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Diagnostic Significance of Cytotoxic Genes Expression by Western blotting of Serum in Helicobacter pylori Infection (Helicobacter pylori 감염 환자에서 Western blot 법에 의한 혈청내 세포독성 유전자의 발현에 관한 연구)

  • Kim, Dae-In;Lee, Goo;Shu, Jung-Ill;Lee, Chang-Woo;Kim, Jung-Ran;Ha, Gyoung-Yim;Lee, Kyu-Chun;Nam, Kyung-Soo;Yang, Chang-Heon
    • Journal of Life Science
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    • v.10 no.6
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    • pp.630-639
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    • 2000
  • The gastric pathogen Helicobacter pylori(H. pylori) establishes long-term chronic infection that can lead to atrophic gastritis, intestinal metaplasia, and gastric cancer. H. pylori, which express cytotoxic genes is now recohnized as a cause of peptic ulcer and is also a major risk factor for the development of gastric adenocarcinoma. We performed this study 1) to assess the detection rate of H. pylori according to direct investigation of bacteria of gastric biopsy specimen and two serologic tests of GAP test and Helico blot 2.0 system in the symptomatic and non-symptomatic group 2) to evaluate and compare the efficacy of two serologic tests of GAP test and Helico blot 2.0 system for the diagnosis of H. pylori infection. Forty-nine patients were positive for H pylori infection based on direct investigation of bacteria by histology. The detection rates of H. pylori infection based on direct investigation of bacteria by histology. The detection rates of H. phlori were significantly lower in gastric cancer than in other gastroduodenal disease(p<0.05). The concordance of two serologic tests of GAP test and Helico blot 2.0 system is poor. There was no statistically significant difference between the expression rate of CagA and VacA in the symptomatic and non-symptomatic group. Although Helico blot 2.0 system may not displace GAP test, it was a very sensitive serologic test for the diagnosis of H. pylori infection and it was used to detect IgG antibodies to H. pylori-specific antigens, including CagA, VacA and the various urease subunit. Our data suggest that further investigation is needed to determine whether or not the serologic expression of cytotoxic gene may be clinical usefulness of diagnostic methods in the gastroduodenal disease.

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Effects of Fermented Milk Containing Herb Extract from Acanthopanax divaricatus var. albeofructus and Codonopsis Ianceolata on the Immune Status of Mouse (흰털오가피와 더덕 추출물을 첨가한 발효유 급여가 마우스의 면역기능에 미치는 영향)

  • Lim, Sang-Dong;Seong, Ki-Seung;Kim, Kee-Sung;Han, Dong-Un
    • Food Science of Animal Resources
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    • v.27 no.1
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    • pp.95-101
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    • 2007
  • We have investigated the immunomodulatory activity of water extracts of Acanthopanax divaricatus var. alveofructus in male ICR mice. Mice were administrated with fermented milk containing freeze-dried extract 3 mg/Kg (A), 9 mg/kg (B), 27 mg/Kg (C) per body weight with A. divaricatus var. alveofructus (loots : leaves : stem) : Codonopsis lanceolata = (5 : 2 : 1.5) : 1.5 for 7 and 10 weeks, respectively. Body weight, relative organ weight, cellularity of lymphoid organs, plaque- forming cell (PFC) assay, agglutination (AGG) test and lymphoproliferation were examined in various groups of animals. Any significant differences of body weight gain were not recorded in the tested ICR mice. There was significant different (p<0.05) in the spleen index in B group of 10 weeks and C group of 7 weeks fed mouse. The thymus gain weight was increased during administration of the extract, but there was no significant increase on other organs gain. Humoral immunity as measured by PFC showed more decreased PFC level in 10 weeks than in 7 weeks. In the HT, A. divaricatus var. albeofructus extract also showed a significant increase (p<0.05) in C group of 10 weeks. Administration of extracts from A. divaricatus var. albeofructus increased significantly in the production of IgG antibodies on the mice immunized with SRBC in B group of 7 and 10 weeks (p<0.05).

