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The influence of cytomegalovirus carrier status on lymphocyte subsets and natural immunity.

The influence of the cytomegalovirus (CMV) carrier status on peripheral lymphocyte subsets was studied in 70 healthy individuals. IgG-class antibodies against CMV late antigen were used as markers for the presence of CMV in those individuals. The 39 CMV-seropositive individuals had significantly higher numbers of CD3+ (P = 0.009), CD8+ (P = 0.005) and HNK1+ (P = 0.002) cells than the 31 CMV-seronegative individuals. Two-colour immunofluorescence studies revealed that the HNK1+ cells coexpressing CD4 or high density CD8 were particularly increased in the number under the influence of CMV, but not the HNK1+ cells coexpressing CD16. Since HNK1 and CD16 are markers associated with natural killer (NK) activity and antibody-dependent cellular cytotoxicity (ADCC), we investigated the influence of the CMV carrier status on those functions. The NK and ADCC functions of peripheral blood mononuclear cells (PBMC), HNK1+ and HNK1- cells were correlated with the percentages of CD16+ cells among those cells. Although CMV-seropositive individuals had significantly less CD16+ cells among their HNK1+ cells than CMV-seronegative individuals (mean and s.d.: 39 and 19%, versus 58 and 11%, P = 0.003), the NK and ADCC functions of the HNK1+ cells were similar in both groups. Also, the CMV carrier status did not influence significantly those functions of PBMC and HNK1- cells. We conclude that the CMV carrier status, i.e. CMV-seropositivity, is associated with a significant increase in the numbers of HNK1+ lymphocytes coexpressing T cell markers. That situation may reflect the continuing interaction between CMV and the immune system of its host.

Adult↗

[Analysis of peripheral lymphocyte subsets and changes due to erythromycin therapy in patients with diffuse panbronchiolitis].

It is well known that patients with diffuse panbronchiolitis (DPB) have various immunological abnormalities. The authors evaluated the peripheral lymphocyte subsets using two-color analysis by FACS. Convening percentages of activated CD3+, CD8+, and CD4+ cells in DPB patients, all, especially that of CD8+ cell, were greater in number before therapy compared to healthy controls. After erythromycin therapy, all three parameters decreased in all patients. Therapy with other antibiotics did not obtain any effects. These results suggested that patients with DPB had some immunological hyperreactivity and that erythromycin was effective in these patients by suppressing this activity. From these results, together with the fact that DPB is a type of sino-bronchial syndrome, the possible pathogenesis of DPB was thought to be related to some immunological defect and hyperreactivity in the respiratory system.

Adult↗

Lymphocyte subsets in the lymph nodes of rats with autoimmune orchitis.

We determined temporal variations of cell subpopulations in the immunization draining lymph nodes during the development of an experimental autoimmune orchitis (EAO) induced in Wistar rats. A phenotypic characterization of T cells and their subsets (CD4+ and CD8+), B, and Ia+ cells was performed by immunofluorescent technique. At the end of the immunization period (30 days), rats injected with testicular homogenate plus adjuvants presented a considerable increase in absolute cell number but normal lymphocyte subset percentages. Testicular damage became evident at 50 days after the first immunization and increased its severity at 80 days: animals that developed EAO presented a lower number of CD8+ cells as compared with undamaged rats. This latter group showed a low CD4/CD8 ratio due to the high proportion of CD8+ cells, which could probably have a suppressor function. At 80 days massive testicular infiltration and decreased absolute cell number in lymph nodes suggest the possible migration of specific lymphocytes to the target organ.

Adjuvants, Immunologic↗

Effects of herpes virus carrier status on peripheral T lymphocyte subsets.

