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PubMed1995

Differential cell count and lymphocyte subsets in bronchoalveolar lavage during pneumonia with and without peripheral neutropenia.

One hundred immunocompromised HIV negative patients with microbiologically positive pneumonia underwent bronchoalveolar lavage (BAL) studies. Thirty cases showed peripheral neutropenia (< 1000 neutrophils/microL), 70 did not. The total cell number in BAL, the differential cell counts, and the lymphocyte subsets (CD4, CD8, CD19, CD57) were measured. Patients with pneumonia and normal or elevated peripheral neutrophils had a significantly increased total number of cells in BAL compared to patients with peripheral neutropenia (3.2 +/- 2 vs 1.3 +/- 0.6 x 10(5) cells/ml2 lavage fluid, p < 0.01). Ninety percent of the BAL differential cell counts obtained in patients exceeding 1000 neutrophils/microL showed a lymphocytic and/or neutrophilic alveolitis, whereas only 54% of patients with peripheral neutropenia displayed abnormal counts (p < 0.01). Yet the typical pattern of neutrophilic alveolitis was found more often for peripheral neutrophil counts over 1000/microL with high significance (p < 0.0001). Abnormal BAL cell patterns for neutropenic patients uniformly showed a lymphocytic alveolitis, only 10% additionally conformed with the pattern of neutrophilic alveolitis. Patients with pneumonia with and without peripheral neutropenia had similar findings in BAL lymphocyte subsets and exhibited a reduced CD4/CD8 ratio compared to controls (p < 0.05). The high susceptibility of severe neutropenic patients to pulmonary, especially fungal infections may be explained by the local lack of neutrophils.

Adolescent↗
PubMed1997

Distribution of lymphocyte subsets in bone marrow and peripheral blood is associated with haptoglobin type. Binding of haptoglobin to the B-cell lectin CD22.

Haptoglobin is an acute phase protein showing a genetic polymorphism with 3 major types: Hp 1-1, Hp 2-1, and Hp 2-2. In this study, flow cytometric analysis demonstrated that all three haptoglobin types bind to CD22 on human B-lymphocytes with equal affinity. Comparison of reference values for lymphocyte subsets in peripheral blood and bone marrow showed significant differences between haptoglobin types. Haptoglobin 2-2 is associated with higher peripheral B-lymphocyte counts (P < 0.001) and CD4+ T-lymphocyte counts (P < 0.05). In bone marrow, CD4+ T-cell percentages were highest (P < 0.001) but B-cell percentages were lowest (P < 0.001) in haptoglobin 2-2 type. A negative correlation between serum haptoglobin 1-1 concentration and peripheral B-cell counts was observed (r = -0.663). Our results suggest that haptoglobin is involved in lymphocyte distribution. The present findings are a potential cause of over- or underestimation of lymphocyte subset counts in the clinical staging of immunodeficiency diseases.

Adult↗
PubMed1989

[Study of IgE and T lymphocyte subsets in patients with serious bronchial asthma].

In the present study, 30 patients with serious asthma were selected for the investigation of IgE and T lymphocyte subsets in PBL. By the indirect McAb immuno-SPA assay, it was observed that the percentages of OKT3 and OKT8 but the percentage of OKT4 subset and T4/T8 ratio were significantly higher than those of normal persons (P less than 0.01). The total IgE in serum of patients (performed by ELISA) was also much higher than that of normal persons (P less than 0.05). In addition, there was a closed relationship between the level of IgE and the percentage of OKT of the patients (r = 0.37369, P less than 0.05). The results indicated that one of possible reasons of higher IgE in serum of asthma patients was the abnormality of T lymphocyte subsets in vivo.

Adult↗
PubMed1994

Immunopathological patterns in the rectal mucosa of patients with shigellosis: expression of HLA-DR antigens and T-lymphocyte subsets.

