• Title/Summary/Keyword: ELISA method

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Anti-inflammation and hangover relief effects of Schisandra chinensis (SC) and Lycium chinense (LC) water extracts depending on drug processing and fermentation (포제 및 발효 가공에 따른 오미자와 구기자 물 추출물의 항염증 및 숙취해소 효과)

  • Kim, Ha-Rim;Kim, Sang-Jun;Kim, Sol;Kim, HongJun;Jeong, Seung-Il;Yu, Kang-Yeol;Kim, Seon-Young
    • Herbal Formula Science
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    • v.26 no.4
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    • pp.295-306
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    • 2018
  • Schisandra chinensis (SC) and Lycium chinense (LC) were widely distributed in Asia and the fruit has been used traditionally for medicinal herbs. The processing method was solid-state fermentation using Aspergillus oryzae for 48 h after stir-frying treatment at $220^{\circ}C$ for 12 min. In this study, in vitro the anti-inflammatory effect and in vivo hangover reduction were compared to unprocessed SC and LC water extract. Anti-inflammatory effects have been evaluated in pro-inflammatory mediators which were secreted by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Nitric oxide (NO) was determined using Griess reaction. Proinflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$ and interleukin $(IL)-1{\beta}$ were measured by enzyme-linked immunosorbent assays (ELISA). Alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were compared to processed SC or LC and mixtures thereof (1:1). In vivo study was compared to hangover relief in alcohol-fed mice. After administering a mixture of SC and LC (300 mg/kg) water extract (1:1), mice were fed 3 g/kg of ethanol. Serum was collected at 1, 3, and 5 h intervals to analyze ethanol and acetaldehyde levels using a colorimetric assay kit. The processed SC and LC water extracts compared to raw materials significantly inhibited LPS-induced NO and inflammatory cytokine production in RAW 264.7 cells. The results of the hangover mouse model are also consistent with anti-inflammatory effects. These results suggest that processed SC and LC extracts may be functional materials for the treatment of inflammation and hangover.

Selection of Vaccinia Virus-Neutralizing Antibody from a Phage-Display Human-Antibody Library

  • Shin, Yong Won;Chang, Ki-Hwan;Hong, Gwang-Won;Yeo, Sang-Gu;Jee, Youngmee;Kim, Jong-Hyun;Oh, Myoung-don;Cho, Dong-Hyung;Kim, Se-Ho
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.651-657
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    • 2019
  • Although smallpox was eradicated in 1980, it is still considered a potential agent of biowarfare and bioterrorism. Smallpox has the potential for high mortality rates along with a major public health impact, eventually causing public panic and social disruption. Passive administration of neutralizing monoclonal antibodies (mAbs) is an effective intervention for various adverse reactions caused by vaccination and the unpredictable nature of emerging and bioterrorist-related infections. Currently, vaccinia immune globulin (VIG) is manufactured from vaccinia vaccine-boosted plasma; however, this production method is not ideal because of its limited availability, low specific activity, and risk of contamination with blood-borne infectious agents. To overcome the limitations of VIG production from human plasma, we isolated two human single-chain variable fragments (scFvs), (SC34 and SC212), bound to vaccinia virus (VACV), from a scFv phage library constructed from the B cells of VACV vaccine-boosted volunteers. The scFvs were converted to human IgG1 (VC34 and VC212). These two anti-VACV mAbs were produced in Chinese Hamster Ovary (CHO) DG44 cells. The binding affinities of VC34 and VC212 were estimated by competition ELISA to $IC_{50}$ values of $2{\mu}g/ml$ (13.33 nM) and $22{\mu}g/ml$ (146.67 nM), respectively. Only the VC212 mAb was proven to neutralize the VACV, as evidenced by the plaque reduction neutralization test (PRNT) result with a $PRNT_{50}$ of ~0.16 mg/ml (${\sim}1.07{\mu}M$). This VC212 could serve as a valuable starting material for further development of VACV-neutralizing human immunoglobulin for a prophylactic measure against post-vaccination complications and for post-exposure treatment against smallpox.

