• Parasites, Hosts and Diseases
• /
• 제52권3호
• /
• pp.251-256
• /
• 2014

A novel recombinant Bacille Calmette-Guerin (rBCG) vaccine co-expressed Eimeria tenella rhomboid and cytokine chicken IL-2 (chIL-2) was constructed, and its efficacy against E. tenella challenge was observed. The rhomboid gene of E. tenella and chIL-2 gene were subcloned into integrative expression vector pMV361, producing vaccines rBCG pMV361-rho and pMV361-rho-IL2. Animal experiment via intranasal and subcutaneous route in chickens was carried out to evaluate the immune efficacy of the vaccines. The results indicated that these rBCG vaccines could obviously alleviate cacal lesions and oocyst output. Intranasal immunization with pMV361-rho and pMV361-rho-IL2 elicited better protective immunity against E. tenella than subcutaneous immunization. Splenocytes from chickens immunized with either rBCG pMV361-rho and pMV361-rho-IL2 had increased $CD4^+$ and $CD8^+$ cell production. Our data indicate recombinant BCG is able to impart partial protection against E. tenella challenge and co-expression of cytokine with antigen was an effective strategy to improve vaccine immunity.

• 한국어병학회지
• /
• 제24권2호
• /
• pp.153-160
• /
• 2011

본 구에서는 백신을 넙치 양식 현장에 적용하기 위한 기초연구로서 항원의 투여방법 및 항원 투여 후 사육환경 변화에 따른 넙치의 특이 항체 반응에 대한 영향을 조사하였다. 항원의 투여방법에 따른 항체 반응을 조사하기 위해 넙치에 BSA를 복강 및 근육 주사한 후 주기적으로 혈액을 취해 BSA에 대한 항체기를 측정한 결과, 근육 주사법이 복강 주사법보다 항체형성 초기시점에서 약간 높은 항제기를 나타났으나 그 이후의 항체 반응에는 유의적인 차이가 관찰되지 않아, 백신을 투여하는 방법으로 근육주사법 뿐만 아니라 복강 주사법도 유용하게 사용될 수 있을 것으로 사료되었다. 사육환경 변화에 따른 백신의 영향을 조사하기 위해 BSA를 면역시킨 넙치에 사육수온의 급하강, handling 및 중유에 노출시킨 후 항체기를 측정한 결과, 수온변화 및 handling 실험구가 양성 대조구보다 항체형성 초기시점에서 약간 높은 항체기를 나타났으나, 그 이후의 항체 반응에는 서로간의 유의적인 치아는 관찰되지 않아, 위의 환경 변화는 넙치의 특이 항체반응에 영향을 거의 끼치지 않는 것으로 확인되었다. 이상의 결과, 넙치 양식현장에서 백신은 일시적인 수온변화, 중유노출 및 handling에 의한 환경변화에서도 유용하게 사용 될 수 있을 것으로 사료되었다.

• Parasites, Hosts and Diseases
• /
• 제58권2호
• /
• pp.185-189
• /
• 2020

Immunogenicity of dendritic cell-derived exosomes stimulated with Toxoplasma gondii lysates (TLA exo), mixed with cholera toxin as an adjuvant, was investigated in mice immunized via 2 mucosal routes (ocular vs intranasal). BALB/c mice were injected 3 times with TLA exo vaccine at 2 week interval, and the levels of IgG in serum and IgA in tear, saliva, feces, and vaginal wash were measured. To observe the expression of T. gondii-specific B1 gene, mice infected with ME49 T. gondii cysts were immunized with TLA exo or PBS exo (not stimulated with TLA), and their brain tissues were examined. The mice vaccinated via intranasal route elicited significantly higher humoral and mucosal immune responses compared with mice treated with PBS alone. Also, mice immunized via ocular route (by eyedrop) induced significantly higher T. gondii-specific IgG in serum and IgA in tear and feces in comparison with PBS controls. B1 gene expression was significantly lower in TLA exo vaccinated mice than in PBS or PBS exo vaccinated mice. These results demonstrated that ocular immunization of mice with TLA exo vaccine has the potential to stimulate systemic or local antibody responses. This study also highlighted an advantage of an eyedrop vaccine as an alternative for T. gondii intranasal vaccines.