Expression of HLA-DR antigens and infiltration of T-lymphocyte subsets (CD4, CD8), cell activation marker (CD25), B cells (CD20), macrophages (CD68 and Ber-Mac 3) and natural killer cells (CD56) in the rectal mucosa of patients with bacillary dysentery and in healthy controls were studied in an effort to interpret the immunopathological changes taking place in the rectal mucosa during the acute phase of shigellosis. The epithelium of the rectal mucosa from 21 of 32 patients was HLA-DR+. Conventional histology showed acute inflammation in 16 of these patients, chronic inflammation in 3, and in 2 histology was normal. In 7 of 20 controls the epithelium was HLA-DR+; 4 of these 7 were found to suffer from chronic inflammation, whilst in 3 the mucosa was normal. The number of HLA-DR+ intraepithelial lymphocytes in biopsies from patients with Shigella infection was significantly higher (p = 0.005) than in controls. The infiltration of CD8+ cells in the surface epithelium and in the lamina propria, and of CD4+ cells in the lamina propria alone, was significantly higher (p < 0.001) in patients than in controls. The results demonstrate that infiltration of T cells with suppressor/cytotoxic or helper/inducer phenotype in the epithelium and in the lamina propria in Shigella-infected patients may be related to the induction of HLA-DR expression in non-lymphoid cells during acute Shigella infection.

Acute Disease↗
PubMed2004

[Preliminary study on lymphocyte subsets of sentinel lymph nodes in breast cancer patients].

OBJECTIVE: To study the lymphocyte subsets of sentinel lymph nodes (SLN) in breast cancer patients. METHODS: Flow cytometry was used to analyze the markers on the surface of lymphocytes such as CD3, CD4, CD8, CD16 and CD19 in the sentinel lymph node of breast cancer. RESULTS: When lymph node metastasis did not occur, there was no significant difference in the number of CD3(+) T, CD4(+) T, CD8(+) T, CD16 NK and CD19 B cells between SLN cells and non-SLN cells. With lymph node metastasis, the proportion of CD4(+) and CD8(+) T cells was significantly changed, CD8(+) T cells (66.15 +/- 5.97) were the predominant cells instead of CD4(+) T cells (69.07 +/- 5.02), whereas no significant difference in CD3(+) T, CD16 NK and CD19 B cells. CONCLUSION: The CD4(+) to CD8(+) T cell ratio in sentinel lymph nodes with metastasis is reversed in breast cancer patients. This might results from changes in microenvironment due to tumor cell invasion.

Breast Neoplasms↗
PubMedDate not supplied

A reduced CD8+ lymphocyte subset distinguishes patients with polymyalgia rheumatica and temporal arteritis from patients with other diseases.

OBJECTIVE: To determine the diagnostic and screening test qualities of concentrations of the CD8+ lymphocyte subset in peripheral blood to discriminate patients with giant cell arteritis (GCA) from patients with other diseases. METHODS: A CD8+ lymphocyte test was performed in 454 patients from the Department of Medicine, Randers Central Hospital. The sensitivity, specificity and predictive values of the test were calculated and presented as receiver operating characteristic (ROC) curves. RESULTS: The median percentage and numbers of CD8+ cells were significantly reduced in 227 patients with active untreated GCA compared with 227 in-patients of similar age and sex (GCA vs in-patients CD8% : 12.0 vs 20.0, CD8+ x 10(9)/l : 0.195 vs 0.374, p < 0.05). Identical ROC curves were obtained when patients with GCA were tested against various subgroups of patients. The sensitivity and specificity of the test for GCA at an optimal cutoff point were 71% and 80%, respectively, while the positive predictive value was 80%. CONCLUSION: At an optimal cutoff point, concentrations of the CD8+ lymphocyte subset in peripheral blood discriminate patients with GCA from patients with other diseases. The sensitivity and specificity of the test for GCA are equal to those of other tests used in rheumatology.

Blood Cells↗
PubMed2003

Development of early apopotosis and changes in lymphocyte subsets in lymphoid organs of mice orally inoculated with nivalenol.

