ABSTRACT
Chronic lymphocytic leukemia (CLL) is characterized by the peripheral accumulation of neoplastic B cells and is frequently complicated by the systemic immunosuppression associated with an impairment in B and T lymphocyte activation. We hypothesized that the expression of immune checkpoint suppressors B and T lymphocyte attenuator (BTLA) and cytotoxic T lymphocyte antigen (CTLA-4) is disturbed in both lymphocyte subpopulations in CLL. The expression of CTLA-4 and BTLA mRNA was determined by real-time PCR, while CTLA-4 protein expression (surface or intracellular) was estimated in BTLA+ lymphocytes by flow cytometry. In CLL patients, we observed a higher gene transcript level of BTLA and CTLA-4 than in healthy individuals in both freshly isolated and PMA stimulated B and T cells. Remarkably, lower amounts of both inhibitory proteins were found in peripheral blood (PB) CLL B cells, whereas normal BTLA and elevated CTLA-4 were found in T cells. Consistently, there was a prevalence of CTLA-4+ cells within circulating BTLA+ T cells cells of patients confronting PB healthy cells. After in vitro stimulation, the only change found in CLL patients was a decrease in BTLA expression in B and T lymphocytes. In contrast, healthy lymphocytes responded more vigorously as regards the BTLA and CTLA expression with substantially higher frequency of CD69+ cells under the stimulating condition compared to corresponding cells from the CLL group. Our results indicate that CLL development is associated with the affected expression of BTLA and CTLA-4 checkpoint receptors in PB and its impaired expression might be associated with lowering of the threshold for B cell activation and proliferation, while upregulated CTLA-4 expression in CLL peripheral BTLA+ T cells may contribute to suppressed T cell effector functions. This hypothesis needs to be validated in future studies, which would allow us to explain how the increased or decreased expression of these molecules affects the cell function.
Subject(s)
CTLA-4 Antigen/metabolism , Immune Checkpoint Proteins/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , Receptors, Immunologic/metabolism , Adult , Aged , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/metabolism , B-Lymphocytes/metabolism , Cell Proliferation/physiology , Cells, Cultured , Female , Humans , Lectins, C-Type/metabolism , Leukocytes, Mononuclear/metabolism , Lymphocyte Activation/physiology , Male , Middle Aged , RNA, Messenger/metabolism , T-Lymphocytes/immunology , Up-Regulation/physiologyABSTRACT
BACKGROUND: The prevalence of cardiovascular and metabolic disorders in paediatric patients with psoriasis is not well established. AIM: To conduct a meta-analysis of previously published studies dealing with the occurrence of metabolic disorders in children with psoriasis. METHODS: Data from 7 studies with a total of 965 children with psoriasis were analysed using a random effects model. RESULTS: Prevalence of metabolic syndrome (MetS) was significantly higher in patients with psoriasis than in healthy controls (HCs). In most studies, significantly decreased levels of high-density lipoprotein (HDL) cholesterol were found in children with psoriasis. Mean level of HDL cholesterol in patients with psoriasis was 2.05 mg/dL lower than in HCs. Patients with psoriasis and HCs did not differ significantly in their mean triglyceride levels, although the difference was at a threshold of statistical significance. Mean level of fasting glucose in children with psoriasis was 5.75 mg/dL higher than in HCs (P < 0.01). The two groups did not differ significantly in mean waist circumference or in systolic and diastolic arterial pressures. CONCLUSIONS: Decreased levels of HDL cholesterol and increased concentrations of fasting glucose may represent very early stages of MetS in children with psoriasis. However, a large population-based study is needed to establish the relationship between psoriasis and MetS in children, including the environmental, genetic and immunological factors leading to their co-occurrence.