• 생명과학회지
• /
• 제11권4호
• /
• pp.385-391
• /
• 2001

Filamentous hemagglutinin (FHA) is considered as an essential immunogenic component for incorporation into acellular vaccines against Bordetella pertussis, the causative agent of whooping cough. Classically, antipertussis vaccination has employed an intramuscular route. An alternative approach to stimulate mucosal and systemic immune responses is oral immunization with recombinant live vaccine carrier strains of Salmonella typhimurium. An attenuated live Salmonella vaccine sgrain($\Delta$cya $\Delta$crp) expressing recombinant FHA(rFHA) was developed. Stable expressionof rFHA was achieved by the use of balanced-lethal vector-host system. which employs an asd deletion in the host chromosome to impose in obligate requirement for diaminopimelic acid. The chromosomal $\Delta$asd mutation was complemented by a plasmid vector possessing the asd$^{+}$ gene. A 3 kb DNA fragment encoding immuno dominant regionof FHA was subcloned in-frame downstream to the ATG translation initiation codon in the multicopy Asd$^{+}$ pYA3341 vector to create pYA3457. Salmonella vaccine harboring pYA3457 expressed approximately 105kDa rFHA protein. The 100% maintenance of [YA3457 in vaccine strain was confirmed by stability examinations. Additionally, a recombinant plasmid pYA3458 was constructed to overpress His(8X)-tagged rFHA in Essherichia coli. His-tagged rFHA was purified from the E. coli strain harboring pYA3458 using Ni$^{2+}$-NTA affinity purification system.>$^{2+}$-NTA affinity purification system.

• 대한수의학회지
• /
• 제57권1호
• /
• pp.9-15
• /
• 2017

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

• IMMUNE NETWORK
• /
• 제13권4호
• /
• pp.157-162
• /
• 2013

Application of vaccine materials through oral mucosal route confers great economical advantage in animal farming industry due to much less vaccination cost compared with that of injection-based vaccination. In particular, oral administration of recombinant protein antigen against foot-and- mouth disease virus (FMDV) is an ideal strategy because it is safe from FMDV transmission during vaccine production and can induce antigen-specific immune response in mucosal compartments, where FMDV infection has been initiated, which is hardly achievable through parenteral immunization. Given that effective delivery of vaccine materials into immune inductive sites is prerequisite for effective oral mucosal vaccination, M cell-targeting strategy is crucial in successful vaccination since M cells are main gateway for luminal antigen influx into mucosal lymphoid tissue. Here, we applied previously identified M cell-targeting ligand Co1 to VP1 of FMDV in order to test the possible oral mucosal vaccination against FMDV infection. M cell-targeting ligand Co1-conjugated VP1 interacted efficiently with M cells of Peyer's patch. In addition, oral administration of ligand-conjugated VP1 enhanced the induction of VP1-specific IgG and IgA responses in systemic and mucosal compartments, respectively, in comparison with those from oral administration of VP1 alone. In addition, the enhanced VP1-specific immune response was found to be due to antigen-specific Th2-type cytokine production. Collectively, it is suggested that the M cell-targeting strategy could be applied to develop efficient oral mucosal vaccine against FMDV infection.

• 약학회지
• /
• 제54권6호
• /
• pp.494-499
• /
• 2010

We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.

• 대한수의학회지
• /
• 제8권2호
• /
• pp.125-146
• /
• 1968

Throughout the studies the following experimental results were summarized. 1. It was impossible to infect and kill the mice, weighing 10 to 12 gm, by inoculating 0.2ml of virulent Cl. chauvoei, diluted 1 to 10 with physiological saline, via subcutaneous, intramuscular, intraperitoneal or intraveonus, route. 2. The mice which were inoculated in brain with 0.03ml of Cl. chauvoei diluted 1 : 5120 with physiological saline were resulted in all death after infection, but not in case of attenuated strain even in dilution of one to five. 3. Virulent Cl. chauvoei were diluted with each of those of whole blood, erythrocytes and serum of horse, calf, swine, sheep, rabbit, guinea pig, chicken and duck, human plasma and 2% CaCl solution, and inoculated subcutaneously 0.25 to 0.5ml in mice, weighing 12 to 15gm. It was resulted in significant increase in virulence as comparing with the case of physiological saline solution except when horse and pig sera were used. Such a phenomena were not seen in attenuated strain. 4. Virulence of virulent Cl. Chauvoei could be increased significantly in rat, as the procedures used in mice, by suspending in whole blood, erythrocytes, serum, or plasma of various animals, or 2% $CaCl_2$ solution and by inoculating subcutaneously 0.5 to 10ml in rat, weighing 30 to 60 gm, as compared with those of control group which used physiological saline solutionos diluent. 5. Mice resisted 100 and 80 percent against challenge of $10^3$ and $10^4$ M.L.D.. respectively, 24 hours after inoculation of 0.5ml black leg antiserum. 6. Immune response to the black leg living vaccine in mice could be obtained more favorably in the group of respected vaccination rather than those of single inoculation and the most profitable inoculm size of the vacine was 0.5 to 1.0ml. 7. Challenge for the immunized mice could be carried out effectively 3 weeks after first vaccination. 8. Satisfactory results could be obtained by inoculating subcutaneously for the immunization and intracerebrally or subcutaneously for the challenge. 9. Mice which were inoculated with 0.5ml of black leg living vaccine via subtaneucously two times at seven days interval and 21 days after first inoculation and challenged with 5 and 10 M.L.D. of virulent strain, resited 100 and 70 to 80 percent respectively. Same results were obtainable in black leg killed vaccine as the procedures used in living vaccine. 10. There were significantly different resistances against the definite challenge does between the mice groups which were immnuized with the living vaccine diluted five or 10 times and the undiluted. 11. For the biological assay of black leg living vaccine and antiserum, satisfactory results could be obtained using mice.

