Immunogenicity Monitoring Cell Chip Incorporating Finger-Actuated Microfluidic and Colorimetric Paper-Based Analytical Functions

이경원,Yang Eun Kyeong,Oh Yujeong,박은혜,정관영,Yoon Hyun C. 한국바이오칩학회 2023 BioChip Journal Vol.17 No.3

Recently, with the development of microfl uidic chips, attempts to integrate cell culture and biomedical material testing functions into a single chip have increased to supplement experimental animal models. Among the evaluations of biomaterials, immunogenicity testing, a current priority, is attracting attention. In this study, we developed a simple and easy-to-handle immunogenicity-testing cell chip to evaluate the immunogenicity of biomaterials. On this chip, macrophages were introduced as immunogenicity indicators, and a micro-paper-based analytical device (μPAD) was used for optical analysis. Macrophages are present in all parts of the body and mediate immune reactions against body implants or bio-derived substances, resulting in the production of hydrogen peroxide. In the cell chamber of the developed cell chip, macrophages grow and react to immunogenic materials. Activated macrophages secrete hydrogen peroxide, which is then transferred to the PAD with single- fi nger actuation. The hydrogen peroxide molecules reaching the PAD detection zone react with the colorimetric detection substrate, resulting in a color that corresponds to the hydrogen peroxide concentration. With the developed testing chip, the immunogenicity of biomaterials can be determined before administration.

Immunogenicity and Safety of Diphtheria-tetanus Vaccine in Adults

최정현,추은주,허애정,최수미,엄중식,이진서,박선희,강진한 대한의학회 2010 Journal of Korean medical science Vol.25 No.12

This study was conducted to evaluate the immunogenicity and safety of diphtheria–tetanus (Td) vaccine in adults over 40 yr old who had never received a diphtheria-tetanuspertussis (DTP) vaccination. A total of 242 subject completed three-doses of Td vaccination and subsequent assays for immunogenicity. Before vaccination, 33.9% and 96.7%participants showed antibody levels of diphtheria and tetanus, respectively, which were below protective level (<0.1 U/mL). After the first dose of Td vaccine, 92.6% and 77.6%of subjects gained protective antibody concentrations (≥0.1 U/mL) for diphtheria and tetanus, with an increase to 99.6% and 100% after the third dose. Local and systemic adverse events occurred in 37.9% and 15.5% of the subjects. No serious adverse event requiring an unscheduled hospital visit occurred. In conclusion, three-doses of Td vaccination to unimmunized adults are safe and effective in inducing protective immunity against diphtheria and tetanus.

Effect of Smoking on Influenza Illness and Vaccineinduced Immune Response in Mice

이진수,기세윤,황인숙,박신구,김루시아,정문현,김우주,정희진 대한감염학회 2010 Infection and Chemotherapy Vol.42 No.6

Background: Since cigarette smoking is an important risk factor for respiratory infections and alters the immune response, the severity of influenza illness and the immunogenicity of influenza vaccination may differ between cigarette smokers and non-smokers. This study investigated the effect of cigarette smoke exposure on the severity of influenza illness and vaccine-induced antibody production in mice. Materials and Methods: Cigarette smoke exposed (CE) and non-cigarette smoke exposed (NCE) mice were infected with mouse-adapted influenza A/PR8/8/34(H1N1). Influenza virus was quantified in bronchoalveolar lavage fluid by real-time polymerase chain reaction and the lung pathology was examined to investigate the influence of smoke exposure on the severity of illness. To assess immunogenicity,hemagglutination inhibition antibodies were measured in pre- and post-influenza vaccination blood samples from CE and NCE mice. Results: Influenza viral proliferation was higher and inflammatory changes such as macrophage infiltration in the alveolar space and necrotizing bronchitis were more pronounced in CE mice, compared with controls. Vaccine-induced immunogenicity was achieved in both CE and NCE mice. Conclusions: Cigarette smoke exposure enhanced influenza viral replication and the inflammatory changes associated with influenza illness, but had no significant effect on vaccine-induced immunogenicity.

