• Title/Summary/Keyword: ELISA antibody

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Cell-mediated immunity in mice infected with Acanthumoeba culbertsoni (Acanthamoeba culbertsoni 감염에 있어서 세포 매개성 면역)

  • Kim, Myeong-Jun;Sin, Ju-Ok;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.28 no.3
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    • pp.143-154
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    • 1990
  • Observations were made on the differences of cell-mediated responses in mice of three infectiorl groups di여erently scheduled in their severity with pathogenic Acanthamoeba culbertseni. Infections were done by dropping $5{\;}{\mu}l$ saline suspension containing $3{\times}10^3,{\;}1{\times}10^4,{\;}or{\;}1{\times}10^5$ trophosoites, respectively. Amoebae were cultured anenically in CGV medium and inoculated into the right nasal cavity of CSH/HeJ mice aging around 6∼8 weeks, under the anesthesia by intraperitoneal injection of secobarbital. Delayed type hypersensitivity (DTH) responses in footpad and blastogenlc responses of mouse spleen cells using ($^3H$)-thymidine and the serum antibody titer were measured up to day 14 after infection, and natural killer cell activities were measured up to day, i after infection. The results obtained in this study were as follows: 1. The mice infected with $3{\times}10^3$ trophosoites showed mortality rate of 17%, and 345 in the mice infected with $1{\times}10^4$ trophozoites and 65% with $1{\times}10^5$ trophozoites. 2. In regard to DTH responses in all experimental groups, the level increased on day 7 and declined on day 14 after infection, but their differences could not be noted between infected and control groups. 3. The blastogenic responses of splenocytes treated with amoeba Iysates and lipopolysaccharides (LPS) showed no difference from the control group. The blastogenic responses of splenocytes treated with concanavalin A were declined significantly in the experimental group as compared with the control group, but the blastogenic responses of splenocytes treated with polyinosinic acid were not different from the control group. There was also no difference among three infected groups. 4. The cytotoxic activity of the natural killer cells was activated on day 1 after infection and declined to the level of control group on day 2 in all experimental groups. On day 5 after infection, the natural killer cell cytotoxicity was significantly suppressed as compared with the control groups. 5. The serum antibody titers of the infected mice increased after day 7, but there was no statistical difference between the three infected groups. In summary of the results, there was no difference in cell-mediated immune responses of three experimental groups scheduled with different infection intensities. But there was a significant difference in cell$.$mediated immune responses between infected and control mice. It is considered that cell-mediated immune responses should be involved in murine model infected with A. culbertsoni.

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Adjuvant Effect of PAMAM Dendrimer on the Antigenicity of Keyhole Limpet Hemocyanin in Balb/c Mice (Balb/c 마우스에서 Keyhole limpet hemocyanine (KLH)의 항원성에 대한 PAMAM dendrimer 의 면역증강 효과)

  • Lee, Ga-Young;Kim, Min Jee;Kim, So Yeon;Lee, Kyung Bok;Oh, Dong Hyun;Cho, Young Ho;Yoo, Yung Choon
    • Journal of Life Science
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    • v.30 no.10
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    • pp.905-911
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    • 2020
  • The adjuvant effect of PAMAM dendrimer G4 (PAMAM) on the induction of humoral and cellular immune responses against keyhole limpet hemocyanin (KLH) was examined. Mice were immunized subcutaneously twice at two-week intervals with KLH, with or without PAMAM dendrimer (100 ㎍/mouse), and the mice immunized with KLH+PAMAM showed significantly higher antibody titers against KLH than those immunized with KLH alone. The assay for determining the isotypes of the antibodies showed that PAMAM augmented the KLH-specific antibody titers of IgG1, IgG2a, IgG2b, IgG3, and IgM. In addition, mice immunized twice with KLH+PAMAM followed by a subcutaneous injection of KLH (20 ㎍/site) 7 weeks after the primary immunization exhibited a higher delayed-type hypersensitivity (DTH) reaction than those treated with KLH alone. In an in vitro analysis of T lymphocyte proliferation in response to KLH in week 8, the splenocytes of mice treated with KLH+PAMAM showed significantly higher proliferating activity than those treated with KLH alone, and the culture supernatants of cell cultures from mice immunized with added PAMAM dendrimer showed higher levels of KLH-specific cytokine (IL-4 and IFN-r) production. These results suggest that PAMAM dendrimer G4 possesses a potent immune-adjuvant activity for enhancing both humoral and cell-mediated immunity specific to foreign antigens.