• 한국가금학회지
• /
• 제8권2호
• /
• pp.77-89
• /
• 1981

The experimental study was undertaken to confirm the effect of vaccination of birds with Newcastle disease (ND) vaccines on the Market by use of th. various vaccination programs. Sixteen groups of birds varying from 2 to f days of age, which were originated from hyper-immunised hens against ND were immunised by three different ways, a live vaccine only, a killed vaccine only, and the combination of a live and killed vaccine according to the each schedule of employed programs. In the administration of a live vaccine only, birds were immunized by one of following methods, the combination of intranasal and intraocular inoculation, intramuscular inoculation, via drinking water and the double inoculation by spray and drinking water application. Except for the double application, all the birds were vaccinated 2,3 or 4 times with two volumes of the virus dose (drinking water application) instructed by the commercial vaccine laboratory, until 21, 28 or 30 days of age, and all the immunized birds 19, 21 or 28 days postvaccination were challenged intramuscularly with 1.0$m\ell$ of 10,000 MLD per $m\ell$ of a virulent ND virus. In the administration of the combination of a live and killed vaccine, birds were immunized 2 or 3 times intranasally at first until 14 or 28 days of age with the same dose of the above experiment of a live vaccine, and then inoculated intramuscularly 1 or 2 times until 60 days of age with 1.0 $m\ell$ of a killed vaccine. And all immunized birds 11 days postvaccination were challenged with the same procedure of the above experiment. In the administration of a killed vaccine only, birds were immunized 3 times intramuscularly until 28 days of age with varied dose (0.2-0.5 $m\ell$) of a killed vaccine and all immunized birds 33 days postvaccination were challenged with the same procedure of the above experiment. The results obtained are summerised as follows: All birds vaccinated by using the combination of a live and killed vaccine program or a killed vaccin only appeared to be refractory. without any sign of illness, to the challenge exposure with 1.0$m\ell$ of 10,000 MLD per $m\ell$ of a virulent ND virus. On the other hand, the survival rates of birds of live vaccine groups immunized by a number of vaccine program such as Salsbury's day old program, 3-3-3 program, the Institute of Veterinary Reserch program and Multiple inoculation program, were 39.58%, 43.7%, 43.75% and 47.80%, respectively. And the survival rates of birds vaccinated with a live vaccine by 4 different ways of administration, i.e., double inoculation by water and aerosol application, intramuscular injection, intranasal instillation and via 4.inking water were 87.50%, 64.06%, 42.18% and 25.00%, respectively.

• 한국가금학회지
• /
• 제8권2호
• /
• pp.69-75
• /
• 1981

본 연구는 오늘날 뉴캣슬병(ND) 예방접종 목적에 널러 쓰여지는 Adjuvant가 불활화예방약과 약독주 생독예방약(LV)의 두가지 중 특히 논의가 많은 LV의 접종경로에 따르는 면역효과를 밝혀. 그 결과를 일반에게 명확히 제시하여 본병방과에 도움을 주는 목적으로 시행하였다. 이 연구를 위해서 먼저 LV생산재료로 사용되는 $B_1$ 접종 발육계묘에서 얻은 장요막질액의 접종경로별 면역력가를 30일영추를 공시하여 측정한 후, 그 유효량을 사용하여 분무, 비강내적하 근육주사, 음수투여의 4가지 경로를 통해서 상기시험일때와 동일하게 30일추에 각각 접종, 접종경로별로 면역효과를 비교 검토하였다. 그리하여 얻어진 결과를 요약하면 다음과 같다. 1. $10^{8.5}$ EI $D_{50}$$m\ell$의 역가를 가진 $B_1$독주장요막강액재료의 10진희척 각단계액을 30일추에 대하여 업종경로별 각소정량을 분무, 비강내적하, 근육주사하였을 때 상기재료의 $10^{-2}$ 희척($10^{6.5}$ EI $D_{50}$$m\ell$) 응용은 만족한 면역성을 부여하였으나 $10^{-3}$희척($10^{5.5}$ EI $D_{50}$$m\ell$)에서는 만족스럽지 못하였으며 I $D_{50}$(-log)는 근육주사=2.8, 분묘＞4.1, 비강내적하＞4.2이었다. 2. $10^{-2}$ 희포장요막강($10^{6.5}$ EI $D_{50}$$m\ell$)의 접종경로별 각소정량을 30일령 공시추에 접종경로를 달리하여 접종하였을 때 10,000MLD/$m\ell$ ND 병독/$m\ell$의 근육내접종공격에 대한 내과율은 분무가 93.75%, 비강내적하가 95.3%, 근육내접종이 92.6%의 좋은 성적이었는데 반하여 식수투여는 47.18%로 극히 불량하고, 시험한 4가지 접종경로 중 가장 낮은 성적이었다. 낮은 성적이었다.은 성적이었다.