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Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P＜0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P ＜ 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.
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ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44％±0.04％,which was higher than those to other four HBV antigen peptides (0.68％±0.08％ of FLLTRILTI,1.06％±0.09％ of FLPSDFFPSV,0.56％ ±0.04％ of FLLSLGIHL,and 0.46％ ±0.08％ of WLSLLVPFV) (t=0.001,P＜0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.
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ObjectiveTo investigate the effect of combined spinal epidural analgesia (CSEA) on immune function by observing the changed level of T lymphocyte subsets in maternity sera in labor.MethodsFifty healthy primipara with single birth,vertex present and ASA I between July 2007 and Dec 2007 at the first Affiliated Hospital of Nangchang University,who were in spontaneous labor,were randomly divided into two subgroups when their rerviral dilations were in 2～3 cm.In interfering subgroup( n =25),the puncture point of CSEA was at L3-4 interspace,the fentanyl (20 μg) was used in lumbar anesthesia,the ropivaraine (0.1％) rombined with fentanyl (2 μg/ml) was used in epidural analgesia.Blood samples were taken from the mother vein at cervical dilation in 2 ～ 3 m (T1),fetal disengagement(T2),24 hrs after childbirth ( T3 ).Flow cytometry was used to measure T lymphocyte subsets,Radioimmunoassay (RIA) was used to measure cortisol.In addition,Data on labor progress,VAS score,and neonatal Apgar score were recorded for each patient.Results(1)The active phase in the first stage of labor after analgesia in the CSEA group [ ( 177.64 ± 67.98 ) min ] was significantly lower than that in control group [ (219.40 ± 67.37) min ].No significant difference was found for the active phase in the second stage and the third stage,and for the neonatal Apgar score between the CSEA group [ (32.92 ± 11.59 ) min,( 7.56 ± 2.47 )min,9.20 ± 0.82,respectively ] and the control group [ ( 31.44 ± 13.93 ) min,( 7.28 ± 2.25 ) min,8.84± 1.31,respectively ].(2)The level of cortisol in A group [ ( 548.11 ± 75.67) ng/ml ] was significantly lower than that in C group[ (789.32±96.07) ng/ml] at T2.(3) In two groups,the levels of the CD3+,CD4+,CD4+/CD8 + degraded in different degree at each point,more significantly decreased at T3,and these in C group[ (48.43 ± 6.46) ％,( 31.35 ± 8.93 ) ％,(0.96 ± 0.21 ) ％,respectively ] were significantly lower than those in A group [ (52.3 ± 5.62 ) ％,( 36.90 ± 7.91 ) ％,( 1.16 ± 0.25 ) ％,respectively ].ConclusionsCSEA could shorten the active delivery phase in the first stage of labor,and did not affect the neonatal Apgar score.It can alleviate the inhibitory effect of pain stress response on the immune function.
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Objective To study on the changes of the DNT cells and T lymphocyte subtype in children with infectious mononucleosis (IM) and its clinical significance.Methods The flow cytometry (FCM) was used to detected the DNT cells and other T lymphocyte subtype in 48 cases of IM.Results The study showed that DNT cells( 9.39 ± 4.89 )% were greatly increased in comparison with normal controls (NC) (4.26 ± 1.68)% ( P ＜0.01 ).CD4 cells(21.45 ±9.87)% were decreased ( P ＜0.01 ) and CD8 cells increased in comparison with NC (32.43 ± 5.07) % ( P ＜ 0.01 ).Conclusion DNT cells and T lymphocyte subtype can be used to evaluate the immune function of children with infectious mononucleosis (IM) and provide guidance for adoptive immunotherapy.
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Objective To investigate.the co-stimulatory pathway of gastric cancer patients by examining the expression of co-stimulatory molecules CD28,CTLA.4(CDl 52)and B7 molecules on peripheral blood lymphocytes(PBLC)in patients with gastric cancer,and to discuss the role of costimulatory pathway in the pathogenesis of gastric cancer.Methods The expression of CD28,CD152,CD3,CD4,CD8 and B7 molecules on peripheral blood lymphocytes and T cell subpopulation were measured by flow cytometry in 56 patients with gastric cancer,and the data was compared and analyzed with that in gastric benign tumor group and healthy control group.Results The expression levels of CD3+,CD4+,CD3+ CD28+,B7-1(CD80),B7-2(CD86)and CD41/CD8+on PBLC in patients with gastric cancer was significantly lower than that of benign stomach tumor group and healthy control group.The expression levels of CD3+ CDl52+ and CD8+ on PBLC in patients with gastric cancer were significantly higher than those of gastric benign tumor group and healthy control group.Conclusion Peripheral lymphocytcs in gastric cancer patients express low level of CD28 and B7 molecules and high level of CDI 52 molecules,which results in dysfunction of B7:CD28/CTLA-4 co-stimulation pathway and in turn affects the capacity of T cell to eliminate tumor cells,which makes tumor cells evading the immune surveillance and metastasizing.
