Immunological regulation of CD70/CD27 pathway and the expression of CD70 in tumor / 国际肿瘤学杂志

CD70/CD27 pathway plays an important role in human immune regulation.The role of CD70/CD27 pathway in immune regulation is mainly to promote the activation and proliferation of T cells,induce the differentiation and formation of effector T cells and memory T cells,and interfere with regulatory T cells.In addition,the high level of CD70 expression in some tumor cells provides a new way for tumor immunotherapy.

Changes in natural killer cell subsets defined by CD11b and CD27 in spleen of septic mice / 中华麻醉学杂志

Objective To evaluate the changes in natural Killer (NK) cell subsets definetd by CD11b and CD27 in the spleen of septic mice.Methods A total of 168 pathogen-free healthy male C57BL/6 mice,weighing 20-30 g,aged 8-10 weeks,were divided into 2 groups (n=84 cach) using a random number table:sham operation group (Sham group) and sepsis group (Sep group).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized mice.Thirty mice in each group were selected to assess the survival within 4 days after operation,and the survival rate was calculated.At 2,4,6,24,48 and 72 h after operation (T1-6),6 mice were randomly selected,and blood samples were taken from the eyeballs for determination of serum tumor necrosis factor-alpha and interleukin-10 concentrations by enzyme-linked immunosorbent assay.Six mice were randomly selected at T4-6 and sacrificed,and the spleens were removed for measurement of the percentage of NK cells and their subsets by flow cytometry.Results Compared with Sham group,the survival rate was significantly decreased at different time points,the serum necrosis factor-alpha concentration at T1-3,6 and serum interleukin-10 concentration at T3-6 were increased,the percentage of NK cells was decreased at T4-6,the percentage of CD27-CD11b+NK cells was decreased and the percentage of CD27+CD11b+ and CD27+CD11b-NK cells was increased at T5,the percentage of CD27-CD11b-NK ceils was decreased at T4,5,and the percentage of CD27-CD11b-NK cells was increased at T6 in Sep group (P＜0.05).Conclusion Changes in NK cell subsets defined by CD11b and CD27 in the spleen play an important role in the development of sepsis in mice.

Changes of B lymphocyte subsets in peripheral blood of patients with rheumatoid arthritis and their ;significance / 中华微生物学和免疫学杂志

Objective To investigate the changes of B lymphocyte subsets ( naive B cells, memory B cells and plasmablasts) in peripheral blood of patients with rheumatoid arthritis ( RA) and their correla-tions with the clinical manifestation and laboratory indexes.Methods Sixty-six patients with RA were di-vided into two groups including the group with active RA and the group with inactive RA according to the dis-ease activity score in 28 Joints (DAS28).A control group with healthy subjects was set up accordingly.The distributions of B lymphocyte subsets in peripheral blood samples were detected with flow cytometry analysis and their correlations with clinical manifestations and laboratory indicators were analyzed.Results ( 1 ) Compared with healthy subjevts, the mean fluorescence intensities ( MFIs) of CD19 and the percentages of memory B cells in peripheral blood of the patients with RA were significantly decreased, while the percenta-ges of naive B cells were increased (P<0.05).The percentages of plasmablasts in the patients with active RA were significantly increased as compared with those of healthy subjects and the patients with inactive RA (P<0.05).(2) The percentages of plasmablasts in peripheral blood of the patients with RA were positively correlated with the joint tenderness count, joint swelling count and joint swelling index (P<0.05).(3) A positive correlation was found between the MFIs of CD19 and the erythrocyte sedimentation rates ( ESRs ) among the patients with RA.The percentages of plasmablasts were positively correlated with C reaction pro-tein (CRP) and anti-cyclic citrullinated peptide (anti-CCP) antibody (P<0.05).(4) The percentages of plasmablasts were also positively correlated with the DAS28 among the patients with RA ( R2=0.343, P<0.01).Conclusion The distributions of B lymphocyte subsets varied among the patients in different stages of RA, which were related to the patient′s clinical symptoms and laboratory indexes.The study suggested that different subsets of the B lymphocytes might play an important role in the pathological process of RA.

