• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.1
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• pp.295-301
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• 2017

Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

• Parasites, Hosts and Diseases
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• v.27 no.2
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• pp.79-86
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• 1989

A pathogenic free-living amoeba, Naegleria fowleri, causes primary amoebic meningoencephalitis to human and experimental animals. This infection is rare, but the mortality is very high. Nowadays, drug treatment or active immunization of human or mice are being tried with partial effectiveness. This study shows passive immunization effect by transfer of immunized spleen cells, serum, or milk from immunized mother in mouse experimental model. Young BALB/c mice were immunized intraperitoneally with $2~3{\times}10^{6}$ trophozoites of N. fowleri, and spleen cells and sera were collected for injection to recipient mice. There were seven transfer groups, i.e., immunized mouse serum, spleen cells, serum and spleen cells, normal mouse serum, spleen cells, serum and spleen cells, and control group. Three days later, BALB/c mice were inoculated with $1{\times}10^{4}$ trophozoites of N. fowleri intranasally. After infection, decreased mortality ana prolonged survival time of mice were noted in immunized Bloops compared with non.immuniBed control group. The groups Injected with immunized spleen cells or normal serum shewed lower moltality than that of controls bult showed no changes of Serum IgG level. The groups injected with immunized serum or normal spleen cells showed increased serum IgG level after immunization but hundred percent mortality was observed. Mother mice were ifnfnunised increperitqneeliy with $2~3{\times}10^{6}$ trephozoites of N. fowleri at the end of pregnancy and weaning Period. Soon after the delivery, Jitters born of non-immunszed mother were matched with immunized mother for feeding immune milk. After three weeks, the litters were infected with $1{\times}10^{4}$ trophozeites of N. fowleri or sacrificed for serum collection to measure the IgG levels. The results show that anti-JV. fowleri IgG from mother was transferred to litter through milk but this IgG did not inauence the mortality or survival time of the infected mice.

• Archives of Pharmacal Research
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• v.14 no.3
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• pp.217-224
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• 1991

One of water and/or methanol extracts from 14 herbal deugs which were screened using murine splenocytes showed immunosuppressive activities previously. After water extract from Xanthium strumarium was treated with chloroform. $100 \mu{g/ml}$ of water layer (XS-WCI) has very strong immunosimulating activities tested by $^3H$-thmidine incorporation (control as $100 \mu{g/ml}$, 26345 cpm was 69515 cpm). MLR also appears to be simulated strongly (control vs $100 \mu{g/ml}$, 4962 cpm vs 78688 cpm). When $100 \mu{g/ml}$ of XS-WCI and $0.8 \mu{g/ml}$ of concanavalin a (ConA) were added. more $^3H$-thymidine were incorporated significantly, compared with $0.8 \mu{g/ml}$ of ConA only. In contrast with ConA. results from $5 \mu{/ml}$ of lipopolysaccharide (LPS) and $100 \mu{g/ml}$ of XS-WCI were not different. compared with $5\mu{/ml}$of LPS only. These results indicated the responses of XS-WCI to B cell and T cell may be different. XS-WCI was injected intraperitoneally (10 mg/kg. 50mg/kg/ 100 mg/kg) for 4 days or 10 days and tested secretion of IgM or IgG by direct and indirect hemolytic plaque-forming cell assays, respectively. Numbers of hemolytic plaques for both IgM and IgG were increased significantly. Especially, secretion of IgGs was increased more than 10 times. After administration of XS-WCI for 7 days (50 mg/kg. 100 mg/kg) splenomegaly deu to graft vs host reaction was observed. Human lymhocytes separated from whole blood by Ficoll-Hypaque method were also proliferated after treatment of $10 \mu{g/ml}$ and $50 \mu{g/ml}$ of XS-WCI. As seen in murine lymphocytes, human lymphocyte proliferation was increased synergistically after treatment with both of XS-WCI and phytohemagglutinin (PHA). It appears that XS-WCI may have potential immunosimulating activities and that it remains to be purified further for isolation of active components.