Purification of cystic fluid antigen of Taenia solium metacestodes by affinity chromatography using monoclonal antibody and its antigenic characterization (단세포군항체에 의한 유구낭미충 낭액 특이항원의 순수분리 및 항원특성 관찰)

  • Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull;Hwang, Eung-Soo;Cha, Chang-Yong
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.145-158
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    • 1986
  • This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatogaphy using specific monoclonal antibody(McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secreted antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen (A-Ag) and unbound pool CF (U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitivity was about 70% of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis.

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Mumps & Rubella-specific IgG in MMR Vaccinees (소아의 연령증가에 따른 볼거리 및 풍진 항체)

  • Cheon, Haewon;Sin, Yeong-Gyu;Lee, Kangwoo;Lee, Youngkyoo;Chung, Jitae;Tockgo, Youngchang
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.175-184
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    • 1996
  • Purpose : This study was intended to measure seropositivities and the levels of mumps- and rubella-specific IgG of MMR vaccinees over 17 months of age in Korea. Materials and Methods : From June 1994 to April 1995 we obtained sera from visitors of well baby clinic and patients in Korea University Hospital, who were MMR vaccinees over 17 months of age and had no evidence of immunodeficiency. These 275 study population include 145 males and 130 females. Mumps- and rubella-specific IgG antibody levels were measured by ELISA. Cut-off values for seropositivity were 20 GU(Gamma Unit) in mumps and 0.17 in rubella. Results : 1) As age increased, seropositivities of mumps-specific IgG increased significantly, being 69.0% in 1.5~2 year, 75.0% in 3~4 year, 76.0% in 5~6 year, 90.0% in 7 year, 100% in 8 year, 96.9% in 9 year, 97.4% in 10 year, 97.4% in 11 year, and 96.6% in 12 year of age(p<0.001). 2) As age increased, the levels of mumps-specific IgG antibody(mean${\pm}$standard deviation, GU) increased significantly, being $64.9{\pm}66.5$ in 1.5-2 year, $117.7{\pm}126.4$ in 3~4 year, $152.3{\pm}147.1$ in 5~6 year, $194.3{\pm}168.2$ in 7 year, $258.1{\pm}190.6$ in 8 year, $193.1{\pm}130.1$ in 9 year, $225.7{\pm}119.6$ in 10 year, $220.7{\pm}114.3$ in 11 year, and $222.3{\pm}127.1$ in 12 year of age(p<0.001). There was positive correlation between age and mumps-specific antibody level (r=0.3282, p<0.001). 3) As age increased, seropositivities of rubella-specific IgG decreased significantly, being 72.4% in 1.5~2 year, 75% in 3~4 year, 72% in 5~6 year, 60% in 7 year, 44.4% in 8 year, 40.6% in 9 year, 28.2% in 10 year, 23.1% in 11 year, and 17.2% in 12 year of age(p<0.001). 4) As age increased, rubella-specific IgG decreased significantly, being $0.462{\pm}0.356$ in 1.5~2year, $0.438{\pm}0.306$ in 3~4 year, $0.287{\pm}0.179$ in 5~6 year, $0.204{\pm}0.139$ in 7 year, $0.189{\pm}0.153$ in 8 year, $0.124{\pm}0.121$ in 9 year, $0.093{\pm}0.114$ in 10 year, $0.104{\pm}0.135$ in 11 year, and $0.080{\pm}0.001$ in 12 year of age(p<0.001). There was negative correlation between age and rubella-specific IgG titer (r=-0.551, p<0.001). Conclusions : Eventhough seropositivities and the level of mumps-specific IgG increased as age increased, they are not enough to prevent mumps infection in 1.5 to 6 years of age. Seropositivities and the level of rubella-specific IgG decreased as age increased. Appropriate change in vaccine schedule may be needed to decrease the risks of mumps and rubella infection.