Development of early apoptosis and changes in lymphocyte subsets were examined in lymphoid organs of female BALB/c mice after oral administration of 15 mg/kg b.w. of nivalenol (NIV), the major type B trichothecene mycotoxin, by FACS analysis. Judging from the results of viable cell count and apoptotic cell index, NIV attacked Peyer's patches first and thymus most severely. In thymus, selective damage in CD4(+)CD8(+) cells was observed at 12 and 24 h after inoculation (HAI), following the peak of apoptosis at 9 HAI. CD4(+) cells were clearly suppressed at 3 HAI in Peyer's patches, at and after 9 HAI in mesenteric lymph nodes, and 3 to 12 HAI in spleen, respectively. CD8(+) cells were also suppressed at 24 HAI in mesenteric lymph nodes and at 12 HAI in spleen, respectively. As to changes in B cell subsets, IgG(+) cells significantly decreased from 3 to 12 HAI and all B cell subsets at 24 HAI in mesenteric lymph nodes. In spleen, IgM(+) cells were suppressed at 9 HAI. On the other hand, in Peyer's patches, following clear decrease in the numbers of pan-T and pan-B cells and viable cells at 3 HAI, all B cell subsets, especially IgA(+) cells, showed a significant increase in their numbers at 9 HAI, and the numbers of IgA(+) and IgM(+) cells remained higher values than controls thereafter. Taken together, in the course of recovery from NIV-induced prominent damage in Peyer's patches at 3 HAI, interaction of NIV with Peyer's patches might result in in vivo stimulation of interleukin production at this site and result in increased proliferation and differentiation of IgA-secreting B cells at and after 9 HAI.

Administration, Oral↗
PubMed1995

Lymphocyte subsets in bronchoalveolar lavage after exposure to Actinobacillus pleuropneumoniae in pigs previously immunized orally or by aerosol.

Young pigs were immunized with the lung-pathogenic bacterium Actinobacillus (Haemophilus) pleuropneumoniae by aerosol or orally using viable and inactivated bacteria. The cellular changes in the bronchoalveolar lavage (BAL) were studied in repeated lavages after the pigs were infected with live bacteria. The nucleated cells in the BAL were differentiated and lymphocyte subsets determined. There were no major differences between the two routes of immunization or between viable and inactivated bacteria. The immunization induced an increase in all lymphocyte subsets studied and in the appearance of plasma cells and lymphoid blasts. The infection did not cause a further increase except in granulocytes. The lack of a booster-type increase in lymphocytes in the BAL might indicate a different immunologic reaction of the lung or that lymphocytes of the BAL do not represent lung lymphocytes in general. The protective effect of the immunization might be deduced from the increase in lymphocytes after immunization but not from the reaction pattern after infection.

Actinobacillus Infections↗
PubMed2004

Effect of freezing of sputum samples on flow cytometric analysis of lymphocyte subsets.

Sputum samples should be processed shortly after induction to prevent cell degradation. For intermediate storage, freezing of homogenised samples or immediate fixation have been shown to be suitable for cytospins. The aim of this study was to investigate whether freezing or immediate fixation of sputum affect the analysis of lymphocyte subsets by flow cytometry. Selected plugs from 24 sputum samples were homogenised. One aliquot was processed immediately and analysed by flow cytometry. A second aliquot was homogenised, frozen at -20 C after addition of dimethylsulfoxide and stored for a median time of 6 days. In six samples a third aliquot was fixed in formalin after induction and stored for up to 72 h before further processing. Compared to immediate processing, percentages of total lymphocytes and T-suppressor cells were elevated after being frozen, with a minor decrease in the T4/T8 ratio. Proportions of total lymphocytes, T-helper and T-suppressor cells correlated between native and frozen samples, intra-class correlation coefficients being 0.74, 0.85 and 0.70, respectively. The formalin-fixed aliquots could not be analysed with the antibodies used. In conclusion, freezing seems to be a suitable technique to store sputum samples for flow cytometry of CD3, CD4 and CD8 lymphocyte subsets. Its effects were minor compared to the variation between subjects.

Adult↗
PubMed2005

Lymphocyte subsets and cytokines in women with gestational diabetes mellitus and their newborn.

This study aimed to identify potential immunological markers for predicting type 1 diabetes in patients with gestational diabetes mellitus (GDM) and any immunological impairment in their newborn. In 62 GDM patients and 74 women with normal glucose tolerance (NGT), and their babies, we assessed total lymphocytes, T lymphocyte subsets CD3 and CD8 expressing T cell receptor (TCR) alpha/beta or gamma/delta, CD16 and CD19, pancreatic autoantibodies and cytokines (IL-5, IL-2, soluble receptor IL-2). At delivery, umbilical cord blood samples were taken for lymphocyte subpopulations and cytokine measurements. GDM mothers had higher levels of total lymphocytes, CD8 expressing TCR gamma/delta, and lower levels of CD3 expressing TCR alpha/beta than NGT controls. Insulin-treated GDM mothers had lower CD4 and CD4/CD8 ratios, and higher CD8 and IL-5 than diet-treated GDM or controls. Five women were positive for pancreatic autoantibodies, with lower CD4 (p<0.01) and CD4/CD8 ratios (p<0.05), and higher CD8 (p<0.03) and CD19 than GDM and control mothers negative for autoantibodies. GDM newborn had higher CD8 gamma/delta and lower CD16 than NGT babies. There were no significant differences in TNF-alpha concentrations in the cord blood obtained from the GDM and NGT newborn. In conclusion, GDM women and their newborn have lymphocyte subset impairments, which are more important in patients positive for autoantibodies and/or treated with insulin.

