黃기에 의한 Helper T 세포의 생존 및 활성 증가

강희,배현수,안규석 대한동의병리학회 2001 동의생리병리학회지 Vol.15 No.4

Astragali Radix(AR), one of the strong tonic herbs, is known to improve immunological responses in mice and humans. In this study, AR's Qi-reinforcing effect was examined in the context of CD4+ T cells' survival and TCR/CD3 induced activation responses. Spleen cells from 8 week BALB/c mice were cultured in AR containing medium without activation for 24, 48 h. The MTT assay and flow cytometry revealed that AR treated lymphocytes were more viable and showed the higher percentage of CD4+ T cells than the control cells. Furthermore, the expression of LKLF, a transcription factor that presumably regulates the survival of resting T cells. was increased in AR treated cells. Subsequently CD4+ T cells were isolated and cultured in AR containing medium. AR also increased survival of sorted CD4 T cells without any involvement of APC. In order to evaluate the direct effect of AR on helper T cells's proliferative capacity prior to activation, CD4+ T cells are isolated after 24 h of culture in AR containing medium and activated with and without anti-CD3/anti-CD28 activation for 48h. A higher level of CD69 was observed in AR treated cells using flow cytometry. Results presented here suggest that AR may be desirable for the maintenance of resting lymphocytes and CD4+ T cells activity in immune responses.

Resveratrol의 CD4+ T 세포 활성과 분화 억제 효과

서동원,이영주,이상명 한국자원식물학회 2014 한국자원식물학회지 Vol.27 No.5

Resveratrol is a naturally occurring stilbene which is safe and well-described compound with a potent anti-inflammatory activity. Recent studies suggested that resveratrol suppressed various inflammation mediated diseases such as asthma, chronic colitis, rheumatoid arthritis, and type 1 diabetes. These studies indicated that resveratrol might directly modulate CD4 + helper T cells (Th cells)-mediated immune responses. However, it is not fully elucidated whether resveratrol directly regulates CD4 + Th cell activation and differentiation. In the present study, CD4 + Th cells were purified from C57BL/6 and treated with various concentrations of resveratrol. We found that resveratrol directly suppressed CD4 + Th cells activation, leading to a defect in T cell proliferation. When CD4 + Th cells were treated with resveratrol, cytokine production was also significantly reduced in a dose dependent manner. In accordance with these results, resveratrol even inhibited CD4 + Th cells differentiation into Th1, Th2 or Th17, which produces IFN-γ, IL-4 or IL-17 respectively. We also found that resveratrol could induce apoptosis of CD4 + T cells at a high concentration. Our data demonstrated that resveratrol inhibited directly CD4 + Th cells activation and differentiation. It suggests that resveratrol could be an efficient therapeutic strategy for autoimmune diseases in which CD4 + Th cells play a critical role. Resveratrol은 천연 stilbene으로 안전성 있는 항염증 활성을 가진 화합물로 알려져 있다. 최근의 연구들에서 resveratrol이 천식, 만성 대장염, 류마티스성 관절염과 같이 염증에 의해발생하는 다양한 질병을 억제한다고 보고되었다. 이러한 연구들은 resveratrol이 CD4+ helper T cells (Th cells)에 의한 면역반응을 조절할 것이라고 제시하였다. 그러나 resveratrol이 직접적으로 Th cells의 활성화와 분화를 조절하는지 완전히 밝혀지지 않았다. C57BL/6에서 Th cells을 분리하여 다양한 농도의resveratrol을 세포에 처리하였다. 본 연구에서는 resveratrol이 직접적으로 Th cells의 활성화와 증식을 억제하는 것을 확인하였다. Th cells에 resveratrol을 처리하였을 때 IFN-γ, IL-4,IL-17 사이토카인 생성이 농도에 따라 유의하게 감소하였고 또한 Th cells이 이러한 사이토카인들을 분비하는 Th1과 Th2과Th17으로 분화되는 것이 억제되었다. 그리고 고농도의 resveratrol이 Th cells의 세포사멸을 유도하는 것으로 확인되었다. 본 연구에서는 resveratrol이 Th cells의 활성화와 분화를 직접적으로억제하는 것을 확인하였으며, 이는 resveratrol이 CD4+ Thcells에 의해 발생되는 자가면역질환의 효과적인 치료법이 될수 있을 것이라고 제시한다.