유전자 재조합 B형간염 백신의 10세이하 소아에서의 면역원성 및 안전성

김명아,최은하,장미숙,동은실,장성희,안영민,윤희상,손영모,Kim, Myoung Ah,Choi, Eun Ha,Jang, Mee Suk,Dong, Eun Sil,Jang, Seong Hee,Ahn, Young Min,Youn, Hee Sang,Sohn, Young Mo 대한소아감염학회 1997 Pediatric Infection and Vaccine Vol.4 No.1

목 적 : B형 간염 바이러스 보균자가 많은 우리 나라에서는 모든 신생아에게 B형 간염에 대한 능동면역이 시행되고 있다. 혈장 백신은 공급원에 한계가 있고 전염성 질환이 전파될 가능성이 있으며 값이 비싸다는 문제점이 있어 이와는 다른 유전자 재조합 공법으로 생산된 HG-II$^{(R)}$백신의 면역원성과 안전성을 조사하고 BCG 선행군과 그렇지 않은 군에서의 면역원성을 비교하고자 본 연구를 시행하였다. 방 법 : 1995년 4월부터 1996년 6월까지 경상대학병원(Group A), 지방공사 강남병원 (Group B)과 영동 세브란스병원(Group C)에서 10세이하의 소아를 대상으로 하여 유전자 재조합 B형간염 백신(HG-II$^{(R)}$)$10{\mu}g$의 양을 0, 1, 6개월 접종 방식으로 3회 근주 하였다. 4군는 BCG를 간염 2차 접종하기 1주전에, B군은 1주후에 접종하였다. C군에서는 두가지 예방접종의 선후관계를 알 수 없었다. 3회 접종후 1개월에 채혈하여 anti-HBs Ab를 측정하였으며 부작용을 조사하였다. 결 과 : 1) 총 114례중 신생아는 104례였으며 이 중 55례는 간염 2차 접종하기 1주전에 BCG를 접종하였고 43례는 간염 2차 접종후에 BCG를 접종하였으며, 6례는 선후관계를 알 수 없었다. 2) 항체양전율은 99.1%였고 기하 평균 항체가는 131.2mIU/ml였다. 3) BCG를 간염 2차 접종하기 1주전에 접종한 군과 1주후에 접종한 군에서의 기하 평균 항체 가는 각각 105.5mIU/ml, 162.8mIU/ml였다(p<0.025). 4) 부작용은 국소 반응이 1.4%, 전신 반응이 7.8%였다. 결 론 : 유전자 재조합 간염 백신은 부작용이 적고 면역원성이 우수하였다. 간염 2차 접종 전에 BCG를 접종한 군에서의 기하 평균 항체가가 낮게 관찰된 바 향후 이에 대한 연구가 더 필요할 것으로 사료된다. Objective : To evaluate the immunogenicity and safety afforded by the HG-II$^{(R)}$ recombinant hepatitis B vaccine given to healthy neonates and children and to find the influence of preceding BCG vaccination on immunogenicity. Methods : Three doses of recombinant hepatitis B vaccine with a dose of $10{\mu}g$ were given at birth, 1 and 6 months of age. This study was conducted in three hospitals (Gyeongsang National University Hospital(Group A), Kangnam General Hospital(Group B) and Younsei University Hospital(Group C)) from April, 1995 to June, 1996. Group A and Group B received 2nd dose of hepatitis B vaccine at 1 week after and before BCG vaccine, respectively. Antibidy levels, at 1 month after the 3rd dose of hepatitis B vaccine were determined by a radioimmunoassay. Results : 1) One hundred four infants and ten children were enrolled : 55 infants and 43 infants received 2nd dose of hepatitis B vaccine at 1 week after( After BCG Group) and before BCG vaccine(Before BCG Group), respectively. 2) The seropositive rate was 99.1%, and geometric mean anti-HBs titer was 131.2mIU/ml. 3) The geometric mean titers were 105.5mIU/ml and 162.8mIU/ml in After BCG and Before BCG Group, respectively(p<0.025). 4) Among 359 episodes of vaccination, the occurrence of systemic and local side reaction were reported in 7.8% and 1.4%, respectively. Conclusion : Recombinant hepatitis B vaccine(HG-II$^{(R)}$))was highly immunogenic and safe. The significantly lower geometric mean antibody titer in the BCG preceding group was observed. Well-designed controlled study with the large number of sample size will be required to show the influence of preceding BCG vaccination.