Subject(s)
Cholesterol, HDL/blood , Metabolic Syndrome/complications , Psoriasis/complications , Blood Glucose , Child , Cholesterol, HDL/analysis , Humans , Metabolic Syndrome/epidemiology , Prevalence , Psoriasis/blood , Triglycerides/bloodABSTRACT
Interaction between TL1A and death receptor 3 (DR3) co-stimulates T cells, induces production of several pro-inflammatory cytokines and has been linked to pathogenesis of inflammatory bowel disease (IBD). This study aimed to establish a link between expression of TL1A and selected TL1A-induced pro-inflammatory cytokines involved in IBD pathogenesis (IL-4, IL-13, IL-17A and IFN-γ) and to investigate a connection between serum concentration of TL1A in patients with IBD and activation of peripheral blood T cells. Elevated levels of IL-4 (2.91-fold) and IL-13 (4.05-fold) mRNA were detected in the inflamed colon mucosa of patients with ulcerative colitis (UC), IFN-γ mRNA was upregulated (3.23-fold) in the inflamed colon mucosa of patients with Crohn's disease (CD), whereas upregulation of IL-17A and TL1A mRNA was present in the inflamed colon mucosa of patients with both CD and UC (IL-17A: 4.48-fold and 2.74-fold, TL1A: 3.19-fold and 3.22-fold, respectively) vs. control subjects. We did not detect any changes in DR3 mRNA expression in the investigated groups of patients. TL1A mRNA level in colon mucosa of patients with IBD correlated only with the level of IL-17A mRNA but no other investigated cytokines. In colon mucosa, expression of TL1A and DR3 was localized to enterocytes and lamina propria mononuclear cells. We did not find any correlation between serum concentrations of TL1A and IL-17A or changes of CD4(+) or CD8(+) lymphocytes phenotype in patients with IBD. Therefore, our data indicate that TL1A may contribute to pathogenesis of IBD via local but not systemic induction of IL-17A but not IL-4, IL-13 or IFN-γ.
Subject(s)
Colitis, Ulcerative/metabolism , Colon/metabolism , Crohn Disease/metabolism , Interleukin-17/metabolism , Intestinal Mucosa/metabolism , Tumor Necrosis Factor Ligand Superfamily Member 15/metabolism , Adult , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Enterocytes/metabolism , Female , Humans , Interferon-gamma/metabolism , Interleukin-13/metabolism , Interleukin-17/biosynthesis , Interleukin-17/blood , Interleukin-4/metabolism , Male , Middle Aged , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Receptors, Tumor Necrosis Factor, Member 25/genetics , Receptors, Tumor Necrosis Factor, Member 25/metabolism , Tumor Necrosis Factor Ligand Superfamily Member 15/blood , Tumor Necrosis Factor Ligand Superfamily Member 15/genetics , Up-RegulationABSTRACT
The aim of the current study was to investigate the incidence of mtDNA(4977) deletion in peripheral blood leukocytes of patients diagnosed with premature ovarian failure (POF) and primary infertility. The study group consisted of 17 patients with POF, 32 women with primary infertility, and 31 fertile women with the prevalence of the mtDNA(4977) deletion using the reverse transcription-polymerase chain reaction (RT-PCR) based technology. None of the patients affected by POF revealed mtDNA(4977) deletion. This deletion was detected only in one 26-year-old infertile patient. No significant difference in relation to mtDNA(4977) deletion was reported between the groups investigated (p > 0.05). In conclusion, mtDNA(4977) deletion is not a common finding in peripheral blood leukocytes of women affected by POF and primary infertility. The occurrence of mtDNA(4977) deletion in women between 20 and 39 years of age may not increase with increasing patients' age, independently of their fertility status.
