• 대한수의학회지
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• 제10권1호
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• pp.11-23
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• 1970

Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.

• Journal of Radiation Protection and Research
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• 제30권4호
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• pp.197-208
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• 2005

Although little information is available on the underlying mechanisms, various genetic factors have been associated with tissue-specific responses to radiation. In the present study, we explored the possibility whether organ specific gene expression is associated with radiosensitivity using samples from brain, lung and spleen. We examined intrinsic expression pattern of 23 genes in the organs by semi-quantitative RT-PCR method using both male and female C57BL/6 mice. Expression of p53 and p21, well known factors for governing sensitivity to radiation or chemotherapeutic agents, was not different among the organ types. Both higher expression of sialyltransferase, delta7-sterol reductase, leptin receptor splice variant form 12.1, and Cu/Zn superoxide dismutase (SOD) and lower expression of alphaB crystalline were specific for spleen tissue. Expression level of glutathione peroxidase and APO-1 cell surface antigen gene in lung tissue was high, while that of Na, K-ATPase alpha-subunit, Cu/ZnSOD, and cyclin G was low. Brain, radioresistant organ, showed higher expressions of Na, K-ATPase-subunit, cyclin G, and nucleolar protein hNop56 and lower expression of delta7-sterol reductase. The result revealed a potential correlation between gene expression patterns and organ sensitivity, and Identified genes which might be responsible for organ sensitivity.

• 동의생리병리학회지
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• 제25권1호
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• pp.29-36
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• 2011

Nardostachyts chinensis (N. chinensis) belonging to the family Valerianaceae has been used to elicit stomachic and sedative effects. The MAPKs are serine/threonine kinases involved in the regulation of various cellular responses, such as cell proliferation, differentiation and apoptosis. The PKC also plays a key role in regulating the response of hematopoietic cells to both physiological and pathological inducers of proliferation and differentiation. This study investigated the signaling pathways on the U937 cell differentiation induced by N. chinensis. N. chinensis induced the differentiation of U937 cells, as shown by increased of differentiation surface antigen CD11b. Activation of ERK increased time-dependently in differentiation of U937 cells induced by N. chinensis, but activations of JNK and p38 were unaffected. Inhibitor of ERK (PD98059) significantly reduced CD11b expression induced by N. chinensis in U937 cells. In addition, N. chinensis increased protein level of PKC ${\beta}$I and PKC ${\beta}$II isoforms, but the protein level of PKC ${\alpha}$ and PKC ${\gamma}$was constant. PKC inhibitors (GF 109203X and H-7) inhibited U937 cell differentiation and the ERK activation induced by N. chinensis. These results indicated that PKC and ERK may be involved in U937 cell differentiation induced by N. chinensis.

• Clinical and Experimental Pediatrics
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• 제48권12호
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• pp.1348-1353
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• 2005