건강한 한국 소아에서 HM175주 A형 간염 불활화 백신의 면역원성 및 이상반응에 관한 연구

김창휘,편복양,홍영진,강진한,Kim, Chang Hwi,Pyun, Bok Yang,Hong, Young Jin,Kang, Jin Han 대한소아감염학회 2000 Pediatric Infection and Vaccine Vol.7 No.1

목 적 : 환경적 변화에 잘 적응되는 특성을 지닌 A형 간염 바이러스는 환경과 위생이 개선된 현재에도 전 세계적으로 연간 140만명 정도가 발생되고 있는 중요한 전염성 감염 질환으로서 국내에서도 A형 간염 항체 보유율이 역학적 변화 상태에서 1996년 이후부터 10세에서 20세 사이의 연령에서 발생이 현저히 상승되는 양상을 보이고 있다. 국내에서 1996년부터 A형 간염 발생의 장기적 조절을 위하여 A형 간염 백신이 선별접종으로서 활용되고 있는 상황에서 저자들은 1세에서 15세 사이의 건강한 소아를 대상으로 A형 간염 백신에 대하여 면역원성 및 이상반응에 관한 기본적 임상연구를 실시하게 되었다. 대상 및 방법 : 본 연구는 1999년 2월부터 1999년 3월까지 가톨릭대학 성모자애병원, 인하대학병원, 순천향대학병원을 방문한 1세에서 15세 사이의 기저질환이 없으며 과거력상 A형 간염의 기왕력이나 A형 간염백신의 접종력이 없고 진찰상 건강한 소아들을 대상으로 국내에서 1997년부터 사용되고 있는 HM175주 A형 간염 백신(Havrix)을 1회 기초 접종하고 6개월 후에 추가 접종하여 접종때마다 단기간 내의 국소 및 전신적 이상반응을 확인하고 각 접종 1개월 후에 채혈 분리된 혈청에서 A형 간염 항체를 ELISA법으로 측정하여 면역원성을 평가하였다. 결 과 : 128명(남아 65명, 여아 63명; 평균 연령, 6.0세)이 1차 접종시부터 면역원성 및 이상반응에 대한 연구 대상아로 참여되었고 120명(남아 60명, 여아 60명; 평균 연령 6.0세)이 최종 연구기간까지 참여하였다. 면역원성 연구 결과 일차 접종 1개월 후 A형 간염 항체가 전체에서 양성으로 전환되었으며 평균 농도는 $389.2{\pm}392.7mIU/mL$, 기하평균치(GMT)는 266.4이었다. 추가 접종 1개월 후 면역혈청검사에서 역시 대상아 전체에서 양성을 보였으며 평균 농도가 $3,709{\pm}3,270mIU/mL$, 기하평균치가 2,502로 일차 접종 때보다 상승되었다. 한편 이차접종 1개월 후 면역검사에서 영아의 평균 A형 간염항체가는 남아의 평균 농도보다 유의하게(P=0.0017) 높았다. 이상반응 평가에서는 주사부위의 동통, 발적, 부종, 소양감, 발열감의 순으로 국소적 이상반응이 나타났고 권태감, 식욕부진, 두통, 발열, 오심의 순으로 이상반응이 일차 접종 후에 발현되었고 이차 접종 후에 발열은 전례에서 관찰되지 않았으며 권태감, 두통, 식욕부진, 오심, 구토, 발진이 관찰되었다. 이러한 이상반응들은 모두 3일 이내에 특별한 조처없이 소실되었다. 결 론 : 건강한 소아를 대상으로 국내에서 사용되고 있는 HM175주 A형 간염 백신에 대한 면역원성 및 이상반응에 관한 연구를 실시한 결과 1회 기초 접종 후 전례에서 방어항체가로 양전되고, 추가접종 후 방어 항체가가 매우 유의하게 상승되고 면역원성을 확인하였고, 국소 및 전신 이상반응은 모두 경미한 정도로 별다른 조치 없이 3일 내에 소실되어 안전한 백신임을 알 수 있었다. Purpose : Active immunization against hepatitis A with an inactivated vaccine reveals excellent immunogenicity, tolerability and protective efficacy. Inactivated hepatitis A vaccines have been selectively used since 1996 in Korea to prevent hepatitis A. This study was performed to assess the immunogenicity and reactogenicity after two doses of HM175 strain hepatitis A vaccine in healthy Korean children. Methods : 128 healthy children(M/F; 65/63) aged 1 to 15 years, who were seronegative for hepaitatis A, participated in this study. A alum-adsorbed vaccine containing 720 EL.U of antigen form HM175 hepatitis A strain per 0.5 mL dose was injected intramuscularly on the deltoid area. The second dose was given 6 months later, Anti-HAV antibodies were measured by ELISA before and 1 month after each vaccination to assess the immunogenicity. Any local and general adverse events were reported by patients parents with the prepared questionnaire after each vaccination. Results : 120 volunteers(M/F; 60/60) completed the whole series of the study. Seroconversion occurred in all cases after primary and booster vaccination. The mean anti-HAV antibody titer after primary vaccination was 389.2mIU/mL, and 3,609mIU/mL after booster vaccination. And levels of anti-HAV antibodies after booster immunization were significantly higher in female children. The most common local adverse event was soreness on the injection site, but it was mild and resolves within 3 days. Fever was not reported after booster vaccination. Conclusion : Based on these data, we conclude that the inactivated HM175 strain hepatitis A vaccine is highly immunogenic and tolerable in healthy Korean children.

