• Parasites, Hosts and Diseases
• /
• 제62권1호
• /
• pp.53-63
• /
• 2024

The intracellular parasite Babesia microti is among the most significant species causing human babesiosis and is an emerging threat to human health worldwide. Unravelling the pathogenic molecular mechanisms of babesiosis is crucial in developing new diagnostic and preventive methods. This study assessed how priming with B. microti surface antigen 1 (BHSA 1) and seroreactive antigen 5-1-1 (BHSA 5-1-1) mediate protection against B. microti infection. The results showed that 500 ㎍/ml rBMSA1 and rBMSA5-1-1 partially inhibited the invasion of B. microti in vitro by 42.0±3.0%, and 48.0±2.1%, respectively. Blood smears revealed that peak infection at 7 days post-infection (dpi) was 19.6%, 24.7%, and 46.7% in the rBMSA1, rBmSA5-1-1, compared to the control groups (healthy mice infected with B. microti only), respectively. Routine blood tests showed higher white blood cell, red blood cell counts, and haemoglobin levels in the 2 groups (BMSA1 and BMSA5 5-1-1) than in the infection control group at 0-28 dpi. Moreover, the 2 groups had higher serum interferon-γ, tumor necrosis factor-α and Interleukin-17A levels, and lower IL-10 levels than the infection control group throughout the study. These 2 potential vaccine candidate proteins partially inhibit in vitro and in vivo B. microti infection and enhance host immunological response against B. microti infection.

• KSBB Journal
• /
• 제22권6호
• /
• pp.371-377
• /
• 2007

나노기술과 바이오기술의 융합연구에 의해 나노바이오기술이 발전되고 있다. 나노바이오기술의 중요한 응용연구 중의 하나로서, 진단이나 바이오센서 분야에서 단백질-단백질 및 단백질-바이오물질간의 상호작용을 연구하기 위한 단백질 센서 칩이 개발되어 왔다. 본 논문에서는 단백질의 선택적 고정화를 위한 새로운 생체고분자 기질로 PHA를 이용하는 첫 번째 예로서, 단백질-단백질 및 항원-항체 반응의 구현을 나타내고자 하였다. 본 시스템은 PHA 표면 위에서 PHA depolymerase의 SBD와의 선택적 결합에 기반한 것으로, PHA depolymerase의 SBD와 융합된 단백질이 PHA가 코팅된 표면 위에 spotting 될 수 있고 미세접촉인쇄방법에 의해 PHA 위에 미세패턴이 제조되어지는 것을 알 수 있었다(52, 53). 이러한 새로운 전략이 PHA depolymerase의 SBD와 다른 단백질을 융합함으로서 미세 spotting과 미세패터닝이 가능하게 되었고 항원-항체의 생물학적 반응을 통해 많은 바이오센서 칩 연구에 응용될 수 있음을 확인하였다. 또한, PHA 마이크로 비드에도 PHA depolymerase의 SBD와 융합된 단백질을 고정시킴으로서 항원-항체 반응을 유도할 수 있음을 확인하였다(54). PHA의 구조를 변경하여 PHA 기판, PHA 필름, PHA 미세패턴, PHA 마이크로 비드 등을 이용할 수 있으며 multiplex assay를 동시에 진행할 수 있는 다양한 융합 단백질을 사용할 수 있을 것이다. 생분해성 플라스틱으로서 성공적으로 개발된 PHA를 이용한 새로운 플랫폼 기술이 PHA depolymerase의 SBD를 이용함으로서 특이적이고 선택적인 단백질의 고정화에 이용될 수 있음을 확인하였다. 본 전략이 다양한 단백질-단백질 및 단백질-바이오물질 반응을 이용한 바이오칩 및 바이오센서의 응용연구에 유용하게 사용될 것이다.

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

• Journal of Periodontal and Implant Science
• /
• 제29권2호
• /
• pp.333-350
• /
• 1999