Autoantibodies↗
PubMed1980

Peanut agglutinin. VI. Identification of murine T lymphocyte subsets during ontogenesis by use of peanut agglutinin and Ly-6.2 antiserum.

Fluoresceinated peanut agglutinin (PNA-FTC), Thy-1.2, and Ly-6.2 antisera were used to identify various murine T lymphocyte subpopulations during fetal and perinatal life. By means of double immunofluorescence labeling, we identified three T lymphocyte subsets: PNA+Ly-6.2+, PNA+Ly-6.2-, PNA-Ly-6.2+ having distinct distribution in the adult thymus and spleen. The study of the appearance of these three subsets in fetal liver, thymus, and spleen has shown that they did not occur similarly. The PNA+Ly-6.2+ subset was present in all three organs during fetal life and at a level rather similar to that of adult life; the PNA+Ly-6.2- subset was also present in the three fetal organs; conversely, the percentage of the PNA-Ly-6.2+ spleen cells was very low in fetal and neonatal life and increased significantly between 1 and 2 wk of age. Our results demonstrate that during fetal life, lymphocyte subsets are phenotypically differentiated as in adult life and that differentiation of T lymphocytes occurs preferentially through various lineages present during early ontogenesis. Moreover, quantitative modulation of some of these subsets is shown from fetal to adult life and may explain some of the immunologic immature features of young murine spleen.

Adrenal Cortex Hormones↗
PubMedDate not supplied

Effect of two-round Ficoll-Hypaque density gradient centrifugation on lymphocyte subsets and natural killer activity of umbilical cord blood mononuclear cells.

Umbilical cord blood (CB) mononuclear cells (MNCs) obtained from Ficoll-Hypaque density gradient centrifugation (FDGC) are frequently contaminated with erythrocytes and nucleated erythroid precursors. The authors investigated the effect of two-round (2-r) FDGC on lymphocyte subsets and natural killer activity of CB and adult peripheral blood (APB) MNCs, in comparison with those obtained from conventional one-round (1-r) separation. The percentage of CD45-expressing CB MNCs was greatly increased after the second density step (p < .01), indicating the efficacy in purification. The percentages of CD3/CD4, CD3/CD8, and CD16/56 double-positive staining CB MNCs were significantly increased after 2-r FDGC, as compared to those after 1-r separation. However, the percentages of CD34+ stem cells and CD19+ B cells were not affected by 2-r FDGC. MNCs obtained from 2-r FDGC had higher natural killer (NK) activity than did MNCs obtained from 1-r separation (p < .01). In contrast, 2-r FDGC did not affect lymphocyte subsets and NK activity of APB MNCs as compared to 1-r FDGC. Thus, 2-r FDGC are recommended to obtain CB MNCs for flow cytometric analysis and NK cytotoxicity assays.

Cell Survival↗
PubMed1991

Differential depression of lymphocyte subsets according to stage in stomach cancer.

Lymphocyte surface markers were determined in the peripheral blood lymphocytes (PBL) in 31 stomach cancer patients (15 males and 16 females) and 47 controls (20 males and 27 females) using an indirect immunofluorescence technique. The monoclonal antibodies used were Leu 2a (CD8, suppressor/cytotoxic T cells), Leu 3a (CD4, inducer/helper T cells), Leu 4 (CD3, pan T reagent), Leu 11 (CD16, natural killer cells) and Leu 12 (CD19, B cells). The numbers of PBL, CD3+, CD4+, CD8+, CD16+ and CD19+ cells significantly decreased and the CD4:CD8 value increased in patients with stomach cancer compared to those in healthy volunteers. In stage I, PBL, none of the PBL subsets nor the CD4:CD8 value were significantly different from those of the controls. In stage II, the numbers of PBL, CD3+, CD4+ and CD8+ cells decreased. In stage III, the CD19+ cells decreased in addition to the decreased subsets in stage II. In stage IV, PBL and all subsets measured decreased. The CD4:CD8 value showed significant increases in stages II, III and IV, because the CD8+ cells decreased to a greater extent than did the CD4+ cells. Changes in the subsets were analyzed with regard to age, sex, performance status and smoking history, no significant relation being observed between these factors and lymphocyte subsets. From the present study, we have demonstrated that lymphocyte subsets were differentially depressed in the order of T cells, B cells and natural killer cells, with progression of the stage of disease.