Significantly Higher Percentage of Circulating CD27(high) Plasma Cells in Systemic Lupus Erythematosus Patients with Infection than with Disease Flare-Up

PURPOSE: To distinguish lupus flare-up from infection in systemic lupus erythematosus (SLE), we analyze the expression of circulating CD27(high) plasma cells in SLE patients with and without infection, in comparison to non-SLE patients with infection. MATERIALS AND METHODS: The percentage of circulating CD27(high) plasma cells was measured by flow cytometry in the following four groups: 36 SLE patients without infection, 23 SLE patients with infection, eight non-SLE patients with infection, and 26 healthy controls. RESULTS: The frequency of CD27(high) plasma cells had a correlation with the SLE disease activity index (SLEDAI) (r = 0.866, p < 0.05), level of anti-dsDNA (r = 0.886, p < 0.05), C3 (r = - 0.392, p < 0.05), and C4 (r = - 0.337, p < 0.05) in SLE patients without infection, but there was no correlation with disease activity in SLE patients with infection. Among three groups in particular-SLE without infection, SLE with infection, and non-SLE with infection-the percentages of CD27(high) plasma cells were elevated. The percentage of CD27(high) plasma cells was higher in SLE patients with infection, when compared to SLE patients without infection. CONCLUSION: The percentage of CD27(high) plasma cells is a biomarker for disease activity of SLE without infection, under correlation with SLEDAI, anti-dsDNA, and C3 and C4 level. However, when the SLE patients have an infection, the percentage of CD27(high) plasma cells is not an adequate biomarker for the survey of disease activity. The percentage of CD27(high) plasma cells may serve as a potential parameter to distinguish a lupus flare-up from infection.

Analysis of CD27 and CD28 expression in antigen-specific CD4~+T cells of patients with pulmonary tuberculosis / 中国医师杂志

Objective To study the expression of CD27 and CD28 in antigen-specific CD4~+T cells in patients with pulmonary tuberculosis and healthy people, and understand the role of differentiated stages of CD4~+T cells in the pathogenesis of tuberculosis. Methods The expression of CD27 and CD28 was analyzed by CD4, CD154, CD27 and CD28 staining and flow cytometry. The distributions of CD27 and CD28 in antigen-specific CD4~+T cells were compared between patients with pulmonary tuberculosis and healthy controls. Results In patients of pulmonary tuberculosis, the frequencies of CD27 + CD28 + (early differentiated stage), CD27~- CD28~+ and CD27~+ CD28~- (intermediate differentiated stage), CD27~- CD28~-(fully differentiated stage) T cell subsets in antigen specific CD4~+T cells were (49. 55 ±6. 15)%, (26. 85 ±3. 87)% ,(7. 2 ± 1.37)% and ( 16. 35 ±3.97)%, respectively. In healthy controls, the frequencies of the four subsets in antigen-specific CD4~+T cells were ( 51.81 ± 4. 94 ) %, ( 29. 83 ± 5.33 ) %, ( 12. 65 ±4. 48)% and (5.7±2)%, respectively. The early differentiated CD4~+T cell was the major subset both in patients and healthy people, however, which had significant difference compared with the fully differentiated subset ( t = 2. 26, P < 0. 05 ). Conclusion The population frequency of the fully differentiated CD4~+T cells in patients with pulmonary tuberculosis was significantly higher than that in healthy people. This suggested that the differentiation degree of the antigen-specific CD4~+T cell might be related with pulmonary tuberculosis.

Imunodeficiência comum variável: distúrbio de diferenciação dos linfócitos B ou distúrbio de ativação dos linfócitos T? / Common Variable Immunodeficiency: disturbance of differentiation of B lymphocytes or disorder of activation of T lymphocytes?

