• Title/Summary/Keyword: Antibody specificity

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Bone Marrow Immunoscintigraphy for the Detection of Skeletal Metastasis in Malignant Tumors: A Comparison with $^{99m}Tc$-MDP Bone Scan (악성종양에서 골수면역신티그라피를 이용한 골수전이의 평가 : $^{99m}Tc$-MDP 뼈스캔과의 비교)

  • Lee, Kyung-Han;Choi, Chang-Woon;Bang, Yung-Jue;Chung, Jun-Key;Chung, Hong-Keun;Lee, Myoung-Chul;Kim, Byoung-Kook;Kim, Noe-Kyeong;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.1
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    • pp.89-97
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    • 1994
  • Although bone scan is a highly sensitive test for detecting bone metastasis, its findings are often limited in specificity and cannot be used for assessing the bone marrow. Bone marrow scintigraphy may provide useful information but previous experience with radiolabelled colloid has been disappointing. Recently, $^{99m}Tc$ labeled anti-granulocyte monoclonal antibody (anti-NCA-95 MAb) has been introduced as a new bone marrow imaging agent. To evaluate the usefulness of $^{99m}Tc$ anti-NCA MAb bone marrow scans for detecting skeletal metastasis, bone marrow scans of 44 malignant tumor patients were evaluated and compared with bone scan findings. Bone scan showed abnormal lesions in 26(59%) cases, and 18 of these patients also had an abnormal bone marrow scan. Seven of the 8 patients who had normal bone marrow scan despite bone scan lesions were confirmed to be free from metastasis. There was one case with a marrow defect despite normal bone scan but the presence of metastasis was not determined due to loss of follow up. Bone scan demonstrated a total of 64 lesions while bone marrow scan showed 38 lesions. Fifty percent (32/64) of the bone scan lesions had matching marrow defects while the remaining 50% did not. Most of these non matched lesions were suggested to be nonspecific lesions such as rib fractures or degenerative change. Meanwhile bone marrow scan was able to detect 6 new lesions not detected by bone scan, bit metastasis in each lesion was not confirmed. Bone marrow scan was also helpful in assessing equivocal bone scan lesions to be of metastatic nature in 10 patients by demonstrating a matched marrow defect. Thus $^{99m}Tc$ anti-NCA MAb bone marrow scan can help exclude metastasis in patients with nonspecific bone scan lesions and may be able to detect metastatic lesions not seen with bone scan. It appears useful as a complementary study to bone scan in evaluating malignant tumor patients.

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Development of an Indirect Enzyme-Linked Immunosorbent assay for Rapid Detection of Adulteration of Food Allergen Mackerel in Processed Marine Foods (수산가공식품 중 알레르겐 고등어 혼입여부 신속 검출을 위한 간접효소면역분석법의 개발)

  • Lee, Jeong-Eun;Kim, Ah-Yoon;Kim, Sol-A;Kim, Hyo-In;Park, Ji-Hye;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.185-192
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    • 2018
  • The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ${\geq}0.3$ O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.

Studies on the Rapid Plasma Reagin(RPR) Card Test for the Diagnosis of Syphilis (매독진단(梅毒診斷)을 위(爲)한 신속(迅速)한 혈장항체검사법(血漿抗體檢査法)에 관(關)한 연구(硏究))

