1.Apoptosis in human germinal centre B cells by means of CC chemokine receptor 3 expression induced by interleukin-2 and interleukin-4.
Qiu-ping ZHANG ; Luo-kun XIE ; Li-jun ZHANG ; Jin-quan TAN
Chinese Medical Journal 2005;118(8):665-670
BACKGROUNDCC chemokine receptor 3 (CCR3), expressed on some inflammatory cells, is a member of the chemokine receptor family. Its ligand is eotaxin/CCL11. In this research, we studied the expression and function of CCR3 induced by interleukin-2 (IL-2) and interleukin-4 (IL-4) on human germinal centre (GC) B cells.
METHODSCells isolated from human tonsils were stimulated with IL-2 or/and IL-4 followed by bonding with eotaxin/CCL11. Flow cytometry was used to detect expression of CCR3 on GC B cells and apoptosis of GC B cells. Real time quantitative reverse transcription polymerase chain reaction and Northern blot assays were used to analyse the CCR3 mRNA expressed in the GC B cells. Chemotaxis and adhesion assays were used to determine the effect of eotaxin/CCL11 ligand bonded to CCR3 on GC B cells.
RESULTSThere was no CCR3 expression on human freshly isolated GC B cells. The combination IL-2 and IL-4 could upregulate CCR3 mRNA and protein expression on GC B cells. Eotaxin could not induce GC B cell chemotaxis and adhesion but triggered apoptosis of GC B cells.
CONCLUSIONIL-2 and IL-4 together induced expression of CCR3 on GC B cells, and the receptor acted as a death receptor.
Apoptosis ; B-Lymphocytes ; metabolism ; pathology ; Cell Adhesion ; Chemotaxis, Leukocyte ; Germinal Center ; metabolism ; pathology ; Humans ; Interleukin-2 ; pharmacology ; Interleukin-4 ; pharmacology ; RNA, Messenger ; analysis ; Receptors, CCR3 ; Receptors, Chemokine ; genetics
2.Study of hemophagocytic lymphohistiocytosis in children.
Wen LIN ; Yan XIAO ; Run-ming JIN
Chinese Journal of Pediatrics 2003;41(10):792-794
3.Serum interleukin-6 in Kawasaki disease.
Yonsei Medical Journal 1992;33(2):183-188
Kawasaki disease (KD) is an acute febrile illness of infancy and early childhood. In spite of extensive studies, the cause of KD is not known. Interleukin 6 (IL-6) has manyfold biological functions involved in the immune or inflammatory responses of the host to various stimuli. Here the author investigated whether IL-6 might be responsible for manifestations of KD, such as immunoglobulin hypersecretion, lymphocyte activation and systemic vasculitis. Serum IL-6 levels in KD were determined by ELISA. Usually sera from healthy children contained only negligible levels of IL-6. Serum IL-6 was markedly elevated in all patients with acute KD, which gradually decreased during the course of the disease. Serum IL-6 correlated with serum concentration of C-reactive protein and with serum soluble interleukin-2 receptor level, but did not show any correlation with peak platelet count during subacute phase of the disease. Increased serum IL-6 level did not show any relation to development of coronary aneurysms and dilatation. Further studies will be needed to examine the source and the pathogenetic roles of increased serum IL-6 in KD.
Child
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Child, Preschool
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Female
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Human
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Infant
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Interleukin-6/*blood
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Male
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Mucocutaneous Lymph Node Syndrome/*blood
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Receptors, Interleukin-2/analysis
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Thrombocytosis/etiology
4.Value of serum soluble interleukin-2R, interleukin-6 and C-reactive protein in the early diagnosis of Kawasaki disease.