We studied the effects of herpes virus carrier status on peripheral blood T lymphocyte subsets in 334 healthy individuals. IgG-class antibodies against cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus (HSV), and varicella-zoster virus (VZV) were used as markers for the carrier status of those viruses. CMV carrier status was associated with significant increases in the numbers of some T cell subsets, whereas the carrier status of EBV, HSV, and VZV had no significant effects. The 159 CMV-seropositive individuals had higher numbers of HNK1+ T cells than did the 175 CMV-seronegative individuals [mean (SD), 292 (196)/microL v 164 (89)/microL, respectively], including the CD4+HNK1+ T cells [38 (48)/microL v 9 (13)/microL, respectively] and the CD8+HNK1+ T cells [166 (146)/microL v 73 (54)/microL, respectively]. Morphological and cytochemical studies showed that the expression of HNK1 by the CD4+ and CD8+ T cells was associated with the occurrence of azurophilic cytoplasmatic granules and a loss of nonspecific esterase activity. The numbers of CD4+HNK1+ and CD8+HNK1+ T cells increased proportionally to the levels of the IgG-class CMV antibody titers. We suggest that the increased numbers of CD4+HNK1+ and CD8+HNK1+ granular T cells in CMV carriers reflect the persistent interaction between CMV and the immune system of its hosts.

Age Factors↗

Peripheral blood lymphocyte subset distribution and function in patients with Alzheimer's disease and other dementias.

BACKGROUND: Until recently, new data on immune aspects of Alzheimer's disease (AD) have suggested that some facets of AD pathogenesis may be immune related. However, the effects of dementia itself on immune function have not been considered. AIM: To compare the distribution of peripheral blood lymphocyte subsets and their function in patients with AD and other dementias. METHODS: Peripheral blood lymphocyte numbers, T cell subset distribution, proliferative responses to mitogens and suppressor cell assay were studied in a well characterised group of patients with AD, and compared to patients with other forms of dementia. Age and sex matched elderly controls were screened to exclude dementia, and young controls were medical, paramedical and laboratory staff. Analysis of variance (ANOVA) and student's test were used for statistical analysis. RESULTS: The CD8+ lymphocyte population was reduced in AD and in other forms of dementia, when compared with non-demented elderly and young controls. Concanavalin A induced lymphocyte transformation was reduced in all dementia groups and in elderly compared with young controls. The changes in T cell numbers and function were not specific for Alzheimer's disease, but were found also in other forms of dementia.

Age Factors↗

Interleukin 2 production and its relationship with T lymphocyte subsets in patients with obstructive jaundice.

Sepsis remains a major risk in the high mortality and morbidity after surgery for obstructive jaundice. The reasons for the increased susceptibility to infection are unknown. This study examined interleukin 2 (IL-2) production and the lymphocyte response to PHA mitogen in 31 patients with obstructive jaundice. Among them, 18 patients were simultaneously investigated by enumeration of T lymphocyte subsets in peripheral blood with APAAP technique. The results showed that the patients had significantly decreased IL-2 production and lymphocyte response to PHA mitogen. The percentage of Leu 3a (helper/inducer T cell) in the patients was significantly lower than that in normal controls. Leu 3a/Leu 2a (suppressor/cytotoxic T cell) ratio was significantly lower in these patients. The reduction of IL-2 production correlated significantly with the suppression of lymphocyte proliferation but not with the percentage of Leu 3a cells. From these results, it may be suggested that the reduction of IL-2 production in the patients with obstructive jaundice is an important reason for the suppression of T lymphocyte proliferative response, not merely a reflection of the decrease of helper T cells.

Adult↗

Effect of nickel chloride on mouse T-lymphocyte subsets.

The study was carried out to observe the effect of nickel chloride (NiCl2) on T-lymphocyte subsets, CD4, CD8 and CD4/CD8 ratios in vivo. BALB/C mice were injected i.p. with NiCl2 every other day for 2 w. The NiCl2 doses were 1, 3, 6, 9 or 12 mg/kg. Lymphocytes were obtained from mouse spleen and cultured in the presence of monoclonal antibody FITC conjugated, anti-CD4 or anti-CD8. Samples were determined with a flow cytometer. A suppressive effect on CD4-positive cells was found in the 6 mg/kg group, but this subset was restored to the control level in the 9 and 12 mg/kg groups, suggesting that there was an "immune tolerance" effect. No significant changes were found in CD8-positive cells and CD4/CD8 ratios. The results suggested the NiCl2 had a deleterious effect on mouse T-lymphocytes in short-term in vivo exposure, but the effects may depend on the dose and cell subset.

Animals↗

[Flow cytometric analysis of T lymphocyte subsets after human bone marrow transplantation using monoclonal antibodies and dual immunofluorescence].