  • Kim, Joo-Deuk;Lew, Joon;Kim, Hyun-Joo
    • The Journal of the Korean Society for Microbiology
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    • v.3 no.1
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    • pp.15-23
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    • 1968
  • For the effective control of Syphilis, many investigators have developed a more rapid, simple and economical screening serological test which is adequately sensitive and specific. To fulfill the requirements of a more rapid serologic test for syphlis, a substitute for the conventional serum specimen was needed since considerable time and labor are involved in the processing of blood to serum. Burdon suggested the use of plasma in the serologic tests for syphilis as a substitute for serum. He noticed that plasma was more sensitive than serum in the Kline and Kahn tests, and attributed this to the presence of more antibody-like substance, "reagin" in plasma than in serum. However, to make plasma sufficiently sensitive, it was necessary to inactivate plasma by heating at a temperature of $56^{\circ}C$ for about 30 minutes. Heating of plasma resulted in the precipitation of fibrinogen which made centrifugation necessary to obtain dear plasma. Since the chief disadvantage to the use of unheated plasma(or serum) was a reduction in sensitivity of results-which probably was due to a labile factor such as complement-Portnoy et al began to consider rapid chemical methods of inactivation of plasma(or serum). They experienced that choline chloirde was shown to be anticomplementary which suggested its use as an inactivating agent for unheated plasma(or serum). In 1959 Portnoy et al reported the Rapid Plasma Reagin(RPR) Test for syphilis which is a more rapid, economical and simple. But still this test has many disadvantages as a rapid performing, field and office procedure, because it requires the usual laboratory equipments such as centrifuge, rotating machine, microscope etc. To substitute these disadvantages of the RPR test, in 1962, Portnoy et al developed the Rapid Plasma Reagin(RPR) card test for syphilis, which has the following advantages: a) Simplicity and rapidity of performance, b) Requires no laboratory equipments, c) Stable antigen suspension, d) Adequate sensitivity and specificity. This RPR card test can be used as a rapidly performing and screening test in field investigation, outpatient clinics, small laboratories and hospitals doing limited syphilis serology, and predonor in blood bank. Private clinic which has limited laboratory equipment and technic for syphilis serology can also use this RPR card test as a tool in the rapid diagnosis of syphilis. It was thought that this RPR card test is a useful tool in Korea for private physician and mass survey for syphilis diagnosis. But Portnoy patented the reagents needed for the performing the RPR card test. Therefore authors developed newly the reagents and according to Portnoy's method evaluated the newly developed. RPR card test compared with the VDRL, Kolmer CF, and RPCF tests. The RPR card and VDRL tests were performed plasma and serum from the total 1,132 cases. Among these 1,131 cases, 521 were syphilis suspected laboratory specimens, and 611 were syphilis unsuspected healthy young men. After screening with these two tests, the RPR card and VDRL tests, reactive specimens to the above one or both tests were retested by the Kolmer CF and RPCF tests.

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Immunogenicity of staphylococcal enterotoxin C mutant antigen in mice and dairy cows (포도상구균 장내 C 형 변이독소 (SEC mutant)의 면역원성에 대한 연구)

  • Chang, Byoung-sun;Joo, Yi-seok;Moon, Jin-san;Seo, Keun-seok;Yang, Soo-jin;Kim, So-hyun;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.177-188
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    • 2001
  • Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

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A Clonorchis sinensis-specific antigen that detects active human clonorchiasis (간흡충 현증감염 특이항원)

  • 김석일
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.37-46
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    • 1998
  • A Czonorchis sinensis-specific antigen in excretory-secretory product of C. sinensis (CsE) was assessed in human clonorchiasis by immunoblot. Thirty and 7 kDa antigens of CsE2, one of four different batches of CsEs reacted strongly with infection sera from clonorchiasis patients; however, the antigens reacted weakly with 6-month post- treatment sera from praziquantel-cured cases, but were still highly detected by the sera from praziquantel∼failed patients, indicating that the 30 and 7 kDa antigens can detect antibodies during an active infection. The 30 kDa antigen showed some cross reactions with sera from patients with Pcragonimus westemani and Metcfonimw vokogcujci, while the 7 kDa antigen did not, suggesting that the 7 kDa antigen has high specificity. The 30 kDa antigen reacted with some past clonorchiasis sera, whereas the 7 kDa antigen did not, supporting that antibodies to the 7 kDa antigen are not present in sera from past clonorchiasis patients. In an endemic area, 92% (23/25) of active clonorchiasis patients and 91% (10/11) of mixed infection patients with C. sinensis and M. Wokosawai had IgG antibodies to the 7 kDa antigen, while 40% (6/15) of past clonorchiasis individuals and 43% (3/7) of metagonimiasis patients cross-reacted to the antigen. These data suggest that the 7 kDa antigen in an excretory-secretory antigen may serve as a marker of an active clonorchiasis with reliable specificities in past clonorchiasis, paragonimiasis and metagonimiasis.