Subject(s)
DNA, Mitochondrial/genetics , Infertility, Female/genetics , Primary Ovarian Insufficiency/genetics , Sequence Deletion , Adult , DNA, Mitochondrial/blood , Female , Humans , Leukocytes/chemistry , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
INTRODUCTION: Pre-eclampsia (PE) is a common obstetric syndrome affecting about 5-10% of pregnant women. The etiology and pathogenesis of this syndrome are not fully understood. There are many studies describing alterations in the innate and adaptive immune system which may have an influence on the onset of this disorder. It was suggested that the activation of cell-mediated immunity may play the key role in the etiology of pre-eclampsia. It was proposed that inappropriate activation of the immune system can lead to pre-eclampsia. OBJECTIVES: The aim of our study was to estimate the surface expressions of CD95(APO-1/Fas) antigen and the intracellular expressions of anti-apoptotic proteinBcl-2 and pro-apoptotic proteinBax in CD4(+)CD25(+)FoxP3(+) T regulatory lymphocytes (Tregs) as well as the percentage of CD8(+)CD28(+) T cytotoxic cells in peripheral blood of patients with pre-eclampsia in comparison with healthy pregnant women in the third trimester of physiological pregnancy. METHODS: Twenty-four women with pre-eclampsia and twenty normal third trimester pregnant women were included in the study. The lymphocytes were isolated from peripheral blood samples and labelled with monoclonal antibodies. The expressions of surface antigens and intracellular proteins were estimated using flow cytometry. RESULTS: The population of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower in peripheral blood of patients with pre-eclampsia when compared to normal third trimester pregnant women. The percentages of CD4(+)CD25(+)FoxP3(+) Treg cells that express Bcl-2 protein were significantly lower in peripheral blood of patients with pre-eclampsia when compared to healthy pregnant women, whereas the percentages of CD4(+)CD25(+)FoxP3(+) Treg cells with the expressions of Bax protein did not differ in both groups. Moreover, the mean fluorescence intensity (MFI) of Bcl-2 protein in CD4(+)CD25(+)FoxP3(+) Treg cells was significantly lower and MFI of Bax protein significantly higher in pre-eclampsia when compared to the control group. The percentage of CD8(+)CD28(+) T cells did not differ in both studied groups but MFI of CD28 antigen on T CD8(+) cells was significantly higher in pre-eclampsia when compared to the control group. CONCLUSION: The obtained results suggest that the deficit of CD4(+)CD25(+)FoxP3(+) Treg lymphocytes which is observed in pre-eclampsia maybe associated with alterations in apoptosis markers.
ABSTRACT
INTRODUCTION: Pre-eclampsia (PE) is a common obstetric syndrome affecting about 5-10% of pregnant women. The syndrome is a leading cause of maternal and fetal morbidity and mortality, even in developed world. The etiology and pathogenesis of this syndrome are not fully understood. There are many studies describing alterations in the innate and adaptive immune system which may have an influence on the onset of this disorder. It was suggested that activation of cell-mediated immunity may play the key role in the etiology of pre-eclampsia. OBJECTIVES: The purpose of our study was to estimate the expressions of B7-H1 and B7-H4 costimulatory molecules on myeloid and lymphoid DCs (CD1c(+), BDCA-2(+)) in the peripheral blood of patients with pre-eclampsia and normal pregnant women in the third trimesters of physiological pregnancy. METHODS: Thirty three patients with pre-eclampsia and 26 normal pregnant women were included in the study. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies against blood dendritic cell antigens and B7-H1 and B7-H4 molecules and estimated using flow cytometry. RESULTS: The expressions of B7-H1 molecule on CD1c(+) myeloid DCs and B7-H4 molecule on BDCA-2(+) lymphoid DCs did not differ in pre-eclampsia and healthy third trimester pregnant women. The expressions of B7-H4 molecule on CD1c(+) myeloid DCs and B7-H1 molecule on BDCA-2(+) lymphoid DCs were significantly higher in peripheral blood of patients with pre-eclampsia in comparison with the control group. CONCLUSION: It seems possible that higher expressions of B7-H4 molecule on CD1c(+) myeloid DCs and B7-H1 molecule on lymphoid BDCA-2(+) DCs in pre-eclampsia may be the tolerogenic mechanism secondary to the pro-inflammatory response which is observed in this syndrome.
ABSTRACT
Little is known about effects of alcohol consumption on dendritic cell (DC) function and resultant immune response. However, quantitative and qualitative disturbances of DCs are speculated to be involved in alcohol-related as well as in other liver pathology. The present study aimed to evaluate changes in circulating DC subsets in alcoholic liver disease (N = 43), autoimmune hepatitis (N = 26) and primary biliary cirrhosis (N = 20). DCs isolated from the peripheral blood of recruited participants were stained with monoclonal antibodies against blood dendritic cell antigens (BDCAs) and estimated using the flow cytometry. Myeloid DCs were defined as BDCA-1(+)/CD19(-) cells, and lymphoid DCs as BDCA-2(+)/CD123(+) cells. Total numbers of circulating DCs in subjects with some liver diseases were markedly lower than in the healthy participants (p = 0.03). There was a significantly lower percentage of circulating BDCA-2(+)/CD123(+) (p = 0.02), and a tendency for the percentage of circulating BDCA-1(+)/CD19(-) cells to decrease in patients with liver diseases compared to the controls (p = 0.09). These results may suggest that decreased numbers of DCs may be responsible for reduced adaptive immune responses and increased susceptibility to infections and cancer development observed in patients exposed to alcohol. Moreover, numerical abnormalities of DCs may contribute to the breakdown of self-tolerance, a feature of autoimmune diseases.