Adult↗
PubMed1999

Lymphocyte subset analyses in blood, spleen and lymph nodes of female Sprague-Dawley rats after short or prolonged exposure to a 50 Hz 100-microT magnetic field.

Based primarily on the results of in vitro studies, it has been suggested that power-line (50 or 60 Hz) magnetic fields (MFs) may reduce immune function, which could lower resistance to infection or cancer. This study was conducted to evaluate the influence of acute and chronic in vivo exposure to a linearly polarized 50 Hz MF on immune function in female Sprague-Dawley rats. Groups of rats were exposed continuously to the MF at a flux density of 100 microT for periods of 3 days, 14 days or 13 weeks. For each exposure period, one control group of rats was sham-exposed together with each MF-exposed group. Experimental end points included analyses of T-lymphocyte subsets as well as other immune cells involved in cell-mediated immune responses, i.e. natural killer (NK) cells, B lymphocytes, macrophages, and granulocytes in blood, spleen and mesenteric lymph nodes. In addition, immunohistochemical methods were used to detect proliferating and apoptotic cells in the various compartments of spleen tissue. The results obtained failed to demonstrate a significant effect of short or prolonged MF exposure on different types of leukocytes, including lymphocyte subsets. Furthermore, the experiments on the in vivo proliferation activity of lymphocytes and the extent of apoptosis in spleen samples did not indicate a difference between the MF-exposed and sham-exposed groups, indicating that MF exposure does not affect the mechanisms involved in the control of lymphocyte homeostasis. The lack of MF effects in the immune tests used in the present in vivo study makes it highly unlikely that MF exposure induces immunotoxicity, at least under the experimental conditions used. However, the data do not exclude the possibility that functional alterations in T-cell responses to mitogens and in NK cell activity as recently described for MF-exposed rodents may be one mechanism involved in the carcinogenic effects of MF exposure observed in some models of co-carcinogenesis.

Animals↗
PubMed2006

Intestinal lymphocyte subsets and turnover are affected by chronic alcohol consumption: implications for SIV/HIV infection.

We recently demonstrated that simian immunodeficiency virus (SIV) viral loads were significantly higher in the plasma of rhesus macaques consuming alcohol compared with controls following intrarectal SIV infection. To understand the possible reasons behind increased viral replication, here we assessed the effects of chronic alcohol consumption on distribution and cycling of various lymphocyte subsets in the intestine. Macaques were administered alcohol (n = 11) or sucrose (n = 12), and percentages of memory and naive and activated lymphocyte subsets were compared in the blood, lymph nodes, and intestines. Although minimal differences were detected in blood or lymph nodes, there were significantly higher percentages of central memory (CD95+CD28+) CD4+ lymphocytes in the intestines from alcohol-receiving animals before infection compared with controls. In addition, higher percentages of naive (CD45RA+CD95-) as well as CXCR4+CD4 cells were detected in intestines of alcohol-treated macaques. Moreover, alcohol consumption resulted in significantly lower percentages of effector memory (CD95+CD28-) CD8 lymphocytes as well as activated Ki67+CD8 cells in the intestines. A subset (7 receiving alcohol and 8 receiving sucrose) were then intrarectally inoculated with SIV(mac251). Viral RNA was compared in different tissues using real-time PCR and in situ hybridization. Higher levels of SIV replication were observed in tissues from alcohol-consuming macaques compared with controls. Central memory CD4 lymphocytes were significantly depleted in intestines and mesenteric lymph nodes from all alcohol animals at 8 weeks postinfection. Thus, changes in the mucosal immune compartment (intestines) in response to alcohol are likely the major reasons behind higher replication of SIV observed in these animals.