A imunodeficiência comum variável (ICV) é uma imunodeficiência primária de origem heterogênea, definida como uma diminuição de pelo menos dois isótipos de imunoglobulinas, a falta de resposta anticórpica a imunizações e a exclusão de outras causas primárias de hipogamaglobulinemia. A ausência de níveis adequados de anticorpos em pacientes com ICV resulta em infecções bacterianas recorrentes, mais frequentes no trato respiratório e digestivo, que podem levar a sequelas sinusais e pulmonares. Nos últimos 6 anos iniciou-se a descoberta de genes relacionados à causa de doenças com o fenótipo de ICV, como os genes de TACI, BAFF-R, CD19 e ICOS. Dentre as alterações imunológicas, podemos também relatar deficiência de células B de memória (CD19+IgM-IgD-CD27+), levando a distúrbio de comutação isotípica e redução da secreção de imunoglobulinas. Atualmente tal característica vem sendo utilizada para classificar a ICV. No decorrer do presente trabalho pudemos observar que pacientes com ICV apresentam alterações na expressão de CD27 não somente em células B, mas também em células T, além de resposta linfoproliferativa ao estímulo de PHA reduzida. O CD27 consiste em uma molécula da família TNF presente constitutivamente em células T e após ativação em células B. Sua atuação na resposta imune está relacionada com a proliferação e co-ativação de células T específicas que atuam na interação T-B, na resposta de células B dependente de T. Dessa forma deficiências na via de CD27 podem resultar em defeitos nos mecanismos de comutação isotípica e de diferenciação de células B do centro germinativo, assim como de células de memória. Essas características podem ser observadas em modelos murinos de deficiências de CD27/CD70. Nossos achados permitem que uma nova janela se abra para o estudo da ICV. A avaliação dos distúrbios associados a defeitos de sinalização de CD27/CD70 em humanos pode se tornar uma nova ferramenta para a compreensão de uma deficiência tão pouco esclarecida...

Common variable immunodeficiency (CVID) is a primary immunodeficiency disorder of heterogeneous origin, defined by a decrease of at least two immunoglobulin isotypes, lack of antibody response to immunization and the exclusion of other causes of primary hypogammaglobulinemia. The absence of adequate levels of antibodies in patients with CVID results in recurrent bacterial infections, most frequently in the respiratory and digestive tract, which can lead to sinusal and lung sequels. Over the past six years the discovery of genes related to the phenotype of CVID began, such as the genes of TACI, BAFF-R, CD19 and ICOS. Among the immunological changes, there is impairment of memory B cells (CD19+/IgM-IgD-CD27+), leading to disturbance of isotypic switching and reduced secretion of immunogobulins. Currently this feature has been used to classify CVID. During the present study we observed that patients with CVID present changes in the expression of CD27 not only in B cells, but also in T cells, and reduced lymphoproliferative response to PHA. CD27 molecule is a member of the TNF family present constitutively in T cells, and after activation in B cells. Its importance in the immune response is related to the proliferation and co-activation of specific T cells that act in T-B interaction, in the T cell dependent B cells response. Thus disturbances in the CD27 pathway can result in defects in isotypic switch and differentiation of germinal center B cells, as well as memory cells. These characteristics can be observed in murine models of CD27/CD70 deficiency. Our findings allow a new approach for the study of CVID. The evaluation of defects in CD27/CD70 signaling in humans might become a new tool for understanding an incompletely understood disease. Such an approach may contribute to the development of new treatments, acting directly on the molecule in question. In addition, we also suggest the use of phenotyping of CD27 molecules on B and T lymphocytes, in addition to...

Expression of B7-related protein-1 on B lymphocytes in peripheral blood from patients with systemic lupus erythematosus and its relationship with disease activity / 中华皮肤科杂志

Objeetive To explore the differentiation of B lymphocytes and expression of B7-related protein-1 (B7RP-1)on B lymphocytes in patients with systemic lupus erythematosus(SLE).Methods Three-color immunofluorescent staining and flow cytometric assay were used to analyze the frequency of three types of B lymphocytes,I.e.,plasma cells,memory B lyphocytes and naive B lymphocytes,as well as the expression of B7RP-1 on these cells in peripheral blood from 23 patients with SLE and 16 normal human controls.Clinical data of these patients with SLE were collected.and SLE disease activi index(SLEDAI)was also evaluated.The relationship was assessed between the expression of B7RP-1 and SLEDAI.Results The frequency of plasma cells was highest in patients with active SLE.followed by patients with inactive SLE and normal human controls(P＜0.01).A significant decrease was observed in the frequency of memory B lymphocytes in patients with active SLE compared with normal controls (P＜0.01),but no significant difference was found between patients with inactive SLE and those with active SLE(P＞0.05).Regarding the frequency of naive B lymphocytes,there was no significant difierence among the three groups.Increased frequency of plasma cells was also noted in patients with lupus nephritis(LN)compared with those without LN [(6.15±3.12)%vs(3.31±1.41)%,P＜0.05 ],but no significant difierence was found with regard to the frequency of memory or naive B lymphocytes between these two groups.The expression rate Of B7RP-1 was significantly lower on total lymphocytes from patients with SLE than from normal human controls (46.51%vs 63.75%,P＜0.05),which was the case with B7RP-1 on plasma cells,memory B lyphocytes and naive B lymphocytes (all P＜0.01),whereas no significant difierence was found between patients with inactive SLE and active SLE or between patients with and without LN.In addition.no correlation was found between the expression of B7RP-1 and SLEDAI(r=0.035,P＞0.05).Conclusions In peripheral blood of patients with SLE,the frequency of plasma cells is increased,while the expression of B7RP-1 on lymphocytes is decreased,which may be relevant to the pathogenesis of SLE.

