• Title/Summary/Keyword: secretory IgA

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Immunization with a Genetically Engineered Uropathogenic Escherichia coli Adhesin-Escherichia coli Enterotoxin Subunit A2B Chimeric Protein

  • Lee, Yong-Hwa;Kim, Byung-O;Pyo, Suhk-Neung
    • Biomolecules & Therapeutics
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    • v.13 no.2
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    • pp.101-106
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    • 2005
  • The generation of secretory IgA antibodies (Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the heat-labile Escherichia coli enterotoxin A2B (ltxa2b) gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ltxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and westem blotting using antibodies to the maltose binding protein (MBP) or the heat labile E. coli subunit B (LTXB), plus the N-terminal amino acid sequence was analyzedd. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/LTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of LTXB. thisstudy also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA (sIgA) and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked LTXA2B acts as a useful mucosal adjuvant, and that adhesin/LTXA2A chimeric protein might be a potential antigen for oral immunization against UPEC.

Enhancement of Mucosal Immune Functions by Dietary Spirulina platensis in Human and Animals

  • Osamu Hayashi;Kyoko Ishii;Chinami Kawamura;Hei, Shi-Yen;Bao, Ning-Ye;Tomohiro Hirahashi;Toshimitsu Katoh
    • Nutritional Sciences
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    • v.7 no.1
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    • pp.31-34
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    • 2004
  • This paper reviews the effects of Spirulina platensis and its extracts and phycocyanin, a blue photosynthetic pigment protein in Spirulina on the mucosal immune functions in humans and animals as follows: TEX>$\bullet$ IgA antibody response and other classes in mucosal immunity of mice treated with Spirulina platensis and its extract. $\bullet$ Effect of Spirulina phycocyanin ingestion on the mucosal antibody responses in mice. - Distinct effects of phycocyanin on secretory IgA and allergic IgE antibody responses in mice following oral immunization with antigen-entrapped biodegradable microparticles. $\bullet$ Influence of dietary Spirulina platensis on IgA level in human saliva. $\bullet$ A study on enhancement of bone-marrow cell-proliferation and differentiation by Spirulina platensis in mice: in vivo and in vitro study

Construction of a live attenuated Salmonella strain expressing FanC protein to prevent bovine enterotoxigenic Escherichia coli and evaluation of its immunogenicity in mice

  • Won, Gayeon;Kim, Hee Jung;Lee, John Hwa
    • Korean Journal of Veterinary Research
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    • v.57 no.1
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    • pp.9-15
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    • 2017
  • To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

Degradations of human immunoglobulins and hemoglobin by a 60 kDa cysteine proteinase of Trichomonas vaginalis (질편모충의 60 kDa 시스테인 단백분해효소의 인체 면역글로불린 및 헤모글로빈 분해능)

  • Duk-Young MIN;Keun-Hee Hyun;Jae-Sook Ryu;Myoung-Hee AHN;Myung-Hwan CHO
    • Parasites, Hosts and Diseases
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    • v.36 no.4
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    • pp.261-268
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    • 1998
  • The present study was undertaken to investigate the role of cysteine proteinase of Trichomonas vaginalis in escaping from host defense mechanism. A cysteine proteinase of T. vaginalis was purified by affinity chromatography and gel filtration. Optimum pH for the purified proteinase activity was 6.0. The proteinase was inhibited by cysteine and serine proteinase inhibitors such as E-64, NEM, IAA, leupeptin. TPCK and TLCK, and also by $Hg^{2+}$, but not affected by serine-, metallo-, and aspartic proteinase inhibitors such as PMSF, EDTA and pepstatin A. However, it was activated by the cysteine proteinase activator, DTT. The molecular weight of a purified proteinase was 62 kDa on gel filtration and 60 kDa on SDS-PAGE. Interestingly, the purified proteinase was able to degrade serum IgA, secretory IgA, and serum IgG in time- and dose-dependent manners. In addition, the enzyme also degraded hemoglobin in a dose-dependent manner. These results suggest that the acidic cysteine proteinase of T. vaginalis may play a dual role for parasite survival in conferring escape from host humoral defense by degradation of immunoglobulins, and in supplying nutrients to parasites by degradation of hemoglobin.

