• IMMUNE NETWORK
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• 제2권1호
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• pp.41-48
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• 2002

Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

• 대한수의학회지
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• 제33권4호
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• pp.657-664
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• 1993

우리나라 육계산업을 저해하는 중요한 질병중의 하나가 닭의 맹장콕시듐증이다. 그 병원체는 Eimeria tenella로서 병아리에 감염하면 증체저하와 폐사율이 높기 때문에 양계농가에 커다란 경제적 손실을 준다. 닭의 맹장콕시듐증을 효과적으로 관리하기 위하여 몇가지 예방백신의 개발을 시도하여 왔으며 근래에는 재조합 DNA기법을 이용, Escherichia coli를 매체로 제조한 항원으로 닭에 방어력을 부여시키고자 하는 방법과 조기발육형원충의 작출법에 의하여 생산된 약독원충으로 면역을 부여하는 방법 등에 관한 연구를 진행하고 있으나 아직 만족할만한 성과를 이루지 못하였다. 본 시험에서는 약독원충을 이용하여 닭 맹장콕시듐증에 대한 저항성을 부여하고자 하였다. 콕시듐증의 병원체인 E tenella에 Cobalt-60 감마선을 조사한 원충 또는 그 원충을 SPF병아리에 계대증식시켜 회수한 오오시스트로 병아리에 접종하고 체내발육상태와 병원성을 조사하였으며, 면역접종한 후 병원성이 강한 원충의 공격접종에 대한 방어효과를 관찰한 성적은 다음과 같다. 원충감염 닭의 생잔율, 증체량, 혈변도 및 병변도 등에 대한 효과에서 감마선 100Gy로 조사한 E tenella를 접종한 것이 가장 좋았으며, 접종량은 1대 계대한 오오시스트 약 10,000개를 접종한 것이 좋았다. 면역접종한 후 공격접종한 결과 계대원충접종군은 혈변도 및 병변도가 낮게 나타났으며, 최대 오오시스트 배설수(OPG)는 103,900으로서 공격접종대조군의 1,658,900에 비하여 현저하게 낮았으나 적혈구용적율은 계대원충접종군이 가장 높았다. 공격접종한 닭의 1주 및 2주 후의 증체량은 계대원충접종군이 2.6g 및 155.4g으로서 공격접종대조군의 -85.8g 및 63.6g보다 좋았으며, 사료요구율도 계대원충접종군이 3.28로서 공격접종대조군의 5.60보다 낮았다. 생잔율은 계대원충접종군이 100%로서 공격접종대조군의 87.0%와 비교하면 방어효과가 있었다. 계대원충접종군의 항콕시듐 지수는 70.5로서 공격접종대조군의 -81.9에 비하면 현저하게 높았다. 그러므로 맹장콕시듐 원충은 감마선 조사가 병원성을 저하시키고 계대수가 증가할수록 병원성은 약간 증가하지만 공격접종에 대한 방어효과가 높아지는 것을 볼 수 있었다. 이는 병원성이 있는 원충이 면역원성도 좋으며 앞으로 면역원성이 유지되며 병원성이 없는 원충의 선발이 필요할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권5호
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• pp.671-676
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• 2008

