Expression Of Activation Antigens Research Articles

Donor cell induced CD69 expression and intracellular IL-2 and IL-4 production by peripheral blood lymphocytes isolated from kidney transplant recipients

Flow cytometry assays, which measure CD69 activation and intracellular cytokine production, have been used to measure peripheral blood lymphocyte (PBL) responses to in vitro antigen exposure. In the present study, we show that, in healthy individuals and immunosuppressed kidney transplant recipients, CD69 expression and intracellular cytokine production by peripheral blood T cells compare favorably to thymidine uptake as a measure of PBL response to alloantigen in mixed leukocyte culture (MLC). Heparinized whole blood from 23 healthy individuals was incubated for 24–48 h with 3rd party allogeneic monocytes; blood from twelve kidney transplant recipients was incubated with monocytes from their kidney donor and with monocytes from unrelated individuals. The percentage of T cells expressing surface CD69 or intracellular IL-2 or IL-4 was determined by 3-color flow cytometry. We identified 5 donor-specific response patterns in our kidney transplant group. One transplant recipient was hyporesponsive; his cells did not express CD69 or produce IL-2 in response to either donor or 3rd party allogeneic cells. All other transplant recipients expressed CD69 and IL-2 in response to 3rd party allogeneic cells. Two had no response to donor cells (donor-specific hyporesponsiveness), three had donor-specific anergy (CD69 expression without cytokine production in response to donor cells), five had a donor-specific Th1 response (CD69 expression and IL-2 production in response to donor cells), and one had a donor-specific Th2 response (CD69 expression and IL-4 but not IL-2 production in response to donor cells). Rapid measures of donor-specific hyporesponsiveness such as CD69 activation antigen expression and intracellular cytokine production may prove valuable in monitoring lymphocyte function and aid in the long-term management of kidney transplant recipients.

The influence of surgical stress on T cells: enhancement of early phase lymphocyte activation.

For the control of postoperative infection, it may be important to understand the possible influences of surgical stress on the host immune system. To this end, we examined how the early phase of lymphocyte activation was affected in patients after major surgery (eight patients with esophageal carcinoma and six undergoing cardiac surgery) using a flow cytometric assay based on expression of the early activation antigen, CD69. Freshly isolated T cell in preoperative and postoperative samples did not express CD69. When peripheral blood mononuclear cells were stimulated in vitro, the expression of CD69 was greatly enhanced in both CD4 and CD8 T cells, compared with the preoperative samples. The proportion of de novo CD69-expressing cells in the CD4 subset was approximately 3 times (Postoperative Day 1) and 4 times (Postoperative Days 2, 3, 5, and 7) greater than those preoperatively, whereas the proportion of de novo CD69-expressing cells in the CD8 subset was approximately 1.5 times (Postoperative Days 2 and 5) and 2 times (Postoperative Day 3) greater than those preoperatively. The proportion of CD69+ cells was significantly greater in the CD4+ subset than in the CD8+ subset during the postoperative period. Our results show that major surgical stress enhances the early phase of lymphocyte activation. The augmentation of activation was greater in CD4 (helper) T cells than in CD8 (cytotoxic) T cells.

Dermal Dendritic Cells Associated with T Lymphocytes in Normal Human Skin Display an Activated Phenotype

The CMRF-44 and CD83 (HB15) antigens are associated with functional maturation and activation of blood dendritic cells (DC). We describe the expression of these antigens on freshly isolated epidermal Langerhans cells and dermal DC as well as the distribution of CD83+/ CMRF-44++-activated DC within sections of normal human skin. Fresh Langerhans cells were prepared by standard techniques and large numbers of enriched (25%-55%), viable dermal DC were obtained using an improved collagenase treatment protocol with density gradient enrichment. Freshly isolated Langerhans cells and dermal DC had similar costimulator and activation antigen expression, and both stimulated moderate levels of allogeneic T lymphocyte proliferation as determined in the 7 d mixed leukocyte reaction. In situ labeling of DC within skin sections revealed a population of CD83 and CMRF-44 positive dermal cells of which most (approximately 75%) were in intimate contact with CD3+ T lymphocytes, especially in the adnexal regions. In contrast, only 25%-30% of the more numerous CD1a++ dermal DC population were directly apposed to T lymphocytes. The CMRF-44++ dermal DC population stimulated an allogeneic mixed leukocyte reaction, confirming their identity as DC. These data, plus comparative data obtained for migratory dermal DC, suggest that only a small proportion of dermal DC have been triggered to a more advanced state of differentiation or activation. The striking association of the activated dermal DC population with T lymphocytes suggests that communication between these two cell types in situ may occur early in the immune response to cutaneous antigen.

