Profile Of Peripheral Blood Lymphocytes Research Articles

Phenotypic features of innate and adaptive immunity in patients with chronic hepatitis C and end‐stage renal disease

The knowledge of the immunological profile of patients with chronic hepatitis C (CHC) and end-stage renal disease (ESRD) on haemodialysis (HD) is still limited. This study investigated the immune response profile in HCV patients with concomitant ESRD focusing on the influence of the renal disease on the phenotypic profile of peripheral blood lymphocytes. Immunophenotypic features of peripheral blood leucocytes were assessed by flow cytometry in two distinct groups: HCV patients with ESRD (CHC+ESRD, n=16) and HCV patients with normal renal function (CHC, n=20). Two control groups that were included were as follows: HCV negative blood donors (BD, n=15) and HCV negative patients with ESRD (ESRD, n=19). Higher frequency of macrophage-like and pro-inflammatory monocytes along with enhanced frequency of CD3(-) CD16(-) CD56(+) , mainly CD56(dim) NK-cells, were the hallmark of CHC+ESRD patients. Lower frequency of B cells with significant decreased of B1 and CD23(+) B-cells were associated with ESRD, regardless the HCV infection. Although higher rates of activated CD4(+) and CD8(+) T cells were observed in the CHC and CHC+ESRD groups, the chemotaxis of T-cell subsets, based on their chemokine receptor expression, was affected by ESRD. Chronic hepatitis C patients with ESRD on HD exhibit distinctive phenotypic profile of circulating leucocytes. It may be implicated in the natural history of HCV infection in this particular group of patients and warrants further investigation.

Gene Expression Profile of Peripheral Blood Lymphocytes from Renal Cell Carcinoma Patients Treated with IL-2, Interferon-α and Dendritic Cell Vaccine

Lymphocytes are a key component of the immune system and their differentiation and function are directly influenced by cancer. We examined peripheral blood lymphocyte (PBL) gene expression as a biomarker of illness and treatment effect using the Affymetrix Human Gene ST1 platform in patients with metastatic renal cell carcinoma (mRCC) who received combined treatment with IL-2, interferon-?-2a and dendritic cell vaccine. We examined gene expression, cytokine levels in patient serum and lymphocyte subsets as determined by flow cytometry (FCM). Pre-treatment PBLs from patients with mRCC exhibit a gene expression profile and serum cytokine profile consistent with inflammation and proliferation not found in healthy donors (HD). PBL gene expression from patients with mRCC showed increased mRNA of genes involved with T-cell and TREG-cell activation pathways, which was also reflected in lymphocyte subset distribution. Overall, PBL gene expression post-treatment (POST) was not significantly different than pre-treatment (PRE). Nevertheless, treatment related changes in gene expression (post-treatment minus pre-treatment) revealed an increased expression of T-cell and B-cell receptor signaling pathways in responding (R) patients compared to non-responding (NR) patients. In addition, we observed down-regulation of TREG-cell pathways post-treatment in R vs. NR patients. While exploratory in nature, this study supports the hypothesis that enhanced inflammatory cytotoxic pathways coupled with blunting of the regulatory pathways is necessary for effective anti-cancer activity associated with immune therapy. This type of analysis can potentially identify additional immune therapeutic targets in patients with mRCC.

Clinical Features and Immunological Markers of Atopic Dermatitis in Elderly Patients

Background: Atopic dermatitis (AD) is increasing among the elderly. Staphylococcus aureus colonization is one of the most important aggravating factors in AD. Objective: This study analyses the clinical features of AD in elderly patients and determines the pathogenic roles of staphylococcal superantigens in AD with low and high total IgE levels. Methods:S. aureus enterotoxin genes were evaluated using PCR. Additionally, S. aureus-specific IgE levels and peripheral blood lymphocyte profiles were assessed. Results: A total of 44 women and 77 men diagnosed with AD with a mean age of 68.92 ± 6.51 years were evaluated. In 17 (68%) patients with AD and low levels of total IgE, there was a positive correlation between the positive results for enterotoxin B, S. aureus-specific IgE antibodies and Th1 cytokine profiles (Spearman’s rank correlation test, r = 0.89, p < 0.05). Conclusion: Our results indicate that AD patients with low total IgE levels differ in immunopathogenesis from AD patients with high circulating levels of total IgE AD.