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Evaluation of a Serodiagnostic Method for Tuberculosis by Using Secreted Protein Antigens of Mycobacterium Tuberculosis (결핵균 분비항원을 이용한 결핵의 혈청학적 진단 방법에 대한 평가)

  • Bai, Gill-Han;Park, Eun-Mi;Kim, Sang-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.315-323
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    • 2000
  • Background : An immunochromatographic assay (ICT Diagnostics) which facilitates the diagnosis of tuberculosis(TB) by detecting serum antibodies mainly directed against specific 38KDa of Mycobacterium tuberculosis has come into the market. The test consists of a cardboard folding device containing nitrocellulose strip and absorbent pads. The whole procedure is completed within 15 min and does not require any additional equipment. The test has been reported to be sensitive and specific in diagnosing active TB. Thus the test had been evaluated with sera from TB patients and TB-free subjects. Method : Sera from patients with active pulmonary tuberculosis(40 sputum positives for Mycobacterium tuberculosis, 79 sputum negatives, and 3 extrapulmonary tuberculosis) were obtained from the Double-Cross Chest Clinic of the Korean National Tuberculosis Association (KNTA) in Seoul. The control group consisted of TB-free 68 subjects(21 children under 7 years old and 47 healthy staff members of KNTA). Results : Nine out of 68(13.2%) TB-free controls had positive antibody response. Total 106 of 122(86.9%) radiologically active patients had positive antibodies while 16 (13.1%) showed negative reaction. Antibody was detected in 38 of 40(95.0%) sputum positive patients and 68 of 82(82.9%) sputum negative patients who were under the antituberculosis chemotherapy. The sensitivity and specificity were all 87% and the positive predictive value was 92.2% while the negative predictive value was 78.7%, when the prevalence of TB in the sample was 64.2%. Our results clearly show that the detection of antibodies which mainly react with the 38KDa antigen of M. tuberculosis is not suitable as the first-line method of diagnosis but considered only as an adjunctive test to standard techniques of tuberculosis diagnosis. when considering its high false positivity.

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Diagnostic Value of Serum CYFRA 21-1 in Lung Cancer (폐암에서 혈중 CYFRA 21-1의 진단적 가치)

  • Yoon, Hyun-Dae;Kim, Ki-Deok;Chung, Jin-Hong;Lee, Hyung-Woo;Lee, Kwan-Ho;Lee, Hyun-Woo;Cho, Ihn-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.2
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    • pp.149-155
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    • 1995
  • Background: Cytokeratin 19 is 40KD acidic molecule whose distribution is restricted to simple or pseudo-stratified epithelia, such as the epithelial layer of the bronchial tree. Immunohistochemical study have shown that cytokeratin 19 is overexpressed in lung carcinoma tissue. An immunoradiometric assay, CYFRA 21-1 has been developed using two monoclonal antibody, BM 19-21 and KS 19-1, reactive to different epitopes on cytokeratin 19. We studied the diagnostic value of CYFRA 21-1 in lung cancer. Method: The serum CYFRA 21-1 level using immunoradiometric kit(ELSA-CYFRA 21-1) was measured in 54 patients who admit to Yeungnam University Hospital from April, 1993 to August, 1994. Lung cancer group was 39 primary lung cancer patients(19 patients with squamous cell carcinoma, 11 patients with adenocarcinoma and 9 patients with small cell carcinoma). Control group was 15 patients with non malignant lung diseases(8 patients with pulmonary tuberculosis, 3 patients with chronic obstructive pulmonary disease, 2 patients with pneumonia and 2 patients with chronic obstructive pulmonary disease combined with pulmonary tuberculosis). Results: The mean serum value of CYFRA 21-1 was $20.2{\pm}4.7ng/ml$ in squamous cell carcinoma, $7.2{\pm}1.6ng/ml$ in adenocarcinoma and $15.5{\pm}4.7ng/ml$ in non-small cell lung cancer. The serum value of CYFRA 21-1 in control group was $1.7{\pm}0.5ng/ml$. All of the serum values of 3 histologic types were significantly higher than that of control group(p<0.01). The serum value of CYFRA 21-1 of squamous cell carcinoma was significantly higher than that of adenocarcinoma(p <0.05). Serum value of CYFRA 21-1 in small cell lung cancer was $2.9{\pm}0.9ng/ml$ and not significantly different compared with control group. Using cut off value of 3.3ng/ml, sensitivity and specificity was 11.1%, 65.2% in small cell lung cancer, 70.0%, 62.5% in non-small cell lung cancer, 73.7%, 75% in squamous cell carcinoma and 63.6%, 78.9% in adenocarcinoma, respectively. Conclusion: The serum levels of CYFRA 21-1 may be useful in diagnosis of non-small cell lung carcinoma, especially in squamous cell carcinoma with its high specificity.