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Effect of Antibody Titer on Xenograft Survival in Pig-To-Dog Heterotopic Cardiac Xenotransplantation -Opening of Xenotrasplantation Era- (돼지\longrightarrow개 이소이종심장 이식모델에시 생존에 미치는 항체 역가의 영향 -이종이식시대의 개막-)

  • 이정렬;김희경;김지연;최대영;이재형;위현초;강희정;김영태;강병철
    • Journal of Chest Surgery
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    • v.37 no.5
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    • pp.391-400
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    • 2004
  • Xenotransplantation in discordant species results in immediate and irreversible hyperacute rejection due to natural antibodies, IgM. With this, antibody depletion is one option to reduce hyperacute rejection, we investigated the effect of PCPP (postcentrifugal plasmapheresis) on the depletion of natural antibodies and the effect of antibody titer on xenograft survival. Material and Method: Outbred swines (n=4) weighing 10∼20 kg were used as donors and mongrel dogs (n=4) weighing 25∼30 kg were used as recipients. Recipient canines underwent plasmapheresis (COBE TPE Laboratories, Lakewood. CO, USA). Pre-transplantation PCPP was peformed on day -2 and day 0. There were three groups (Group 0: no PCPP, Group 1: 1 pla sma-volume (PV) at day -2 and 2 PV at day 0, Group 2: 2 PV at day -2 and 2 PV at day 0). A swine heart was heterotopically transplanted into a recipient's abdominal infrarenal aorta and inferior vena cava. Mean percent depletion of total IgM and IgG in plasma of the recipients was calculated. Serum albumin, electrolyte, complement activity and coagulation factors were measured. Histopathologic examination of heart specimens was performed. Result: Mean percent depletion of IgM and IgG were 95.7$\pm$1.2%, 80.5$\pm$2.4% in the group 2 at the end of PCPP. The percent depletion of serum albumin concentration was decreased from 2.8 to 1.4 g/㎗ in the group 1 and 3.0 to 1.5 g/㎗ in the group 2. Complement hemolytic activity was decreased in group 1 and 2, but returned to normal level within 24 hours. Complement hemolytic activity was reduced to 10% of pre-PCPP level in group 2. Serum fibrinogen decreased to 20% or less and was recovered within 24 hours in group 2. Antithrombin III decreased but less than fibrinogen. PT and aPTT were sometimes but not always prolonged during plasmapheresis. After plasmapheresis, PT and aPTT were prolonged beyond the measurable level. D-dimer was not found during PCPP, but appeared and maintained from 10 minutes after trasplantation. Graft Survival time was 5 min in group 0, and it was 90$\pm$0 min in the group 2. Histopathologic changes were more typically characterized by edema, hemorrhages, thrombosis in all groups at the end of experiment. Conclusion: PCPP effectively removed immuoglobulins and reduced the titer of natural antibodies, as a result, significantly prololonged swine heart xenograft survival.

Mumps- and Rubella-specific IgG Levels in Adolescents (청소년기의 연령증가에 따른 볼거리 및 풍진 항체가 변동)