Low Number of Peripheral Blood B Lymphocytes in Patients with Pulmonary Tuberculosis

The cellular immune response plays a critical role in the containment of persistent Mycobacterium tuberculosis infection; however, the immunological mechanisms that lead to its control are not completely identified. The goal of this study was to evaluate B (CD19+) and T (CD3+) peripheral blood lymphocyte profiles and T-cell subsets (CD4+ and CD8+) in patients with pulmonary tuberculosis (TB). Percentages (p = 0.02) and absolute numbers (p = 0.005) of B cells were significantly lower in patients with pulmonary TB than in healthy donors. In contrast, percentages (p = 0.12) and absolute numbers (p = 0.14) of T cells were similar in TB patients and healthy donors. No significant differences in percentages of CD4+ (p = 0.19) or CD8+ (p = 0.85) T cells between patients and healthy donors were observed. In summary, patients with pulmonary tuberculosis had a lower number of peripheral blood B lymphocytes than healthy controls.

T lymphocyte subset abnormalities in the blood and lung in pulmonary arterial hypertension

Mounting data suggest that immune cell abnormalities participate in the pathogenesis of pulmonary arterial hypertension (PAH). To determine whether the T lymphocyte subset composition in the systemic circulation and peripheral lung is altered in PAH. Flow cytometric analyses were performed to determine the phenotypic profile of peripheral blood lymphocytes in idiopathic PAH (IPAH) patients (n=18) and healthy controls (n=17). Immunocytochemical analyses of lymphocytes and T cell subsets were used to examine lung tissue from PAH patients (n=11) and controls (n=11). IPAH patients have abnormal CD8+ T lymphocyte subsets, with a significant increase in CD45RA+ CCR7- peripheral cytotoxic effector-memory cells (p=0.02) and reduction of CD45RA+ CCR7+ naive CD8+ cells versus controls (p=0.001). Further, IPAH patients have a higher proportion of circulating regulatory T cells (T(reg)) and 4-fold increases in the number of CD3+ and CD8+ cells in the peripheral lung compared with controls (p<0.01). Alterations in circulating T cell subsets, particularly CD8+ T lymphocytes and CD4+ T(reg), in patients with PAH suggest that a dysfunctional immune system contributes to disease pathogenesis. A preponderance of CD3+ and CD8+ T lymphocytes in the peripheral lung of PAH patients supports this concept.

Constitutive gene expression profile segregates toxicity in locally advanced breast cancer patients treated with high-dose hyperfractionated radical radiotherapy

Breast cancer patients show a wide variation in normal tissue reactions after radiotherapy. The individual sensitivity to x-rays limits the efficiency of the therapy. Prediction of individual sensitivity to radiotherapy could help to select the radiation protocol and to improve treatment results. The aim of this study was to assess the relationship between gene expression profiles of ex vivo un-irradiated and irradiated lymphocytes and the development of toxicity due to high-dose hyperfractionated radiotherapy in patients with locally advanced breast cancer. Raw data from microarray experiments were uploaded to the Gene Expression Omnibus Database (GEO accession GSE15341). We obtained a small group of 81 genes significantly regulated by radiotherapy, lumped in 50 relevant pathways. Using ANOVA and t-test statistical tools we found 20 and 26 constitutive genes (0 Gy) that segregate patients with and without acute and late toxicity, respectively. Non-supervised hierarchical clustering was used for the visualization of results. Six and 9 pathways were significantly regulated respectively. Concerning to irradiated lymphocytes (2 Gy), we founded 29 genes that separate patients with acute toxicity and without it. Those genes were gathered in 4 significant pathways. We could not identify a set of genes that segregates patients with and without late toxicity. In conclusion, we have found an association between the constitutive gene expression profile of peripheral blood lymphocytes and the development of acute and late toxicity in consecutive, unselected patients. These observations suggest the possibility of predicting normal tissue response to irradiation in high-dose non-conventional radiation therapy regimens. Prospective studies with higher number of patients are needed to validate these preliminary results.