Three Cases of Pulmonary Paragonimiasis in a Family after Ingestion of Raw Fresh-water Crayfishes caught in a Stream of Wolchulmountain (월출산 민물가재 생식에 의한 한 가족내 폐흡충증 3례)

  • Lee, Hyun Jung;Sun, Gyu Geun;Na, Kyung Hee;Park, Sun Young;Kim, Eun Young;Kim, Kyoung Sim;Kim, Yong Wook;Kim, Suk Il
    • Pediatric Infection and Vaccine
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    • v.9 no.2
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    • pp.222-229
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    • 2002
  • Human infection with the lung fluke Paragonimus westermani has become rare in Korea. Human paragonimiasis is caused by eating raw fresh-water crayfishes or crabs infected with larval metacercariae. Recently, we experienced three cases of pulmonary paragonimiasis in a family. They ate raw fresh-water crayfishes that lived in a stream in Wolchulmountain. All the parients had hypereosinophilia and pulmonary infiltrates with pleural effusion or hydropneumothorax, which did not improve on antibiotics. Ingestion of raw crayfishes was a clue for paragonimiasis. Positive results were shown both on intradermal skin test and ELISA for Paragonimus westermani specific IgG. After treatment with praziquantel, the patients showed an improvement. This is the first familial human paragonimiasis, reported from Wolchulmountain in Chonnam Province where there had been no previous cases of paragonimiasis.

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Antibody changes in paragonimiasis patients after praziquantel treatment as observed by ELISA and immunoblot (폐흡충증에서 치료 후 혈청내 특이 항체의 변화)

  • 조승열;김석일
    • Parasites, Hosts and Diseases
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    • v.27 no.1
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    • pp.15-22
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    • 1989
  • To observe antibody changes after praziquantel treatment in paragonimiasis, a total of 46 serum samples from 13 serologically diagnosed patients was collected for 4~28 months. The specific antibody (IgG) levels were measured by enzyme- linked immunosorbent assay (ELISA). All but one patient who needed retreatment became symptom-free within a week. Antibody levels were dropped near to or below a cut-off absorbance (abs.) of 0.25 in varying intervals from 4 to 18 months. Of 9 patients who were retested within 3 months, 5 revealed temporary elevation of antibody level. After the elevation, the levels be낙an to decline slowly to negative ranges. If treated earlier after symptoms developed, the temporary elevation did not occur and intervals to negative conversion were shorter. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) /immunoblot, antigen-antibody reactions in individual patient faded gradually without significant changes in reacting antigen bands.

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Immunoblot observation of antigenic protein fractions in Paragonimus tvestermani reacting with humall patients sera (폐흡충 항원단백질에 대한 폐흠충증 한자 혈청의 반응 양상)

  • Kim, Sung-Hwan;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.26 no.4
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    • pp.239-244
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    • 1988
  • In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot

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Variation of antigenic proteins of eggs and developmental stages of Paragonimus westermani (폐흡충 발육 단계에 따른 항원 단백질의 변화)

  • Yoon KONG;Joon-Yong CHUNG;Doo-Hee YUN;Lee-Su KIM;Shin-Yong KANG;Akira ITO;Liang MA;Seung-Yull CHO
    • Parasites, Hosts and Diseases
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    • v.35 no.3
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    • pp.197-202
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    • 1997
  • Diagnosis of early paragonimiasis is difficult because parasitological evidence is not easily obtained. Antibody tests have been proposed as a good substitute for classical diagnostic techniques. Using the crude extracts of Parnsonimus westermnni eggs, metacercariae. 4- and 7-week Juveniles, and 16-week adults as antigens, we observed the early antibody responses. Sera were obtained from 4 experimental cats, fed 50 metacercariae each, at intervals until 13 weeks post-infection. Antibody (IgG) responses were identified by ELISA using extracts of 4-week juveniles, followed by those of 7- and 16-week worms. Antibody responses were minimal against the metacercarial extracts. Antibodies to p. westemoni egg extracts were elevated after 10 weeks post-infection. In immunoblot analysis, more than nine protein bands in 4-week juveniles reacted with the early infection sera. Antigenic proteins in adult worms were different from those of juveniles. After four weeks of infection, 32 and 35 kDa bands in the adult extracts were increasingly reactive. Egg specific proteins at 28, 46 and 94 kDa were reactive only after 10 weeks. Antigenic components reactin료 to the early infection sera changed during the maturation stages of P. westermani; almost all juvenile antigens were replaced by adult antigen components.