Qian PENG ; Qing WU ; Chang-Hui CHEN ; Hua HONG ; Ling-Ying ZHANG
Chinese Journal of Contemporary Pediatrics 2006;8(3):208-210
OBJECTIVEImmunovasculitis is a pathologic process of Kawasaki disease (KD) in the early stage and it is more likely to result from abnormal immunoactivation. It is thus speculated that the serum levels of some cytokines have changed before immunovasculitis occurs, suggesting the cytokines may be useful markers for the early diagnosis of KD. In this study, we measured the serum levels of soluble interleukin-2 receptors (sIL-2R), interleukin-6(IL-6) and high-sensitive C-reactive protein (hs-CRP) in patients with KD to evaluate the significance of these cytokines in the early diagnosis of KD.
METHODSSerum levels of sIL-2R and IL-6 were measured by rapid one-step sandwich enzyme immunoassay and the serum hs-CRP level was measured by Dade Behring BN ProSpec in 32 KD patients before and after intravenous immunoglobulin (IVIG) therapy. Twenty healthy children were used as the controls.
RESULTSBefore IVIG therapy serum levels of sIL-2R (9253.41 +/- 2568.38 pg/mL vs 2161.53 +/- 696.92 pg/mL; P < 0.05), IL-6 (57.19 +/- 45.78 ng/mL vs 7.04 +/- 1.69 ng/mL; P < 0.05) and hs-CRP (117.69 +/- 42.05 mg/L vs 1.15 +/- 0.54 mg/L; P < 0.05) in KD patients were significantly higher than those in healthy controls. After IVIG therapy in KD patients serum IL-6 levels returned to normal and sIL-2R and hs-CRP levels decreased significantly but remained significantly higher than controls (P < 0.05). There was a positive correlation between sIL-2R and hs-CRP levels (r=0.60, P < 0.01). IL-6 levels positively correlated with hs-CRP levels in KD patients before IVIG therapy (r=0.68, P < 0.01).
CONCLUSIONSIL-2R, IL-6 and hs-CRP are activated in the development of KD, and they may be of important value in the early diagnosis of KD.
C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Infant ; Interleukin-6 ; blood ; Male ; Mucocutaneous Lymph Node Syndrome ; blood ; therapy ; Receptors, Interleukin-2 ; blood
5.Effect of activation of cellular immunity on p58+ cells expressing killer-cell-inhibitory receptor cells.
Xing-Hua PANG ; Rong-Qing PANG ; Kun-Yuan GUO ; Jiu-Gang SONG ; Jiang-Qi LI ; Yu-Jin ZHANG ; Xiao-Fen YANG
Journal of Experimental Hematology 2003;11(1):70-73
UNLABELLEDThe purpose of this study was to evaluate the effects of cellular immunity activation on P58(+) cells expressing killer cell inhibitory receptor (KIR) and their regulatory function on cellular immunity, and provid theoretical data for preventing graft-vers-host disease (GVHD) in stem cell transplantation therapy. The mononuclear cells from human peripheral blood were incubated with IL-2, Con A and Lipostin (LP) for 72 hours. The KIR expressing cells, P58.1(+) and P58.2(+) cells, were analyzed by flow cytometry. The results showed that the percentages of CD3(+), CD4(+), CD8(+), CD16(+)CD56(+), P58.1(+) and P58.2(+) cells were greatly increased after treated with IL-2, Con A and LP, separately or in combination, and the percentages of above cells in combined treatment groups were higher than those of single stimulated groups, especially the percentage of cells in the IL-2 + LP group was significantly higher than those in IL-2 and LP singly treated groups.
IN CONCLUSIONIL-2, Con A and LP possess the ability to induce the expression of KIR and stimulate proliferation of P58.1(+) and P58.2(+) cells while to activate the celluar immunity response, the expression of P58 gene may be regulated by the activation of cellular immunity.
Adult ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD56 Antigen ; analysis ; CD8 Antigens ; analysis ; Cell Count ; Cell Division ; drug effects ; Concanavalin A ; pharmacology ; Flow Cytometry ; Humans ; Interleukin-2 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Receptors, IgG ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR ; Receptors, KIR2DL3
6.Effects of humanized recombinant CD25 monoclonal antibody on activation and proliferation of t lymphocytes.