The modulation of leukocyte cell surface adhesion molecules may influence the development of cellular events that determine the course of the inflammatory process. Direct interaction between activated T cells and monocytes resulted in a large production of $IL-1{\beta}$ by monocytes. In this reactions, adhesion molecules play an important part, yet the role of them in Tmonocytes interaction remain unclear. This study was undertaken in an effort to elucidate, 1) the influence of 1.25(OH)$_2D_3-induced$ differentiation on the monocyte responsiveness to direct contact with T lymphocytes, and 2) the role of adhesion molecules on the T-monocyte direct interaction. Initially, I observed that direct contact of monocyte cell line THP-1 with stimulated fixed T cell line HuT78 markedly induces IL-1${\beta}$ production by THP-1. $IL-1{\beta}$ production was higher when THP-1 had been previously exposed to 1.25(OH)$_2D_3$ as compared to control, with ${\alpha}$- 1.25(OH)$_2D_3$ dose-dependent and exposure time-dependent manner. It was shown that 1.25(OH)$_2D_3$ also increased the expression of ${\beta}_2$ integrin adhesion receptor Mac-1(CD11b/CD18) dose- and timedependently, but did not increase the expression of human leukocyte antigen- D(HLA-D) and intercellular adhesion molecule-1(ICAM-1). The $IL-1{\beta}$ producing activity of THP-1 cells correlated well with the ability to induce the Mac-1 expression on THP-1 surface. Monoclonal antibody raised against relevant cell surface glycoproteins on THP-1 were tested for their ability to block the response of THP-1 to T cells. Antibody to Mac-1 only partially blocked $IL-1{\beta}$ production by THP-1, whereas antibodies to ICAM-1 and HLA-D did not. These data indicate that regulation of Mac-1 expression on THP-1 cells can alter the responsiveness of these cells to contact by activated T cells, however other unknown structures on the THP-1 cells may be involved in this process also.

• Parasites, Hosts and Diseases
• /
• 제55권5호
• /
• pp.473-480
• /
• 2017

Plasmodium vivax merozoite surface protein-1 (PvMSP1) gene codes for a major malaria vaccine candidate antigen. However, its polymorphic nature represents an obstacle to the design of a protective vaccine. In this study, we analyzed the genetic polymorphism and natural selection of the C-terminal 42 kDa fragment within PvMSP1 gene ($PvMSP1_{42}$) from 77 P. vivax isolates, collected from imported cases of China-Myanmar border (CMB) areas in Yunnan province and the inland cases from Anhui, Yunnan, and Zhejiang province in China during 2009-2012. Totally, 41 haplotypes were identified and 30 of them were new haplotypes. The differences between the rates of non-synonymous and synonymous mutations suggest that $PvMSP1_{42}$ has evolved under natural selection, and a high selective pressure preferentially acted on regions identified of $PvMSP1_{33}$. Our results also demonstrated that $PvMSP1_{42}$ of P. vivax isolates collected on China-Myanmar border areas display higher genetic polymorphisms than those collected from inland of China. Such results have significant implications for understanding the dynamic of the P. vivax population and may be useful information towards China malaria elimination campaign strategies.

• Archives of Pharmacal Research
• /
• 제25권6호
• /
• pp.895-902
• /
• 2002

Monocytes and macrophages playa major role in defense mechanism of the host response to tumor, in part through the secretion of several potent products and macrophage cytokines. Monocytes and tissue macro phages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1) and tumor necrosis factor (TNF). Recent studies emphasizes that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. In this study, our work is directed toward studying the in vitro effects of Korean propolis on the ability to induce cellular and secretory responses in murine macrophage cell line, RAW 264.7. It was found that Water Extract of Korean Propolis (WEP) could activate macro phages by producing cytokines. The production of the macrophage cytokines, IL-1 and TNF-$\alpha$, by RAW 264.7 treated with WEP was examined from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml with dose dependent manner. Nitric oxide (NO) production was also increased when cells were exposed to combination of LPS and WEP from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml. At high dose of WEP (50 to 100 $\mu\textrm{g}$/ml) used to prescribe for anti-inflammatory and analgesic medicine showed inhibition of NO production in LPS-stimulated macrophage. Besides cytokine production, NO release, surface molecule expression and cell morphologic antigen expression were increased in response to the stimulation by WEP. These results suggested WEP may function through macrophage activation.

• 한국산학기술학회논문지
• /
• 제14권7호
• /
• pp.3419-3424
• /
• 2013

B형간염표면 항원 및 항체(HBsAg와 anti-HBs)에 대한 검사 방법으로 화학발광면역검사법(CIA)을 포함한 면역검사법을 주로 사용하고 있으나 최근 간편한 면역크로마토그래피법(ICA)의 사용이 늘고 있는 추세이다. 본 연구에서는 CIA법을 이용한 정량검사법과 비교하여 신속검사인 ICA법의 민감도를 평가하고자 하였다. 대학병원에 간염검사를 의뢰한 검체 중 CIA법에서 양성 및 음성으로 나온 120 검체를 선정하여 ICA법으로 분석하였다. ICA법과 CIA법의 일치율은 항원 97%와 항체 90%로 나타났다. 항원검사의 ICA법 민감도, 특이도, 양성예측도, 음성예측도는 각각 97%, 100%, 100% 및 96.8%로 나타났으며, 항체검사는 각각 90%, 93.3%, 93.1% 및 90.3%로 나타났다. ICA법을 임상에서 사용 시 결과판독에 주의가 필요하며, 낮은 역가의 경우 검출되지 않는 제한점이 있으므로 예민도가 높은 다른 검사법을 이용하여 이중검사를 시행해야 할 것으로 사료된다.