Preventive Effect of the Water Extract of Agrimonia pilosa Ledeb and Micronucleus Assay-Based Evaluation of Genotoxicity in Gastritis Animal Models (동물모델을 이용한 짚신나물 물 추출물의 위염 예방효과 및 유전독성 평가)

  • Nho, Jong Hyun;Jang, Ji Hun;Lee, Hyun Joo;Yang, Beodul;Woo, Kyeong Wan;Kim, A Hyeon;Seo, Jae Wan;Hwang, Tae Yeon;Cho, Hyun Woo;Jung, Ho Kyung
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.2
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    • pp.136-142
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    • 2019
  • Background: Agrimonia pilosa Ledeb has been used as a traditional medicine for the treatment of hematuresis and uterine bleeding in Korea. It has been reported to have anti-obesity, anti-diabetes and anti-inflammaotry effect by regulating the inflammatory signaling pathway. However, the preventive effect of Agrimonia pilosa Ledeb on gastritis has not been elucidated. Thus, in the present study, we evaluated the effects of the water extract of Agrimonia pilosa Ledeb (APW) using HCl/EtOH-induced gastritis rat models. Method and Results: Gastritis was induced in rats by HCl/EtOH administration. The rats in each group were orally administered with two doses of APW (100 and 500 mg/kg). Omeprazole was used as a positive control drug. An enzyme-linked immunosorbent assay (ELISA) was used to measure the prostaglandin $E_2$ ($PGE_2$) levels in stomach. The treatment with 500 mg/kg APW reduced the gastric ulcer area. The APW treatment prevented a decreased in $PGE_2$ concentration induced by HCl/EtOH in rats. In the micronucleus test, the ratio of micronucleated polychromatic erythrocytes to polychromatic erythrocytes showed no significant change in the APW-treated group compared with the control group. Conclusions: These results indicate that APW could be used to prevent the gastritis caused by the HCl/EtOH-induced damage to stomach lining. In addition, the APW treatment showed no significant change in results of the micronucleus test. However, further experiments are required to determine how APW influenced the secretion of mucus and gastric acid using the chromosome aberration test and bacterial reverse mutation assay.

Antimicrobial resistance studies in staphylococci and streptococci isolated from cows with mastitis in Argentina

  • Elisa, Crespi;Ana M., Pereyra;Tomas, Puigdevall;Maria V., Rumi;María F., Testorelli;Nicolas, Caggiano;Lucia, Gulone;Marta, Mollerach;Elida R., Gentilini;Mariela E., Srednik
    • Journal of Veterinary Science
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    • v.23 no.6
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    • pp.12.01-12.10
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    • 2022
  • Background: Staphylococcus aureus and Streptococcus agalactiae are the main cause of clinical mastitis in dairy cattle in Argentina, whereas coagulase-negative staphylococci (CNS) and environmental streptococci are the main cause of subclinical mastitis. Bacteria isolated from infected animals show increasing antimicrobial resistance. Objectives: This study aims to determine the antimicrobial resistance of staphylococci and streptococci isolated from milk with mastitis, and to genotypically characterize the methicillin-resistant (MR) staphylococci. Methods: Isolation was performed on blood agar and identification was based on biochemical reactions. Antimicrobial susceptibility was according to the Clinical and Laboratory Standards Institute guidelines. The antimicrobial resistance genes, SCCmec type and spa type were detected by the polymerase chain reaction method. Results: We isolated a total of 185 staphylococci and 28 streptococci from 148 milk samples. Among the staphylococcal isolates, 154 were identified as CNS and 31 as S. aureus. Among the 154 CNS, 24.6% (n = 38) were resistant to penicillin, 14.9% (n = 23) to erythromycin, 17.5% (n = 27) to clindamycin, 6.5% (n = 10) to cefoxitin and oxacillin. Among the S. aureus isolates, 16.1% (n = 5) were resistant to penicillin, 3.2% (n = 1) to cefoxitin and oxacillin (MRSA). Six MR isolates (5 CNS and 1 MRSA) were positive to the mecA gene, and presented the SCCmec IVa. The MRSA strain presented the sequence type 83 and the spa type 002. Among the 28 streptococcal isolates, 14.3% (n = 4) were resistant to penicillin, 10.7% (n = 3) to erythromycin and 14.3% (n = 4) to clindamycin. Conclusions: The present findings of this study indicate a development of antimicrobial resistance in main bacteria isolated from cows with mastitis in Argentina.