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Objective To observe the changes of PECAM-1,Bcl-2 and Bax expressed by ischemic cerebrum of adult rats after focal cerebral ischemia. Methods Middle cerebral artery occlusion (MCAO) models were made by operation with Longa suture method in Wistar rats. The expression levels of PECAM-1,Bcl-2 and Bax at the 6th,12th,18th,24th,48th,72nd hour after MCAO were detected by immunohistochemistry staining. Results The expression levels of PECAM-1,Bcl-2 and Bax in cerebrum were significantly increased after MCAO. The expression levels of Bcl-2 we、re up to the peak at the 12th hour after MCAO, while the levels of Bax and PECAM-1 were up to the peak at the 24th and 48th hour after MCAO. At the 72nd hour after MCAO, the expression levels of PECAM.1.Bcl-2 and Bax were still higher than that in the control group(all P＜0.001).Conclusions PECAM-1,Bcl-2 and Bax participate in the different pathological stages of focal cerebral ischemia.
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The incidence of posttransplantation lymphoproliferative disorders (PTLDs) has increased in recent years. Although rare, various types of T-cell lymphoma have been reported and their association with Epstein-Barr virus (EBV) has been compared with B-cell PTLDs. We report a case of splenic peripheral T-cell lymphoma occurring in a 47-yr-old male patient 7 yr after renal allograft transplantation. The spleen showed sinusoidal proliferation of focal CD30 positive, large, atypical lymphoid cells. Positivity for CD3 and cytolytic granule-associated proteins was also demonstrated in the tumor cells, while anaplastic large cell lymphoma kinase (ALK) and CD8 were not expressed. Strong nuclear signals for EBV mRNA were noted by EBER1 in situ hybridization. A molecular genetic study demonstrated a rearrangement of the gamma T-cell receptor gene. To our knowledge, this case is unique in terms of a posttransplant T-cell lymphoma that shows focal CD30, cytolytic granule-associated proteins, and EBV positivity.
Subject(s)
Humans , Male , Middle Aged , Ki-1 Antigen/genetics , Ki-1 Antigen/metabolism , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/metabolism , Kidney Transplantation , Lymphoma, T-Cell, Peripheral/pathology , Lymphoma, T-Cell, Peripheral/virology , Membrane Proteins/metabolism , RNA, Viral , RNA-Binding Proteins/metabolism , Serine Endopeptidases/metabolism , Splenic Neoplasms/pathology , Splenic Neoplasms/virologyABSTRACT
To investigate the role of Fas/FasL system in pathogenesis of hemodialysis(HD) and its correlation with clinical status, plasma levels of sFas/sFasL and the expression of Fas antigen on peripheral blood lymphocytes(PBL) membrane were assayed by ELISA and flow cytometry respectively. The CD3 + Fas + of PBL cells was singnificantly higher( P
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To investigate the changes in the activated T-lymphocyte CD3/HLA-DR and CD3/CD(16+56) populations in peripheral blood of the patients with allogeneic hand transplantation, lymphocytes from peripheral blood of the patients at different time points were immunologically labeled with dual color fluoresecent monoclonal antibodies CD3/CD(16+56) and CD3/HLA-DR, mono-color fluoresecent monoclonal antibody CD25. CD25, CD3/CD(16+56), and CD3/HLA-DR were determined with flow cytometry (FCM). The levels of activated T-lymphocyte (CD25 +,CD3 +/HLA-DR + ), silent T-lymphocyte [CD3 +/CD(16+56) -,CD3 +/HLA-DA - ] decreased significantly during the first week after transplantation and then increased gradually to the pre-operafive level. Nature killer cells [CD3 -/CD(16+56) +] increased significantly at the first day after transplantation, then decreased sharply and maintained a lower level. The results suggest that immunosuppressive agents have significantly effects on lymphocyte subsets in allogenaic hand transplanted patients, and dynamic determination of HLA-DR, CD3 /CD(16+56) could be valuable in immunomonitoring after allogeneichand transplantation.
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Objective To discuss the relationship between disease progress with the change of cell im munological function as well as the effect of immunological function in severe a cute respiratory syndrome. Methods Retrospective study was designed to analyze the relationship of disease progres s with the change of immunological function. According to the disease outcomes, the patients are divided into two groups: died group and survival group. The dif feren ce of immunological function in groups was performed statistics analysis. Results Immunological function (CD3 +, CD4 +, CD8 +) descended to the lowest level in period of fastigium in SARS patient, and then recovery quickly. Comparison of died grou p and survival group was found that Immunological function (CD3 +T, CD4 +T, CD 8 +T) were lower in period of fastigium and recovery in died group than in survival g roup(P
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Objective To study the percentage of HBV specific CD3 + T cells in the peripheral blood of chronic hepatitis B patients. Methods HLA A 2 patients were screened by MHC:Ig dimer reagent. HLA A 2 restricted HBV core, polymerase, and surface peptides were added to T cells of chronic hepatitis B patients, followed by double staining with anti CD3 antibodies and antibodies reactive with the heavy chain of Ig. FACS was used for monitoring HBV specific T cell percentage. Results 23/54 (42.6%) patients belonged to HLA A 2. Fifteen of them were reacted with HBV peptides. CD3 + T cells of five patients showed higher percentage of binding with the HBV peptides than that in the controls. Four patients showed CD3 + T cells binding with S peptide, one with P peptide and one binding with both S and P peptide. Conclusions The percentage of T cells which could bind with HBV peptide was low in chronic hepatitis B and the binding efficiency was low as well. The MHC:Ig dimer reagent is convenient for counting the number of HBV specific T cells, but the nonspecific staining background seems to be relatively high. However, this technology is useful to monitor changes of immune responses in chronic hepatitis B patients during immuno therapy.