia. marcescens as (gram-negative bacteria) to test the hypothesis that H. illucens is protected from infection by its immune-related gene expression repertoire. To identify the inducible immune-related genes, we performed and cataloged the transcriptomes by RNA-Seq analysis. We compared the transcriptomes of whole larvae and obtained a DNA fragment of 465 bp including the poly (A) tail by RACE as a novel H. illucens immune-related gene against bacteria. A novel target mRNA expression was higher in immunized larvae with E. faecalis and S. marcescens groups than non-immunized group. We expect our study to provide evidence that the global RNA-Seq approach allowed for the identification of a gene of interest which was further analyzed by quantitative RT-PCR, together with genes chosen from the available literature.

Isolation and Purification of Novel Anti-Fungal Peptides from Hemolymph of Immunized Larvae of Housefly, Musca domestica (집파리유충 hemolymph 중신형의 anti-fungal peptides의 분리정제)

  • Gu Li-Juan;Wu Jian-Wei;Su Xiao-Qing;Sung Chang-Keun
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.387-395
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    • 2006
  • To isolate and purify anti-fungal active substances from immunized housefly (Musca domestica), low dose of Candida albicans was injected into the larvae of the housefly to induce the appearance of potent anti-fungal active substances in the hemolymph. This purification work was performed by the routine isolation and purification processes of protein, namely, solid phase extraction (SPE), SDS-PACE electrophoresis, HPLC purification. Three 4-16 kDa peptides which exhibited antifungal activity against Candida albican and other fungi were isolated from induced hemolymph. Consequently, further anti-fungal activity study showed that these three peptides were different either in molecular weight or in anti-fungal activity. All isolated substances were proved to be active and resistant to high-temperature. It was deduced that these peptides isolated from induced housefly were novel members of the insect defensin family and they were inducible.

Modulation of Antibody Responses against Gnathostoma spinigerum in Mice Immunized with Crude Antigen Formulated in CpG Oligonucleotide and Montanide ISA720

  • Intapan, Pewpan M.;Hirunpetcharat, Chakrit;Kularbkaew, Churairat;Yutanawiboonchai, Wiboonchai;Janwan, Penchom;Maleewong, Wanchai
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.637-644
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    • 2013
  • This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.

The Antimicrobial Activity of Bacterial-challenged Black Soldier Fly, Hermetia illucens (세균에 의해 면역이 유도된 동애등에의 항균활성)

  • Park, Kwanho;Yun, Eun-Young;Park, Seung-Won;Goo, Tae-Won
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1409-1414
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    • 2016
  • In the larvae of the black soldier fly, Hermetia illucens, innate immunity mechanisms are activated in response to various pathogens and stimulants, resulting in the expression of antimicrobial peptides (AMPs). To induce the mass production of AMPs, H. illucens fifth instar larvae were immunized with five different kinds of bacteria. We isolated from the hemolymph of the H. illucens larvae after bacterial challenge, and their antimicrobial activities against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) were measured using the inhibition zone assay. Among these five different kinds of bacteria, the hemolymph of Bacillus subtilis-challenged H. illucens larvae showed the strongest antimicrobial activity against both Gram-positive bacteria and Gram-negative bacteria. The antimicrobial activity of the hemolymph of $1{\times}10^9cfu/ml$ B. subtilis-challenged H. illucens peaks at 24 hr at 48 hr post-infection and gradually declines with time. Moreover, the immunized hemolymph also showed strong antimicrobial activity against various poultry pathogens such as S. enteritidis, S. typhimurium, and S. pullorum. These results suggest that the expression of AMP genes in B. subtilis-challenged H. illucens is up-regulated by innate immune responses, and that B. subtilis-challenged H. illucens overexpressing AMPs may be useful as a feed additive in livestock diets to reduce the need for antibiotics.

Efficacy of recombinant enolase as a candidate vaccine against Haemaphysalis longicornis tick infestation in mice

  • Md. Samiul Haque;Mohammad Saiful Islam;Myung-Jo You
    • Parasites, Hosts and Diseases
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    • v.61 no.4
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    • pp.439-448
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    • 2023
  • Tick infestation causes a significant threat to human and animal health, requiring effective immunological control methods. This study aimed to investigate the potential of recombinant Haemaphysalis longicornis enolase protein for tick vaccine development. The exact mechanism of the recently identified enolase protein from the H. longicornis Jeju strain remains poorly understood. Enolase plays a crucial role in glycolysis, the metabolic process that converts glucose into energy, and is essential for the motility, adhesion, invasion, growth, and differentiation of ticks. In this study, mice were immunized with recombinant enolase, and polyclonal antibodies were generated. Western blot analysis confirmed the specific recognition of enolase by the antiserum. The effects of immunization on tick feeding and attachment were assessed. Adult ticks attached to the recombinant enolase-immunized mice demonstrated longer attachment time, increased bloodsucking abilities, and lower engorgement weight than the controls. The nymphs and larvae had a reduced attachment rate and low engorgement rate compared to the controls. Mice immunized with recombinant enolase expressed in Escherichia coli displayed 90% efficacy in preventing tick infestation. The glycolytic nature of enolase and its involvement in crucial physiological processes makes it an attractive target for disrupting tick survival and disease transmission. Polyclonal antibodies recognize enolase and significantly reduce attachment rates, tick feeding, and engorgement. Our findings indicate that recombinant enolase may be a valuable vaccine candidate for H. longicornis infection in experimental murine model.

The Localization of the Excretory, Purified and Infected Antigenic Protein in the Tissue of Trichinella spiralis Larval Worm (선모충(Trichinella spiralis) 유충의 조직 내 배설, 분리 및 감염항원 단백의 분포)

  • Kim, Soo-Jin;Joo, Kyoung-Hwan;Chung, Myung-Sook;Rho, Young-Bok
    • Applied Microscopy
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    • v.37 no.1
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    • pp.43-52
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    • 2007
  • In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.