Quantitation of CD38 activation antigen expression on CD8+ T cells in HIV-1 infection using CD4 expression on CD4+ T lymphocytes as a biological calibrator.

For some membrane-associated antigens, the number of molecules expressed per cell carries information about the cell's differentiation and activation state. Quantitating antigen expression by flow cytometry has immediate application in monitoring CD38 expression on CD8+ T cells in human immunodeficiency virus 1-associated disease, where elevated CD38 antigen expression is a marker of CD8+ T-cell activation and a poor prognostic indicator. Reproducible methods are needed in order to quantify such antigens. Here we describe a reproducible method for quantitative fluorescence cytometry (QFCM) that depends on the tightly regulated expression of CD4 antigen on human CD4+ T lymphocytes, which we estimated in a study of 57 normal donors to have an interperson coefficient of variation of 4.9%. Using phycoerythrin (PE)-conjugated CD4 monoclonal antibody (mAb) with a nominal fluorochrome to protein ratio of 1:1 and a nominal published value of approximately 50,000 CD4 antibody molecules bound per CD4+ T lymphocyte, we estimated the number of PE molecules detected per relative fluorescence intensity (RFI) unit on our flow cytometer to be 41 (19, 20). This value is called the "RFI multiplier." To estimate the number of CD38 antibodies bound per CD8+ T cell (CD38-ABC) on patient samples, we multiply the measured CD38 RFI value of CD38 staining using a nominal 1:1 conjugate of CD38-PE by the "RFI multiplier." The measurements for CD4 and CD38 were stable for 2 years despite the use of different mAb lots and the potential for drift in instrumentation. We used this approach in a study of nine flow cytometers in which the interinstrument interlaboratory coefficients of variation for CD3-ABC ranged from 3.3% to 5.8% and those for CD38-ABC ranged from 9.8% to 13.8%. These data indicate that CD4 expression can serve as a biological calibrator to standardize fluorescence intensity measurements in longitudinal and multicenter studies.

Quantitative fluorescence: To count or not to count. Is that the question?