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Immunoelectron microscopic localization of partially purified antigens in adult Paragonimus iloktsuenensis

  • Lee, Ok-Ran;Chung, Pyung-Rim
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.119-132
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    • 2001
  • An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of Dl fraction (DIA) among 5 antigenic protein fractions partially purified by DEAE- anion exchange chromatography from water- soluble crude antigen (PIWA) of adult Paragonimus iloktsuenensis. Immune reactions of adult worm tissues with rabbit serum immunoglobulin immunized with crude antigen (PI-Ig) and D1 antigen (D1-Ig), as well as rat serum immunoglobulin infected with P. iloktsuenensis were observed. DlA showed strong antigenicity in the intestinal epithelium of the worms during the early infection period of 2-4 weeks after infection. The vitellaria also showed stronger antigenicity than the other tissue sites in immune reaction of tissues against all immunoglobulins from 4 to 33 weeks after vitelline development. Therefore, it is suggested that DlA was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immune-reactivity of two immunoglobulins (PI-Ig, Dl-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with Dl-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in Dl antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue .

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Protection Against Salmonella Typhimurium, Salmonella Gallinarum, and Salmonella Enteritidis Infection in Layer Chickens Conferred by a Live Attenuated Salmonella Typhimurium Strain

  • Lee, John Hwa
    • IMMUNE NETWORK
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    • v.15 no.1
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    • pp.27-36
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    • 2015
  • In the present study, we investigated the protection conferred by a live attenuated Salmonella enterica serovar Typhimurium (ST) strain against Salmonella Typhimurium, Salmonella Gallinarum (SG), and Salmonella Enteritidis (SE) infection in layer chickens. Birds were orally primed with the attenuated ST strain at 7 days of age and then boosted at 4 weeks post prime immunization (PPI). Sequential monitoring of plasma IgG and mucosal secretory IgA (sIgA) levels revealed that inoculation with ST induced a significant antibody response to antigens against ST, SE, and SG. Moreover, significant lymphoproliferative responses to the 3 Salmonella serovars were observed in the immunized group. We also investigated protection against virulent ST, SE, and SG strain challenge. Upon virulent SG challenge, the immunized group showed significantly reduced mortality compared to the non-immunized group. The reduced persistence of the virulent ST and SE challenge strains in the liver, spleen, and cecal tissues of the immunized group suggests that immunization with the attenuated ST strain may not only protect against ST infection but can also confer cross protection against SE and SG infection.

A seroepidemiological survey for toxocariasis in apparently healthy residents in Gangwon-do, Korea

  • Park, Hyun-Young;Lee, soo-Ung;Huh, Sun;Kong, Yoon;Magnaval, Jean-Francois
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.113-117
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    • 2002
  • We investigated the sero-prevalence of toxocariasis among healthy Korean adults in 1999. A total of 314 sera from normal inhabitants in Whachon-gun, Gangwon do, Korea was examined for specific antibody levels against excretory-secretory products of second stage larvae of Toxocara (TES). The presence of cross-reactions with other helminthiases such as cysticercosis, paragonimiasis, sparganosis or clonorchiasis was also checked by specific IgG ELISA. Sera showing positive reaction against TES were also tested by IgG immunoblot and by IgE ELISA. Out of 314 subjects, 16 was found to be positive by TES IgG ELISA and immunoblot, among whom 12 were also positive by TES IgE ELISA. Among the 16 seropositive samples, two sera showed positive reaction against Paragonimus and sparganum antigen, respectively. These results inferred that cross-reactions were negligible between toxocariasis and other helminthiases. Toxocariasis seroprevalence among Korean rural adults was detected to be approximately 5%.

Adherence of Salivary Proteins to Various Orthodontic Brackets (다양한 교정용 브라켓 표면에 부착하는 타액단백질에 관한 연구)

