• The Plant Pathology Journal
• /
• v.38 no.1
• /
• pp.12-24
• /
• 2022

In this study, we conducted whole-genome sequencing with six species of Pectobacterium composed of seven strains, JR1.1, BP201601.1, JK2.1, HNP201719, MYP201603, PZ1, and HC, for the analysis of pathogenic factors associated with the genome of Pectobacterium. The genome sizes ranged from 4,724,337 bp to 5,208,618 bp, with the GC content ranging from 50.4% to 52.3%. The average nucleotide identity was 98% among the two Pectobacterium species and ranged from 88% to 96% among the remaining six species. A similar distribution was observed in the carbohydrate-active enzymes (CAZymes) class and extracellular plant cell wall degrading enzymes (PCWDEs). HC showed the highest number of enzymes in CAZymes and the lowest number in the extracellular PCWDEs. Six strains showed four subsets, and HC demonstrated three subsets, except hasDEF, in type I secretion system, while the type II secretion system of the seven strains was conserved. Components of human pathogens, such as Salmonella pathogenicity island 1 type type III secretion system (T3SS) and effectors, were identified in PZ1; T3SSa was not identified in HC. Two putative effectors, including hrpK, were identified in seven strains along with dspEF. We also identified 13 structural genes, six regulator genes, and five accessory genes in the type VI secretion system (T6SS) gene cluster of six Pectobacterium species, along with the loss of T6SS in PZ1. HC had two subsets, and JK2.1 had three subsets of T6SS. With the GxSxG motif, the phospholipase A gene did locate among tssID and duf4123 genes in the T6SSa cluster of all strains. Important domains were identified in the vgrG/paar islands, including duf4123, duf2235, vrr-nuc, and duf3396.

• Molecules and Cells
• /
• v.46 no.11
• /
• pp.688-699
• /
• 2023

We set up this study to understand the underlying mechanisms of reduced ceramides on immune cells in acute rejection (AR). The concentrations of ceramides and sphingomyelins were measured in the sera from hepatic transplant patients, skin graft mice and hepatocyte transplant mice by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Serum concentrations of C24 ceramide, C24:1 ceramide, C16:0 sphingomyelin, and C18:1 sphingomyelin were lower in liver transplantation (LT) recipients with than without AR. Comparisons with the results of LT patients with infection and cardiac transplant patients with cardiac allograft vasculopathy in humans and in mouse skin graft and hepatocyte transplant models suggested that the reduced C24 and C24:1 ceramides were specifically involved in AR. A ceramide synthase inhibitor, fumonisin B₁ exacerbated allogeneic immune responses in vitro and in vivo, and reduced tolerogenic dendritic cells (tDCs), while increased P3-like plasmacytoid DCs (pDCs) in the draining lymph nodes from allogeneic skin graft mice. The results of mixed lymphocyte reactions with ceranib-2, an inhibitor of ceramidase, and C24 ceramide also support that increasing ceramide concentrations could benefit transplant recipients with AR. The results suggest increasing ceramides as novel therapeutic target for AR, where reduced ceramides were associated with the changes in DC subsets, in particular tDCs.

• IMMUNE NETWORK
• /
• v.21 no.5
• /
• pp.34.1-34.16
• /
• 2021

Sjögren's syndrome (SS) is an autoimmune disease characterized by dryness of the mouth and eyes. The glandular dysfunction in SS involves not only T cell-mediated destruction of the glands but also autoantibodies against the type 3 muscarinic acetylcholine receptor or aquaporin 5 (AQP5) that interfere with the secretion process. Studies on the breakage of tolerance and induction of autoantibodies to these autoantigens could benefit SS patients. To break tolerance, we utilized a PmE-L peptide derived from the AQP5-homologous aquaporin of Prevotella melaninogenica (PmAqp) that contained both a B cell "E" epitope and a T cell epitope. Repeated subcutaneous immunization of C57BL/6 mice with the PmE-L peptide efficiently induced the production of Abs against the "E" epitope of mouse/human AQP5 (AQP5E), and we aimed to characterize the antigen specificity, the sequences of AQP5E-specific B cell receptors, and salivary gland phenotypes of these mice. Sera containing anti-AQP5E IgG not only stained mouse Aqp5 expressed in the submandibular glands but also detected PmApq and PmE-L by immunoblotting, suggesting molecular mimicry. Characterization of the AQP5E-specific autoantibodies selected from the screening of phage display Ab libraries and mapping of the B cell receptor repertoires revealed that the AQP5E-specific B cells acquired the ability to bind to the Ag through cumulative somatic hypermutation. Importantly, animals with anti-AQP5E Abs had decreased salivary flow rates without immune cell infiltration into the salivary glands. This model will be useful for investigating the role of anti-AQP5 autoantibodies in glandular dysfunction in SS and testing new therapeutics targeting autoantibody production.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.15 no.1
• /
• pp.113-116
• /
• 2011