CytometryVolume 34, Issue 4 p. 203-204 PerspectiveFree Access Quantitative fluorescence: To count or not to count. Is that the question? Janet K. A. Nicholson, Corresponding Author Janet K. A. Nicholson [email protected] Division of AIDS, STD, and TB Laboratory Research, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia1–1202 (A25), 1600 Clifton Rd., N.E., Atlanta, GA 30333Search for more papers by this authorMaryalice Stetler-Stevenson, Maryalice Stetler-Stevenson Flow Cytometry Unit, Department of Pathology, Division of Clinical Sciences, National Cancer Institute, Bethesda, MarylandSearch for more papers by this author Janet K. A. Nicholson, Corresponding Author Janet K. A. Nicholson [email protected] Division of AIDS, STD, and TB Laboratory Research, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia1–1202 (A25), 1600 Clifton Rd., N.E., Atlanta, GA 30333Search for more papers by this authorMaryalice Stetler-Stevenson, Maryalice Stetler-Stevenson Flow Cytometry Unit, Department of Pathology, Division of Clinical Sciences, National Cancer Institute, Bethesda, MarylandSearch for more papers by this author First published: 26 December 2002 https://doi.org/10.1002/(SICI)1097-0320(19980815)34:4<203::AID-CYTO5>3.0.CO;2-GCitations: 2AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article. Literature Cited 1 Borowitz MJ, Bray R, Gascoyne R, Melnick S, Parker JW, Picker L, Stetler-Stevenson M: U.S.-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: Data analysis and interpretation Cytometry 30: 236– 244, 1997. 2 Kuchnio M, Sausville EA, Jaffe ES, Greiner T, Foss FM, McClanahan J, Fukushima P, Stetler-Stevenson M: Flow cytometric detection of neoplastic T cells in patients with mycosis fungoides based upon levels of T-cell receptor expression. Am J Clin Pathol 102: 856– 860, 1994. 3 Stelzer GT, Marti G, Hurley A, McCoy P, Lovett EJ, Schwartz A: U.S.-Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry: Standardization and validation of laboratory procedures Cytometry 30: 214– 230, 1997. 4 Lui Z, Cumberland WG, Hultin LE, Prince HE, Detels R, Giorgi JV: Elevated CD38 antigen expression on CD81 T cells is a stronger marker for the risk of chronic HIV disease progression to AIDS and death in the Multicenter AIDS Cohort Study than CD41 cell count, soluble antigen activation markers, or combinations of HLA-DR and CD38 expression. J Acquir Immune Defic Syndr Hum Retrovirol 16: 83– 92, 1997. 5 Hultin L, Matud JL, Giorgi JV: Quantitation of CD38 activation antigen expression on CD81 T cells in HIV-1 infection using CD4 expression on CD41 T lymphocytes as a biological calibrator. Cytometry 1998, in press. 6 Davis KA, Abrams B, Iyer S, Hoffman RA, Bishop JE: Determination of CD4 antigen density on cells: Role of antibody valency, avidity, clone, and conjugation. Cytometry 1998, in press. Citing Literature Volume34, Issue415 August 1998Pages 203-204 ReferencesRelatedInformation

Increased platelet responsiveness following coronary stenting. Heparin as a possible aetiological factor in stent thrombosis.

Platelet activation may be a determinant of thrombotic and restenotic complications following intracoronary stenting. In order to measure the effect of stenting on platelet activation antigen expression we used whole blood flow cytometry in 18 patients undergoing Palmaz-Schatz stenting (treated with full anticoagulation) and compared these with a group of 18 patients undergoing elective angioplasty. The effects of low molecular weight heparin and unfractionated heparin on platelet behaviour were also studied, both in vitro and in vivo to determine the contribution of prolonged heparin therapy to platelet activation following stenting. Fibrinogen binding to activated GPIIb-IIIa, and surface expression of P-selectin, GPIb and GPIIb-IIIa antigens were measured in unstimulated peripheral blood samples (rest) and on stimulation with adenosine diphosphate (0.1-10 micromol x 1(-1)) and thrombin (0.02-0.16 U x ml(-1)). No changes were seen in resting samples following angioplasty or stenting. Agonist responsiveness was unaltered after angioplasty, but in stented patients antigen expression in response to thrombin was significantly reduced (P< or =0.04), whilst the adenosine diphosphate response was significantly increased (P=0.01). Similar effects were observed in patients with unstable angina treated with either low molecular weight heparin or unfractionated heparin in vivo. In vitro, both unfractionated and low molecular weight heparin inhibited thrombin-induced platelet activation, but stimulation of adenosine diphosphate responses was more marked with unfractionated than low molecular weight heparin. There was a significant increase in platelet responsiveness to adenosine diphosphate following intracoronary stenting in patients treated with conventional anticoagulants. This was probably a consequence of treatment with heparin. Activation of platelets by heparin may explain the increased rate of stent thrombosis in patients treated with anticoagulant therapy. Low molecular weight heparins stimulate platelets less than unfractionated heparin.