Effects of Aflatoxin on the Proportions of Peripheral Blood Leukocytes and Alpha-Naphtyl Acetate Esterase (ANAE) Positive Lymphocytes in the Mouse

Summary In this study detrimental effects of aflatoxin (AF) on the peripheral blood formula and the alpha naphthyl acetate esterase (ANAE) - positivity profile of peripheral blood lymphocytes (PBL) on the mice were determined. A total of 40 white mice were randomly divided in to 5 groups each having 8 animals. The first group served as a control and received standard diet. The remaining 4 groups were fed diets containing 200, 400, 800 and 1600 ppb AF, respectively. The results of this study have revealed that AF given by the food caused significant declines both in the proportion of PBL and in the percentages of ANAEpositive peripheral blood lymphocytes (T-lymphocytes) in a dose dependent manner. These results may be concerned as a haematological evidence for immuno-suppressive effect of AF on cell-mediated immunity.

Changing survival, memory cell compartment, and T-helper balance of lymphocytes between severe and mild asthma

BackgroundAsthma is a complicated network of inflammatory reactions. It is classified into mild, moderate, and severe persistent asthma. The success of asthma therapy relies much on understanding the underlying mechanisms of inflammation at each stage of asthma severity. The aim of this study was to explore the differences in apoptotic potential, CD4/CD8 ratio, memory compartment, and T- helper (Th) 1 and 2 profile of peripheral blood lymphocytes (PBL) in patients with mild intermittent asthma and severe persistent asthma during exacerbation periods.ResultsFour research lines were investigated and compared among mild asthmatics, severe asthmatics, and healthy groups by applying immunocytochemical staining of PBL. Antiapoptotic and proapoptotic proteins with Bcl-2/Bax ratio, CD4, CD8 markers with CD4+/CD8+ ratio, CD45RO+, CD45RA+ markers with memory/naïve ratio (CD45RO+/CD45RA+). Th2/Th1 cytokines balance represented by IL-4/IFN-γ ratio was measured by enzyme-linked immunosorbent assay (ELISA) for in vitro PBL cytokine synthesis. It was found that Bcl-2/Bax ratio was higher in severe than in mild asthmatics which in turn was higher than in healthy group. And memory/naïve ratio of PBL was higher in severe than in mild asthmatics. Moreover, memory cells, CD45RO+ and CD45RO+/CD45RA+ ratio were correlated directly with Bcl-2/Bax, in severe and mild asthma patients. In contrast, CD4+/CD8+ ratio was not changed significantly among healthy group, mild and severe asthmatics. However, CD8+ cells were correlated directly with memory cells, CD45RO+, in severe asthmatics only. Interestingly, the dominant profile of cytokines appeared to change from T helper 2 (Th2) in mild asthmatics to T helper 1 (Th1) in severe asthmatics where the lowest in vitro IL-4/IFN-γ ratio and highest IFN-γ were found.ConclusionIt was concluded that the underlying mechanisms of inflammation might vary greatly with asthma stage of severity. Mild intermittent asthma is mainly Th2 allergen-oriented reaction during exacerbations with good level of apoptosis making the inflammation as self-limiting, while in severe persistent asthma, the inflammatory reaction mediated mainly by Th1 cytokines with progressive loss of apoptosis leading to longer exacerbations, largely expanded memory cells, CD45RO+, leading to persistent baseline inflammation.

