• 융합정보논문지
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• 제10권6호
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• pp.164-176
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• 2020

인체 적응 면역 반응을 일으키는데 중요한 항원 특이적 T 세포를 활용한 면역 세포 치료에서 T 세포를 체외에서 배양하고 클론 확장시키는 과정은 매우 섬세하고 복잡하여 조절하기가 쉽지 않아 T 세포의 활성화와 클론 확장을 유도하면서도 조절 및 취급이 용이한 인공 항원제시세포 개발의 필요성이 대두되고 있다. 인공 항원제시세포는 인체의 항원제시세포의 세포 표면 분자와 작용을 모방하게 되는데, 기본적인 신호 분자인 MHC-항원 복합체, 공동 자극 분자, 그리고 용해성 면역 조절 분자를 필수적으로 발현하여야 한다. 또한 T 세포가 항원과 접촉할 때, 이들 분자들이 잘 조직화되어 작용하는 것이 효과적인 T 세포 활성화에 중요하다. 본 논문에서는 여러 인공 항원제시세포 제작 방법과 세포 표면 분자들의 결합 방법과 물리적인 특성이 T 세포와의 상호작용에 중요함을 고찰하였으며, 효과적인 T 세포 활성화를 유도하며 면역세포치료에 적용 가능한 인공항원제세세포의 제작 방법을 살펴보았다.

• 약학회지
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• 제40권5호
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• pp.599-607
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• 1996

Acetylarsonate was prepared for testing antitumor and immunological effects. It showed cytotoxicity directly on Sarcoma 180. L1210 and MOLT-4 by MTT assay. It did not seemed to trigger the mitosis of human lymphocytes in culture, but that showed the cytotoxicity with higher dose. The rosette formation and spleen weight of mouse which acetylarsonate was administered to for 2 weeks were increased. Furthermore, peripheral helper T- and cytotoxic/suppressor T-lymphocytes were increased in acetylarsonate-injected-mice significantly when it was estimated with simultaneous 2 color analysis using anti Lyt2-FITC and L3T4-PE monoclonal antibody by Flow cytometer.

• Clinical and Experimental Pediatrics
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• 제49권5호
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• pp.545-551
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• 2006

목 적 : Cytotoxic T lymphocyte-associated antigen-4(CTLA-4)를 표현하는 유전자는 IgE 조절과 T 세포의 활성에 중요한 역할을 하는 후보유전자로 알려져 있다. 본 연구에서는 CTLA-4 exon 1(＋49 position)과 promoter 유전자(-318 position)의 다형성과 한국 소아의 아토피피부염의 유전적 감수성 및 아토피피부염의 임상양상의 관계에 대해 알아보고자 하였다. 방 법 : 아토피성 습진 145명, 비아토피성 습진 69명과 대조군 96명을 대상으로 restriction fragment length polymorphism방법으로 CTLA-4 promoter(-318 T/C)와 exon 1(＋49 A/G)의 다형성을 조사하였다. 결 과 : CTLA-4 exon 1과 promoter의 유전자 다형성이 아토피성 습진 환아군이나 비아토피성 습진 환아군에서 대조군에 비해 의의있는 분포의 차이를 보이지 않았으며, 아토피피부염의 중증도, IgE 농도, 호산구의 수와도 유의한 관계가 없없다. 결 론 : 본 연구에서는 CTLA-4 유전자 다형성이 한국 소아의 아토피피부염의 유전적 감수성 및 아토피피부염의 임상양상에 관여하지 않는 것으로 생각된다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.44-52
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• 2004

Background: As a potent antigen presenting cell and a powerful inducer of antigen specific immunity, dendritic cells (DCs) are being considered as a promising anti-tumor therapeutic module. The expected therapeutic effect of DCs in renal cell carcinoma was tested in the mouse model. Established late-stage tumor therapeutic (E-T) and minimal residual disease (MRD) model was considered in the in vivo experiments. Methods: Syngeneic renal cell carcinoma cells (RENCA) were inoculated either subcutaneously (E-T) or intravenously (MRD) into the Balb/c mouse. Tumor cell lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started 3 week (E-T model) or one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, the tumor growth and the systemic immunity were observed. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with RENCA cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor growth (E-T model) or formation (MRD model) was suppressed in pulsed-DC treated group. RENCA specific lymphocyte proliferation was observed in the RENCA tumor-bearing mice treated with pulsed-DCs. Primary cytotoxic T cell activity against RENCA cells was increased in pulsed-DC treated group. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs in established or minimal residual disease/metastasis state of renal cell carcinoma. Systemic tumor specific immunity including cytotoxic T cell activity was modulated also in pulsed-DC treated group.