Study on activation of CD27-CD70 co-stimulatory pathway in patients with inflammatory bowel disease / 中华消化杂志

Objective To investigate the expression of CD27-CD70 co-stimulatory pathway in peripheral circulation and intestinal mucosa of patients with inflammatory bowel disease, and to find the difference between the expression of CD27-CD70 in patients with inflammatory bowel disease and in healthy controls. Methods A total of 62 patients with Crohn's disease, 64 patients with ulcerative colitis and 56 healthy controls were enrolled. Enzyme-linked immunosorbent assay was applied to evaluate plasma CD27-CD70 protein expression in patients with inflammatory bowel disease and healthy controls. SYBR-green real time PCR was applied to access CD27-CD70 mRNA expression in peripheral blood mononuclear cells in patients with inflammatory bowel disease and healthy controls.And CD27-CD70 protein expression in intestinal mucosa was determined by immunohitochemistry.Results Plasma levels of CD27 (P=0. 025) and CD70 (P=0. 000) were significantly higher in patients with Crohn's disease than in healthy controls. However, CD27 (r= 0. 055, P= 0. 673) and CD70 (r= 0. 024, P = 0. 852) were not significantly associated with endoscopic disease activity in patients with Crohn's disease. Similarly, CD27 (P=0. 001) and CD70 (P=0. 000) were significantly higher in patients with ulcerative colitis than in healthy controls. And CD27 (r=0. 077, P=0. 547)and CDT0 (r=0.021, P=0. 869) were not significantly associated with endoscopic disease activity in patients with ulcerative colitis. Moreover, CD27 and CD70 mRNA expression in peripheral blood mononuclear cells were significantly higher in patients with Crohn's disease and ulcerative colitis than in healthy controls (all P=0. 000), and immunostaining indicated that CD27 and CD70 expression in intestinal mucosa were significantly higher in patients with Crohn's disease and ulcerative colitis than in healthy controls (all P=0. 000). Conclusions CD27-CD70 pathway activated in plasma, peripheral blood mononuclear cells and intestinal mucosa of patients with inflammatory bowel disease. However,plasma levels of CD27 and CD70 can not reflect endoscopic disease activity.

Down-regulation of Membrane Bound CD27 Expression Mediated by Protein Kinase C-beta and Protein Kinase C-delta in B Cells / 대한류마티스학회지

OBJECTIVE: CD27 is a member of the tumor necrosis factor receptor (TNFR) superfamily and is expressed on T, B, and NK cells. The signaling via CD27 plays pivotal roles in T-T and T-B interaction. CD27 is a useful marker in assessing the number of circulation B cells and B cell subsets because it permits one step identification of the major B cell compartments, CD27- naive and CD27+ memory B cells as well as CD27high plasma cells. We have analyzed the mechanisms underlying the regulation of CD27 expression. METHODS: Isolation B cells and Raji cells were cultured with PMA. The levels of cell surface CD27 and CD 27 mRNA were analyzed by FACs staining and RT-PCR. Raji cells were cultured with phorbol 12-myristate 13-acetate (PMA), with or without pretreated shedding inhibitor BB3103 and TAPI-1. sCD27 was measured in culture supernatant by ELISA. Cell lysates were analyzed for PKC isotype activation by Western blot. We used PKC inhibitor Ly379196 and rottlen. RESULTS: Membrane expression of CD27 was down-regulated after PMA stimulation without cytotoxic effect in B cells. Furthermore, PMA treatment could directly reduce CD27 mRNA without intermediate protein synthesis in B cells. In contrast, PMA treatment induced soluble form of CD27 (sCD27), which was shed from the cell surface and was found in PMA treatment B cell culture supernatant. PMA-induced sCD27 proteins were decreased with shedding inhibitor BB3103 and TAPI-1. PMA-induced down regulation of CD27 expressions were quenched with protein kinase C (PKC) inhibitor Staurosporin, PKC-beta inhibitor rottlerin and PKC-delta inhibitor Ly379196. CONCLUSION: These data suggest that PMA-induced activation of PKC plays a crucial role in down-regulation of the expression of the CD27 and up-regulation of the shedding of the CD27 in human B cells.