High Concentrations of Thrombopoietin Activate Platelets In Vitro

Thrombopoietin (TPO), the ligand for the proto- oncogene c-mpl, has been cloned and expressed from both human and murine sources. Thrombopoietin increases platelet counts when given in vivo and acts on progenitor cells to in crease their proliferation and maturation into megakaryocytes. The effects of TPO on mature platelets were investigated by evaluating platelet aggregation and platelet activation- dependent antigen expression. Platelet aggregation score, a quantitative representation of aggregation, showed potentiation of response to ADP-induced aggregation but no direct agonist response to TPO alone. Soluble c-mpl blocked the effect of TPO on the platelet aggregation score. Flow cytometry showed that TPO at concentrations &gt;250 U/ml (50 ng/ml) caused a minority population of platelets to express the activation mark ers CD62, CD63 and activated glycoprotein IIb/IIIa. While stem cell factor and interleukins-3 and -6 did not affect platelet activation antigen expression, interleukin-11 increased CD62 expression on platelets in vitro. The effects of TPO on antigenic expression and aggregability were partially inhibited in vitro by preincubation with aspirin. We conclude that high concentra tions of TPO promote platelet activation antigen expression on a proportion of platelets and potentiate platelet aggregability to ADP in vitro by a process that is partially inhibited by aspirin.

Activated lymphocytes (CD25+ CD69+ cells) and decreased CD19+ cells in well-nourished chronic alcoholics without ethanol-related diseases.

To assess lymphocyte subsets and expression of activation antigens in peripheral blood lymphocytes (PBLs) in chronic alcoholism, a cross-sectional study with 30 well-nourished chronic alcoholics and 30 controls was performed. Studies included detailed clinical and laboratory evaluation, nutritional status assessment, and determination of lymphocyte subpopulations, as well as activation antigens. A significant decrease of B cells (CD19+) was observed in chronic alcoholics, compared with controls (p < 0.001). A significant increase of PBLs expressing CD69 and CD25 (p < 0.01, both) in chronic alcoholics was also detected, whereas CD71 expression was unaffected. In addition, T lymphocytes expressing HLA-DR were significantly higher in chronic alcoholics than controls (p < 0.05). The serum level of soluble interleukin-2 receptor was also significantly higher in the alcoholic group, compared with controls (p = 0.04). Moreover, the estimated total lifetime dose of ethanol consumed correlated positively with the percentage of PBLs expressing CD25 (r = 0.48; p = 0.01) and negatively with PBLs expressing CD71 (r = -0.39; p = 0.04). By contrast, the changes were not related to age, nutritional status, or the presence of other ethanol-related diseases. In conclusion, chronic alcoholics present a significant decrease of B cells and an "incomplete activation state" of PBLs that depends on the dose of ethanol consumed.

Effect of ex vivo storage on human peripheral blood neutrophil expression of CD11b and the stabilizing effects of Cyto-Chex™

CD11b is the α sub-unit of the CD11b/CD18 heterodimeric complex that has a key role in neutrophil–endothelial cell interactions and early events in inflammatory responses. Accurate assessment of CD11b expression by neutrophils can be problematic since the procedures that isolate cells from whole blood can increase antigen expression. This study used whole blood flow cytometry to monitor neutrophil CD11b expression following 0, 1, 2, 3 and 4 h of ex vivo incubation at room temperature and examined the effects of storage at 4°C and the cell stabilization solution, Cyto-Chex™, on antigen expression. Cyto-Chex™-treated samples were also re-analyzed after 7 days storage at 4°C. Neutrophil CD11b expression was high (>90%) in five of the seven samples studied and incubation at room temperature induced a progressive upregulation which was significant after 2, 3, and 4 h ( p<0.05). Storage at 4°C tempered this effect, although an increased expression (from baseline) was still observed after 4 h ( p<0.05). Cyto-Chex™ had no effect on the light scatter characteristics of treated cells and prevented CD11b upregulation in samples stored at room temperature. Furthermore, expression in Cyto-Chex™-treated samples after 7 days was not significantly different from that observed in baseline (time 0) samples. In two samples demonstrating low neutrophil CD11b expression, a temperature-independent increase in the proportion of CD11b + cells was observed over time. However, this increase was attenuated by treatment with Cyto-Chex™. These findings indicate that sample storage significantly affects CD11b expression and caution should be exercised when interpreting data. Given that Cyto-Chex™ had no effect on the light scatter properties of cells and prevents antigen upregulation, this reagent may be useful for studies involving analysis of neutrophil activation antigen expression.