CD70-expressing CD4 T cells produce IFN-γ and IL-17 in rheumatoid arthritis

Park, Jin Kyun,Han, Bobby Kwanghoon,Park, Ji Ah,Woo, Youn Jung,Kim, So Young,Lee, Eun Young,Lee, Eun Bong,Chalan, Paulina,Boots, Annemieke M.,Song, Yeong Wook Oxford University Press 2014 Rheumatology Vol.53 No.10

<P><B>Objective.</B> CD70-expressing CD4 T cells are enriched in RA and promote autoimmunity via co-stimulatory CD70-CD27 interaction. This study aimed to explore the phenotype and cytokine production of CD70<SUP>+</SUP> CD4 T cells in RA.</P><P><B>Methods.</B> Peripheral blood mononuclear cells from 32 RA patients were isolated and frequencies of CD70<SUP>+</SUP> cells within different CD4 T subsets were analysed using flow cytometry. IFN-γ and IL-17 production were compared between the CD70<SUP>+</SUP> and CD70<SUP>−</SUP> cells. Expression of master transcription factors T-bet, GATA3 and retinoic acid–related orphan receptor gamma t (RORγt) were examined by real-time PCR. Results are presented as mean (<SMALL>s.e.m</SMALL>.).</P><P><B>Results.</B> CD4 T cells of healthy controls rarely expressed CD70 as compared with CD4 T cells of RA patients [mean 0.9% (<SMALL>s.e.m</SMALL>. 0.3%) <I>vs</I> 7.6 (0.6), <I>P</I> < 0.001]. In RA, CD70<SUP>+</SUP> cells were present within all CD4 T cell subsets, i.e. CD45RA<SUP>+</SUP>CCR7<SUP>+</SUP> naive, CD45RA<SUP>−</SUP>CCR7<SUP>+</SUP> central memory, CD45RA<SUP>−</SUP>CCR7<SUP>−</SUP> effector memory and CD45RA<SUP>+</SUP>CCR7<SUP>−</SUP> terminally differentiated effector memory T cells with a mean frequency of 3.9% (<SMALL>s.e.m</SMALL>. 1.1%), 4.0 (0.5), 4.2 (0.7) and 9.4 (4.3), respectively. As compared to CD70<SUP>−</SUP> CD4 T cells, CD70<SUP>+</SUP> CD4 T cells produced significantly more IFN-γ and IL-17 after short activation. CD70<SUP>+</SUP> CD4 T cells preferentially expressed transcription factor RORγt.</P><P><B>Conclusion.</B> CD70<SUP>+</SUP> CD4 T cells are enriched in RA and may directly contribute to RA pathogenesis by producing IFN-γ and IL-17. Targeting CD70<SUP>+</SUP> CD4 T cells might offer new therapeutic opportunities in RA.</P>

주조직적합항원이 불일치하는 마우스 동종 조혈모세포이식에서 IL-2로 유도된 CD4+CD25+ T세포를 이용한 이식편대숙주병의 억제

현재호,정대철,정낙균,박수정,민우성,김태규,최병옥,김원일,한치화,김학기,Hyun, Jae Ho,Jeong, Dae Chul,Chung, Nak Gyun,Park, Soo Jeong,Min, Woo Sung,Kim, Tai Gyu,Choi, Byung Ock,Kim, Won Il,Han, Chi Wha,Kim, Hack Ki 대한면역학회 2003 Immune Network Vol.3 No.4

Background: In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. Methods: The splenocytes (SP) were obtained from 6 week-old BALB/c mice ($H-2^d$) and irradiated as a single cell suspension. The donor mice (C3H/He, $H-2^k$) received $5{\times}10^6$ irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with $1{\times}10^7$ donor BM and $5{\times}10^6$ CD4+CD25+ cells. The other recipient mice received either $1{\times}10^7$ donor BM with $5{\times}10^6$ CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. Results: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was $30.0{\pm}13%$ compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells ($5.4{\pm}1.5%$) than the untreated mice SP ($1.4{\pm}0.3%$)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells ($61.1{\pm}6.1%$), but a marked proliferation in the CD4+CD25- cells ($129.8{\pm}65.2%$). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). Conclusion: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