Genipin-Crosslinked, Immunogen-Reduced Decellularized Porcine Liver Scaffold for Bioengineered Hepatic Tissue

Xiujuan Wu,Yujia Wang,Qiong Wu,Yi Li,Li Li,Jing Tang,Yujun Shi,Hong Bu,Ji Bao,Mingjun Xie 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.6

Liver disease affects millions of patients each year worldwide. Decellularized biologic matrices are plausible biomedical materials for bioengineered replacement hepatic tissue. However, one of the concerns for its safe medical application is the lack of objective assessment of the immunogen within the materials and in vivo immune responses to the matrices. The purpose of this study was to produce immunogen- reduced and biocompatible matrices from porcine liver. Whole porcine livers were perfusion decellularized and cross-linked with glutaraldehyde (GA) or genipin (GP). Proteins were extracted, and the migratory response of human leukocytes toward protein extracts was examined using an in vitro migration chamber. In addition, biopsy specimens of decellularized scaffolds were implanted subcutaneously into rodents to investigate scaffold immunogenicity. Histological staining confirmed cellular clearance from pig livers, with removal of nuclei and cytoskeletal components and widespread preservation of structural extracellular molecules. Polymerase chain reaction analysis showed that galactose-alpha-1,3-galactose-beta-1,4-N-acetylglucosamine (1,3 gal), swine leukocyte antigen, and porcine endogenous retrovirus were completely removed in the matrices. Decellularization significantly reduced the migration of monocytes compared with native porcine tissue. Although the proportion of transmigrating lymphocytes was much lower, repeating the cross-linking procedure reduced the migratory response. After implantation for 4 weeks, the decellularized and native samples were degraded, and the GA-treated group demonstrated a severe inflammatory reaction; however, minimal inflammatory cell infiltration was seen in the GPtreated group during the 8-week investigation period. In conclusion, our study provided evidence that GP crosslinking could significantly reduce the immunogenicity of decellularized liver biomaterials.

Evaluation of Immunogenicity and Safety of the New Tetanus-Reduced Diphtheria (Td) Vaccines (GC1107) in Healthy Korean Adolescents: A Phase II, Double-Blind, Randomized, Multicenter Clinical Trial

임정우,김황민,이경일,김상용,김종현,김현희,최영윤,마상혁,김동호,안동호,강진한 대한의학회 2013 Journal of Korean medical science Vol.28 No.4

This phase II clinical trial was conducted to compare the immunogenicity and safety of a newly developed tetanus-reduced diphtheria (Td) vaccine (GC1107-T5.0 and GC1107-T7.5)and control vaccine. This study was also performed to select the proper dose of tetanus toxoid in the new Td vaccines. Healthy adolescents aged between 11 and 12 yr participated in this study. A total of 130 subjects (44 GC1107-T5.0, 42 GC1107-T7.5 and 44 control vaccine) completed a single dose of vaccination. Blood samples were collected from the subjects before and 4 weeks after the vaccination. In this study, all subjects (100%) in both GC1107-T5.0 and GC1107-T7.5 groups showed seroprotective antibody levels ( ≥ 0.1 U/mL)against diphtheria or tetanus toxoids. After the vaccination, the geometric mean titer (GMT) against diphtheria was significantly higher in Group GC1107-T5.0 (6.53) and GC1107-T7.5 (6.11) than in the control group (3.96). The GMT against tetanus was 18.6 in Group GC1107-T5.0, 19.94 in GC1107-T7.5 and 19.01 in the control group after the vaccination. In this study, the rates of local adverse reactions were 67.3% and 59.1% in GC1107-T5.0 and GC1107-7.5, respectively. No significant differences in the number of adverse reactions, prevalence and degree of severity of the solicited and unsolicited adverse reactions were observed among the three groups. Thus, both newly developed Td vaccines appear to be safe and show good immunogenicity. GC1107-T5.0, which contains relatively small amounts of tetanus toxoid, has been selected for a phase III clinical trial.