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Serodiagnosis of canine toxoplasmosis by latex agglutination and indirect fluorescent antibody test (Latex 응집반응과 간접형광항체법을 이용한 개 톡소플라즈마병의 혈청학적 진단)

  • Lee, Byung-hoon;Lee, Eung-goo;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.641-647
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    • 1992
  • This study was conducted to determine the serum antibodies against toxoplasma in the artificially infected dogs, pet and street dogs by latex agglutination (LA) and indirect fluorescent antibody (IFA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical Co.) and IFA test was carried out with rabbit-anti-dog IgG labelled with FITC (Cappel Co.) and toxo-antigen slides prepared in laboratory. The results obtained were as follows ; 1. Antibodies to Toxoplasma gondii in the artificially infected dogs were detected firstly at the Day 8 in IFA and Day 9 in LA test after inoculation. Positive antibody reactions by these tests were declined gradually afterward but maintained up to 12 weeks. 2. In LA test serum antibody titers in 310 test sera were shown as 10 cases(32%) in 1 : 32.5(1.0%) in 1 : 64, 4(1.3%) in 1 : 128 and 2(0.7%) in 1 : 256. In IFA test serum antibody titers 310 test sera were shown as 17 cases(5.5%) in 1 : 64, 8(2.6%) in 1 : 128 and 5(1.6%) in 1 : 256. 3. In the total of 310 sera from pet and street dogs toxoplasma antibody positive rates were 21 cases (6.8%) by LA and 30 cases (9.7%) by IFA test and the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 4. In the total of 115 sera from pet dogs toxoplasma antibody positive rates were 12 cases(10.4%) by LA and 15(13.0%) by IFA test. And in the 195 street dogs the positive rates were 9 cases(4.6%) by LA and 15(7.7%) by IFA test. Also, the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 5. Agreement of reactivity between LA and IFA test for 310 sera was 91.3% in total of 283 cases consisting of 12 cases(3.9%) of both LA and IFA positive and 271 cases(87.4%) of LA and IFA negative. 6. LA test was almostly equivalent to the IFA test in producibility and proved to be a simple tool for the screening of toxoplasma antibody in laboratory.