Malignant plasma cell lines express a functional CD28 molecule.

The function of CD28 molecules that are present on malignant plasma cells of human myeloma cell lines (HMCL) was studied. First, myeloma cells expressed a similar density of CD28 antigen to that of normal T cells. The myeloma CD28 molecules were able to bind B7-Ig molecules as well as L cells transfected with a B7-1 cDNA, and anti-CD28 mAb inhibited the binding. Myeloma cells did not express B7-1 antigens but a low density of B7-2 antigens. The myeloma B7-2 molecules of two HMCL were able to bind CTLA-4 protein. No autocrine CD28:B7-2 activation could be evidenced as we found no spontaneous binding of the p85 subunit of PI-3 kinase to CD28 molecules. In addition, a blocking anti-CD28 mAb did not affect the IL-6-dependent or autonomous proliferation of the HMCL. The activation of myeloma CD28 molecules with or without TPA stimulation did not affect the proliferation, survival, differentiation, expression of activation antigens and cytokine receptors or cytokine production of myeloma cells. However, the triggering of myeloma CD28 molecules by B7-1 transfectant cells resulted in binding of the p85 subunit of PI-3 kinase to CD28 molecules as previously shown for T cell CD28 molecules. This expression of a large density of CD28 molecules able to bind B7 molecules might contribute to a downregulation of the immune control of myeloma cells.

Characteristic changes of large granular lymphocytes that strongly express CD56 in endometrium during the menstrual cycle and early pregnancy.

Large granular lymphocytes that strongly express CD56 (CD56++ LGL) constitute a major population of leukocytes in the secretory endometrium and pregnancy decidua and are considered to be involved in reproductive immunity and in maintaining the pregnancy. The present study aimed to reveal the relationship between the characteristic changes of CD56++ LGL and altered hormonal environment and/or trophoblast invasion in the endometrium. Cell surface markers of CD56++ LGL obtained from the endometrium during the menstrual cycle and early pregnancy were analysed using flow cytometry. The percentages of both CD56++ LGL that express activation antigens (CD69, HLA-DR) and those that express lymphocyte function associated antigen-1 (LFA-1) (CD11a/CD18) were highest in the proliferative phase and decreased gradually throughout the menstrual cycle. Expression of these antigens was further suppressed in the late secretory phase, as well as in the early stage of pregnancy. However, the percentage of CD56++ LGL that express these antigens was significantly higher in spontaneous abortions than in normal pregnancies. On the other hand, the percentage of CD56++ LGL that express CD45RA was lower during normal pregnancy than during the menstrual cycle. The present results suggest that characteristics alterations of CD56++ LGL are regulated by altered hormonal environment and by trophoblast invasion.

Protein A immunoadsorption in chronic refractory ITP reverses increased platelet activation but fails to achieve sustained clinical benefit.

Adults with chronic relapsing ITP present a difficult therapeutic challenge. The ongoing antibody-mediated platelet destruction in this group might be expected to be associated with increased expression of platelet surface membrane activation antigens. We have studied a group of 10 patients with refractory ITP and 35 healthy controls. Using an immediate, sensitive, unfixed, whole blood, flow cytometric method to detect platelet surface P-selectin and GP53, we have detected markedly increased platelet activation in the ITP group compared with the controls (P-selectin; patient median 24.5% v control median 2.0%. GP53 median 6.5% v 2.1%, P < 0.01 for both). Five patients underwent protein A immunoadsorption therapy. The effect of protein A immunoadsorption on platelet activation before, during and after 18 treatments in these patients was studied and patients were followed-up to assess clinical outcome. Platelet-associated immunoglobulin measurements were made before and at the end of six treatments. Platelet activation decreased after immunoadsorption. P-selectin expression fell significantly; pre- and post-treatment median values differed by 15.5%, P < 0.01, for GP53 the difference was 2.5%, P = NS. A reduction in both platelet-associated IgG (median reduction of 11.8 ng/10(6) platelets, P = 0.08) and IgM (7.6 ng/10(6) platelets, P = 0.06) was recorded.