Forty-eight Naemi lambs (avg. BW 31.7 kg) were transported by truck for a distance of 1,450 km from Al-Jouf to Riyadh, Saudi Arabia. On arrival day, the lambs were randomly allocated to four groups receiving diets supplemented with 0.0, 0.3, 0.6 and 0.9 ppm organic chromium (Cr). Each group consisted of four separately housed replicates of three lambs each. The animals were fed ad libitum on a grower diet for 84 days. Blood samples were obtained shortly before transportation, upon arrival and at weekly intervals thereafter from all lambs for analysis of plasma and serum. Plasma glucose and serum cortisol, total protein, albumin, urea-N and total cholesterol concentrations were determined. A cursory clinical examination of the lambs, along with rectal temperature, was undertaken at different intervals during the experiment. The lambs were inoculated each with 2 ml i.v. chicken red blood cells (CRBC) on days 0, 21, and 42. Serum total, IgG and IgM antibody titers were determined at weekly intervals post-immunization. An in vivo intradermal hypersensitivity test was carried out on 6 lambs from each group on days 10 and 70. Transportation of the lambs resulted in a significant (p<0.001) elevation of serum cortisol, total protein and albumin levels, as well as increased plasma glucose concentration, with corresponding decrease in total cholesterol, while blood urea-N remained largely unchanged. These constituents returned to normal levels during subsequent weeks, with no significant differences in their concentrations being observed between the Cr-supplemented groups and controls. Rise in rectal temperature after transportation was reduced to a greater extent (p<0.05) in Cr-supplemented versus control lambs. Total, IgG and IgM antibody titers against CRBC rose significantly (p<0.05) during immunizations in all groups, with significantly and linearly higher (p<0.05) total and IgG titers in Cr-supplemented versus control lambs. By contrast, no significant effect due to Cr supplementation was recorded in IgG titers, which increased equally in Cr-fed and control groups. Skin thickness in response to intradermal inoculation of phytohaemagglutinin (PHA) was also significantly (p<0.01) increased as a result of Cr supplementation. These results indicate that dietary Cr supplementation might be useful during stress especially for enhancing immune responses in transport-stressed lambs.

• 동의생리병리학회지
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• 제21권1호
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• pp.39-49
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• 2007

Rheumatoid arthritis is an autoimmune disease involving multiple joint. In order to access the suppressive effects of JTT on rheumatoid arthritis and it's effects on immune system we investigated whether JTT could suppress the disease progression of collagen-induced arthritis. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with DW, JTT (200 or 400 mg/kg) or methotrexate (MTX, 30 mg/kg) as a positive control. Oral administration of JTT significantly suppressed the progression of CIA, which extend is comparable to that of MTX. Histological examination reveled that JTT inhibited infiltration of inflammatory cells into affected paw joint and bone erosion and cartilage destruction were greatly reduced compared with control. Total cell number of spleen, lymph node and peripheral blood were significantly reduced. The absolute number of CD19$^+$, CD3$^+$/CD69$^+$, CD4$^+$/CD25$^+$ cell in spleen from JTT treated mice were significantly decreased. The absolute number of CD19$^+$, CD3$^+$, CD3$^+$/CD69$^+$, CD4$^+$, CD4$^+$/CD25$^+$ CD8$^+$, CD49b, CD3/CD49b cells in draining lymph node were significantly increased compared with control. In peripheral blood mononuclear cells of JTT treated mice, the absolute number of CD4$^+$, CD4$^+$/CD25$^+$, CD3$^+$/CD69$^+$ cells were significantly decreased compared with control, while that of CD49b$^+$ was slightly increased. Infiltration of CD3$^+$ cells and CD11b$^+$/Gr-1$^+$ cells into paw joint was significantly reduced in JTT treated mice. The levels of pathologic cytokines including TNF-a and IL-6 in serum were significantly decreased by oral treatment with JTT The levels of IFN-g in the culture supernatant of splenocyte stimulated with CD3$^+$/CD28$^+$ or collagen were dramatically decreased, while the levels of IL-4 was increased under CD3$^+$/CD28$^+$ or collagen stimulation. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of JTT treated mice than control. Taken together, JTT has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use anti-rheumatic arthritic agent in human.