  • Ahn, Sug-Joon;Ihm, Jong-An;Nahm, Dong-Seok
    • The korean journal of orthodontics
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    • v.32 no.6 s.95
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    • pp.443-453
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    • 2002
  • The principal aims of this study were to identify the composition of salivary pellicles formed on various orthodontic brackets and to obtain a detailed information about the protein adsorption profiles from whole saliva and two major glandular salivas. Four different types of orthodontic brackets were used. All were upper bicuspid brackets with a $022{\times}028$ slot Roth prescription; stainless steel metal, monocrystalline sapphire, polycrystalline alumina, and plastic brackets. Bracket pelicles were formed by the incubation of orthodontic brackets with whole saliva, submandibular-sublingual saliva, and parotid saliva for 2 hours. The bracket pellicles were extracted and confirmed by employing sodium dodecyl sulfatepolyacrylamide gel electrophoresis, Western transfer methods, and immunodetection. The results showed that low-molecular weight salivary mucin, ${\alpha}-amylase$, secretory IgA (sIgA), acidic proline-rich proteins, and cystatins were attached to all of these brackets regardless of the bracket types. High-molecular weight mucin, which promotes the adhesion of Streptococcus mutans, did not adhere to uy orthodontic brackets. Though the same components were detected in all bracket pellicles, however, the gel profiles showed qualitatively and quantitatively different pellicles, according to the origins of saliva and the bracket types. In particular, the binding of sIgA was more prominent in the pellicles from parotid saliva and the binding of cystatins was prominent in the pellicles from the form plastic brackets. This study indicates that numerous salivary proteins adhere to the orthodontic brackets and these salivary proteins adhere selectively according to bracket types and the types of the saliva.

The Relationship between the Salivary IgA against AgI/II of S. mutans and Dental Caries Experience among Children and Adults (소아와 성인의 타액 내 AgI/II 특이 IgA 와 우식경험도의 관계)

  • Lim, Su-Min;Kim, Jae-Gon;Baik, Byeong-Ju;Yang, Yeon-Mi;Kim, Su-Kyung;Lee, Kyung-Yol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.4
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    • pp.671-676
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    • 2008
  • AgI/II of Streptococcus mutans(S. mutans) is an important virulence factor that contributes to the pathogenesis of S. mutans-induced dental caries. In oral cavity, salivary IgA antibodies act as safeguards against enormous challenges from oral bacteria. IgA antibodies inhibit adherence of cariogenic microorganisms to hard surfaces. Analysis of salivary IgA against AgI/II can be very useful diagnostic and powerful communication tools to the dental caries The purpose of this study was to investigate correlation between salivary AgI/II specific IgA and incidence of dental caries among children and young adults. Subjects consisted of 28 children and 18 adults. They were assigned to four groups : Group I deft index $\leq$3), Group II(deft index $\geq$4), Group III(DMFT index $\leq$3), Group IV(DMFT index $\geq$4) and they was divided two groups into caries resistant group and caries susceptible group. The study group were examined caries activity and their salivary IgA was evaluated by enzyme-linked immunosorbent assay. The results are as follows : 1. There was a positive correlation between the number of S. mutans and caries activity. 2. The titer of salivary IgA against the AgI/II was significantly higher in caries resistant group than caries susceptible group(p<0.01). 3. The titer of salivary IgA against the AgI/II in Group III was significantly higher than Group II(p<0.05).

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Study on Intestinal Flora and IgA Concentration Analysis in Newborn Mice by Stress and Diet in Pregnant Mice to Investigate the Effect of Taedok on the Intestinal Flora (태독이 장내 세균총에 미치는 영향을 규명하기 위한 임신쥐의 스트레스 및 식이에 따른 신생쥐의 장내 세균총 및 IgA 농도 분석 시험)

  • Jeong, Jieun;Choi, Yoomin;Jeong, Minjeong
    • The Journal of Pediatrics of Korean Medicine
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    • v.35 no.4
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    • pp.96-111
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    • 2021
  • Objectives The purpose of this study is to look for pathological mechanism of disease development caused by Taedok, by studying whether stress and diet in pregnant ICR mice affect the intestinal flora and IgA (Immunoglobulin A) concentration. Methods The mice were divided into 4 groups (n=5 per group) based on the concept of Taedok: the control group (G1), stress group (G2), capsaicin diet group (G3), high fat diet group (G4). We collected and analyzed intestinal flora from maternal feces and cecal flora from neonatal mice by group. Then, IgA concentration in the maternal feces and sIgA (secretory Immunoglobulin A) concentration in the cecal contents of newborn mice were analyzed. In addition, serum corticosterone was analyzed before and after stress application. Results Changes in maternal intestinal flora and neonatal mice cecal flora by stress and diet were observed. There were no significant changes in the IgA concentration in maternal feces and the sIgA concentration in the cecal contents of neonatal mice. No significant changes compared to the control group were observed between groups before and after applying stress. However, when comparing within one subject, a significant increase was confirmed after stress application in the stress group (G2). Conclusions Based on the results, we observed stress and diet in pregnant mice affect the intestinal flora of maternal and neonatal. We were able to interpret the pathological mechanism of Taedok based on the principle of interaction between mother and newborn intestinal flora.