Methimazole therapy in Graves' disease influences the abnormal expression of CD69 (early activation antigen) on T cells.

At present, the in vivo response of T, B and natural killer (NK) cells to antithyroid drug therapy remains largely unknown. In the present study, we have prospectively analyzed the in vivo effects of methimazole treatment on a large number of circulating T and NK cell subsets, some of them expressing cell surface activation antigens involved in the very early phase of the immune response, in a group of 17 hyperthyroid, untreated patients with Graves' disease (GD). As one of the first events during T cell activation is the expression of interleukin (IL) receptors, we also studied the binding of IL-2 and IL-6 to T cells. Patients with Graves' disease were sequentially studied at diagnosis/before treatment (day 0) and 7, 14, 30, 90 and 180 days after methimazole therapy. The results were compared with both a group of 19 age- and sex-matched control volunteers and a group of 20 untreated/euthyroid patients with Graves' disease in long-term remission. The combination of flow cytometry and three-color immunofluorescence revealed a clear (P < 0.01) decrease in the percentage of NK cells before and during the whole course of therapy with respect to both controls and patients with Graves' disease who were in long-term remission. Before therapy, a marked increase (P < 0.001) in the ratio of B to NK cells was also observed; thereafter, a slight decrease in this ratio was observed, although normal values were detected only in patients in long-term remission. Expression of the CD69 early activation antigen in the hyperthyroid untreated patients with Graves' disease was clearly increased (P < 0.01) with respect to both controls and patients with Graves' disease who were in long-term remission. This abnormal CD69 expression was found to be significantly reduced (P < 0.001) by methimazole therapy, and this represents a new effect of the drug. Expression of the low-affinity receptor for IL-2 (CD25)--another early T cell activation marker--was not altered in Graves' disease, but the binding of IL-2 and IL-6 to T cells exhibited a progressive and parallel increase during the first 30 days of therapy, decreasing thereafter. Our results show that methimazole therapy downregulates the abnormally high expression of the CD69 early activation antigen on T cells, being less effective on inducing changes in other T cell activation markers and in NK cells.

Cytotoxic T-lymphocyte responses in cattle infected with bovine viral diarrhea virus

A system for a reproducible in vitro restimulation of bovine viral diarrhea virus (BVDV)-specific cytotoxic T-cells (CTL) was developed. Lymphocyte cultures of BVDV-immunised cattle were stimulated with infectious BVDV isolate PT810 and recombinant bovine interleukin-2 for 12 to 25 days. A specific lysis of Concanavalin A-stimulated BVDV-infected autologous target cells was observed, whereas allogeneic BVDV-infected target cells were only marginally lysed as detected by flow cytometry. BVDV-specific lymphocyte transformation was further characterised by the expression of bovine lymphocyte activation antigens and bovine MHC class-II molecules. Secondary stimulation of CTL was influenced by in vitro production of BVDV-specific neutralising antibodies, which were secreted exclusively in BVDV-inoculated lymphocyte cultures of immunised cattle. These results demonstrate the presence of CTL in peripheral blood mononuclear cells (PBMC) of immunised cattle which can kill autologous BVDV-infected antigen-presenting cells after in vitro restimulation.

Analysis of T-cell immune response in renal cell carcinoma: polarization to type 1-like differentiation pattern, clonal T-cell expansion and tumor-specific cytotoxicity.