Journal of Experimental Hematology 2007;15(1):134-137
The study was purposed to investigate the effects of humanized recombined CD25 monoclonal antibody (rhCD25MAb) on activation and proliferation of T lymphocytes in vitro. Peripheral blood mononuclear cells (PBMNC) were incubated with phytohemagglutinin (PHA). Before or after T cell activation, the cells were cultured with or without rhCD25MAb or cyclosporine A (CsA) in vitro. After 72 hours incubation, the proliferation of lymphocytes was analyzed by MTT assay. The expression of CD3 and CD25 antigens on T lymphocytes were detected by flow cytometry. The levels of sIL-2R in the supernatants were determined with ELISA. The results showed that both rhCD25MAb and CsA could inhibit the proliferation of T lymphocytes significantly in concentration-dependent manner and CsA was more efficient than rhCD25MAb. Both rhCD25MAb and CsA could also decrease the levels of sIL-2R in the supernatant and inhibit the expression of CD25 antigen on T lymphocytes. The level of sIL-2R and the expression of CD25 on T lymphocytes decreases more profoundly in rhCD25MAb group. It is concluded that rhCD25MAb shows strong immunosuppressive activity both before and after T cell activation, suggesting that this agent may be useful in not only prophylaxis but also the treatment of acute graft-versus-host disease.
Antibodies, Monoclonal
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pharmacology
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CD3 Complex
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biosynthesis
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genetics
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Cell Proliferation
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Cells, Cultured
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Cyclosporine
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pharmacology
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Humans
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Immunosuppressive Agents
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pharmacology
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Interleukin-2 Receptor alpha Subunit
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biosynthesis
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genetics
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immunology
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Lymphocyte Activation
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immunology
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Receptors, Interleukin-2
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analysis
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Recombinant Proteins
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immunology
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pharmacology
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T-Lymphocytes
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immunology
7.Expression of killer cell inhibitor receptors on immunocompetent cells with relation to graft-versus-host disease after hematopoietic stem cell transplantation.
Lian-Ning DUAN ; Chun CHEN ; Shao-Liang HUANG ; Jian-Pei FANG ; Jing WEI ; Rong BAO ; Yan LI ; Hong-Xing HAN ; Shu-Nong LI
Journal of Experimental Hematology 2003;11(6):625-632
The study was aimed at the exploration of relationship between T cells expressing killer cell inhibitor receptors (KIR, CD158 and CD94) and graft-versus-host disease (GVHD) after hematopoietic stem cell transplantation. The expression rates of CD158a, CD158b and CD94 on T cells and NK cell were detected by flow cytometry and donor/recipient HLA-Cw was analyzed using PCR after peripheral blood stem cell transplantation (PBSCT) and umbilical cord blood transplantation (UCBT). After both PBSCT and UCBT, the rates of CD3(+)CD158a(+) and CD3(+)CD158b(+) T cells increased, especially the rate of CD8(+)CD158b(+) T cells. In both acute and chronic GVHD groups, the rate of CD3(+)CD158b(+) T cells increased, especially in acute GVHD. The CD94 mainly expressed on CD3(+)CD8(+) T cells. The percentage of the expression of CD94 on CD4(+) and CD8(+) cells after UCBT and PBSCT increased significantly. The expression of KIR in GVHD (early stage of transplantation) increased but the expression of KIR in chronic GVHD (advanced stage of transplantation) decreased. Five patients who HLA-Cw matched had no severe GVHD. In four patients who underwent allo-PBSCT and UCBT from related HLA-matched donors, only 2 patients had no aGVHD. Four patients underwent transplantation from unrelated HLA-matched donors had GVHD. These observations suggested that there is some relationship between GVHD and KIR expression on T cells. CD158b might be an inhibitory molecule of T cell activated at early stage after transplantation. Understanding the mechanism of GVHD with the expression of KIR on T cells, especially those binding the HLA-Cw might shed light on the establishment of the specific immunotolerance for the prevention of GVHD. To pay attention to HLA-Cw typing is very important to reduce GVHD and increase GVL effect in related or unrelated HLA-matched transplantation.