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耐 마모 코우팅 생산을 위한 CVD와 PVD 기술

  • O, Seung-Tak
    • Journal of the Korean institute of surface engineering
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    • v.20 no.1
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    • pp.15-24
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    • 1987
  • 超硬合金과 高速度鋼에 TiC, TiN 그리고 $Al_2O_3$의 耐 마모 코우팅시에 사용되는 CVD와 PVD 기술을 M E Sjostrand와 A G Thelin 등이 본 논문에서 논의하였다. 또한 CVD와 PVD 의 특별한 장점과 응용영역에 대하여도 토의하였다. 수년동안 CVD 기술은 금속질화물, 금속탄화물 그리고 금속산화물 등의 耐 마모 코우팅을 생산하는데 성공적으로 사용되어 왔다. Munster와 Ruppert에서는 1950년대 초기에 TiC와 TiN의 CVD 공정을 연구했는데 이런 코우팅 재료의 높은 화학적 안정성과 더불어 高硬鋼라는 독특한 성질로 인해 그 주요 영역이 명백해졌다. 1968년에 처음으로 CVD법에 의한 보호막 코우팅의 대규모 응용이 이루어졌는데 그것은 超硬合金 절삭공구의 코우팅이었다. (그림1 참고) 이것은 硬금속산업에 있어서 가장 중요한 발전단계였다. TiC, TiN 그리고 $Al_2O_3$로 코우팅된 공구의 생산은 1970년대에 빠른 성장을 보였으며 오늘날 사용되는 절삭날의 50% 이상을 점유하고 있다. (그림2 참고) TiC와 TiN은 현재 이용되고 있는 모든 耐 마모 코우팅 중에서 독보적인 위치를 차지하고 있다. 그 이유는 생산과정이 비교적 단순하기 때문이다. 지난 5년동안 절삭공구와 總形바이트의 코우팅에 대한 PVD기술의 응용이 폭발적으로 증가했다. 증착기술인 이들 CVD와 PVD 각각은 자체의 독특한 장점이 있으므로 그 응용영역은 증착조건, 즉 기지금속의 접착성과 증착온도 그리고 증착속도등에 따라 매우 다를 것이다. PVD 공정으로 인해 고속도 공구강이 급속도로 발전되었고 더우기 PVD공정은 500$^{\circ}C$ 이하의 증착온도에서 brazed carbide 공구의 코우팅을 가능하게 하였다. 따라서 두 증착기술은 서로 상반적이라기 보다는 상호보완적인 것이라고 생각하는 편이 더 좋다.TEMPLA에 비해 CLB의 수가 15.58% 감소되었다. 높은 활성을 나타내었다. 정제된 효소의 최적온도는 40$^{\circ}C$이었으며 20~50$^{\circ}C$에서 비슷한 활성을 나타내었고, 30$^{\circ}C$에서는 60분동안 효소활성이 거의 상실되지 않았다. 정제된 효소는 ethanol과 chloroform 처리에는 안정하였으나 12mM AT 와 0.1mM $NaN_3$ 및 1mM KCN에 의해 90% 이상의 활성이 억제되었다.이에 근거하여 서울시 학생들($7{\sim}18$세)의 만성신부전증 유병률은 1백만명당 5.7명으로 추정되었다. 결론 : 서울시내 학생들 중 11세, 14세, 17세 3개 군에서 한 번 검사로 확인된 무증상 단백뇨의 유병률은 0.28%(약 2.8명/1,000명)이었고 이들중 약 5%만이 3차검사에서 신질환이 의심되었으며 이에 따른 신질환 유병률은 1만명당 1.4명이었다. $7{\sim}18$세 연령층에서 무증상으로 발생하는 사구체 신질환 중에는 IgA 신병증의 유병률이 가장 높아 1만명당 0.64명으로 추정되었고 만성신부전증의 유병률은 1백만명당 5.7명으로 추정되었다. 집단뇨 검사를 통해 확인되는 신질환은 대부분 사구체 질환이기 때문에 집단뇨검사의 의의는 좀더 연구가 필요할 것으로 생각되었다. 오히려 증상을 동반하는 경우보다 빈도가 증가한다는 사실은 집단뇨 검사에서 소변의 이상소견이 발견되어 신장 조직검사를 실시할 경우 혈청 $C_3$치의 감소 여부에 관계없이 MPGN도 진단적 고려 대상이 되어야 한다고 생각한다.신장 조직검사를 시행한 결과 진행성 경과를 취할 수 있는 막 증식성 사구체 신염과 매우 희귀한 증례인 신유전분증 등으로 진단됨으로써 지속성 단백뇨의 경우 정확 진단적 접근이 필수적임을 알 수 있다. 기립성

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The effects of Sakunjatang and Samultang Aqua-acupuncture on immune response (사군자탕(四君子湯) 및 사물탕(四物湯) 약침(藥鍼)이 면역반응(免疫反應)에 미치는 영향(影響))