• Biomolecules & Therapeutics
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• v.7 no.1
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• pp.14-21
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• 1999

The immunogenicity of the recombinant human basic fibroblast growth factor (rh-bFGF) was investigated by tests for active systemic anaphylaxis (ASA), passive cutaneous anaphylaxis (PCA), passive hemagglutination (PHA) and guinea pig maximization test (GPMT) in mice or guinea pigs. Guinea pigs were sensitized with rh-bFGF ($100-1000\;\mu\textrm{g}/kg$) or rh-bFGF-CFA mixture ($1000\;\mu\textrm{g}/kg$). All animals sensitized with rh-bFGF alone or mixture with CFA showed symptoms of anaphylactic shock. IgE antibodies to rh-bFGF were detected in sera obtained from rh-bFGF and rh-bFGF-Alum ($1000\;\mu\textrm{g}/kg$) sensitized mice, indicating that rh-bFGF has immunogenicity eliciting potential. IgG and/or IgM antibodies to rh-bFGF were also detected in all the sera obtained from sensitized mice by PHA. In the GPMT for delayed type skin reaction, no skin reaction was observed in sensitized guinea pigs after intradermal injection and dermal application of 0.01% rh- bFGF. However, these positive reactions were consistent with the results of another rh-bFGF, showing that rh- bFGF is a heterogenous protein to rodents. Considering the fact that rh-bFGF is a genuine human protein of which structure is identical to the endogenous human bFGF, it is thought that rh-bFGF is rarely associated with immunological problems in clinical use.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.447-453
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• 1986

To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

• 한국생활과학회:학술대회논문집
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• 2009.05a
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• pp.165-166
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• 2009

• 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

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.177-184
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• 2006

This study was performed to evaluate the prevalence of high risk Human papilloma virus (HPV), Herpes simplex virus type 1, 2 (HSV-1,2), Hepatitis B virus (HBV) and Human Immuno deficiency virus (HIV) infection with sexual transmitted viral diseases in Busan during 2004 to 2005. six hundred seventy four samples of cervical swabs were tested for sexually transmitted viral diseases. Among the isolated viruses, 23 (3.4%) samples were HPV and 3 (0.4%) and 9 (1.3%) samples were HSV 1 and 2, respectively. Among the 586 serum samples tested for viruses, HSV IgM 121 (3.6%), HSV-1 IgG 487 (83.1%), HSV-2 IgG 135 (23.0%), HBsAg 26 (4.4%), HBeAg 7 (1.2%), and HIV (0%) types were found. HPV genotypes were detected in 16 patients, of which 13 cases were high risk type HPV, 3 cases were low risk type HPV, and multi infection were detected in 7 cases. In the age distribution of the patients, 7.2% of infection tested from cervical swabs occurred in under the age of 20, while 100% of infection was found to occur in those who were 40 years old or older in the serum samples. The outbreak pattern in their occupations was found to be the highest at the health organization (amusement quarter) for the cervical swabs, and at infirmary (commercial sex worker) for the serum samples, respectively.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.441-446
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• 1986

Previous studies have demonstrated that phagocytosis of encapsulated bacteria needs the opsonization of bacteria with immunoglobulin and complement. Several investigators have studied the role of specific antibody to the bacteria. The purpose of this study is to investigate the role of specific anti-Actinobacillus actinomycetemcomitans Y4($A{\alpha}Y4$) antibody, which was obtained from the immunized rabbit serum for phagocytosis of $A{\alpha}Y4$ by PMNL. For this study, specific and nonspecific IgG were separated from the sera of the rabbits and PMNL were isolated from 15 healthy adults. By an enzyme-linked immunosorbent assay, the results showed that the binding capacity of anti-$A{\alpha}Y4$ IgG to $A{\alpha}Y4$ was much higher than that of nonspecific IgG; 0.75 and 0.14(O.D. at 400nm), respectively. The oxygen consumption of PMNL, phagocytizing $A{\alpha}Y4$ which was opsonized with specific $A{\alpha}Y4$ IgG(37.13 nmol/min/$1{\times}10^7$ PMNL), was significantly higher than that with nonspecific IgG(27.95 nmol/min/$1{\times}10^7$ PMNL, p<0.01). In immunofluorescence microscopic examination, the difference between the numbers of the ingested $A{\alpha}Y4$ opsonized with specific anti-$A{\alpha}Y4$ IgG and nonspecific IgG reached to statistically significant level; $184{\pm}11.4$ and $133.2{\pm}8.3$ per 100 PMNL, p<0.05. These results suggest that specific anti-$A{\alpha}Y4$ IgG has a significant role in PMNL phagocytosis of encapsulated $A{\alpha}Y4$ and also it can be available to adopt this system to develop anti-capsular antibody to $A{\alpha}Y4$ for enhancing and emphasizing the phagocytic activity against this bacterium.

• Parasites, Hosts and Diseases
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• v.24 no.1
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• pp.25-41
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• 1986

The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.