• 한국동물위생학회지
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• 제30권2호
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• pp.275-281
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• 2007

Neospora caninum (N caninum) is an intracellular protozoa parasite, and its infection is one of the important diseases because it can cause abortion in cattle. This study was conduct to gain epidemiological data for more effective control of the disease. Bloods were collected from 2,162 cattle on 90 farms from February to September 2006. Serums were tested for antibodies to N caninum using ELISA CherdCheck, IDEXX). The dairy breed presented a higher proportion of seropositive results than that of Hanwoo. The estimated proportion of N caninum-positive dairy farms was 71.1 %, but the overall seroprevalence was estimated as 22.8%. The seroprevalence of dairy cattle and Hanwoo were 31.0% and 4.9%, respectively. There was no significant difference in the seroprevalence by age, but the not-intensive managed farms had a high seroprevalence (OR=1.91, p-value<0.01). The antibody rate of cattle with dog(s) was greater than that of those without dogs (OR=2.13, p-value<0.01). There was a significant difference in abortion rate between seropositive cattle and seronegative ones (OR=6.2, p-value<0.01).

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.320-325
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• 2012

Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1194-1199
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• 1997

The effect of hot taste preference on selected immune responses was investigated in human peripheral immunocompetent cells. Human lymphocytes and natural killer(NK) cells were prepared at a concentration of 2$\times$$10^{6}$ cells/ml in RPMI-1640 containing 10% fetal bovine serum. Lymphocytes proliferation was determined with the [$^{3}H$]-thymidine pulse for 18hrs after concanavalin A, phytohemagglutinin, Salmonella typhimurium mitogen, or media alone. NK cell activity was measured by cytolysis of $^51Cr$-labeled target cells K562. Serum antibodies levels such as IgM, IgG, IgA were also measured by ELISA method. There was no difference of serum IgM level among the groups, but IgG and IgA levels were greater in the group with hot taste preference than those of the group without hot taste preference. In lymphocytes of the group with hot taste preference there was a greater mitogen-induced lymphocyte proliferative responses compared to the group without hot taste preference. In addition, NK cell activity in group with hot taste preference was lower than that of the group without hot taste preference. These results suggest that the eating habit of spicy food containing hot components may affect immune status by modulating selective immunocompetent cells function.

• Journal of Veterinary Science
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• 제22권3호
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• pp.29.1-29.6
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• 2021

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

• 한국어병학회지
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• 제32권1호
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• pp.45-48
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• 2019

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.

• 한국어병학회지
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• 제33권2호
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• pp.171-175
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• 2020

We developed and subsequently characterized mouse monoclonal antibodies (mAbs) against marine birnavirus (MABV). Eight hybridoma clones secreting mAbs against MABV were established. All eight mAbs (8G6, 11C3, 15E3, 17H6, 32A6, 35A7, 38B5, and 47E3) were reacted with viral protein 3 of MABV in MABV-infected CHSE-214, whereas, no reactivity was observed in normal CHSE-214 by western blot analysis. Moreover, these eight mAbs were strongly reacted with MABV, and no cross-reactivity has been observed against other five fish viruses (hirame rhabdovirus, infectious hematopoietic necrosis virus, nervous necrosis virus, spring viraemia of carp virus, and viral hemorrhagic septicemia virus), although five mAb (11C3, 15E3, 17H6, 32A6, and 38B5) reacted with both MABV and infectious pancreatic necrosis virus by enzyme linked immunosorbent assay (ELISA). These results indicate that the mAbs can be of value in MABV detection.

• IMMUNE NETWORK
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• 제2권2호
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• pp.102-108
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• 2002

Background: This study was aimed to differentiate two forms of CTLA-4 (CD152) in activated peripheral blood lymphocyte and clarify the mechanism how cytoplasmic form of this molecule is targeted to cell surface. Methods: For this purpose we generated 2 different anti-human CD152 peptide antibodies and 5 different N'-terminal deletion mutant CTLA4Ig fusion proteins and carried out a series of Western blot and ELISA analyses. Antipeptide antibodies made in this study were anti-CTLA4pB and anti-CTLA4pN. The former recognized a region on extracellular single V-like domain and the latter recognized N'-terminal sequence of leader domain of human CD152. Results: In Western blot, the former antibody recognized recombinant human CTLA4Ig fusion protein as an antigen. And this recognition was completely blocked by preincubating antipeptide antibody with the peptide used for the antibody generation at the peptide concentration of 200 ug/ml. These antibodies were recognized human CD152 as a cytoplasmic sequestered- and a membrane bound- forms in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocyte (PBL). These two forms of CD152 were further differentiated by using anti-CTLA4pN and anti-CTLA4pB antibodies such that former recognized cytosolic form only while latter recognized both cytoplasmic- and membraneforms of this molecule. Furthermore, in a transfection expression study of 5 different N'-terminal deletion mutant CTLA4Ig, mutated proteins were secreted out from transfected cell surface only when more than 6 amino acids from N'-terminal were deleted. Conclusion: Our results implies that cytosolic form of CTLA-4 has leader sequence while membrane form of this molecule does not. And also suggested is that at least N'-terminal 6 amino acid residues of human CTLA-4 are required for regulation of targeting this molecule from cytosolic- to membrane- area of activated human peripheral blood T lymphocyte.