Differential Signaling via Tumor Necrosis Factor-Associated Factors (TRAFs) by CD27 and CD40 in Mouse B Cells

BACKGROUND: CD27 is recently known as a memory B cell marker and is mainly expressed in activated T cells, some B cell population and NK cells. CD27 is a member of tumor necrosis factor receptor family. Like CD40 molecule, CD27 has (P/S/T/A) X(Q/E)E motif for interacting with TNF receptor-associated factors (TRAFs), and TRAF2 and TRAF5 bindings to CD27 in 293T cells were reported. METHODS: To investigate the CD27 signaling effect in B cells, human CD40 extracellular domain containing mouse CD27 cytoplamic domain construct (hCD40-mCD27) was transfected into mouse B cell line CH12.LX and M12.4.1. RESULTS: Through the stimulation of hCD40-mCD27 molecule via anti-human CD40 antibody or CD154 ligation, expression of CD11a, CD23, CD54, CD70 and CD80 were increased and secretion of IgM was induced, which were comparable to the effect of CD40 stimulation. TRAF2 and TRAF3 were recruited into lipid-enriched membrane raft and were bound to CD27 in M12.4.1 cells. CD27 stimulation, however, did not increase TRAF2 or TRAF3 degradation. CONCLUSION: In contrast to CD40 signaling pathway, TRAF2 and TRAF3 degradation was not observed after CD27 stimulation and it might contribute to prolonged B cell activation through CD27 signaling.

Study on CD19 and CD27 of B lymphocyte subsets in peripheral blood of patients with rheumatoid arthritis and its correlation / 中华风湿病学杂志

Objective To study B lymphocyte subsets(na(?)ve B cells,memory B cells and plas- mablasts)of peripheral blood in patients with rheumatoid arthritis(RA)and its relationship with autoantibod- ies and clinical manifestation.Methods Blood samples and clinical data of 60 patients with RA were enrolled into this study.They were divided into three groups:active,inactive and refractory RA based on clinical mani- festations and 24 healthy controls were included.CD19 and CD27 of B cells in peripheral blood of RA patients and healthy controls were detected using flow cytometry at single-cell level.Frequence of na(?)ve B cells (CD19~+CD27~-),memory B cells(CD19~+CD27~(dim)),plasmablasts(CD19~+CD27~(high))and average fluorescence in- tensity of CD19 were analyzed,and their relationship with clinical manifestations and rheumatoid factor(RF), anti-typeⅡcollagen(anti-CⅡ),anti-cyclic citrullianted peptide(CCP)antibodies were investigatied.Results Frequence of na(?)ve B cells and plasmablasts in peripheral blood of patients with RA was increased compared with normal control.In contrast,memory B cells in patients with RA were decreased.The na(?)ve B cells subset in inactive and refractory RA was higher than that of healthy controls(P＜0.05),and the memory B cells subset in those groups was lower than that of healthy controls(P＜0.05).The plasmablasts in active and refractory groups of RA were higher than those of healthy controls(P＜0.05).The average fluorescence intensity of CD19 in peripheral blood in patients with RA was positively correlated with ESR,C-reactive protein(CRP),healthy assessment questionaire(HAQ),and plasmablasts was positively correlated with arthrocele index.Na(?)ve B cells,memory B cells and plasmablasts subsets had no relation with RF,anti-CⅡand anti-CCP antibodies. Conclusion B cell subsets in peripheral blood of patients with RA are significantly abnormal,characterized by expanded naive B cells and plasmablasts but diminished memory B cells.Plasmablasts are increasesd in active and refractory groups of RA,and have positive correlation with swollen joint index.B cells may play an important rote in the pathogenesis of RA.