• 한방재활의학과학회지
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• 제20권1호
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• pp.37-59
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• 2010

Objectives : The object of this study was to observe the favorable anti arthritic effects of Euiiin-tang($y{\grave{i}}y{\breve{i}}r{\acute{e}}n-t{\bar{a}}ng$) aqueous extract(EIITe), has been traditionally used in Korean medicine for treating rheumatoid arthritis on collagen induced arthritic(CIA) DBA/1 mice. Methods : In the present study, effects of EIITe on the releases of human tumor necrosis factor(TNF)-${\alpha}$, interleukin(IL)-$1{\beta}$, matrix metalloproteinase(MMP)-13 and production of Nitric oxide(NO) were observed by in vitro. In addition, to observe the effects on the CIA mice, three different dosages of EIITe, 300, 150 and 150 mg/kg were orally administered once a day for 18 days from 24hrs after antigen challenges(type II collagen) on 21 days after immunization using Type II collagen Freund's complete adjuvant. Six groups, each of 8 DBA/1 mice per group were used in the present study as follows. Changes on the body weights, macroscopic arthritis scores, splenic weights, splenic TNF-${\alpha}$ and IL-6 contents, articular cartilage(femur and tibia) collagen and glycosaminoglycans-chondroitin sulphate, sulphate and hyaluronic acid contents, histopathological observations(microscopic arthritis scores, thicknesses of femur and tibia cartilage thicknesses were monitored, compared to that of dexamethasone, a potent anti inflammatory agents, 1 mg/kg treated mice. Results : As results of collagen challenges, marked decreases of body weights and gains, articular cartilage collagen and glycosaminoglycan - chondroitin sulphate, sulphate and hyaluronic acid contents were observed with increases of macroscopic arthritis scores, splenic weights, splenic TNF-${\alpha}$ and IL-6 contents, articular cartilage(in the both femur and tibia) loss and damages. However, these CIA signs were significantly and dosages dependently inhibited by treatment of EIITe 300 and 150 mg/kg as compared with CIA control, respectively. In addition, the releases of TNF-${\alpha}$, IL-$1{\beta}$, NO and MMP-13 were markedly and dose dependently inhibited by treatment of EIITe, invitro. Although CIA were more favorably inhibited by treatment of dexamethasone 1 mg/kg as compared with EIITe 300 mg/kg, marked decreases of body weights were detected in dexamethasone 1 mg/kg treated mice. Conclusions : The results obtained in this study suggest that over 150 mg/kg of EIITe showed favorable anti arthritic effects on the CIA mediated by immunomodulatory and/or anti oxidative effects. However, detail mechanism studies should be conduced in future with the screening of the biological active compounds in this herb. lthough CIA were more favorably inhibited by treatment of dexamethasone 1 mg/kg as compared with EIITe 300 mg/kg, marked decreases of body weights were detected in dexamethasone 1 mg/kg treated mice, in the present study.

• 한방재활의학과학회지
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• 제20권4호
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• pp.33-49
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• 2010

Objectives : This study was designed to elucidate the anti-pathogenetic and curative effects of Gyehyeoldeungbokhap-bang(Jixietengfuge-fang, GCP) on Type II collagen-induced arthritis(CIA) in mice. Methods : In experiment, twenty four mices were divided into non-treated normal group(n=6), bovine type II collagen-induced control group(n=6), collagen immunized and treated two group medicated with extract of GCP(concentration of extraction: 200 mg/kg n=6, 400 mg/kg n=6) for 4 weeks after collagen immunization, Various experimental such as arthritis, incidence, index, total cell number of spleen, total cell number of peripheral lymph node(PLN), paw joint total cell number, analysis on the percentage of immunofluorescent cells of spleen in CIA induced mice, effects of inflammatory gene expression in spleen, PLN and paw joint of CIA mice, production of cytokine(IFN-${\gamma}$, IL-4, TNF-${\alpha}$, IL-6), analysis of rheumatoid factor(anti-collagen lgG, lgM level in serum) and histopathological study on the paw joint. The arthritis index, incidence were measured a week over 4 weeks after second boosting. Total cell number of spleen, peripheral lymph node, paw joint were measure after performed experiment over 7 weeks. Concentration of cytokine and rheumatoid factor in serum were measured after experiment finished. Histopathological study on the paw joint was measured after 40 days medicated with extract of GCP. Results : 1. Incidence of arthritis and index of arthritis were significantly decreased in treated group with 400 mg/kg. 2. Total cell number of spleen, PLN and paw joint were significantly decreased in treated group. 3. Analysis on the percentage of immunofluorescent cells of spleen in CIA induced mice were significantly controled compare with control group. 4. Effects of inflammatory gene expression in spleen, PLN and paw joint of CIA induced mice were significantly controled compare with control group. 5. IFN-${\gamma}$, IL-6, TNF-${\alpha}$, concentration($pg/m{\ell}$) in serum of treated group was significantly decreased compare with control group. But IL-4 was significantly increased. 6. lgM and lgG concentration($pg/m{\ell}$) in serum of treated group was significantly decreased compare with control group. 7. Histopathologically, suppurative and destructive lesion of synovial membrane, articular cartilage and subchondral bony tissue in treated group were alleviated compare with those of control group. Conclusions : Based on the results described above, it might be consider that Gyehyeoldeungbokhap-bang(Jixietengfuge-fang, GCP) has antiarthritic and analgesic effects and also inhibitory effects on the progression of collagen-induced arthritis mice.