  • Cheon, Hae Won;Shin, Young Kyoo;Lee, Kang Woo;Choung, Ji Tae;Tockgo, Young Chang
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.128-135
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    • 1998
  • Purpose : This study was intended to measure seropositivities and the level of mumps- and rubella-specific IgG of MMR vaccinees from 12 to 17 years of age in Korea. Materials and Methods : From May 1996 to July 1996 we obtained sera from students of 1 middle and 2 high schools in Seoul, who were MMR vaccinees from 12 to 17 years of age and had no evidence of immunodeficiency. These 216 study population include 110 males and 106 females. Mumps- and rubella-specific IgG antibody levels were measured by ELISA. Cut-off values for seropositivity were 20 U(Gamma Unit) in mumps and over 0.17 in rubella. Results : 1) As age increased, seropositivities to mumps increased, being 68.4% in 12 year, 79.3% in 13 year, 72.2% in 14 year, 82.0% in 15 year, 87.5% in 16 year, 87.0% in 17 year, which however has no statistical significance. 2) As age increased, the level of mumps-specific IgG antibody(mean+standard deviation, GU) increased, being $52.0{\pm}49.2$ in 12 year, $65.9{\pm}51.4$ in 13 year, $71.1{\pm}66.0$ in 14 year, $67.8{\pm}53.6$ in 15 year, $82.8{\pm}67.8$ in 16 year, $92.0{\pm}68.9$ in 17 year, which however has no statistical significance. 3) As age increased, seropositivities of rubella-specific IgG increased significantly, being 26.3% in 12 year, 20.7% in 13 year, 50.0% in 14 year, 67.2% in 15 year, 66.7% in 16 year, 65.2% in 17 year(P<0.001). 4) As age increased, rubella-specific IgG increased significantly, being $0.13{\pm}0.145$ in 12 year, $0.087{\pm}0.101$ in 13 year, $0.194{\pm}0.168$ in 14 year, $0.260{\pm}0.187$ in 15 year, $0.305{\pm}0.213$ in 16 year, $0.325{\pm}0.221$ in 17 year(P<0.001). There was positive correlation between age and rubella-specific IgG titer(rubella-specific $IgG=0.0517{\times}age-0.5586$, r=0.3752, P<0.001). Conclusion : In adolescent, seropositivities and the level of mumps-specific IgG remained relatively high, but approximately 20% of study population showed seronegativity. Seropositivities and the level of rubella-specific IgG showed the lowest level at 13 years of age and were increased with age after 14 years of age. Further evaluation may be needed to elucidate the cause of these changes of rubella-specific IgG.

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IL-23 P19 Expression Induced by IL-17 and $IL-1{\beta}$ in Rheumatoid Arthritis Synovial Mononuclear Cells (류마티스관절염 환자의 활액 세포에서 IL-17과 $IL-1{\beta}$에 의한 IL-23p19의 발현 증가)

  • Cho, Mi-La;Heo, Yu-Jung;Oh, Hye-Jwa;Kang, Chang-Min;Lee, Seon-Yeong;Hong, Yeon-Sik;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.29-37
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    • 2008
  • Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.

Protective Efficacy of Recombinant Proteins Adenylate Kinase, Nucleoside Diphosphate Kinase, and Heat-Shock Protein 70 against Mycobacterium tuberculosis Infection in Mice (유전자 재조합 단백질 Adenylate Kinase, Nucleoside Diphosphate Kinase와 Heat-Shock Protein 70의 결핵균에 대한 방어면역효능 분석)

  • Lee, Seung-Heon;Lee, Eun-Gae;Kim, Su-Yeon;Cho, Sang-Nae;Park, Young-Kil;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.2
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    • pp.142-152
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    • 2005
  • Background : Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method : M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and $IFN-{\gamma}$ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result : Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and $IFN-{\gamma}$ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion : Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.

A Case of Paragonimiasis in the Abdominal Subcutaneous Tissue with Pleural Effusion (흉막 삼출을 동반한 복부 피하 지방 조직의 폐흡충증 이소기생 1례)