  • Lee, Byung-Ruyl;Ahn, Byoung-Choul
    • The Journal of Korean Medicine
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    • v.18 no.1
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    • pp.357-374
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    • 1997
  • In order to investigate the effects of Sakunjatang(四君子湯) and Samultang(四物湯) aqua-acupuncture on immune response, Sprague-Dawley male rats were used and randomly divided into four group. Normal group was normal control, Control group was injected i.v. with 2mg/kg MTX on 9th and 11th day after sensitization with SRBC on 5th day, Sakunjatang aqua-acupunctured group(Sample A) was aqua-acupunctured daily for 18 days into the locus corresponding Bisu(B20) locus of the human body to Rat and MTX was administered 9th and 11th experimental day, Samultang aqua-acupunctured group(Sample B) was aqua-acupunctured daily for 18 days into the locus coresponding Bisu(B20) locus of the human body to Rat and MTX was administered 9th and 11th experimental day. In the 9th day and the 11th day after aqua-acupuncture, MTX was injected to reduce immune function in the tail of rat. leukocyte count, lymphocyte ratio, lymphocyte count of spleen, lymphocyte count of bonemarrow, contact hypersensitivity to DNFB, morphologic change of thymus cell, and electropherogram of serum protein. The result were summarized as follows: 1. Before MTX injection, leukocyte count had no significant difference in Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group compared to control group and after MTX injection, leukocyte count had no significant difference Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group. 2. Before MTX injection, lymphocyte ratio was decreased significantly Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group compared to control group. 3. After MTX injection, lymphocyte ratio was increased significantly Sakunjatang aqua-acupunctured group, Samultang aqua-acupunctured group had no significant difference compared to control group. 4. The lymphocyte count of spleen was increased significantly Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group compared to control group. 5. The lymphocyte count of bonemarrow was increased significantly Sakunjatang aqua-acupunctured group, Samultang aqua-acupunctured group had no significant difference compared to control group. 6. Contact hypersensitivity was no significant difference in Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group compared to control group. 7. In the morphologic change of thymus cell, control group compared to normal group had a indistinct boundary between cortex and medulla, and lymphocyte cell density of thymus was low, Sakunjatang aqua-acupuryctured group had a distinct boundary between cortex and medulla, and lymphocyte cell density of thymus was high. 8. In the SDS-PAGE electrophorogram of serum protein, Sakunjatang aqua-acupunctured group and Samultang aqua-acupunctured group had a wide band of nearby 50,000 and 25,000 Dalton which meant IgG generate more activity.

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Fusobacterium nucleatum modulates serum binding to Porphyromonas gingivalis biofilm (Porphyromonas gingivalis biofilm에 대한 면역혈청의 침투력에 대한 Fusobacterium nucleatum의 조절효과)

  • Choi, Jeom-Il;Kim, Sung-Jo;Kim, Soo-Jin
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.661-668
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    • 2001
  • Anti-P. gingivalis immune sera were obtained from mice immunized with either P. gingivalis alone, or F. nucleaturm followed by P. gingivalis. Two groups of immune sera were examined for binding capacity to P. gingivalis biofilm by confocal laser scanning microscope, Antibody avidity index was also determined for each immune sera. The results indicated that prior immunization of mice with F. nucleaturm impaired P. gingivalis-specific immune sera in binding capacity to biofilm and antibody avidity to P. gingivalis. Elevated antibody responses in patients with destructive periodontal disease has often been related to suboptimal level of protective antibody $(opsonophagocytosis)^{1-3)}$ while post-immune sera obtained with experimental animals using a single periodontal pathogen demonstrated satisfactory levels of protective function against the homologous bacterial $challenge^{4,5)}$.The reason is unclear why elevated IgG responses in periodontal patients to periodontal pathogens do not necessarily reflect their protective function. Such an immune deviation might be derived from the fact that destructive periodontal disease is cumulative result of immunopathologic processes responding to an array of different colonizing microorganisms sequentially infecting in the subgingival environmental niche. Fusobacterium nucleaturm is one of the key pathogens in gingivitis, in the transitional phase of conversion of gingivitis into destructive periodontitk, and in adult $periodontitis^{6-8)}$. It also plays a central role in coaggregation with other important microbial species in subgingival $area^{6,9,10)}$ as well as in $biofilm^{11)}$, especially with Porphyromonas gingjvalis in synergism of virulence in human periodontal disease or in animal $models^{12-14)}$. This organism has also been reported to have immune modulating activity for secondary immune response to Actinobacillus $actinomycetemcomitans^{15)}$. It is presumed that sequential colonization and intermicrobial coaggregation between intermediate and late colonizers could potentially modulate the immune responses and development of specific T cell phenotypes in periodontal lesions. We have recently demonstrated the skewed polarization of P. gingivalis-specific helper T cell clones in mice immunized with F. nucleaturm followed by P. $gingivalis.^{16)}$. Consequently F. nucleaturm may initially prime the immune cells and modify their responses to the successive organism, P. gingivalis. This could explain why one frequently observes non-protective serum antibodies to P. gingivalis in periodontal patients in contrast with those obtained from animals that were immunized with $P.gingivalis\;alone^{17)}$. The present study was performed to investigate the immune modulating effect of F. nucleatum on serum binding to experimental biofilms and the avidity of anti-P. gingivalis antibody.

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