• 대한수의학회지
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• 제41권2호
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• IMMUNE NETWORK
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• 제11권3호
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• pp.182-189
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• 2011

Background: Cytotoxic T lymphocytes (CTLs) appear to play an important role in the control and prevention of human cytomegalovirus (HCMV) infection. The pp65 antigen is a structural protein, which has been defined as a potential target for effective immunity against HCMV infection. Incorporation of an 11 amino acid region of the HIV TAT protein transduction domain (Tat) into protein facilitates rapid, efficient entry into cells. Methods: To establish a strategy for the generation of HCMV-specific CTLs in vitro, recombinant truncated N- and C-terminal pp65 protein (pp65 N&C) and N- and C-terminal pp65 protein fused with Tat (Tat/pp65 N&C) was produced in E.coli system. Peripheral blood mononuclear cells were stimulated with dendritic cells (DCs) pulsed with pp65 N&C or Tat/pp65 N&C protein and immune responses induced was examined using IFN-${\gamma}$ ELISPOT assay, cytotoxicity assay and tetramer staining. Results: DCs pulsed with Tat/pp65N&C protein could induce higher T-cell responses in vitro compared with pp65N&C. Moreover, the DCs pulsed with Tat/pp65 N&C could stimulate both of $CD8^+$ and $CD4^+$ T-cell responses. The T cells induced by DCs pulsed with Tat/pp65 N&C showed higher cytotoxicity than that of pp65-pulsed DCs against autologous lymphoblastoid B-cell line (LCL) expressing the HCMV-pp65 antigen. Conclusion: Our results suggest that DCs pulsed with Tat/pp65 N&C protein effectively induced pp65-specific CTL in vitro. Tat fusion recombinant protein may be useful for the development of adoptive T-cell immunotherapy and DC-based vaccines.

• Journal of Veterinary Science
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• 제22권3호
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• pp.30.1-30.15
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• 2021

Background: New-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects. Objectives: In this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles. Methods: The mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination. Results: AEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4⁺ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months. Conclusions: These findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.

• IMMUNE NETWORK
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• 제10권4호
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• pp.120-125
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• 2010

Dysfunction of the virus-specific T cells is a cardinal feature in chronic persistent viral infections such as one caused by hepatitis C virus (HCV). In chronic HCV infection, virus-specific dysfunctional CD8 T cells often overexpress various inhibitory receptors. Programmed cell death 1 (PD-1) was the first among these inhibitory receptors that were identified to be overexpressed in functionally impaired T cells. The roles of other inhibitory receptors such as cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and T cell immunoglobulin and mucin domain-containing molecule 3 (Tim-3) have also been demonstrated in T-cell dysfunctions that occur in chronic HCV patients. Blocking these inhibitory receptors in vitro restores the functions of HCV-specific CD8 T cells and allows enhanced proliferation, cytolytic activity and cytokine production. Therefore, the blockade of the inhibitory receptors is considered as a novel strategy for the treatment of chronic HCV infection.

• Archives of Pharmacal Research
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• 제22권4호
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• pp.340-347
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• 1999

Dendritic cells (DCs) are potent professional antigen-presenting cells (APC) capable of inducing the primary T cell response to antigen. Although tumor cells express target antigens, they are incapable of stimulating a tumor-specific immune response due to a defect in the costimulatory signal that is required for optimal activation of T cells. In this work, we describe a new approach using tumor-DC coculture to improve the antigen presenting capacity of tumor cells which does not require a source of tumor-associated antigen. Immunization of a weakly immunogenic and progressive tumor cocultured with none marrow-derived DCs generated an effective tumor vaccine. Immunization with the cocutured DCs was able to induce complete protectiv immunity against tumor challenges and was effective for the induction of tumor-specific CTL (cytotoxic T lymphocyte) activity. Furthermore, high NK cell activity was observed in mice in which tumors were rejected. In addition, immunization with tumor-pulsed DC s induced delayed tumor growth, but not tumor eradication in tumor-bearing mice. Our results demonstrate that coculture of DCs with tumors generated antitumor immunity due to the NK cell activation as well as tumor-specific T cell. This approach would be used for designing tumor vaccines using DCs when the information about tumor antigens is limited.

• 미생물학회지
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• 제47권4호
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• pp.342-347
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• 2011

인체의 바이러스 감염 방어기전에서 T 림프구는 중요한 역할을 한다. 하지만, 만성 C형 간염 바이러스의 일차적 복제장소인 간염 환자의 간에서 분리된 HCV 특이 T 림프구는 심각한 기능결핍을 보인다. 이러한 T 림프구 기능결핍의 이유로는 PD-1, CTLA-4 등 면역억제 물질의 발현, 또는 간에서 특이적으로 유도되는 면역내성(immune tolerance)이 있으나, 간세포(hepatocytes)와 HCV 특이 T 림프구의 상호작용에 대해서는 명확하게 확립되어 있지 않다. 따라서 본 연구에서는 HLA(human leukocyte antigen) A2+ 간암세포주(human hepatoma cell line; huh7.5)가 항원제시(antigen presentation)를 통해 효과적으로 HCV 특이 T 림프구를 활성화시키며 간암세포주(huh7.5) 표면의 PD-L (program death ligand) 1 발현은 T림프구의 활성을 감소시켜 면역조절의 가능성이 있음을 시사하였다. 또한, HCV 특이 tetramer 반응은 T 림프구의 과도한 활성으로 자기사멸(apoptosis)의 경로에 있음을 caspase 3 활성으로 확인하였고, 역시 PD-L1의 발현이 T 림프구를 자기사멸(apoptosis)로부터 구제하여 caspase 3 활성이 감소하는 것을 확인하였다. 이는 PD-L1과 간성(liver) T 림프구 표면의 PD-1 결합이 T 림프구의 자기사멸을 막고, 또한 그 기능을 회복시켜 만성 C형 간염 치료에 응용될 수 있음을 시사한다.