Cleaning Methods to Effectively Remove Peanut Allergens from Food Facilities or Utensil Surfaces (식품 시설 또는 조리도구 표면에서 땅콩 알레르겐을 효과적으로 제거하는 세척 방법)

  • Sol-A Kim;Jeong-Eun Lee;Jaemin Shin;Won-Bo Shim
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.228-235
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    • 2023
  • Peanut is a well-known food allergen that causes adverse reactions ranging from mild urticaria to life-threatening anaphylaxis. Consumers suffering from peanut allergies should thus avoid consuming undeclared peanuts in processed foods. Therefore, effective cleaning methods are needed to remove food allergens from manufacturing facilities. To address this, wet cleaning methods with washing water at different temperatures, abstergents (peracetic acid, sodium bicarbonate, dilute sodium hypochlorite, detergent), and cleaning tools (brush, sponge, paper towel, and cotton) were investigated to remove peanuts from materials used in food manufacture, including plastics, wood, glass, and stainless steel. Peanut butter was coated on the surface of the glass, wood, stainless steel, and plastic for 30 min and cleaned using wet cleaning. The peanut residue on the cleaned surfaces was swabbed and determined using an optimized enzyme-linked immunosorbent assay (ELISA). Cleaning using a brush and hot water above 50℃ showed an effective reduction of peanut residue from the surface. However, removing peanuts from wooden surfaces was complicated. These results provide information for selecting appropriate materials in food manufacturing facilities and cleaning methods to remove food allergens. Additionally, the cleaning methods developed in this study can be applied to further research on removing other food allergens.

Effects of Coptidis Rhizoma on the Anti-inflammation and Motor Recovery in Photothrombotic Brain Infarction Model in Rats (광화학적 뇌경색 백서 모델에서 황련의 항염증 및 운동기능 회복에 미치는 효과)

  • Lee, Su-Kyung;Lee, In;Shin, Sun-Ho;Kim, Eun-Young;Shin, Byung-Cheul
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.179-189
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    • 2009
  • Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

Thymus quinquecostatus Extracts Suppress IFN-γ/TNF-α -induced Inflammatory Cytokine in Lung Fibroblast MRC-5 Cells

  • Young-Jae Song;Sa-Haeng Kang;Jae-Bin Seo;Se-Woong Ko;Ju-Ryoun Soh;Jeong-Hyang Park;Dong-Keun Kim;Tae-Hyun Kim;Hyuck-Se Kwon;Na Young Lee;Jong-Sik Jin
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.81-81
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    • 2020
  • Pneumonia is infection of air sacs in lungs by bacteria, viruses, fungi and abnormal atmosphere condition. That causes to inflammation and fills up with pus and fulid in air sacs. The symptoms of pneumonia are heavy cough, fever, shortness and rapid respiration and sputum with blood. Also, that remains sequela after the recovery. Thymus quinquecostatus (TQ) has a strong antibacterial effect. That has been used a natural medicine for bronchitis, asthma and nervous inflammation. In this study evaluated anti-inflammatory effects of TQ on IFN (interferon)-γ/TNF (tumor necrosis factor)-α (20 ng/ml) stimulation. MRC-5 cells were seeded at 1 × 105 cells/well in a 24-well plate and stabilized overnight at 37℃. The cells were treated with various concentrations of TQ extracts (DW, 30, 50, 70, 95% EtOH, 0.1~100 mg/ml) for 4h, Subsequently IFN-γ/TNF-α (20 ng/ml) was added to each well and incubated over 12h. the production of inflammatory cytokines were measured by Enzyme-linked immunosorbentassay(ELISA) method. The absorbance was measured at 450 nm using microplate reader. The treatment with TQ extract at dose of 0.1 to 100 mg/ml did not show cytotoxicity in MRC-5 cells (showed cytotoxicity at 95% EtOH 100 mg/ml). The results demonstrated that stimulation of TNF-α/IFN-γ significantly increased the production of inflammatory mediators in the MRC-5 cells compared with unstimulated cells. By contrast, treatment with TQ extreats doses of 0.001 to 10 mM significantly suppressed the production of IL-6 and IL-8. Especially. The higher percent of ethyl alcohol extracts suppressed the production of inflammatory cytokines. This results are considered that essential oil of TQ is extracted according to ethyl alcohol percent. Therefore, TQ has potential to mitigate pulmonary disease that can be used a medicine for pneumonia