• IMMUNE NETWORK
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• 제11권5호
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• pp.258-267
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• 2011

Background: Current management strategies attempt to diagnose rheumatoid arthritis (RA) at an early stage. Transcription profiling is applied in the search for biomarkers for detecting early-stage disease. Even though gene profiling has been reported using several animal models of RA, most studies were performed after the development of active arthritis, and conducted only on the peripheral blood and joint. Therefore, we investigated gene expression during the initial phase of collagen-induced arthritis (CIA) before the arthritic features developed in the thymus in addition to the peripheral blood and synovium. Methods: For gene expression analysis using cDNA microarray technology, samples of thymus, blood, and synovium were collected from CIA, rats immunized only with type II collagen (Cll), rats immunized only with adjuvant, and unimmunized rats on days 4 and 9 after the first immunization. Arrays were scanned with an Illumina bead array. Results: Of the 21,910 genes in the array, 1,243 genes were differentially expressed at least 2-fold change in various organs of CIA compared to controls. Among the 1,243 genes, 8 encode T-cell receptors (TCRs), including CD3${\zeta}$, CD3${\delta}$, CD3${\varepsilon}$, CD8${\alpha}$, and CD8${\beta}$ genes, which were down-regulated in CIA. The synovium was the organ in which the genes were differentially expressed between CIA and control group, and no difference were found in the thymus and blood. Further, we determined that the differential expression was affected by adjuvant more than Cll. The differential expression of genes as revealed by real-time RT-PCR, was in agreement with the microarray data. Conclusion: This study provides evidence that the genes encoding TCRs including CD3${\zeta}$, CD3${\delta}$, CD3${\varepsilon}$, CD8${\alpha}$, and CD8${\beta}$ genes were down-regulated during the initial phase of CIA in the synovium of CIA. In addition, adjuvant played a greater role in the down-regulation of the CD3 complex compared to CII. Therefore, the down-regulation of TCR gene expression occurred dominantly by adjuvant could be involved in the pathogenesis of the early stage at CIA.

• 한국가금학회지
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• 제45권1호
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• pp.17-28
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• 2018

전염성기관지염은 전세계적으로 양계산업에 문제시되고 있는 질병으로서 이를 예방하기 위한 연구가 활발히 진행되고 있다(Dhama et al., 2014). 본 연구는 전염성기관지염 백신을 진행한 여러 계군의 혈청을 수집해 국내회사와 다국적기업에서 제조한 ELISA 키트의 검출능 차이를 확인하고자 하였다. 육용원종계부터 산란계까지 총 35개의 계군으로부터 주기적인 채혈을 통해 얻어진 혈청을 사용해 두 가지 키트의 검출능 비교시험을 진행한 결과, 국내회사에서 제조된 ELISA 키트가 더 높은 항체가를 나타냈으며, 이는 미량의 항체도 국내회사 제조 키트를 사용하게 되면 더욱 민감하게 검출될 수 있으며, 농장의 상황 판단에 더욱 도움이 될 수 있을 것임을 시사한다. 이러한 결과를 토대로 비추어 보았을 때, 국내에서 발생하고 있는 전염성기관지염에 맞춰 제작된 국내회사 제조 키트가 더 검출능이 뛰어난 것으로 판단된다. 또한, 주기적인 혈청을 이용한 항체가 검사 결과 전염성기관지염 백신 후 항체가 변화 추이를 관찰할 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2667-2672
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• 2016