We assessed the naturally occurring T-cell immune response in primary renal cell carcinoma (RCC) tumors from 12 unselected patients. A predominance of CD3+ T-cell receptor (TCR)alpha/beta+ T cells was observed in tumor-infiltrating lymphocytes (TILs), in contrast with peripheral blood lymphopenia found in some patients. Activation antigen expression on TILs revealed an imbalance in the activation status, with a significant percentage of CD69+ and HLA-DR+ and a low percentage of CD25+ and CD71+ TILs. The lymphocyte activation gene-3 (LAG-3) was detected in some TIL subpopulations and especially in one patient in whom TILs were predominantly TCR alpha/beta+CD8+DR+LAG-3+. In addition, we found that RCC TILs are polarized to a global type 1-like (Th1/Tc1) differentiation pattern (strong secretion of interferon-gamma and interleukin-2 (IL-2) following CD3/TCR crosslinking) but are under the influence of the down-modulatory cytokines IL-6 (secreted by tumor cells) and IL-10, within the tumor microenvironment. In 3 of 5 patients, clonal T-cell expansion at the tumor site was found for several Vbeta specificities, suggesting that in situ stimulation of specific clonotypes in response to potential tumor antigens is a frequent event in RCC. Furthermore, in one patient, selective intratumor amplification of a Vbeta1 subpopulation (5% of TCR alpha/beta+ cells) corresponding to 2 distinct Vbeta1-Jbeta1.6 and Vbeta1-Jbeta2.3 tumor-specific MHC class I-restricted cytotoxic T lymphocytes supports the view that discrete T-cell subsets contribute readily to in situ immunosurveillance.

A new human cell line with pre-B cell phenotype and t(5;12).

A new cell line (LR10.6) with pre-B cell phenotype has been established from bone marrow cells obtained from a child with B lineage acute lymphoblastic leukemia in complete clinical remission. The line expresses nuclear TdT enzyme, cytoplasmic Ig lambda-chain and membrane mu-chain and other B but no T or myeloid markers. The cells also show activation antigens CD69 and CD71, adhesion molecules CD54, CD50 and CD56 and the tyrosine kinase receptor CD117. No expression of multidrug resistance phenotype MDR-1 is observed on these cells which nevertheless express the transcriptional factor p53 protein in a mutant form. Cytogenetic study shows a translocation t(5;12)(q31;p13) involving breakpoints which contain the growth factor interleukin 3 gene (5q31) and the recently identified TEL/ETV6 gene (12p13). Activation of the cells with phorbol-12 myristate 13-acetate (PMA) up-regulates the expression of the CD69 activation antigen and down-regulates the CD117 molecule. In addition, PMA fails to induce the CD20 B cell antigen.

Abnormalities of Peripheral Blood T Lymphocytes and Natural Killer Cells in Alcoholic Hepatitis Persist after a 3‐Month Withdrawal Period

Present information about the behavior of the different lymphoid subsets in alcoholic hepatitis (AH), including cells displaying cytotoxic activity, is scanty and contradictory. The aim of this study was to gain further insight into knowledge of the immunological abnormalities involved in AH and the possible role of ethanol (EtOH) consumption in these changes. We analyzed the distribution of a wide range of peripheral blood (PB) lymphoid subsets, both during active EtOH intake and after a 3-month withdrawal period, using multiple stainings with monoclonal antibodies and flow cytometry, as well as natural killer (NK) cytotoxic activity. AH patients entering the study were selected strictly; only those undergoing their first episode of AH with no other lesions at liver biopsy were enrolled. Regarding the alcohol intake period, the most striking finding was a significant increase of the absolute number of PB T cells affecting both CD4+ and CD8+ lymphocytes. These changes were associated with a higher expression of T-cell activation antigens, such as HLA DR and CD11c. Simultaneously, a significant increase in both NK cells (CD3-/CD56+) and the cytotoxic T cells coexpressing the CD3 and the CD56 molecules together with an increased NK cytotoxic activity were observed. By contrast, the CD19+/CD5+ B-cell subset was significantly decreased. No significant changes were observed with EtOH withdrawal except in CD5+ B lymphocytes, which returned to normal values. Our results show that, in AH patients, a significant expansion of both activated T cells and NK lymphocytes occurs in the PB, which is associated with an increased NK cytotoxic activity. Interestingly these abnormalities persist during the withdrawal period.