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An Alternative Method for a Rapid Urease Test Using Back-table Gastric Mucosal Biopsies from Gastrectomy Specimen for Making the Diagnosis of Helicobacter pylori Infection in Patients with Gastric Cancer (위암 환자의 헬리코박터 파이로리 감염 진단에 있어서 위절제술 직후 생검된 위점막 조직을 이용한 신속 요소 분해 효소 검사법 도입의 의의)

  • Kim, Sin-Ill;Jin, Sung-Ho;Lee, Jae-Hwan;Min, Jae-Seok;Bang, Ho-Yoon;Lee, Jong-Inn
    • Journal of Gastric Cancer
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    • v.9 no.4
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    • pp.172-176
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    • 2009
  • Purpose: The rapid urease test is a rapid and reliable method for diagnosing Helicobacter pylori infection. However it requires gastric mucosal biopsies during endoscopy, and the test is not covered by national health insurance for patients with gastric cancer. So, we introduced an alternative method for a rapid urease test using back-table gastric mucosal biopsies from gastrectomy specimen. Materials and Methods: Ninety gastric cancer patients underwent an anti H. pylori IgG ELISA test and gastrectomy. Just after gastrectomy, two gastric mucosal biopsies from the prepyloric antrum and lower body of the gastrectomy specimen were taken from the back table in the operative room, and these were fixed immediately with the rapid urease test kit, and the color change was monitored for up to 24 hours. In this study, H. pylori infection was defined as positive when the serology or rapid urease test showed positive results. Results: The positive rate of the rapid urease test and serology was 91.1% and 77.8%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of the rapid urease test and serology were 94.3 and 80.5%, 100 and 100%, 100 and 100%, and 37.5 and 15%, respectively. The accuracy of the rapid urease test was higher than that of serology (94.4 vs. 81.1%, respectively). The rapid urease test showed a higher rate of detecting H. pylori infection than that of serology (McNemar's test, P=0.019). Conclusion: The result of the rapid urease test using back-table gastric mucosal biopsies from a gastrectomy specimen is comparable to the reference data of the conventional rapid urease test using gastric mucosal endoscopic biopsies. Therefore, it can be an alternative diagnostic method for H. pylori infection.

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Immunogenicity and Safety of a Haemophilus influenzae Type b Polysaccharide-Tetanus Toxoid Conjugate Vaccine (PRP-T: HiberixTM) in Korean Infants (우리나라 영아에서 PRP-T(HiberixTM)백신의 면역원성 및 안전성에 대한 연구)

  • Chung, Eun Hee;Kim, Yae Jean;Kim, Yun Kyung;Kim, Dong Ho;Seo, Jeong Wan;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.71-80
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    • 2003
  • Purpose : Four kinds of Haemophilus influenzae type b protein conjugate vaccines, PRPD, PRP-T, PRP-OMP and PRP-CRM197, have been developed, and PRP-T vaccines are currently produced by two manufacturer, $ActHib^{(R)}$ by Aventis and $Hiberix^{TM}$ by GlaxoSmith-Kline Biologicals. The purpose of this study is to evaluate the immunogenicity and safety of $Hiberix^{TM}$ in Korean infants. Methods : Seventy-three healthy infants(43 male infants) were recruited for this study after parental informed consent was obtained. Each infant was vaccinated at 2, 4 and 6 months of age with the study vaccine. At each visit, infants were also immunized with DTaP, trivalent oral polio vaccine and hepatitis B vaccine when indicated. The serum anti-PRP antibody was measured at prevaccination, 2 month later after the 2nd dose, and 1 month later after the 3rd dose by the ELISA method. The local and systemic adverse reactions of vaccination were monitored for 3 consecutive days after each immunization. Immunogenicity of vaccine was evaluated in infants who received all the scheduled immunization and the adverse reactions were evaluated for infants who received at least one dose of the study vaccine. Results : Among seventy three infants, enrolled in this study; sixty three(37 male infants) completed all the scheduled immunizations. The geometric mean titer(GMT) of anti-PRP antibodies at prevaccination was 0.17 ${\mu}g/mL$(95% confidence interval[CI]; 0.13~0.22). The GMT of anti-PRP antibodies increased to 4.14 ${\mu}g/mL$(95% CI; 2.65~6.48) at 2 month later after the 2nd dose of PRP-T and 14.65 ${\mu}g/mL$(95% CI; 10.83~19.81) at 1 month later after the 3rd dose. Anti-PRP antibody ${\geq}0.15$ ${\mu}g/mL$, was observed in 98.4%(95% CI; 91.8~100) after 2 doses and 100%(95% CI; 100~100) after 3 doses. Anti-PRP antibody ${\geq}1.0$ ${\mu}g/mL$, was obtained in 77.8%(95% CI; 67.5~88.0) after 2 doses, and 98.4%(95% CI; 95.3~100) after 3 doses. Most of the adverse reaction after vaccination were mild. Irritability, the most common systemic reaction, was observed in 45.5%, followed by drowsiness(30.5%), poor feeding(26.7%) and fever(5.6%). Among the local reactions tenderness was observed in 7.9%, redness(${\geq}5$ mm) in 2.8% and swelling(${\geq}5$ mm) in 1.8%. Conclusion : The PRP-T vaccine used in this study was highly immunogenic and safe in Korean young infants. The finding that high GMT and high frequency of infants with a protective titer achieved after 2 doses is consistent with the previous studies which were done with a PRP-T vaccine of other manufacturer. This study suggests that the immunization schedule of PRP-T vaccine for Korean infants may need re-evaluation.