Immunological profile of peripheral blood lymphocytes and monocytes/macrophages in Kawasaki disease

Kawasaki disease (KD) is an acute illness of early childhood characterized by prolonged fever, diffuse mucosal inflammation, indurative oedema of the hands and feet, a polymorphous skin rash and nonsuppurative lymphadenopathy. The histopathological findings in KD comprise panvasculitis with endothelial necrosis, and the infiltration of mononuclear cells into small and medium-sized blood vessels. The levels of many proinflammatory cytokines, chemokines and adhesion molecules can be elevated in sera from children with KD at the acute stage. Although many immunological studies on KD involving peripheral blood have been reported, the data obtained remain controversial. This review focuses on the immune response of peripheral blood lymphocytes and monocytes/macrophages during acute KD.

Polymorphism of blood leukocyte acid phosphatase and the profile of peripheral blood lymphocytes in the first of lactation trimester of cows naturally-infected with bovine leukaemia virus

Abstract. This study covered a population of 91 cows aged 3–6 years. Enzootic bovine leukaemia (EBL) was diagnosed with ELISA and PCR tests. The assays were performed in the 1st, 2nd, and 3rd month after calving. Use was made of anti-bovine monoclonal antibodies (Mab), (VMDR Inc. Pullman-USA) and conjugates of FITC and R-PE (Medac – Germany) with an indirect immunofluorescence reaction (IMF). A differentiation observed between the population numbers of CD19 + B lymphocytes and CD2 + T lymphocytes as well as the CD8 + T lymphocyte sub-population was found to be significant between cows with different genotypes of AcP. Moreover, a correlation was found between the polymorphism of AcP and EBL incidence in cows with reference to the number and percentage of CD19 + B lymphocytes (AcP polymorphism x EBL interaction). A significant differentiation in the profile of peripheral blood lymphocytes was observed between EBL-positive and EBL-negative cows as well as over the first three months after calving. The results reported in this study seem to indicate the potential contribution of a genetic predisposition connected with the expression of biological functions of the blood leukocyte acid phosphatase system in the activation and proliferation of these cells.

Gammadelta T cells in Behçet's disease (BD) and recurrent aphthous stomatitis (RAS).

The immunopathogenesis of BD is believed to be T cell-mediated. The objective of this study was to characterize the activation stage and cytokine profile of peripheral blood lymphocytes (PBL), with particular emphasis on gammadelta T cells. Venous blood was collected from 20 patients with BD, and for comparison, from 11 patients with RAS and from 15 healthy controls. Both the expression of activation markers (CD25, CD29, CD40 ligand, CD69 and HLA-DR) on freshly isolated PBL and T cell subsets, and the expression of intracellular cytokines (IL-4, IL-10, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha)) on mitogen-stimulated PBL and T cell subsets were analysed by double immunofluorescent staining and flow cytometry. Significantly decreased proportion of alphabeta T cells and increased proportion of gammadelta T cells, CD56+ cells and CD8+ gammadelta T cells were found in BD patients compared with healthy controls. This was also seen to a lesser extent in patients with RAS. Furthermore, in BD a significantly increased proportion of the gammadelta T cell population expressed CD69 and high levels of CD29 and were induced to produce IFN-gamma and TNF-alpha compared with healthy controls. In contrast, an increased percentage of gammadelta T cells from RAS patients was induced to produce IFN-gamma, but not TNF-alpha. These results indicate that in BD, activated gammadelta T cells, capable of producing IFN-gamma and TNF-alpha, are present in peripheral blood, suggesting that gammadelta T cells are dynamic and may be regulating immunopathogenic events.

Cross-staining of cytotoxic T lymphocyte populations with peptide-MHC class I multimers of natural HIV-1 variant antigens.