말초혈액 CD4+CD25- T 세포로부터 IL-4로 유도된 CD4+CD25+ 면역조절 T 세포의 획득과 특성 규명

허성범 ( Seong Beom Heo ),조미라 ( Mi La Cho ),박미경 ( Mi Kyung Park ),민소연 ( So Youn Min ),주지현 ( Ji Hyun Joo ),박경수 ( Kyung Soo Park ),조영규 ( Young Kyu Cho ),윤종현 ( Chong Hyeon Yoon ),박성환 ( Sung Hwan Park ),김호연 대한류마티스학회 2005 대한류마티스학회지 Vol.12 No.4

Objective: The CD4+CD25+ regulatory T cells (Treg) can be induced by TGFβ and IL-10 in the periphery, and understanding the biological function of cytokine-induced Treg is critically important for the control of autoimmune diseases. We investigated the IL-4-induced CD4+CD25+ regulatory T cells in human PBMCs, which were derived from the CD4+CD25- T cells. Methods: The CD4+CD25- T cells from human PBMC were isolated by MACS and cultured in the presence of IL-4 or absence of IL-4. The presence and phenotype of induced CD4+CD25+ T cells were determined by flow cytometry. Supressive activity of induced CD4+CD25+ T cells were assessed by culturing CD4+CD25- and CD4+CD25+ T cells with anti-CD3 monoclonal antibodies and antigen-presenting cells, followed by proliferation. Results: After 5 days, significant amount of CD4+CD25+ T cells were generated from the CD4+CD25- T cells cultured with anti-CD3 antibody in the presence of IL-4. These IL-4 induced CD4+CD25+ T cells presented with similar phenotype to natural occurred Treg cells, including CD45ROhi, CD45RAlo, CTLA-4hi, OX40hi, CD62Lhi and HLA-DRhi, and also exhibited high expression of Foxp3 molecule. In addition, the IL-4 induced CD4+CD25+ T cells can suppress the proliferative responses against anti-CD3. The regulatory property of IL-4 induced CD4+CD25+ T cell was partially abrogated after treatment with anti-IL-10 and anti-TGFβ antibodies. Conclusion: These data indicate that IL-4 induced CD4+CD25+ Treg cells can be generated from the CD4+CD25- T cells in the peripheral blood, and may contribute to the important immunoregulatory function in human.

인간 CD4+ T 임파구의 활성도와 증식에 대한 노화의 영향

홍명선 ( Myung Sun Hong ),단진명 ( Jin Myung Dan ),이원우 ( Won Woo Lee ),강인수 ( In Soo Kang ) 대한류마티스학회 2009 대한류마티스학회지 Vol.16 No.4

Objective: Alterations in the immune system occur with aging, and these contribute to an increased risk of infection and malignancy. The age-associated changes in T cell immunity range from single cell function to the maintenance of cell populations. We investigated the kinetics of CD4+ T cell activation and proliferation in young and elderly subjects after stimulating their peripheral blood mononuclear cells with anti-CD3 and anti-CD28 antibodies (Abs). Methods: The expressions of the activation markers CD69, CD40L and CD25 on the CD4+ T cells from young (n=14) and elderly (n=19) were analyzed at 6, 24 and 48 hours (hrs) of T cell receptor (TCR) stimulation by using flow cytometry. In the same individuals, the CD4+ T cell proliferation was determined at 48 and 96 hrs of TCR stimulation by using the CFSE dilution method. Results: The elderly had decreased CD69 and CD40L expressions on the CD4+ T cells at 6 hrs of stimulation, as compared to that of the young patients. The elderly also had a decreased CD25 expression on the CD4+ T cells at 24 hrs of stimulation. However, the two groups had similar levels of the CD25, CD69 and CD40L expressions at 48 hrs of stimulation. The elderly had decreased CD4+ T cell proliferation at 96 hrs of stimulation, as compared to that of the young, although both groups had similar levels of CD4+ T cell proliferation at 48 hrs of stimulation. Conclusion: Our findings suggest that the elderly have altered kinetics of CD4+ T cell activation and proliferation in response to anti-CD3 and -CD28 Ab stimulation, and that such an altered response is governed by the duration of stimulation.

