The Journal of the Korean Society for Microbiology
/
v.3
no.1
/
pp.43-49
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1968
Japanese encephalitis(JE) shows its explosive epidemicity in the temperate zone of Asia but little is known on the overwintering mechanism. One of the hypotheses on the overwintering mechanism is that the virus overwinters in the hibernating animals. There has been no report on the proliferation of JE virus(JEV) or antibody formation in the snakes. The purpose of this experiment is to explore the mutual relationship between JEV and snake and to clarify whether JEV proliferates and induce antibody formation in snakes. Three species of non-poisonous common snakes were employed. Precipitation test was carried out after injecting calf serum and, HI and neutralization tests were done by injecting JEV into the snakes. The gamma globulin fraction of pre- and post-injection serum were compared by paper chromatography. According to the results, precipitating antibody reaction to calf serum could be observed only at $4^{\circ}C$. It was failed to demonstrate HI antibody formation but neutralizing antibody could be detected in one of the 9 snakes. Although antibody could not be detected in test-tube, tile result of paper chromatography shows the remarkable increase of gamma globulin fraction after the injection. Above results are strongly indicating the antibody formation in the snakes.
Alloimmunization to red blood cell (RBC) antigens may cause a delayed hemolytic transfusion reactions (DHTR) and a delayed serologic transfusion reactions (DSTR). In the present study, the frequency of alloimmunization and its clinical significance were evaluated. Also, transfusions were correlated with antibody formation. Alloimmunization rate was 0.63%. Alloimmunization rate in multiple transfused patients was 24.5%. The most common clinically significant alloantibodies of alloimmunized patients were found to be Rh antibodies (52.6%). Nine patients out of 38 (23.7%) became undetectable after the first detection. To be positive at antibody screening test after RBC transfusion was mean transfused numbers: 3.7 units, mean transfused periods: 56 days, mean transfused frequencies: 1.7 times. The results from antibody specificity and RBC transfusions were comparatively analyzed and it shows that Rh system antibodies were longer than other antibodies (P<0.05). In case of disease group, malignant diseases was longer than other diseases (P<0.05). In order to prevent the formation of RBC alloimmunization, irregular antibody screening tests were performed at propriety intervals in multiple transfused patients.
With the ongoing rise of coronavirus disease 2019 (COVID-19) pandemic across the globe, interests in COVID-19 antibody testing, also known as a serology test has grown, as a way to measure how far the infection has spread in the population and to identify individuals who may be immune. Recently, many countries reported their population based antibody titer study results. South Korea recently reported their third antibody formation rate, where it divided the study between the general population and the young male youths in their early twenties. As previously stated, these simple point estimates may be misinterpreted without proper estimation of standard error and confidence intervals. In this article, we provide an updated 95% confidence intervals for COVID-19 antibody formation rate for the Korean population using asymptotic, exact and Bayesian statistical estimation methods. As before, we found that the Wald method gives the narrowest interval among all asymptotic methods whereas mid p-value gives the narrowest among all exact methods and Jeffrey's method gives the narrowest from Bayesian method. The most conservative 95% confidence interval estimation shows that as of 00:00 November 23, 2020, at least 69,524 people were infected but not confirmed. It also shows that more positive cases were found among the young male in their twenties (0.22%), three times that of the general public (0.051%). This thereby calls for the quarantine authorities' need to strengthen quarantine managements for the early twenties in order to find the hidden infected people in the population.
The divalent antibody-toxins are expected to have increased binding avidities to target cells because of the two cell-binding domains. However, previous studies showed that the refolding yield of divalent antibody-toxin is very low, and it is assumed that homodimer formation of antibody-toxin is strongly interfered by the repulsion between the two large toxin domains that come close to each other during dimer formation. In this study, B3 antibody was used as a model antibody, and its Fab domain was used to construct three different kinds of Fab divalent molecules, $[B3(Fab)-toxin]_{2}$. The monomer Fab-toxin molecules were made by fusing the Fab domain of monoclonal antibody B3 to PE38, a truncated mutant form of Pseudomonas exotoxin (PE), and a connecting sequence that contained spacer amino acid sequence (G4S)n (n=l, 2, 3) was inserted between Fab and PE38. The prepared divalent molecules were $[Fab-S\;1,\;2,\;3-PE38]_{2}\;(=[Fab-SKPCIST-KAS(G_{4}S)nGGPE-PE38]_{2}\;(n=1,\;2,\;3))$, and they are derivatives of previously studied $[Fab-H2cys-PE38]_{2}\;(=[Fab-SKPCIST-KASGGPE-PE38]_{2})$. In $[Fab-Sl,\;2,\;3-PE38]_{2}$, two Fab-S1, 2, 3-PE38 monomers were covalently linked by the disulfide bond bridge made from cysteine in the -SKPCIST- sequence. The insertion of spacer amino acids after the disulfide bridge resulted in a 12-18 fold higher yield of dimer formation than previously constructed $[Fab-Hlcys-PZ38]_{2}[7]$, 3-4-fold higher than $[Fab-ext-PZ38]_{2}[25]$. These two molecules have less amino acid spacer sequence between the disulfide bridge and PE38 domain. The design of $[Fab-PE38]_{2}$ in this study gave molecules with a higher refolding yield. The results of cytotoxicity assay showed a higher cytotoxic effect of these divalent molecules than that of the monovalent scFv-PE38 molecule.
These mxperiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro fertilization and following development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6mm of diameter. Bovine oocytes were matured in vitro for 24~26hrs in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$ and subsequently cultured in medium containing cumulus cell antibody for 1 hour. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hrs in BO solution 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and then cultured for 7 days. The results obtained in these experiments were summarized as follows : 1. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cells, the fertilization rate of cumulus intact and removed oocytes was ranged to 45.0 to 53.7%. These value is slightly lower than that(64.3%) of follicular oocytes not treated with the antibody, and increased frequency of both male and female pronuclear formation was found in cumulus intact oocytes cultred in medium without the antibody(p<0.05). 2. The fertilization rate of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cells was ranged 45.0 to 52.5%, significantly lowre than that(62.8%) of oocytes cultured in antibody free medium, and increased frequency of ova with male and female pronuclei was found when cumulus cells were present(p<0.05). 3. The rates of cumulus cell intact and removed oocytes developed to 8-, 16-cell and morula or blastocyst after treatment of intact and solubilized cumulus cell antibody were ranged 7.1 to 14.5, 2.9 to 5.9 and 1.5 to 2.9%, respectively, slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results of this stduy indicate that cumulus cells promote not only normal fertilization with proper pronuclear formation, but embryo development and that the beneficial effect of cumulus cell to the pronuclear formation and embryo development is blocked by the action of antibody to cumulus cell.
The Journal of the Korean Society for Microbiology
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v.10
no.1
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pp.1-8
/
1975
Despite a number of recent studies on appendix its function appears to remain unknown. The present studies were undertaken in order to extend and confirm the previous studies concerning the role of appendix in immune response. An early hemagglutinin response of mercaptoethanol sensitive antibody(IgM antibody) in rabbit injected intravenously(i.v.) with 200mcg of bovine gamma globulin(BGG) was abolished by lethal whole body irradiation(900 r), but preserved in animals whose appendix and bone marrow were shielded during irradiation. Late formation of mercaptoethanol resistant antibody(IgG antibody) and the development of memory in bone marrow shielded animals were not affected by irradiation of the appendix. Formation of either IgM or IgG antibody to sheep red blood cells(SRBC) injected i.v. as determined by direct plaque forming cell(DPFC) technique in spleen were effectively abolished by appendectomy, thymectomy, or both followed by irradiation. When bone marrow was shielded in combination with autologous appendix reconstitution, DPFC response was about 5 times greater than the sum of two. Lysed appendix cells failed to restore the response. Lethally irradiated rabbits restored with combination of autologous appendix and thymus cells showed DPFC responses which were essentially normal. Three pools of appendix were obtained by manual separation technique and were stimulated with soluble concanavalin A(Con A), phytohemagglutinin-P(PHA) and pokeweed mitogen(PWM). Rabbit appendix cells responded to Con A, PHA and PWM. Cells of thymus dependent area(TDA) of the appendix were relatively enriched in their response to T cell mitogens compared to dome and follicle cells. The PHA/Con A responsive ratio of appenix TDA subpopulation was high, indicating that Con A responsive cells have a wider distribution among appendix. This finding showed that interfollicular area of the appendix is thymus-dependent. The present studies confirmed other evidence that the rabbit appendix cells itself are unable to form antibody and T lymphocytes in appendix TDA may be heterogenous, and that the appendix cells are synergistic with either bone marrow or thymus cells in the early hemagglutinin on splenic antibody response to BGG or SRBC.
The protein disulfide isomerase (PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody (Mab) refolding and assembly which accompanies disulfide bend formation. The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb in-termediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant fur a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.
The protein disulfide isomerase(PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody(MAb) refolding and assembly which accompanies disulfide bond formation The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb intermediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant for a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.
BALB/C mice were intraperitoneally injected with methamphetamine (5 mg/kg) to observe the effect of methamphetamine on the immune system. Body weights were decreased in both acutely treated group (twice for 2 weeks with 7 days interval) and subchronically treated group (daily injection for 14 days). The relative spleen weights and the numbers of splenocytes were unexpectedly increased (p<0.05) in acutely treated group, but subchronically treated group showed the trend of decrease without significance. But there was no significant effect on antibody formation to hen egg Iysozyme which was immunized during the treatment of methamphetamine and on plaque forming cell number. The relative thymus weights of both groups were significantly decreased by the treatment of methamphetamine (acutely treated group, p<0.05; subchronically treated group, p<0.01). These results suggest that the effect of methamphetamine on the immune system may be caused by thymic dysfunction.
Human growth hormone (hGH) forms antibody by repeated administration. The present study investigated to confirm formation of antibody by repeated subcutaneous administration of hGH for two months in rats and dogs. In this result, hGH-injected sera were significantly higher than control sera by 1:1,000,000 of dilution factor. After antibody formed sera (anti-hGH sera) and control sera were added to 30 $\mu\textrm{g}$/ml hGR, the complex incubated for overnight at $30^{\circ}C$. Anti-hGH sera decreased hGH contents about 90% compared to control sera. Also, body weight gain conducted decreased about 67% compared to control sera in hypophysectomised rat. Inconclusion, repeated administration of hGH formed antibody because hGH was foreign protein to rats and dogs. And formed antibody of hGH was blocked and decreased many efficacy of hGH, the antibody was proved to be neutralizing antibody. Thus, because neutralizing antibodies were decreased pharmacological effects of hGH, administration more than two months were no significance.
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