Subject(s)
Alcohol-Induced Disorders/immunology , Autoimmune Diseases/immunology , Dendritic Cells/immunology , Liver Diseases/immunology , Adult , Alcohol-Induced Disorders/epidemiology , Autoimmune Diseases/epidemiology , Case-Control Studies , Dendritic Cells/cytology , Female , Flow Cytometry , Humans , Liver Diseases/epidemiology , Male , Middle AgedABSTRACT
Most cancers remain incurable. Introduction of novel therapeutic methods, including new cytostatic regimens and targeted therapies, such as monoclonal antibodies and tyrosine kinase inhibitors, have increased remission rates as well as improved patient survival, but the ability to cure many cancer patients remains elusive. It is thus necessary to further develop alternative strategies to improve patient prognosis. The majority of patients who respond to induction therapy inevitably relapse, mainly because of the proliferation of residual malignant cells that have escaped control by induction chemotherapy. Therefore the eradication of minimal residual disease may be crucial to prevent a relapse and achieve a long-term remission. It seems that an advantageous treatment option may be cellular immunotherapy with dendritic-cell vaccines which might induce long-term specific anticancer responses with immune memory cells, which could contribute to effective and lasting elimination of malignant cells.
Subject(s)
Cancer Vaccines , Dendritic Cells/immunology , Cancer Vaccines/immunology , Cancer Vaccines/therapeutic use , Humans , ImmunotherapyABSTRACT
The receptor for hyaluronic acid-mediated motility (RHAMM) is a tumor-associated antigen in chronic lymphocytic leukemia (CLL). CD8(+) T cells primed with the RHAMM-derived epitope R3, which is restricted by human leukocyte antigen (HLA)-A2, effectively lyse RHAMM(+) CLL cells. Therefore, we initiated a phase I clinical trial of R3 peptide vaccination. Six HLA-A2(+) CLL patients were vaccinated four times at biweekly intervals with the R3 peptide (ILSLELMKL; 300 microg per dose) emulsified in incomplete Freund's adjuvant; granulocyte-macrophage colony stimulating factor (100 microg per dose) was administered concomitantly. Detailed immunological analyses were conducted throughout the course of peptide vaccination. No severe adverse events greater than CTC I degrees skin toxicity were observed. Four patients exhibited reduced white blood cell counts during vaccination. In five of six patients, R3-specific CD8(+) T cells were detected with the corresponding peptide/HLA-A2 tetrameric complex; these populations were verified functionally in four of five patients using enzyme-linked immunosorbent spot (ELISpot) assays. In patients with clinical responses, we found increased frequencies of R3-specific CD8(+) T cells that expressed high levels of CD107a and produced both interferon-gamma and granzyme B in response to antigen challenge. Interestingly, vaccination was also associated with the induction of regulatory T cells in four patients. Thus peptide vaccination in six CLL patients was safe and could elicit to some extent specific CD8(+) T-cell responses against the tumor antigen RHAMM.