목 적 : 인플루엔자는 해마다 유행하는 호흡기 질환으로서 만성질환자나 노인 뿐만 아니라 영아나 소아에게도 높은 유병률과 치사율을 나타내어 임상적 중요성이 주목되고 있다. 임상증상만으로는 다른 호흡기 바이러스나 세균감염과 감별이 어렵고, 전통적인 진단방법인 바이러스 배양검사는 소요 시간이 길고 배양방법도 까다로워 진단이 늦어지고 치료에도 적용하지 못하였다. 이에 따라 최근 검사방법이 간편하고 결과를 빨리 알 수 있는 신속검사법들이 소개되어 그 정확성과 임상에의 유용성이 많이 연구되고 있으나 국내에서는 연구례가 드문 실정이다. 이에 저자들은 신속검사의 정확성과 유용성을 평가하여 실질적인 검사법으로 정착시키고자 하였다. 방 법 : 2004년 1월부터 6월, 2005년 2월부터 6월까지 발열, 기침, 인후통 및 근육통 등의 증상으로 외래 내원 및 입원 치료를 받은 408명의 환아를 대상으로, 비인두 가검물을 채취하여 QuickVueR(Quidel Co., San Diego, USA) influenza rapid antigen test와 바이러스 배양을 동시에 시행하였다. 결 과 : 평균 연령은 $2.51{\pm}2.65$(2개월-15세)이고 남아는 239명, 여아는 169명이었다. 바이러스 배양에 양성을 보인 환아는 77명으로 이중 A/H3N2형은 55명(71.4%), B형은 22명(28.5%)이었다. 본 검사의 민감도는 71.4%, 특이도는 95.8%, 양성예측도는 79.7%, 음성예측도는 93.5%였으며 위양성률과 위음성률이 각각 4.2%, 28.5%로 나왔다. 결 론 : 본 검사는 비교적 민감도가 높아 선별 검사로서 적합하다고 생각되며, 특히 특이도가 높고 위양성률이 낮아 결과가 양성으로 나왔을 시 인플루엔자 감염의 가능성이 높아, 치료를 빨리 시작함으로써 증상의 호전을 기대할 수 있겠다. 앞으로 이러한 신속검사가 실제 임상에 널리 적용된다면 환자 침상에서 간편하고 빠르게 검사하여 결과를 바로 알고 치료를 조속히 시작하여 합병증을 줄이고 불필요한 항생제의 사용을 줄일 수 있으리라 기대된다.

• IMMUNE NETWORK
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• 제13권1호
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• pp.34-41
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• 2013

Interleukin-18 (IL-18) has been known to induce interferon-${\gamma}$ (IFN-${\gamma}$) production and promote Th1 immunity. Although mammalian IL-18 has been characterized in great detail, the properties and application of chicken IL-18 remain largely uninvestigated as of yet. In this study, we evaluated the immunomodulatory properties of Salmonella enterica serovar Typhimurium expressing chicken interleukin-18 (chIL-18) on immune responses induced by avian influenza (AI) and Newcastle disease (ND) vaccines. After oral administration of S. enterica serovar Typhimurium expressing chIL-18, chickens were vaccinated intramuscularly with the recommended dose of either inactivated AI H9N2 vaccine or ND (B1 strain) vaccine. Chickens receiving a primary vaccination were boosted using the same protocol 7 days later. Humoral and cell-mediated immune responses were evaluated in terms of HI antibody titers and proliferation and mRNA expression of IFN-${\gamma}$ and IL-4 of peripheral blood mononuclear cells (PBMC) in response to specific antigen stimulation. According to our results, oral administration of S. enterica serovar Typhimurium expressing chIL-18 induced enhanced humoral and Th1-biased cell-mediated immunity against AI and ND vaccines, compared to that of chickens received S. enterica serovar Typhimurium harboring empty vector. Therefore, we conclude that our proposed vaccination regimen using inactivated AI and ND viruses along with oral administration of S. enterica serovar Typhimurium expressing chIL-18 may provide a novel approach in protecting chicken from currently circulating AI and ND virus strains.

• 대한수의학회지
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• 제25권1호
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Tuberculosis and Respiratory Diseases
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• 제44권4호
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• pp.785-795
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• 1997