구강 편평태선에서의 T 림프구 및 랑게르한스 세포의 분포

김수진,조영아,윤혜정,홍성두,이재일,홍삼표 대한구강악안면병리학회 2012 대한구강악안면병리학회지 Vol.36 No.3

Numerous reports on the immunopathogenesis of an oral lichen planus (OLP) have shown some inconsistency in terms of the role and distribution of CD4+ and CD8+ T cells, and Langerhans cells (LC). This has led to the hypothesis that although the clinical and histopathological manifestations of OLPs appear similar, they can be grouped into CD4+ predominant lesions and CD8+ ones on the varied immunopathogenesis or clinical progress. This study investigated the distribution of CD4+ T cells, CD8+ T cells, LC and the ratio of CD4/CD8 and defined the important immunocompetent cells. In addition, the clinicopathological and histopathological correlation with those immunocompetent cells were investigated. Frozen sections of 16 OLPs and 5 normal buccal mucosae were immunostained. Quantification was done using KAPPA Imagebase software and statistical analysis using SPSS 10.0 statistical package. CD8+ T cells were consistently more abundant in the epithelium of the OLP than CD4+ T cells, but no difference between the two cells was detected in the lamina propria. The intra-epithelial CD4+ T cells revealed a positive linear correlation with the intra-epithelial CD8+ T cells. In the lamina propria, the number of LC had a positive linear correlation with CD4/CD8 ratio. The number of LCs was higher in the reticular type of OLP compared to the erosive type. The histopathological features such as hyperkeratosis, acanthosis, the band-like infiltration of mononuclear cells, and liquefaction degeneration did not show significant correlation with the CD4/CD8 ratio. The results suggest that CD8+ T cells play major role in immunopathogenesis of OLP. The immunopathogenesis of OLP appears to vary from lesion to lesion in relation to the clinical progress.

Expression and Function of TLR2 on CD4 Versus CD8 T Cells

이선미,주영돈,서수길 대한면역학회 2009 Immune Network Vol.9 No.4

Background: Toll-like receptors (TLRs) play a fundamental role in innate immunity through their capacity to recognize pathogen-associated molecular patterns. Also, TLRs that are expressed in T cells are reported to function as co-stimulatory receptors. However, the functional capacity of TLRs on CD4 T and CD8 T cells has not been directly compared. Here we compared CD4 and CD8 T cell responses to TLR2 ligand plus TCR-mediated stimulation. Methods: TLR2 expression was analyzed on T cell subsets under naïve and alloantigen- primed conditions. We analyzed the effects of TLR2 co-stimulation on proliferation and survival of T cell subsets in vitro when stimulated with soluble anti-CD3 in the presence or absence of synthetic ligand Pam3CSK4. Results: TLR2 expression on CD8 T cells was induced following activation; this expression was much higher than on CD4 T cells. Thus, the molecule was constitutively expressed on Listeriaspecific memory CD8 T cells. Based on these expression levels, proliferation and survival were markedly elevated in CD8 T cells in response to the TLR2 co-stimulation by Pam3CSK4 compared with those in CD4 T cells. Conclusion: Our data show that TLR2 co-stimulation is more responsible for proliferation and survival of CD8 T cells than for that of CD4 T cells. Background: Toll-like receptors (TLRs) play a fundamental role in innate immunity through their capacity to recognize pathogen-associated molecular patterns. Also, TLRs that are expressed in T cells are reported to function as co-stimulatory receptors. However, the functional capacity of TLRs on CD4 T and CD8 T cells has not been directly compared. Here we compared CD4 and CD8 T cell responses to TLR2 ligand plus TCR-mediated stimulation. Methods: TLR2 expression was analyzed on T cell subsets under naïve and alloantigen- primed conditions. We analyzed the effects of TLR2 co-stimulation on proliferation and survival of T cell subsets in vitro when stimulated with soluble anti-CD3 in the presence or absence of synthetic ligand Pam3CSK4. Results: TLR2 expression on CD8 T cells was induced following activation; this expression was much higher than on CD4 T cells. Thus, the molecule was constitutively expressed on Listeriaspecific memory CD8 T cells. Based on these expression levels, proliferation and survival were markedly elevated in CD8 T cells in response to the TLR2 co-stimulation by Pam3CSK4 compared with those in CD4 T cells. Conclusion: Our data show that TLR2 co-stimulation is more responsible for proliferation and survival of CD8 T cells than for that of CD4 T cells.