Background: Virtually all cases of cervical cancer are caused by persistent infections with a restricted set of human papillomaviruses (HPV). Cancer of the cervix is the third or even the second most common cancer in women worldwide, more than 85% of the cases occurring in developing countries, such as China and India, including the Republic of Kazakhstan. The purpose was to determine the HPV type distribution to evaluate efficacy of vaccination and adjust cancer prevention strategy in Western Kazakhstan in the future. Materials and Methods: A retrospective analysis was conducted of data obtained from PCR laboratories in 4 regional centers for the time period covering 12 months, 2013-2014, using AmpliSens$^{(R)}$ Real-Time PCR kits for HPV testing of 12 genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59). Results: A total of 1,661 persons were HPV tested within 2013-14, but a proprotion examined for 16 and 18 genotypes only (563) was not been included for statistic analysis of distribution and ratio of the most common genotypes. Males accounted for only a small number (N=90 in total). Conclusions: Total number of the HPV-positive appeared to be 26.0%, or 286 of N=1098. Types distribution was as follows: type 16 (10.7%), 39 (5.83%), 51 (5.27%), 31 (4.85%), 56 (4.58%), 18 (3.61%), 59 (2.64%), 58 (2.22%), 35 (1.94%), 33 (1.25%). Overall the HPV infection was highest in 16-29 years old (62.4%) and decreased with age. Total prevalence of the HR-HPVs amongst male population was 21.4% with top five types 16, 18, 39, 51, 31. Trends forcorrelations between Aktau site and type 33 (Cramer's V 0.2029), between Caucasian ethnicity and type 33 (Cramer's V .1716), and between European ethnicities in Uralsk and type 45 (Cramer's V .1752) were found. Of N 563 tested separately for 16 or 18 types, 13.6% were positive. As a whole, the distribution of 16/18 types had a ratio of 3.53:1. Given the vaccine-targeted type 16 is widely spread amongst this regional population, HPV immunization program of adolescent girls 10-13 years should be implemented appropriately.

• Journal of Animal Science and Technology
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• 제46권3호
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• pp.495-502
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• 2004

장출혈성대장균 O157:H7의 eae유전자 산물인 intimin은 장 상피세포의 부착성을 조절하는 단백질로 알려져 있다. Intimin의 C-말단 아미노산 잔기와(His)$_6$이 혼합되어 이루어진 plasmid를 작성하여 재조합 단백질(34kDa)을 발현시켰다. 발현된 재조합 intimin 단백질의 면역원성을 확인하기 위해 토끼와 산란계에서 항혈청 및 난황항체(IgY)를 제조하였다. O157:H7에서 분리한 세포외막단백질(OMPs)과 제조된 항혈청과의 western blot상의 반응에서 약 94kDa의 위치에서 양성반응을 보여 발현된 재조합 단백질의 면역원성 효과가 확인되었다. 면역원성이 확인된 단백질을 산란계에 면역한 결과, 면역 후 4${\sim}$6주 경부터 높은 수준의 항체가 형성되었다. 재조합 intimin 단백질에 대한 난황 항체의 항원결합능력 조사를 위하여 중화반응시험을 수행한 결과, 재조합 intimin 단백질에 대한 난황 항체가 O157:H7과 강하게 결합하는 것으로 나타났다. 따라서 본 연구에서 발현된 재조합 intimin 단백질은 토끼와 산란계의 면역반응을 유도할 수 있는 효율적인 면역원임과 동시에 난황 항체 생산을 위한 항원으로의 이용가능성이 확인되었다.