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Cancer Vaccines/therapeutic use , Extracellular Matrix Proteins/immunology , Hyaluronan Receptors/immunology , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , Peptide Fragments/therapeutic use , T-Lymphocytes, Cytotoxic/immunology , Adult , Aged , Cancer Vaccines/immunology , Cell Proliferation , Epitopes, T-Lymphocyte , Feasibility Studies , Female , Flow Cytometry , HLA-A2 Antigen/immunology , HLA-A2 Antigen/metabolism , Humans , Immunotherapy , Interferon-gamma/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/therapy , Lymphocyte Count , Male , Middle Aged , Peptide Fragments/immunology , T-Lymphocytes, Regulatory/immunology , VaccinationABSTRACT
The aim of our study was to estimate both B7-H1 and B7-H4 molecules on immature myeloid and lymphoid dendritic cells in umbilical cord blood of healthy neonates in comparison with peripheral blood of healthy adults. Thirty nine healthy full-term neonates from physiological single pregnancies and 27 healthy adults were included in the study. The expression of B7-H1 and B7-H4 was revealed using the immunofluorescence method. Statistical analysis was performed using a non-parametric test (Mann-Whitney U-Test). The percentages of BDCA-1+ dendritic cells with B7-H1 and B7-H4 expressions were significantly higher in peripheral blood of healthy adults (p<0.00003). It was either observed that the percentage of BDCA-2+ dendritic cells with the expression of B7-H4 molecules was significantly higher in peripheral blood of healthy adults in comparison with umbilical cord blood (p<0.02). Decreased percentages of dendritic cells and co-stimulatory molecules indicate that neonates have immature immune system. Depletion of co-stimulatory B7-H1 and B7-H4 molecules enable appropriate development of immune response.
Subject(s)
Antigens, CD/metabolism , B7-1 Antigen/metabolism , Dendritic Cells/metabolism , Fetal Blood/metabolism , Lymphocytes/metabolism , Myeloid Cells/metabolism , Adult , B7-H1 Antigen , Dendritic Cells/immunology , Flow Cytometry , Humans , Infant, Newborn , Lectins, C-Type/metabolism , Lymphocytes/immunology , Membrane Glycoproteins/metabolism , Myeloid Cells/immunology , Receptors, Immunologic/metabolism , V-Set Domain-Containing T-Cell Activation Inhibitor 1ABSTRACT
Disturbed immunoregulation and an inappropriate immune response to gut microflora is assumed to be involved in the pathogenesis of inflammatory bowel disease (IBD). Physiologically dendritic cells (DCs) as the professional antigen presenting cells play a crucial role in the control of intestinal inflammation and immune tolerance. In order to evaluate their role in the IBD we analyzed the phenotypic and functional properties of monocyte-derived DCs (MoDCs) generated from UC and CD patients following stimulation with TNF-α, lipopolisaccharide E. coli or hydrocortisone. Thirty seven patients with moderate to severe inflammation (19 UC, 18 CD) were recruited to the study. Monocyte-derived dendritic cell immunophenotypes and their endocytic ability were analysed by flow cytometry and confocal microscopy, IL-6, IL-10, IL-12 and IL-23 secretion were investigated by ELISA. Both unstimulated and stimulated MoDCs generated from IBD patients had more mature phenotype and secreted elevated concentrations of proinflammatory cytokines as compared to a control group. The addition of LPS E. coli to culture media was associated with enhanced dendritic cell activation and maturation as compared to DCs stimulated only with TNF-α. This may suggest altered dendritic cell interactions with intestinal microflora in inflammatory bowel disease. Hydrocortisone decreases the numbers of mature dendritic cells and the proinflammatory cytokine concentrations in all cell culture types that may explain the efficacy of steroid therapy in inflammatory bowel disease.