• 한국식품영양과학회지
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• 제45권2호
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• pp.174-180
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• 2016

본 연구에서는 산양유 발효유(F-GM)의 급여가 면역 활성과 체력증진에 미치는 영향을 조사하였다. F-GM을 경구 투여한 마우스의 비장세포는 림프구 유사분열 촉진인자인 ConA와 LPS에 대한 증식반응의 상승을 나타내었다. ConA를 처리한 비장세포로부터 분비되는 cytokine을 ELISA법에 의해 정량한 결과 F-GM 투여에 의해 IL-2와 IFN-${\gamma}$의 생성은 유의하게 증가하였으나, IL-4 및 IL-10의 생성은 별다른 변화를 보이지 않았다. 또한 RT-PCR을 이용한 이들 cytokine mRNA의 정량실험에서도 F-GM 투여로 IL-2와 IFN-${\gamma}$의 mRNA가 증가하는 것을 확인하였다. 한편 KLH(20 mg/mouse)를 정상 대조군 마우스 혹은 F-GM을 급이한 마우스에 각각 면역하고 5주째에 KLH에 대한 항체가를 측정한 결과 F-GM을 급이한 마우스에서 항체가가 유의하게 상승하는 것으로 나타났다. KLH에 대한 항체의 isotype을 조사한 실험에서는 KLH에 대한 IgG1, IgG2a 그리고 IgM 특이항체의 상승이 관찰되었다. F-GM 경구 투여가 세포성 면역에 미치는 영향을 측정하기 위하여 면역개시 7주째의 마우스를 이용하여 KLH에 대한 지연형 과민반응(DTH)을 측정하였다. 그 결과 F-GM을 경구 투여한 마우스에서 DTH의 유의한 상승효과가 확인되었다. 한편 수영시험을 통해 F-GM의 경구 투여가 체력증강에 미치는 영향을 측정한 결과 F-GM을 투여한 마우스에서 매우 유의한 수영시간의 연장 효과가 관찰되었다. 이들 결과로부터 F-GM은 면역세포의 활성화를 통한 면역력 강화는 물론 체력을 증가시키는 생리활성을 갖는 것으로 확인되었다.

• 생물정신의학
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• 제6권2호
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• pp.202-208
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• 1999

Schizophrenia has many clinical expressions and probably different etiologic factors. Infections, autoimmune mechanism and related neurodevelopmental abnormalities have been suggested as possible etiologic factors of schizophrenia. It has been reported that immunoreactivity to heat shock proteins, which play a protective role against environmental stresses in a cell, might be related to the pathogenesis of schizophrenia. Therefore, we examined the immunoreactivity to heat shock protein 70kDa and 90kDa(HSP70 and 90) in 91 patients with schizophrenia and 83 normal controls. Ig G antibodies to HSP70 and 90 of sera were quantitated by ELISA. The optical density(OD) was measured by an automated microplate reader at a wavelength of 490nm. The amounts of antibodies to HSPs were expressed as arbitrary units(AU)/ml related to a standard serum. The limit for elevated antibody titers(anti-HSPs positive or negative) was set at two standard deviations added to the mean of the normal controls. Twenty nine(31.9%) of the 91 patients showed anti-HSP70 positive and 19(20.9%) of those showed anti-HSP90 positive. On the other hand, only 1(1.4%) of the normal controls and 4(4.8%) of those showed anti-HSP70 positive and anti-HSP90 positive, respectively. The titers of anti-HSP70 positive were related with BPRS scores, while those of anti-HSP90 positive were not. There were no relationship between antibody titers and clinical variables including age at onset, duration of illness, family history of schizophrenia or number of admission. The titers of anti-HSP70 positive were significantly associated with anti-HSP90 positive. Our results suggest the presence of abnormal immune reactivity involving HSP70 and HSP90 in a subset of patients with schizophrenia.

• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1308-1321
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• 2013

The emergence of microcarrier technology has brought a renewed interest in anchorage-dependent cell culture for high-yield processes. Well-known in vaccine production, microcarrier culture also has potential for application in other fields. In this work, two types of microcarriers were evaluated for small-scale monoclonal antibody (mAb) production by CHO-K1 cells. Cultures (5 ml) of microporous Cytodex 3 and macroporous CultiSpher-S carriers were performed in vented conical tubes and subsequently scaled-up (20 ml) to shake-flasks, testing combinations of different culture conditions (cell concentration, microcarrier concentration, rocking methodology, rocking speed, and initial culture volume). Culture performance was evaluated by considering the mAb production and cell growth at the phases of initial adhesion and proliferation. The best culture performances were obtained with Cytodex 3, regarding cell proliferation (average $1.85{\pm}0.11{\times}10^6$ cells/ml against $0.60{\pm}0.08{\times}10^6$ cells/ml for CultiSpher-S), mAb production ($2.04{\pm}0.41{\mu}g/ml$ against $0.99{\pm}0.35{\mu}g/ml$ for CultiSpher-S), and culture longevity (30 days against 10-15 days for CultiSpher-S), probably due to the collagen-coated dextran matrix that potentiates adhesion and prevents detachment. The culture conditions of greater influence were rocking mechanism (Cytodex 3, pulse followed by continuous) and initial cell concentration (CultiSpher-S, $4{\times}10^5$ cells/ml). Microcarriers proved to be a viable and favorable alternative to standard adherent and suspended cultures for mAb production by CHO-K1 cells, with simple operation, easy scale-up, and significantly higher levels of mAb production. However, variations of microcarrier culture performance in different vessels reiterate the need for optimization at each step of the scale-up process.