Enhanced Expression of T-Cell Activation and Natural Killer Cell Antigens Indicates Systemic Anti-Tumor Response in Early Primary Cutaneous T-Cell Lymphoma

Indolent, primary cutaneous T-cell lymphomas (CTCL) are characterized by hyper-proliferation of malignant T-helper cells in the skin with a favorable prognosis in the early stages. Cytotoxic T cells (CTLs) are believed to be of major importance for tumor surveillance, but there is not yet sufficient evidence for a systemic anti-tumor response in mycosis fungoides (MF). On the contrary, there are hints of systemic immunodepression. We wondered whether signs of a systemic anti-tumor response were demonstrable in peripheral blood of patients with MF and CD30+ pleomorphic T cell lymphoma. Using multiparameter flow cytometry, we investigated blood samples from 39 CTCL patients at different stages and compared them with those from patients with psoriasis, atopic dermatitis, and healthy volunteers. In CTCL patients, an elevated number of lymphocytes expressing natural killer cell markers were found, as well as considerable T-cell activation, indicated by increased percentages of T cells expressing HLA-DR, IL-2 receptor alpha-chain, and transferrin receptor. The CD8+ T cells, which were the most strongly activated T-cell subset, were of polyclonal origin, as shown by their usage of different T-cell receptor families. The enhanced expression of activation antigens was associated with an increased proportion of CD8+ T cells with high expression of the adhesion molecule LFA-1, demonstrating the capacity for migration of these cells. These CD8+ effector cells are suspected to be CTLs and may be responsible for the favorable prognosis of indolent, primary CTCL. Interestingly, a stage-dependent decrease in T-cell activation antigen expression was observed, suggesting the development of a lack in tumor surveillance in advanced MF stages. Further investigations are necessary to verify whether any of the parameters determined are of predictive value for prognosis and response to therapy in CTCL.

T cells from individuals in advanced stages of HIV-1 infection do not proliferate but express activation antigens in response to HIV-1-specific antigens.

Like T cells from healthy subjects, those of HIV-1-infected patients are capable of expressing activation antigens on their surface after antigenic or mitogenic stimulation, but their proliferative activity is strongly reduced or even absent, especially in patients with advanced stages of the disease. The characteristic of expressing activation antigens in response to different stimuli in the absence of cell proliferation is shared by CD4+ and CD8+ T-cell subsets from HIV-1-infected patients. The number of T cells capable of expressing CD25 and CD71 in response to HIV-1-related antigens but not of proliferating increased significantly with the progression of the disease, but the number of T cells capable of expressing the two activation antigens in response to the classic tetanus toxoid recall antigen decreased. The higher numbers of T cells capable of responding to HIV-1-related antigens in conjunction with a reduction in the number of T cells responding to recall antigens may explain the occurrence of different infections, including opportunistic microorganisms, during the more advanced stages of HIV-1 infection. Because the increase in the number of HIV-1 antigen-responding T cells (defined by CD25 and CD71 activation antigen expression) is a characteristic of symptomatic HIV-1-infected patients, expression (by flow cytometry) of these activation antigens on T cells in response to HIV-1 antigens could be used as a new marker of disease progression.

Characterisation and expression of the novel lymphocyte activation antigen using the monoclonal antibody 6B2D.

Conference Article| May 01 1997 Characterisation and expression of the novel lymphocyte activation antigen using the monoclonal antibody 6B2D Una O'Shea; Una O'Shea *Molecular Medicine, CSB, St. James's Hospital, Leeds, LS9 7TF, UK Search for other works by this author on: This Site PubMed Google Scholar Denis J. Reen Denis J. Reen +Our Lady's Hospital for Sick Children, Crumlin, Dublin 12, Ireland Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1997) 25 (2): 196S. https://doi.org/10.1042/bst025196s Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation Una O'Shea, Denis J. Reen; Characterisation and expression of the novel lymphocyte activation antigen using the monoclonal antibody 6B2D. Biochem Soc Trans 1 May 1997; 25 (2): 196S. doi: https://doi.org/10.1042/bst025196s Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search Keywords: Mab monoclonal antibody, PHA phytohaemagglutinin, PBMC periperial blood mononuclear cells, Con A concanavalin A, SPA staphyloccal protein A, PWM pokeweed mitogen, PPD tuberculin purified protein derivative, SDS sodium dodecyl sulphate, PAGE polyacrylamide This content is only available as a PDF. © 1997 Biochemical Society1997 Article PDF first page preview Close Modal You do not currently have access to this content.