P045 : The changes of peripheral blood NKG2D+CD4+ T cells in alopecia areata according to disease severity, activity, subtype and treatment: a preliminary study

( Yong Hyun Jang ),( Sang Lim Kim ),( Weon Ju Lee ),( Seok Jong Lee ),( Do Won Kim ) 대한피부과학회 2014 대한피부과학회 학술발표대회집 Vol.66 No.2

Background: NKG2D receptor expressed on CD8+ T cells and NK cells has been reported as important in the initiation of alopecia areata (AA). Autoreactive role for NKG2D+CD4+ T cells has been shown in various autoimmune diseases such as Crohn’s disease. However, in AA, the role of NKG2D+CD4+ T cells is currently not known. Objectives: We investigated the changes of NKG2D+CD4+ T cells in AA and correlation with disease severity, activity, subtype, and treatment. Methods: The proportion of NKG2D+CD4+, NKG2D+CD8+ T cells and NKG2D+ NK cells was analyzed by flow cytometry on peripheral blood samples of patients with AA (n=22) and healthy controls (n=6). For subgroup analysis, AA patients were subdivided according to disease severity, activity, subtype and treatment modalities. In addition, tissue expression and distribution of NKG2D+CD4+ T cells was examined on human AA samples. Results: NKG2D+CD8+T cells and NKG2D+NK cells were increased in AA patients as previously reported. Interestingly, subset of NKG2D+CD4+T cells was also increased in the peripheral blood from AA patients compared with healthy controls (p<0.05). The proportion of NKG2D+CD4+ T cellsshowed a low tendency in AA patients with mild severity and received systemic immunosuppressive therapy. Conclusion: Results of this study suggest that NKG2D+CD4+T cells may involve in the pathogenesis of AA. However, the precise role of NKG2D+CD4+ T cells in AA remains to be elucidated.

Intracellular formyl peptide receptor regulates naïve CD4 T cell migration

Lee, Ha Young,Jeong, Yu Sun,Lee, Mingyu,Kweon, Hee-Seok,Huh, Yang Hoon,Park, Joon Seong,Hwang, Ji Eun,Kim, Kyuseok,Bae, Yoe-Sik Elsevier 2018 Biochemical and biophysical research communication Vol.497 No.1

<P><B>Abstract</B></P> <P>We found that formyl peptide receptor (FPR) 1 and FPR3 were expressed intracellularly and/or the nucleus of naïve CD4 T cell. Activation of naïve CD4 T cells with synthetic intracellular agonists dTAT-WKYMVm and CTP-WKYMVm for FPR members stimulated CD4 T cell migration via pertussis toxin-sensitive manner. Knockdown of FPR1, but not knockdown of FPR3, blocked dTAT-WKYMVm-induced naïve CD4 T cell migration. Stimulation of naïve CD4 T cells with dTAT-WKYMVm elicited the activation of ERK, p38 MAPK, and Akt. Activation of CD4 T cells with anti-CD3 and anti-CD28 antibodies caused surface expression of FPR1 and FPR3, but not FPR2. CD4 T cells isolated from sepsis patients expressed the three members of FPR family on their cell surface. Taken together, our results suggest that intracellular FPR in naïve CD4 T cells and surface FPRs in activated CD4 T cells might regulate immune responses by regulating CD4 T cell activity.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Human naïve CD4 T cells express intracellular FPRs. </LI> <LI> Stimulation of intracellular FPR1 elicits naïve CD4 T cell migration. </LI> <LI> Intracellular FPR1 agonist dTAT-WKYMVm stimulates MAPK and Akt activation. </LI> <LI> Activated CD4 T cells express surface FPRs. </LI> <LI> The finding suggests a novel insight on intracellular FPRs in CD4 T cells. </LI> </UL> </P>