• IMMUNE NETWORK
• /
• 제1권1호
• /
• pp.53-60
• /
• 2001

Background: The human mic2 gene is a pseudoautosomal gene that encodes a cell surface antigen, CD99. High levels of CD99 constitute a tumor marker in Ewing s sarcoma (ES). We have recently demonstrated that CD99-induced apoptosis occurs only in undifferentiated ES cells, not in differentiated ES cells, raising the possibility of the involvement of CD99 in neural ontogeny. Methods: To elucidate the relations between the expression of CD99 and the differentiation of neural cells and the mechanism by which the expression of CD99 is regulated, we analyzed the differential patterns of CD99 expression in SH-SY5Y by treatment of 12-O-tetradecanoyl-13-phorbol acetate (TPA) and retinoic acid. In addition, to explore the transcriptional activity of CD 99 during neural cell differentiation, SH-SY5Y cells were transiently transfected with a CD99 promoter-driven luciferase construct, and treated with the inducers. Results: In immunoblotting and flow cytometry, the expression level of CD99 was increased on differentiated SH-SY5Y cells induced by TPA and retinoic acid. The luciferase activity was elevated by the treatment with TPA, known to mature SH-SY5Y cells toward a sympathetic neuronal lineage, whereas retinoic acid inducing a sympathetic chromaffin lineage displayed little effect. Conclusion: The result indicates that CD99 might be expressed only on cells maturing toward a neuronal lineage among differentiating primitive neuronal cells. In addition, the expression of CD99 seems to be regulated at the transcriptional level during the differentiation.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권2호
• /
• pp.731-736
• /
• 2012

Background and Aims: The National Cancer Screening Program (NCSP) for liver cancer was initiated in 2003 in Korea. The objective of this study was to evaluate the participation rate of the program and to provide preliminary information on its results based on data collected by the NCSP in 2009. Methods: The target population of the NCSP for liver cancer in 2009 was comprised of 373,590 adults aged ${\geq}40$ years at high risk for liver cancer. Participation rates and positivity rates were assessed in this population. Multivariate logistic regression analysis was performed to determine the factors associated with participation in the NCSP for liver cancer. Results: The overall participation rate was 37.9% and 1,126 participants were positive at screening. The highest participation rates were observed in women, those in their 60s, National Health Insurance beneficiaries, and individuals positive for hepatitis B surface antigen. Positivity rates for men, those in their 70s, Medical Aid Program recipients and individuals with liver cirrhosis were the highest in the respective categories of gender, age, health insurance type, and risk factor for liver cancer. Conclusions: The participation rates of the NCSP for liver cancer are still low, despite the fact that the program targets a high-risk group much smaller than the general population. Efforts to facilitate participation and to reduce disparities in liver cancer screening among Korean men and women are needed. These results provide essential data for evidence-based strategies for liver cancer control in Korea.

• Parasites, Hosts and Diseases
• /
• 제55권4호
• /
• pp.385-389
• /
• 2017

Seroprevalence of Toxoplasma gondii infection among the residents of Cheorwon-gun, Gangwon-do, Korea, which partly includes the demilitarized zone (DMZ), were surveyed for 4 years and evaluated by RDT using recombinant fragment of major surface antigen (SAG1A). Sera from 1336, 583, 526, and 583 adult residents were collected on a yearly basis from 2010 to 2013, respectively. The total positive seroprevalence was 19.3, 21.9, 23.4, and 26.8% from 2010 to 2013, respectively. The positive seroprevalence in men (23.6, 27.5, 29.5, 34.6%) was far higher than women (14.1, 18.3, 19.4, 21.4%), from 2010 to 2013, respectively. This high seroprevalence of toxoplasmosis in Cheorwon-gun may have been influenced in part by its geographical locality of the area as it includes the DMZ, where civilian access is strictly limited, thus creating a relatively isolated area that is a well-preserved habitat. Further research is necessary to study the epidemiology of toxoplasmosis in this area.