Evaluation Methods for the Immunogenicity of Varicella and Zoster Vaccines

Park, Hosun 대한미생물학회 2013 Journal of Bacteriology and Virology Vol.43 No.4

Varicella vaccine has been included in the national immunization program for children since 2005 and zoster vaccine has been released since 2012 in Korea. Even though both varicella and zoster are caused by varicella-zoster virus (VZV), pathogeneses are different. In varicella, neutralizing antibody is very important to protect disease because VZV spreads via blood or lymph. In contrast, cell-mediated immunity is more important in zoster because of the neuronal spread of VZV. Therefore, the measurement methods of the immunogenicity against varicella and zoster vaccines are different. Fluorescent antibody to membrane antigen (FAMA) assay is the gold standard method to detect the protective antibody against VZV. It is still used as a reference test for the other methods. However, the fastidious nature required to perform the FAMA assay limits its use as a routine assay for the evaluation of vaccine immunogenicity. Nowadays, glycoprotein ELISA (gpEIA) is used as an alternative method for FAMA assay. However, there is no agreement over the protective level of gpEIA antibody titer with WHO standard international unit. The immunogenicity of zoster vaccine has been evaluated by responder cell frequency assay and IFN-${\gamma}$ ELISpot assay. Nevertheless, skin test is considered to be a more accurate biomarker for cell-mediated immunity against zoster. For the evaluation of varicella vaccine, it is necessary to standardize the FAMA assay and to set the cut-off value for the gpEIA antibody titer through long-term follow-up study. For zoster vaccine, the evaluation of cell-mediated immunity in Korean adults is urgently needed.

Evaluation Methods for the Immunogenicity of Varicella and Zoster Vaccines

박호선 대한미생물학회 2013 Journal of Bacteriology and Virology Vol.43 No.4

Varicella vaccine has been included in the national immunization program for children since 2005 and zoster vaccine has been released since 2012 in Korea. Even though both varicella and zoster are caused by varicella-zoster virus (VZV), pathogeneses are different. In varicella, neutralizing antibody is very important to protect disease because VZV spreads via blood or lymph. In contrast, cell-mediated immunity is more important in zoster because of the neuronal spread of VZV. Therefore, the measurement methods of the immunogenicity against varicella and zoster vaccines are different. Fluorescent antibody to membrane antigen (FAMA) assay is the gold standard method to detect the protective antibody against VZV. It is still used as a reference test for the other methods. However, the fastidious nature required to perform the FAMA assay limits its use as a routine assay for the evaluation of vaccine immunogenicity. Nowadays, glycoprotein ELISA (gpEIA) is used as an alternative method for FAMA assay. However, there is no agreement over the protective level of gpEIA antibody titer with WHO standard international unit. The immunogenicity of zoster vaccine has been evaluated by responder cell frequency assay and IFN-γ ELISpot assay. Nevertheless, skin test is considered to be a more accurate biomarker for cell-mediated immunity against zoster. For the evaluation of varicella vaccine, it is necessary to standardize the FAMA assay and to set the cut-off value for the gpEIA antibody titer through long-term follow-up study. For zoster vaccine, the evaluation of cell-mediated immunity in Korean adults is urgently needed.

PEGylation of bacterial cocaine esterase for protection against protease digestion and immunogenicity

Park, J.B.,Kwon, Y.M.,Lee, T.Y.,Brim, R.,Ko, M.C.,Sunahara, R.K.,Woods, J.H.,Yang, V.C. Elsevier Science Publishers 2010 Journal of controlled release Vol.142 No.2

Enhancing cocaine metabolism by administration of cocaine esterase (CocE) has been considered as a promising treatment strategy for cocaine overdose and addiction, as CocE is the most efficient native enzyme yet identified for metabolizing the naturally occurring cocaine. A major obstacle to the clinical application of CocE, however, lies in its thermo-instability, rapid degradation by circulating proteases, and potential immunogenicity. PEGylation, namely by modifying a protein or peptide compound via attachment of polyethylene glycol (PEG) chains, has been proven to overcome such problems and was therefore exploited in this CocE investigation. The PEG-CocE conjugates prepared in this study showed a purity of greater than 93.5%. Attachment of PEG to CocE apparently inhibited the binding of anti-CocE antibodies to the conjugate, as demonstrated by the enzyme-linked immunosorbent assay (ELISA) assay. In addition, PEGylation yielded protection to CocE against thermal degradation and protease digestion. Furthermore, preliminary in vivo results suggested that, similarly to native CocE, the PEG-CocE conjugates were able to protect animals from cocaine-induced toxic effects. Overall, this study provides evidence that the PEGylation may serve as a tool to prolong CocE functionality in the circulation and reduce its potential immunogenicity.