Antigens, CD
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analysis
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Antigens, Differentiation, T-Lymphocyte
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analysis
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Genotype
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Graft vs Host Disease
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etiology
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HLA-C Antigens
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genetics
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Hematopoietic Stem Cell Transplantation
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Humans
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Lectins, C-Type
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Receptors, Immunologic
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analysis
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Receptors, Interleukin-2
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analysis
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Receptors, KIR
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Receptors, KIR2DL1
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Receptors, KIR2DL3
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T-Lymphocytes
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immunology
8.The changes of soluble interleukin-2 receptor in serum and cerebrospinal fluid of patients with delayed encephalopathy after acute carbon monoxide poisoning.
Renjun GU ; Xiuming ZHANG ; Luxian LU ; Hong LU ; Jinggui SONG ; Xinsheng GUO ; Zhongxing ZHANG ; Maixian LIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(5):350-352
OBJECTIVETo explore the changes of soluble interleukin-2 receptor(sIL-2R) in serum and cerebrospinal fluid (CSF) of patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).
METHODSThere were 31 patients with DEACMP, 32 patients with other encephalopathy and 31 controls in this study. The levels of sIL-2R in serum and CSF were detected by ELISA.
RESULTSSerum sIL-2R in patients with DEACMP[(329.21 +/- 160.99)U/ml] was significantly higher than that in control[(115.67 +/- 89.58) U/ml, P < 0.05)], but not significantly different from that in the other encephalopathy group[(367.50 +/- 123.14) U/ml, P > 0.05)]. CSF sIL-2R in patients with DEACMP[(54.48 +/- 43.04) U/ml] measured a little before discharge was significantly lower than that in patients with the other encephalopathy[(110.24 +/- 76.56) U/ml, P < 0.05)], but not significantly different from that in the control group[(34.96 +/- 22.70)U/ml, P > 0.05)]. At the pre-discharged period, CSF sIL-2R in patients with DEACMP[(100.26 +/- 93.65) U/ml] was significantly higher than that at the early stage of hospitalization[(52.28 +/- 43.31) U/ml, P < 0.05)]. No significant difference in serum sIL-2R was found between early stage of hospitalization[(338.34 +/- 161.53) U/ml] and pre-discharge [(351.31 +/- 175.93) U/ml, P > 0.05)].
CONCLUSIONThe occurrence of DEACMP may be related with immunopathological damage. The sIL-2R levels in serum and CSF may give information about the state of immunological function of the patients with DEACMP and may contribute to determining the patient's condition and prognosis.
Brain Diseases ; cerebrospinal fluid ; immunology ; Carbon Monoxide Poisoning ; cerebrospinal fluid ; immunology ; Enzyme-Linked Immunosorbent Assay ; Hospitalization ; Humans ; Receptors, Interleukin-2 ; analysis ; blood
9.Promotion of the articular cartilage proteoglycan degradation by T-2 toxin and selenium protective effect.
Si-Yuan LI ; Jun-Ling CAO ; Zhong-Li SHI ; Jing-Hong CHEN ; Zeng-Tie ZHANG ; Clare E HUGHES ; Bruce CATERSON
Journal of Zhejiang University. Science. B 2008;9(1):22-33
OBJECTIVETo identify the relationship between T-2 toxin and Kashin-Beck disease (KBD), the effects of T-2 toxin on aggrecan metabolism in human chondrocytes and cartilage were investigated in vitro.
METHODSChondrocytes were isolated from human articular cartilage and cultured in vitro. Hyaluronic acid (HA), soluble CD44 (sCD44), IL-1beta and TNF-alpha levels in supernatants were measured by enzyme-linked immunosorbent assay (ELISA). CD44 content in chondrocyte membrane was determined by flow cytometry (FCM). CD44, hyaluronic acid synthetase-2 (HAS-2) and aggrecanases mRNA levels in chondrocytes were determined using reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemical method was used to investigate expressions of BC-13, 3-B-3(-) and 2-B-6 epitopes in the cartilage reconstructed in vitro.