연구배경 : 종양 표지자는 각종 악성 종양의 진단, 진행정도, 절제 가능성, 예후추정, 치료후 추적 검사 등에도 사용될 수 있으나 민감도와 특이도가 만족스럽지 못한 경우가 많아서 임상에서 보조적인 수단으로 이용되고 있는 경우가 대부분이다. 최근 Cyfra21-1이 비소세포 폐암의 진단에 CEA, SCC보다 우수하다는 보고가 있었고 단일클론 항체를 이용한 TPA-M(Tissue polypeptide antigen)이 비소세포 폐암은 진단하는데 상당히 우수한 결과를 보인다고 보고하고 있다. 이에 폐암환자에서 혈청 CEA, SCC, Cyfra21-1, TPA-M의 임상적 유용성을 서로 비교해 보고자 이 연구를 시행하였다. 방 법 : 1996년 4월 1일 부터 19961건 8월 31일까지 계명대학교 의과대학 병원에 내원한 환자중 조직 생검으로 확진된 비소세포 폐암 37예와 소세포 폐암 12예를 총 49예를 폐암 환자군으로 같은 기간의 입원 환자 중 양성 양성 폐질환으로 확진된 29예를 대조군으로 선정하였고 폐암 환자의 경우 수술, 항암요법 및 방사선치료를 전혀 시행하지 않은 상태에서 혈청을 채취하여 검사를 시행하였다. CEA는 일본 Eiken사의 Ab bead CEA, SCC는 미국 DAINABOT사의 SCC RIA BEAD kit, Cyfra21-1은 일본 TFB사의 CA 21-1 kit, TPA-M은 일본 DAIICHI사의 TPA-M kit를 사용하였다. 결 과 : 전체 폐암환자와 대조군을 비교해보면 CEA가 $10.05{\pm}38.39{\mu}/L$, $1.59{\pm}0.94{\mu}/L$, SCC가 $3.04{\pm}5.79{\mu}/L$, $1.58{\pm}2.85{\mu}/L$, Cyfra21-1이 $8.27{\pm}11.96{\mu}/L$, $1.77{\pm}2.72{\mu}/L$, TPA-M이 $132.02{\pm}209.35U/L$, $45.86{\pm}75.86U/L$으로 나타났으며 Cyfra21-1과 TPA-M은 폐암환자에서 대조군보다 종양 표지자의 측정치가 유의하게 높았다.(p<0.05) CEA, Cyfra21-1, TPA-M, SCC의 cutoff value를 회사에서 권장하는 $2.5{\mu}/L$, $3.0{\mu}/L$, 70.0U/L, $2.0{\mu}/L$로 하였을 때 전체폐암에 대한 민감도와 특이도는 CEA 33.3%와 78.6%, Cyfra21-1 50.0%와 89.7%, TPA-M 52.3%와 89.7%, SCC 23.8%와 89.3%이었으며 비소세포 폐암에 대하여서는 CEA 36.1%와 78.1%, Cyfra21-1 50.1%와 89.7%, TPA-M 53.1%와 89.7%, SCC 33.8%와 89.3% 소세포 폐암의 경우에도 CEA 25.0%와 78.5%, Cyfra21-1 50.0%와 89.6%, TPA-M 50.0%와 89.6%, SCC 0%와 89.2%로 폐암환자에서 Cyfra21-1과 TPA-M이 민감도와 특이도에 있어 더 우수한 결과를 보였고 비소세포 폐암환자에서 SCC도 높은 특이도를 나타내었다. ROC(Receiver operating characteristic) curve에 따라 민감도와 특이도를 얻어 가장 정확도가 높은 cutoff value를 CEA $1.25{\mu}/L$, Cyfra21-1 $1.5{\mu}/L$, TPA-M 35U/L, SCC $0.6{\mu}/L$로 하여 민감도와 특이도, 정확도 및 kappa index를 비교해 보아도 Cyfra21-1과 TPA-M이 더 우수한 결과를 나타내었다. 병기에 따른 비소세포 폐암에서의 종양 표지자를 3A 병기까지와 3B 병기이상으로 나누어 비교해보면 3B 병기이상에서 SCC에서만 통계적 유의성을 보였다. (p<0.05) 그러나 종양크기에 따른 종양표지자의 측정치는 통제적 유의성이 없었다.(p>0.05) 결 론 : 혈청 TPA-M과 Cyfra21-1은 폐암환자에 있어 CEA나 SCC보다 민감도와 특이도가 높으며 SCC의 경우 비소세포 폐암에 있어 높은 특이도를 보였으며 SCC의 측정치가 병기와 유관하게 나타났으나 종양의 크기와 병기의 진행과의 관계에 있어서는 더 많은 환자들을 대상으로 한 연구가 행해져야 할 것으로 생각된다.