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Circulating Cytokine Levels and Changes During the Treatment in Patients with Active Tuberculosis in Korea (결핵 환자의 치료경과 중 혈청 내 Cytokine 분비와 변화)

  • Ryu, Yon-Ju;Kim, Yun-Jung;Kwon, Jung-Mi;Na, Youn-Ju;Jung, Yu-Jin;Seoh, Ju Young;Cheon, Seon Hee
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.2
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    • pp.140-153
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    • 2003
  • Background : The cell-mediated immune reaction to tuberculosis infection involves a complex network of cytokines. The extent of inflammation, tissue damage and severity of the disease suggested to be determined by the balance between extent and duration of the proinflammatory cytokine response versus those of the suppressive cytokines. The systemic cytokine response in pathogenesis of tuberculosis can be assessed by measuring serum cytokine levels. Method : Serum interleukin-1 beta(IL-$1{\beta}$), IL-2, IL-4, IL-6, IL-10, IL-12(p40), tumor necrosis factor-alpha(TNF-${\alpha}$), interferon-gamma(IFN-${\gamma}$) and transforming growth factor-beta(TGF-${\beta}$) levels were measured in 83 patients with pulmonary tuberculosis, 10 patients with endobronchial tuberculosis before treatment and 20 healthy subjects by using a sandwich ELISA. In patients with pulmonary tuberculosis, they were divided into mild, moderate and far advanced group according to the severity by ATS guidelines. To compare with those of pretreatment levels, we measured serum IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-10, IL-12(p40), TNF-${\alpha}$, IFN-${\gamma}$ and TGF-${\beta}$ levels in 45 of 83 patients with pulmonary tuberculosis after 2 and 6 months of treatment. Results : 1) In sera of patients with active pulmonary tuberculosis(n=83), IL-$1{\beta}$, IL-6(p<0.05), TNF-${\alpha}$, and IFN-${\gamma}$ were elevated and TGF-${\beta}$ was decreased comparing to control. IL-2, Il-12(p40), IL-4 and IL-10 were similar between the patients with tuberculosis and control. 2) In endobronchial tuberculosis, IL-6 and TNF-${\alpha}$ were elevated and TGF-${\beta}$ was decreased comparing to control. IL-12(p40) seemed to be elevated comparing to pulmonary tuberculosis. 3) Far advanced tuberculosis showed markedly elevated IL-6 and IFN-${\gamma}$ level(p<0.05). 4) The significant correlations were noted between IL-1, IL-6 AND TNF-${\alpha}$ and between IL-12, Il-2 and IL-4(p<0.01). 5) After 2 and 6 months of standard treatment, the level of IL-6 and IFN-${\gamma}$ was significantly decreased(p<0.05). Conclusion : These results showed that an altered balance between cytokines is likely to be involved in the extent of inflammation, tissue damage and severity of the disease tuberculosis. But, it should be considered diversities of cytokine response according to type of tuberculosis and immunity in clinical application and interpreting future studies.