Multimeric soluble peptide-MHC class I complexes bind to T cell receptors (TCR) on the surface of CD8 cytotoxic T lymphocytes (CTL) with sufficient avidity to enable the labelling of CTL according to their antigen specificity [1]. Visualization of antigen-specific CD8+T lymphocytes with fluorogen-conjugated peptide-MHC class I multimers enables both the physical quantification and the phenotypic characterization of CTL responses directly ex vivo, and these properties have led to the widespread adoption of this technique throughout the field of experimental immunology. However, it is well documented that TCR can exhibit a degree of promiscuity in terms of ligand recognition [2]. We recently demonstrated that the multimerization of peptide-MHC class I can allow binding to TCR with affinities that are too low to generate ligand-induced physiological responses, and result in crossreactive staining of HIV-specific CTL lines and clones by unrecognized natural variant antigens [3]. These results suggest that crossreactivity between peptide-MHC class I multimers and low avidity CTL could be relevant to the interpretation of direct ex-vivo staining. The possibility of such cross-staining is likely to be considerably greater when using peptide-MHC class I multimers to stain CTL specific for epitopes derived from antigenically variable pathogens such as HIV-1 [4]. Here we extend our previous study with the demonstration that the cross-staining of CTL populations with peptide-MHC class I multimers can complicate analyses performed directly ex vivo.

The Follow-Up Study of Skin Reactivity to Recall Antigens and E- and EAC-RFC Profiles in Blood in Asbestos Workers

We have determined cutaneous DTH reactions to SK-SD and PPD and peripheral blood lymphocyte profiles in a group of asbestos workers in two consecutive surveys. It was found that asbestosis and, to a lesser extent, the presence of ANA are significantly correlated with the lack of response to the above antigens. 83% of asbestos workers when tested at a 4 year interval fell into the same two categories of responsiveness (lack of response or response at least to one antigen). The asbestosis cases had lower total lymphocyte count as well as proportions and absolute number of E-RFC as compared to asbestos workers without asbestosis and/or ANA. Furthermore, the latter group showed the lower percentages and absolute number of E-RFC than the matched controls. The presence of ANA is also correlated with lower proportions of E-RFC. However, this is related at least in part to asbestosis.

Ultrastructural alterations in peripheral blood lymphocyte profiles following acute thermal burns

Acute thermal burns result in impaired cellular immunity. This is reflected by an immediate lymphocytopenia, impaired delayed skin hypersensitivity, decreased allograft rejection potential, depletion of thymic dependent areas of lymphoid tissue, failure of peripheral blood lymphocytes (PBL) to respond to PHA, and a defect in both the response and stimulation capacity of PBL in mixed lymphocyte culture. The present study investigates the ultrastructure of the PBL of patients with acute thermal burns and correlates their morphology with the immunological defects previously shown to be present. Based upon their ultrastructural morphology, the PBL were divided into five cell types: small lymphocytes, large lymphocytes with dispersed ribosomes, large lymphocytes with aggregate ribosomes, large lymphocytes with varying amounts of rough endoplasmic reticulum, and plasma cells. The serial studies performed in 14 adult burn patients were compared with similar studies performed in a group of 15 normal controls. On the first day post-burn, the PBL profile did not differ significantly from that of a normal group of adults although the lymphocyte count in peripheral blood was already decreased. By the second day a significant shift in the morphology of the PBL towards cells having larger quantities of rough endoplasmic reticulum had occurred coincident with a marked fall in the percentage of small lymphocytes present. With the passage of time this change became more pronounced. By 6–10 days post-burn, a mean of more than 30% of the PBL could be identified as either immature or mature plasma cells. Following this time of maximal change a return toward normal was seen, so that by Day 21–23 post-burn, the PBL profile had returned to essentially normal values. A comparison of these studies with other studies performed on patients with acute thermal burns makes it apparent that in the immediate post-burn period a marked defect in cellular immunity or T-cell function occurs simultaneously with a marked stimulation in humoral immunity or B-cell function. The latter abnormality is reflected by the development of a polyclonal gammopathy and the development first of a marked B-lymphocytosis followed by a marked plasmacytosis coincident with the return of PBL counts to normal. The mechanisms causing these changes in the burn patient are not known.