  • Im, So Hi;Shin, Sung Hwan;Song, Myung Jun;Kim, Jin Woo;Kim, Seung Joon;Lee, Sook Young;Kim, Young Kyoon;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.5
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    • pp.550-554
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    • 2004
  • A paragonimiasis infestation is caused by the paragonimus species. It is commonly found in the lung but has also been found to exist extrapulmonary infestations including cerebral, spinal, subcutaneous, hepatic, splenic, abdominal, urinary, and gynecologic infestation. On the other hand, a cutaneous infestation is extremely rare. Human infestation is caused by ingesting raw or undercooked intermediate hosts. Because paragonimus westermani larva mature to an adult worm in the lung, the possibility of identifying the adult worm of paragonimus westermani at extrapulmonary region is very rare. Case : After ingesting a fresh-water crab 1 month prior to the hospital visit, a 45-year old female patient was suffering from right pleuritic chest pain during that 1 month. The patient also complained of a palpable mass that was movable and migrating, and it was localized at the right upper quadrant of the abdomen. The eosinophil fraction of the white blood cell of peripheral blood and pleural fluid was elevated to 55.1% and 90%, respectively. Parasite eggs were not found in her sputum and stool examination. By using the enzyme-linked immunosorbent assay (ELISA), the paragonimus-specific IgG antibody titer was elevated to 0.28. During incisional biopsy, we were able to find the young adult worm of paragonimus westermani. We experienced the rare case of ectopic paragonimiasis with pleural effusion that was confirmed by identifying the adult worm of paragonimus westermani within the abdominal subcutaneous tissue. We report a case with brief literature reviews.

Study on Alteration of Interleukin-$1{\beta}$, -2, -6 Production and Serum Level in Schizophrenic Patients (정신분열증 환자에서 Interleukin-$1{\beta}$, -2, -6 생산능과 혈청농도 변화에 관한 연구)

  • Kim, Yong-Ku;Lee, Min-Soo;Suh, Kwang-Yoon
    • Korean Journal of Biological Psychiatry
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    • v.1 no.1
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    • pp.98-108
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    • 1994
  • The etiology and pathophysiology of schizophrenia remain unknown. It has been postulated that infectious-autoimmune process may play a role in the pathogenesis of symptoms in some schizophrenic patients. Findings of altered interleukin(IL) regulation have been regarded as additional proof that schzophrenia has an infectious-autoimmune background. In the present study, we measured mitogen-stimulated production of and serum level of IL-$1{\beta}$, IL-2, IL-6 using ELISA in 16 neuroleptic-free schizophrenic patients and in 16 age, sex matched healthy controls. The results were as follows : 1) There was a significant decrease of IL-2 production in schizophrenic patients than in normal controls(respectively $1.90{\pm}0.13ng/m{\ell}$, $2.79{\pm}0.14ng/m{\ell}$, p<0.001). But there was no significant difference of IL-$1{\beta}$ production and IL-6 production between schizophrenic patients and normal controls. 2) There was a significant increase of serum level of IL-2 in schizophrenic pateitns than in normal controls(respectively $184.8{\pm}12.8pg/m{\ell}$, $104.2{\pm}34.2pg/m{\ell}$, p<0.01). Serum level of IL-$1{\beta}$ was partially detected in both groups and serum level of IL-6 was not detected in both groups. 3) There was no significant differences of IL-$1{\beta}$, -2, -6 production & serum level of IL-2 according to male vs female, paranoid type vs undifferentiated type, drug-naive group vs drug-free group in schizophrenic patients. 4) There was significant correlation between IL-$1{\beta}$ and IL-6 production(r=0.86, p<0.001). No correlation between IL-$1{\beta}$, -2, -6 production, serum level of IL-2 and age, duration of illness, and BPRS score was found. It has been suggested that the low lymphocyte production of IL-2 in the patients with autoimmune disease occurs because the T cells are activated and lymphocyte-derived IL-2 has been released into the serum. The authors suggest that decreased IL-2 production in our schizophrenic patients is due to increased IL-2 serum level in those patients. Thus our finding of low IL-2 production and high serum level of IL-2 in our schizophrenic patients is compatible with the possibility that our patients have an autoimmune process. Further study on relationship between IL alteration and other immunological abnormalities(the presence of serum autoantibody and of anti-brain antibody, $CD4^+$, $CD8^+$ cell index, etc) in schizophrenic patients will be warranted.

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