RESULTST-2 toxin inhibited CD44, HAS-2, and aggrecan mRNA expressions, but promoted aggrecanase-2 mRNA expression. Meanwhile, CD44 expression was found to be the lowest in the chondrocytes cultured with T-2 toxin and the highest in control plus selenium group. In addition, ELISA results indicated that there were higher sCD44, IL-1beta and TNF-alpha levels in T-2 toxin group. Similarly, higher HA levels were also observed in T-2 toxin group using radioimmunoprecipitation assay (RIPA). Furthermore, using monoclonal antibodies BC-13, 3-B-3 and 2-B-6, strong positive immunostaining was found in the reconstructed cartilage cultured with T-2 toxin, whereas no positive staining or very weak staining was observed in the cartilage cultured without T-2 toxin. Selenium could partly inhibit the effects of T-2 toxin above.
CONCLUSIONT-2 toxin could inhibit aggrecan synthesis, promote aggrecanases and pro-inflammatory cytokines production, and consequently induce aggrecan degradation in chondrocytes. These will perturb metabolism balance between aggrecan synthesis and degradation in cartilage, inducing aggrecan loss in the end, which may be the initiation of the cartilage degradation.
Cartilage, Articular ; drug effects ; metabolism ; Cells, Cultured ; DNA ; analysis ; Flow Cytometry ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Interleukin-1beta ; analysis ; Proteoglycans ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Selenium ; pharmacology ; T-2 Toxin ; toxicity ; Tumor Necrosis Factor-alpha ; analysis
10.Differences in immunophenotyping of mucosal lymphocytes between ulcerative colitis and Crohn's disease.
Heung Bum LEE ; Jong Hun KIM ; Chang Yeol YIM ; Dae Ghon KIM ; Deuk Soo AHN
The Korean Journal of Internal Medicine 1997;12(1):7-15
OBJECTIVES: Immunologic studies have characterized the numbers and types of inflammatory cells in diseased inflammatory bowel disease (IBD) mucosa but have yielded conflicting results regarding intestinal lymphocytes activation in IBD. We investigated the levels of lymphocytes subsets, interleukin-2 receptor, transferrin receptor, and T cell receptors in mainly isolated lamina propria lymphocytes. Including intraepithelial lymphocytes of normal colonic mucosa or IBD (ulcerative colitis and Crohn's disease) mucosa to understand the pathogenesis of IBD. We have results from this study. RESULTS: 1) In comparing ulcerative colitis with control, IL-2R (p < 0.05), TR (p < 0.01), and CD3/HLA-DR (<0.05) showed a significant increase. 2) In comparing Crohn's disease with control, CD3 (P < 0.05), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. 3) In comparing Crohn's disease with ulcerative colitis, CD19 (p < 0.01), TR (p < 0.01), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. CONCLUSION: From these results, there are increased T cell markers, IL-2R, TR, and CD3/HLA-DR in UC, but differently, decreased CD3, TCR alpha/beta and TCR gamma/delta in CD compared with control. In addition, definitive differences in lymphocytes markers, CD19, TR, TCR alpha/beta and TCR gamma/delta, which are higher in UC than in CD, may elucidate the different immunopathogenesis between UC and CD.
Adult
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Antigens, CD3/analysis
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Colitis, Ulcerative/pathology
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Colitis, Ulcerative/immunology
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Colitis, Ulcerative/diagnosis*
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Comparative Study
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Crohn Disease/pathology
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Crohn Disease/immunology
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Crohn Disease/diagnosis*
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Diagnosis, Differential
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Female
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HLA-DR Antigens/analysis
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Human
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Immunophenotyping*
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Intestinal Mucosa/pathology
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Intestinal Mucosa/immunology*
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Male
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Middle Age
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Receptors, Antigen, T-Cell/analysis
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Receptors, Interleukin-2/analysis
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Receptors, Transferrin/analysis
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Sensitivity and Specificity
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Tissue Culture