Subject(s)
Colitis, Ulcerative/immunology , Crohn Disease/immunology , Dendritic Cells/immunology , Monocytes/immunology , Adult , Antigens/immunology , Antigens/metabolism , Colitis, Ulcerative/metabolism , Colitis, Ulcerative/pathology , Crohn Disease/metabolism , Crohn Disease/pathology , Cytokines/immunology , Cytokines/metabolism , Dendritic Cells/metabolism , Dendritic Cells/pathology , Escherichia coli/immunology , Female , Humans , Hydrocortisone/immunology , Hydrocortisone/metabolism , Immunophenotyping/methods , Lipopolysaccharides/immunology , Lipopolysaccharides/metabolism , Male , Middle Aged , Monocytes/metabolism , Monocytes/pathology , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolism , Young AdultABSTRACT
Thalidomide represents a promising immunomodulatory drug that targets both leukemia cells and the tumor microenvironment. We treated patients with chronic lymphocytic leukemia (CLL) with a combined thalidomide/fludarabine regimen and monitored cellular and molecular changes induced by thalidomide in vivo before fludarabine treatment. Thalidomide was given daily (100 mg p.o. per day) and fludarabine was administered on days 7-11 (25 mg/m(2) i.v. per day) within each 4-week cycle (maximum of 6 cycles). Twenty patients received thalidomide/fludarabine as first-line therapy and 20 patients were previously treated. Unmutated IgVH mutation status was found in 36 cases and 13 had high-risk cytogenetic aberrations (del17p, del11q). The overall response rate was 80 and 25% for untreated and previously treated patients, respectively. Although thalidomide reduced the number of CLL cells, the number of CD3 lymphocytes showed no significant change, but the number of CD4(+)CD25(hi)FOXP3(+) regulatory T cells (Tregs) was significantly decreased. Gene expression profiling revealed a thalidomide-induced signature containing both targets known to have a function in immunomodulatory drug action as well as novel candidate genes. Combined thalidomide/fludarabine therapy demonstrated efficacy in high-risk patients with CLL. Furthermore, our study provides novel biological insights into thalidomide effect, which might act by enhancing apoptosis of CLL cells and reducing Tregs, thereby enabling T-cell-dependent antitumor effect.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Aged , Biomarkers, Tumor/genetics , Cohort Studies , Enzyme-Linked Immunosorbent Assay , Female , Gene Expression Profiling , Gene Expression Regulation, Leukemic/drug effects , Humans , Immunoglobulin Heavy Chains/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Male , Middle Aged , Neoplasm Staging , Oligonucleotide Array Sequence Analysis , Prognosis , Receptors, Tumor Necrosis Factor/metabolism , Risk Factors , T-Lymphocytes/metabolism , Thalidomide/administration & dosage , Tumor Necrosis Factor-alpha/blood , Vidarabine/administration & dosage , Vidarabine/analogs & derivativesABSTRACT
This study investigated the surface expression of the CD154 antigen (CD40L) on peripheral blood CD4+ T lymphocytes and the sera concentrations of soluble CD40L antigens (sCD40L) in non-pregnant women, normal pregnant women, and patients with pre-eclampsia. Twenty-five patients with pre-eclampsia, 18 healthy pregnant women, and 10 healthy non-pregnant women were included in the study. The expression of the CD154 antigen on CD4+ T lymphocytes was determined using flow cytometry. The serum concentration of sCD40L was measured using an ELISA. Statistical analysis was performed using the Mann-Whitney U and ANOVA tests. The expression of the CD154 antigen on CD4+ T lymphocytes and the serum concentration of soluble CD40L were significantly higher in pre-eclampsia than in normal pregnant women. In normal pregnancy the expression of the CD154 antigen on CD4+ T lymphocytes and the concentration of sCD40L were significantly lower than in non-pregnant women. We conclude that during normal pregnancy the levels of these inflammatory mediators are lower than in non-pregnant women. In pre-eclampsia the levels are higher than those in normal pregnancy, but stable compared with the non-pregnant state. These results suggest that in pre-eclampsia, there are disturbances in the mechanisms responsible for the decrease in innate immunity which occurs in normal pregnancy.
Subject(s)
CD40 Ligand/blood , Health , Pre-Eclampsia/blood , CD40 Ligand/metabolism , Case-Control Studies , Female , Humans , Pregnancy , T-Lymphocytes/metabolismABSTRACT
Differential expression of molecules in chronic lymphocytic leukemia (CLL) may define prognostic markers and suitable targets for immunotherapy. Expression of the tumor-associated antigen (TAA) RHAMM (receptor for hyaluronic acid-mediated motility) as well as RHAMM splicing variants was assessed in series of 72 CLL patients. Quantitative reverse transcriptase PCR showed higher RHAMM expression in high-risk CLL patients, as well as in the advanced stages of the disease. CLL cases with a higher RHAMM expression showed a significantly shorter median treatment-free survival. Among patients with mutated immunoglobulin heavy chain genes, an analysis of RHAMM expression enabled to distinguish subgroup of patients with favorable prognosis. In lymph nodes, RHAMM staining correlated with a higher Ki-67 index and CD40L expression. Functionally, stimulation with CD40L enhanced RHAMM expression in CLL. We further characterized RHAMM-specific CD8(+) T cells in patients with CLL, as the expression of TAAs might influence the clinical outcome by the means of immune reactions. The cytotoxic potential of RHAMM-specific T cells was shown against target cells bearing RHAMM-derived epitope as well as against CLL cells expressing RHAMM. In conclusion, RHAMM expression appears to be of prognostic value, as well as may reflect the proliferative capacity of CLL cells, and might therefore represent interesting target for immunotherapy.