Alcohol Drinking↗
PubMedDate not supplied

Lymphocyte subsets in schizophrenic disorders. Relationship with clinical, neuromorphological and treatment variables.

Peripheral blood lymphocyte subsets and serum immunoglobulin levels were assessed in 42 patients and 37 healthy controls. 24 patients were free from neuroleptic medication and 15 had never been treated with neuroleptics. 31 patients had a diagnosis of schizophrenia (DSM-III criteria) and 11 a diagnosis of a disorder of the schizophrenic spectrum. As compared to healthy subjects, the drug naive schizophrenic patient group showed an increase of T suppressor lymphocytes, while the drug-treated schizophrenic group showed an increase of T helper lymphocytes. The drug-treated schizophrenic group differed from the drug naive one relative to a decrease of T suppressor lymphocytes. As compared to healthy subjects, the drug naive spectrum disorder patients showed an increase of absolute number of lymphocytes, while the drug treated spectrum group showed an increase of B lymphocytes. These findings did not correlate with any clinical or neuromorphological variables taken into account.

Antipsychotic Agents↗
PubMed1993

Expression of CD45R0 antigen on the surface of resting and activated neonatal T lymphocyte subsets.

Expression of CD45R0 antigen has been evaluated on the surface of T lymphocyte subsets obtained from cord blood and on peripheral blood lymphocytes (PBL) from both term and preterm neonates. In some experiments, expression of the same antigen has also been evaluated after in vitro activation of cord blood lymphocytes (CBL) with phytohemagglutinin (PHA) or in mixed lymphocyte culture. The CD45R0 molecule was found to be present on a significantly lower percentage of CD3+, CD4+ and CD8+ CBL as compared with the same subsets evaluated in adult individuals. Moreover, at variance with adult PBL, the great majority of CBL displayed a low level of fluorescence when stained with UCHL1 (CD45R0-specific monoclonal antibody), and both PHA and allogeneic stimuli were able to strongly increase the expression of the CD45R0 molecule on CBL. The percentage of CD45R0+ PBL obtained from both term and preterm neonates on the 1st day of life was comparable to that of CBL, however the expression of this molecule increases remarkably within a few days in the majority of term neonates, while it seems to take more time for the antigen to be fully expressed in preterm infants.

Aging↗
PubMed2001

Decreased CD57 lymphocyte subset in patients with chronic Lyme disease.

BACKGROUND: Chronic Lyme disease (LD) is a debilitating illness caused by tickborne infection with the spirochete Borrelia burgdorferi. Although immunologic abnormalities appear to play a role in this disease, specific immunologic markers of chronic LD have not been identified. METHODS: We evaluated 73 patients with chronic LD for lymphocyte subset abnormalities using flow cytometry. Of these, 53 patients had predominant musculoskeletal symptoms, while 20 patients had predominant neurologic symptoms. The estimated duration of infection ranged from 3 months to 15 years, and all patients had positive serologic tests for B. burgdorferi. Ten patients with acute LD (infection less than 1 month) and 22 patients with acquired immunodeficiency syndrome (AIDS) served as disease controls. RESULTS: All 31 chronic LD patients who were tested prior to antibiotic treatment had significantly decreased CD57 lymphocyte counts (mean, 30+/-16 cells per microl; normal, 60-360 cells per microl, P<0.001). Nineteen of 37 patients (51%) who were tested after initiating antibiotic therapy had decreased CD57 levels (mean, 66+/-39 cells per microl), and all five patients tested after completing antibiotic treatment had normal CD57 counts (mean, 173+/-98 cells per microl). In contrast, all 10 patients with acute LD and 82% of AIDS patients had normal CD57 levels, and the difference between these groups and the pre-treatment patients with chronic LD was significant (P<0.001). Patients with chronic LD and predominant neurologic symptoms had significantly lower mean CD57 levels than patients with predominant musculoskeletal symptoms (30+/-21 vs. 58+/-37 cells per microl, P=0.002). CD57 levels increased in chronic LD patients whose symptoms improved, while patients with refractory disease had persistently low CD57 counts. CONCLUSIONS: A decrease in the CD57 lymphocyte subset may be an important marker of chronic LD. Changes in the CD57 subset may be useful to monitor the response to therapy in this disease.

Adolescent↗