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Measurement of the Levels of IgG Subclasses Reactive to Salmonella typhi in the Sera of Patients with Typhoid Fever (장티푸스환자의 혈청내 Salmonella typhi에 대한 IgG subclass항체의 분포)

  • Kim, Young-Jung;Hwang, Eung-Soo;Kang, Jae-Seung;Cha, Chang-Yong;Chang, Woo-Hyun;Kim, Yoon-Won;Cho, Min-Ki;Min, Chang-Hong
    • 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.

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Overexpression and Activity Analysis of Cystathionine γ-Lyase Responsible for the Biogenesis of H2S Neurotransmitter (새로운 신경전달물질 H2S 발생 효소, cystathionine γ-lyase의 대량발현 조건과 활성측정)

  • Kim, Kyoung-Ran;Byun, Hae-Jung;Cho, Hyun-Nam;Kim, Jung-Hyun;Yang, Seun-Ah;Jhee, Kwang-Hwan
    • Journal of Life Science
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    • v.21 no.1
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    • pp.119-126
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    • 2011
  • There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

Cross-reaction of 6B and 19F Specific Antibodies to Serotypes 6A, 6C, and 19A after Immunization with 7-valent Pneumococcal Conjugate Vaccine in Korean Children Aged 12-23 Months (한국 12-23개월 소아에서 7가 폐구균 단백결합 백신 추가접종으로 유도된 6B와 19F 혈청형 특이 방어항체의 교차혈청형 6A, 6C, 19A에 대한 교차 반응)

  • Kim, Kyung-Hyo;Yang, Joo Yun;Park, In Ho;Lim, Soo Young
    • Pediatric Infection and Vaccine
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    • v.20 no.2
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    • pp.53-62
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    • 2013
  • Purpose: The cross-protection of 7-valent pneumococcal conjugate vaccine (PCV7) against vaccine-related serotypes has been controversial. We investigated the serological properties of cross-protective antibodies against vaccine-related serotypes 6A, 6C, and 19A induced in young children aged 12-23 months after booster immunization of PCV7. Methods: IgG and IgM antibody concentrations and opsonic index (OI) against vaccine serotypes 6B and 19F and vaccine-related serotypes 6A, 6C, and 19A were measured by ELISA and opsonophagocytic killing assay (OPA) in 4 selected immunesera. The serological properties and antigenic specificity of protective antibodies were determined by IgM depletion of immunesera, OPA, and competitive OPA against serogroup 6 and 19 pneumococci. Results: Compared to pre-IgM depleted immunesera, OI of IgM-depleted immunesera against 6B and 19F decreased and OI against 6A, 6C, and 19A decreased, too. In competition OPA, free 6B and 19F polysaccharide completely inhibited the immune protection against vaccine-related serotypes 6A, 6C, and 19A as well as vaccine types 6B and 19F. Conclusions: The booster immunization of PCV7 certainly induced cross-protective antibodies against vaccine-related serotypes 6A, 6C, and 19A with both IgG and IgM isotypes. Furthermore, IgM antibodies are more highly contributed to opsonophagocytic activity against vaccine-related serotypes as well as most of vaccine types than do IgG antibodies. Further studies are needed for the more immunized sera in the children as well as adults.

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