Subject(s)
Extracellular Matrix Proteins/physiology , Hyaluronan Receptors/physiology , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , Neoplasm Proteins/physiology , Adult , Aged , Aged, 80 and over , CD40 Ligand/analysis , CD40 Ligand/pharmacology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/pathology , Cell Division , Cytotoxicity, Immunologic , Disease Progression , Disease-Free Survival , Extracellular Matrix Proteins/analysis , Female , Humans , Hyaluronan Receptors/analysis , Immunoglobulin Heavy Chains/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/mortality , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Male , Middle Aged , Neoplasm Proteins/analysis , Prognosis , Protein Isoforms/analysis , Protein Isoforms/physiology , RNA Splicing , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The immunosuppression accompanies B-cell chronic lymphocytic leukemia (B-CLL) but might be also responsible for disease progression by enabling CLL cells to escape from the immunosurveillance. Some particles involved in the regulation of an immune system might represent prognostic value for B-CLL. Recently we found no correlation between HLA-G on messenger and protein level, suggesting that HLA-G is released in soluble form. To confront this hypothesis we characterized soluble HLA-G (sHLA-G) by the prognostic factors in the first cohort of 34 CLL patients. No correlation was observed between sHLA-G levels in ZAP-70(+) and ZAP-70(-) CLL as well as in CD38(+) CLL and CD38(-) CLL patients. Next, we wondered whether gene expression of HLA-G, which represent the whole HLA-G pool in the cell, posses prognostic value for CLL. In the second cohort of 41 CLL patients we assessed messenger levels of HLA-G by the strongest prognostic factors in CLL including cytogenetics, IgVH mutational status, ZAP-70 as well as CD38. No changes of HLA-G expression levels were found in different CLL groups characterized by IgVH gene mutational status, ZAP-70 as well as CD38. We observed no differences in expression of HLA-G in various cytogenetic groups of CLL including del17p, del13q, del11q, +8q, +3q, del14q and del6q when compared to those with normal karyotype or with 12+. Both, mRNA expression of HLA-G and levels of its soluble form in plasma bring no additional prognostic value for B-CLL patients.
Subject(s)
HLA Antigens/blood , HLA Antigens/genetics , Histocompatibility Antigens Class I/blood , Histocompatibility Antigens Class I/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , RNA, Messenger/genetics , Aged , Aged, 80 and over , Chromosomes, Human, Pair 11 , Chromosomes, Human, Pair 13 , Chromosomes, Human, Pair 17 , Cohort Studies , DNA Primers , Female , HLA-G Antigens , Humans , Karyotyping , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Male , Middle Aged , Mutation , Neoplasm Staging , Prognosis , Sequence DeletionABSTRACT
Recently, we described that vaccination with allogeneic dendritic cells (DCs) pulsed with tumor cell lysate generated specific CD8+ T cell response in patients with B-cell chronic lymphocytic leukemia (B-CLL). In the present study, the potential of autologous DCs pulsed ex vivo with tumor cell lysates to stimulate antitumor immunity in patients with B-CLL in early stages was evaluated. Twelve patients at clinical stage 0-2 as per Rai were vaccinated intradermally up to eight times with a mean number of 7.4 x 10(6) DCs pulsed with B-CLL cell lysate. We observed a decrease of peripheral blood leukocytes and CD19+/CD5+ leukemic cells in five patients, three patients showed a stable disease and four patients progressed despite DC vaccination. A significant increase of specific cytotoxic CD8+ T lymphocytes against the leukemia-associated antigens RHAMM or fibromodulin was detected in four patients after DC vaccination. In patients with a clinical response, an increase of interleukin 12 (IL-12) serum levels and a decrease of the frequency of CD4+CD25(+)FOXP3+ T regulatory cells were observed. Taken together, the study demonstrated that vaccination with autologous DC in CLL patients is feasible and safe. Immunological and to some extend hematological responses could be noted, justifying further investigation on this immunotherapeutical approach.