Purpose: Acetylcholine receptor antibodies cause acetylcholine receptor loss, which is responsible for failure of the neuromuscular junction in the acetylcholine receptor autoantibody. The disease characterized by muscle weakness and fatigue, myasthenia gravis(MG) occurs when the body inappropriately produces antibodies against acetylcholine receptors, and thus inhibits proper acetylcholine signal transmission. And this reason, the measurement of acetylcholine receptor antibodies can be of considerable value in disease diagnosis. Methods: From 2010. August to September, we tested orderd AchRAb 19 samples to get the results. 1. Pipette $5{\mu}{\ell}$ undiluted patient sera and kit control and add 125I AChR $50{\mu}{\ell}$ and incubate at R.T for 2 hours. 2. Pipette $50{\mu}{\ell}$ of anti-human IgG into each tube, and incubate at $2{\sim}8^{\circ}C$ for 2 hours. 3. Pipette $25{\mu}{\ell}$ precipitation enhancer into each tube and add 1mL washing solution into all tubes. 4. Centrifuge each tube for 20minutes at $2{\sim}8^{\circ}C$ at 1500g. 5. Aspirate or decant the supernatant. 6. Pipette 1 mL washing solution into all tubes and resuspend the pellet and repeat centrifugation. 7. Aspirate or decant the supernatant and count all tubes on a gamma counter. Results: Cut off value is 0.2 nmol/L and the results taken below 0.2 nmol/L are negative, the results above that identified as being positive values. We assayed the 19 patients samples and got 7 positive results. Of which, 6 patients were diagnosed as MG.(85.7%). Conclusions: Acetylcholine Receptor autoantibody test is intended for use by persons only for the quantitative determination of it in human serum. Even if measurement of the antibodies is not a routine test, it can be of considerable value in disease diagnosis.

• Journal of Preventive Medicine and Public Health
• /
• v.22 no.2 s.26
• /
• pp.290-301
• /
• 1989

This is preliminary report on anthrax epidemic occurred in an island with about 100 residents. Since 1982 there had been sudden deaths among all kinds of domestic animals including cattle, dogs, ducks, chicken and goat but only a few among cats in an isolated island about three hours distance away by ferry boat from Mokpo city. From 1986 through 1988 nine human deaths and four patients occurred, which made the government intervene for investigation on June 25 1988. The epidemiological investigation consisted of interview survey and medical examination, medical record analysis, laboratory work to isolate the pathogens under the direction of hypothesis derived from the study and further confirmation of the pathogens by international institute. The summarized results are as followings: 1. According to the interview survey there were many deaths among domestic animals usually in cold and dry season such as January through March and September through November; 36 heads of cattle leaving one head, more than 40 hogs(all), hundreds of chicken leaving few alive, goats that had taken home from mountain and two or three cats out of around 40 had sudden deaths from 1982 till 1985, when the residents stopped to purchase and take them into the island anymore. Also there were eleven persons who had experienced the similar syndrome complex to those of admitted and expired patients and four of them revealed typical chest X-ray findings; from one of these four patients(Rho) B. anthracis is isolated. 2. Medical record on patients who had been admitted, showed common characteristics of the disease course. On admission they had either gastrointestinal or upper respiratory infection symptoms which invariably progressed to septicemic nature with pulmonary interstitial infiltration and mediastinal widening/bulging, and then to deadly acute respiratory distress syndrome. At the end stage chest X-ray revealed multiple bullous emphysema. One of another characteristics was oral ulceration with bleeding occurred in about 50% of the patients. Laboratory test results in common were leukocytosis with left shift and abnormal liver and kidney functions, particularly at the later stage of the illness. 3. Epidemiological characteristics was striking in that both mortality and incidence rates were high: the mortality rate was 8.7% average, male being three times higher than females but there was no distictive clustering by age group. The incidence rate for both sexes was 28.2% and there was no sex difference although a tendency of higher incidence among older ages was noticed. The highest mortality and incidence were observed in Won village where the first death of animal occurred and with the highest frequency among three villages of the island. 4. Among twelve bacilli species isolated from various specimens, two strains, one from patient and the other from soil where the recently died cow is hurried, were confirmed as B. anthracis by Pasteur Institute and CDC of USA(strain from soil). CDC reported that the strain did not produce capsule in bicarbonate media but reacted with the bacteriophage and one of five sera taken from the patients. Mode of transmission as well as incubation period of the agent has not been established yet, which needs further investigation in relation to the antigenic structure of the variant when it is confirmed.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.35 no.1
• /
• pp.1-7
• /
• 2002

To study the increased resistance in fish against Vibrio vulnificus known as an important agent of vibrio septicemia in human, we analyzed specific and nonspecific immune response in flounder after administration of V. vulnificus bacterins by oral route. It contained the comparison of antibody concentrations in the sera of flounder after oral administration by two different protocols with uncoated heat killed bacterin of V vulnificus (UHKB, 20 mg/kg body weight), i.e., 4 weeks continuously (group 4W) and taking 2 weeks resting period between the 1st and last week of administration (group 1-2-1W). Even though, 1-2-1W group showed significantly increased level of specific antibody in serum, it did not reach to that of 4W group. Certainly, flounder vaccinated twice a week for four weeks (20 mg/kg b.w.) showed increased concentration of specific antibody against V. vulnificus at week 2 after last administration by oral route and maintained throughout the experimental period. It also was confirmed by the increased numbers of specific antibody secreting cells (SASC) in the leukocytes isolated from the splenocytes of the flounder of 4W group at week 1 after last administration until the end of experimental period. However, enteric, acid-resistant film coated heat killed bacterin (ECHB) did not show both greater immune reaction for antibody production and faster elimination of a challenge dose of V. vulnificus compared with those of the UHKB. These results suggested that UHKB administered by oral route was very effective method to prevent the contamination of V vulnificus in flounder, and did not show the increased antigenicity by coating the surface with acid-resistant film.

• Parasites, Hosts and Diseases
• /
• v.24 no.2
• /
• pp.145-158
• /
• 1986

This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatogaphy using specific monoclonal antibody(McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secreted antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen (A-Ag) and unbound pool CF (U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitivity was about 70% of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis.

• Korean Journal of Animal Reproduction
• /
• v.25 no.2
• /
• pp.119-124
• /
• 2001

This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p＜0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p＜0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

• Korean Journal of Veterinary Research
• /
• v.10 no.1
• /
• pp.11-23
• /
• 1970

Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.