Common phenotypic molecule for classification of NK cell subsets of both human and mouse:CD27 molecule / 中国免疫学杂志

NK cells can be divided into two subsets based on CD27 expression:CD27hiNK cells and CD27loNK cells.NK cells can be further divided into four subsets based on CD27 and CD11b: CD11bloCD27lo,CD11bloCD27hi,CD11bhiCD27hi and CD11bhiCD27lo.These subsets in different development stages have their own phenotypes and functions.In mice,NK cell developing pathway is through CD11bloCD27lo→CD11bloCD27hi→CD11bhiCD27hi→CD11bhiCD27lo.CD27 can also be used to divide human NK cells into two subsets which are comparable to those of mice.

Study on the changes of CD_(28) and soluble CD_(27) and their clinical significance among HIV-1-infected chinese patients receiving highly active antiretroviral therapy / 中国医师杂志

Objective To observe the changes of CD_(28) and soluble CD_(27)(sCD_(27)) among HIV-1-infected Chinese patients receiving highly active antiretroviral therapy(HAART) and to explore their clinical significance.Methods The HIV-RNA viral load(VL) and cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+CD_(62)L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ and the content of sCD_(27) were determined in 28 cases of HIV-1-infected patients at the baseline and 12,24 and 48 weeks respectively after the treatment with three drug regimens based on two nucleoside reverse transcriptase inhibitors(NRTIs) and an Non-NRTI(NNRTI) for 1 year.Results The average of HIV-RNA VL before the treatment was 5.25 logs,and decreased to 3.30,1.92 and 1.74 logs after the treatment for 12,24 and 48 weeks respectively.The cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+62L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ before the treatment were(236?85),(917?345),(58?15),(175?63) and(285?87) cells/?l,and changed into(327?112),(803?343),(114?25),(286?132) and(387?201) cells/?l after the treatment for 48 weeks respectively.The content of sCD_(27) decreased from(475?123) U/ml to(174?77) U/ml after the treatment for 48 weeks.The HIV RNA VL had significantly negative correlation with cell counts of CD_4~+T,CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+,and there was positive correlation between HIV RNA VL and the level of sCD_(27).Conclusion The immune reconstitution and activation depend on the efficacy of antiretroviral therapy.The HIV RNA VL and cell counts of CD_4~+T are the representative parameters of immunologic and viral responses of HAART.The cell counts of CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+ and their percentages show concomitant changes with viral load,and play an effective role in the progression and prognosis of AIDS.The level of sCD_(27) is positively correlated with HIV RNA VL and negatively with counts of CD_4~+T cell.The detection of sCD_(27) by enzyme linked immunosorbent assay(ELISA),which is much more convenient and cheaper than RT-PCR and flow cytometry,may serve as an important parameter to judge the pathogenetic condition of HIV-1-infected patients and to examine the therapeutic effects of HAART in the medical resource-limited area.

Changes of the expression of CD27 and CD28 on cytokine-induced killer cells and natural killer cells during four week cultivation / 中国病理生理杂志

AIM:To investigate the changes of CD27 and CD28 expression on cytokine-induced killer(CIK) cells and natural killer(NK) cells during cultivation.METHODS:The mononuclear cells were treated with interleukin-12(IL-2),IL-7,IL-15,stem cell factor and FLT3 ligand for four weeks. The CD27 and CD28 expression on CD3+CD56+ CIK cells and CD3-CD56+ NK cells were examined by flow cytometric analysis every week during four-week cultivation.RESULTS:The expression of CD27 on CIK and NK cells two weeks after cultivation reached the peak value at(44.57?4.07)% and(51.02?5.20)%,respectively. Then,their expression declined gradually to about 30% in the fourth week. The expression of CD28 on CIK and NK cells reached the peak at the third and the second week,the values were (4.40?0.66)% and (45.14?3.58)%,respectively,decreased rapidly and then were almost completely lost at the fourth week. CONCLUSION:According to the changes of expressions of CD27 and CD28 on CIK/NK cells,optimal harvest time of cultured CIK/NK cells should be at the third week of cultivation.