Subject(s)
Dendritic Cells/transplantation , Leukemia, Lymphocytic, Chronic, B-Cell/therapy , T-Lymphocyte Subsets/immunology , Vaccination/methods , Aged , Antigens, Neoplasm/therapeutic use , Cancer Vaccines , Female , Flow Cytometry , Humans , Interleukin-12/blood , Male , Middle Aged , T-Lymphocyte Subsets/cytology , T-Lymphocytes, Cytotoxic/cytology , T-Lymphocytes, Cytotoxic/immunology , T-Lymphocytes, Regulatory/cytology , T-Lymphocytes, Regulatory/immunology , Transplantation, Autologous , Treatment OutcomeABSTRACT
BACKGROUND: Evaluation of immature myeloid and lymphoid dendritic cells (DCs) in the peritoneal fluid (PF) and peripheral blood (PB) mononuclears of women with ovarian carcinoma (n = 47) and benign ovarian tumors (n = 37). METHODS: Mononuclear cells were isolated from PF and PB, stained with monoclonal antibodies (mAbs) against DC antigens (anti-BDCA-1, anti-BDCA-2), and estimated using flow cytometry. RESULTS: The percentage of PF myeloid DC (MDC) in mononuclears was significantly lower in patients with ovarian cancer in comparison to the group of nonmalignant ovarian tumors (0.65% and 6.95%). The percentage of PF lymphoid DCs (LDCs) was higher in patients with ovarian cancer than in the reference group (0.64% and 0.09%). The percentage of PB MDCs and LDCs did not differ significantly between studied groups. In women suffering from ovarian cancer the percentage of both MDCs and LDCs was higher in the PF than in the PB. In the reference group the percentage of MDCs was higher but that of LDCs was lower in the PF than in the PB. In women with ovarian cancer, PF MDCs/LDCs ratio was lower in comparison to patients with serous cystadenoma. In PB the ratio of MDCs to LDCs did not differ significantly between studied groups. CONCLUSIONS: We concluded that MDCs population may be affected by the presence of malignant disease. LDC subsets may have influence on the local immune response in the PF of women with malignant tumors of the ovary. (c) 2008 Clinical Cytometry Society.
Subject(s)
Ascitic Fluid/cytology , Dendritic Cells/metabolism , Ovarian Neoplasms , Adult , Aged , Aged, 80 and over , Cystadenoma, Serous/immunology , Cystadenoma, Serous/pathology , Disease Progression , Female , Flow Cytometry , Humans , Leukocytes, Mononuclear/immunology , Middle Aged , Neoplasm Metastasis , Ovarian Neoplasms/immunology , Ovarian Neoplasms/pathologySubject(s)
Immunosuppressive Agents/therapeutic use , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , T-Lymphocytes, Regulatory/immunology , Thalidomide/therapeutic use , Adult , Aged , CD4-Positive T-Lymphocytes/metabolism , Female , Forkhead Transcription Factors/metabolism , Humans , Interleukin-2 Receptor alpha Subunit/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , Male , Middle AgedABSTRACT
The expression of HLA-G was reported in certain malignancies and its role in escaping from immunosurveillance in cancers was proposed since HLA-G is a nonconventional HLA class I molecule that protects fetus from immunorecognition during pregnancy. Recent studies proposed HLA-G as novel prognostic marker for patients with B-CLL. HLA-G was showed to bear even better prognostic information compared to Zeta-chain associated protein of 70kDa (ZAP-70) and CD38 although some other authors did not find HLA-G expression in CLL. Therefore in this study we characterized the expression of HLA-G on both RNA and protein level. In most of 20 B-CLL patients we were able to detect signal from HLA-G using flow cytometry analysis. The expression of HLA-G was confirmed on messenger level by real-time RT-PCR experiments. No correlation between HLA-G expression and expression of well established prognostic factors such as ZAP-70 and CD38 was detected. These results confirm that HLA-G is expressed on CLL leukemic cells. Furthermore the expression of HLA-G on CLL cells suggests that this molecule might be involved in escaping of CLL cells from immunosurveillance.