Using monoclonal antibodies and a dual immunofluorescence technique, we studied the abnormalities of T lymphocyte subsets after human marrow transplantation. T lymphocytes bearing HLA-DR antigen increased both in patients with acute and chronic graft-versus-host disease (GVHD). Parallel to a decrease in CD4+ cells, a CD4+ 2H4+ subset decreased and gradually recovered with time while the percentage of CD4+ 4B4+ cells increased. A significant increase of CD8+ cells observed after marrow transplant corresponded mainly to an increase in CD8+ CD11- cells. Patients with chronic GVHD tended to have a higher proportion of CD8+ CD11+ cells. These results indicate abnormalities of T cell subsets among both CD4+ and CD8+ cells after marrow grafting.

Adolescent↗

Therapy-related changes of CD20+ and CD45RO+ lymphocyte subsets in chronic myeloid leukemia (CML): an immunohistochemical and morphometric study on sequential trephine biopsies of the bone marrow.

Little information exists about the amount of CD45RO+-T- and CD20+-B-lymphocytes in the bone marrow of patients with Philadelphia chromosome-positive chronic myelogenous leukemia (Ph1+-CML) at presentation or regarding corresponding changes during therapy. On the other hand, quantification of this cell compartment seems to be imperative for two reasons: first, the presumed association of immunocompetent lymphocyte subsets in the expansion of the leukemic cell clone; and second, a speculated relationship with the complex generation of myelofibrosis. Therefore, an immunohistological and morphometric study was performed on 219 representative trephine biopsies of the bone marrow derived from 70 patients with repeated examinations during the course of Ph1+-CML. For the identification of the different lymphocyte populations, the monoclonal antibodies UCHL-1 (CD45RO) and L26 (CD20) were applied on formaldehyde-fixed and decalcified specimens. In comparison to a control group and calculated per hematopoietic cells, the CML bone marrow showed about a 50% decrease in the total amount of lymphocytes. Determination of CD45RO+ and CD20+ subsets revealed a significant enhancement during treatment. Because of the different intervals (range, 10 to 25 mo) between first and last biopsy in the various therapeutic groups, results had to be modified by considering dynamic features. This calculation included changes of the lymphocyte subpopulations related to time. Contrasting the CD45RO+ lymphocytes, a relevant increase in the CD20+ subset could be observed after interferon-a treatment or corresponding combination regimens. No significant correlations were found between fiber density at onset (first biopsy) or development of fibrosis and lymphocyte proliferations in the course of CML. Our results are in keeping with the finding that a proper immune response consistent with an increased lymphocyte growth seems to be associated with a regression of the clonally-transformed cell population. Opposed to a repeatedly discussed pathomechanism, we failed to demonstrate any quantitative relationships between the extent of lymphocyte proliferations and occurrence or progression of myelofibrosis.

Adult↗

Lymphocyte subsets and plasma IL-1 alpha, IL-2, and TNF-alpha concentrations in acute rheumatic fever and chronic rheumatic heart disease.

The distribution of CD3+, CD4+, CD8+, CD19+, CD16+, and CD25+ lymphocyte populations in peripheral blood as well as the plasma concentrations of interleukin-1 alpha (IL-1 alpha), and IL-2 and tumor necrosis factor alpha (TNF-alpha) were investigated in 25 children with acute rheumatic fever (ARF) at the time of admission and after 3 months and in 15 children with chronic rheumatic heart disease (CRHD) and in 15 children with streptoccocal pharyngitis (SP) in order to determine changes in lymphocyte subsets and cytokine concentrations occurring during different stages of the disease. The percentages and absolute counts of CD4+, CD16+, CD25+ cells, the ration of CD4/CD8 and plasma concentrations of IL-1 alpha and IL-2 in patients with ARF were significantly higher at admission than 3 months later. These levels were also significantly higher than in patients with CRHD, SP, or normal controls. Production of IL-2 in ARF and CRHD patients directly correlated with the percentages of CD4+ and CD25+ cells. According to our results, the evidences of increased cellular immune response in ARF are increased percentages CD4+ and CD25+ cells, CD4/CD8 ratio, and increased plasma concentrations of IL-1 alpha and IL-2. Furthermore, activation of cellular immune response was not present throughout all stages of rheumatic heart disease and also in SP.

Adolescent↗

Lymphocyte subsets in distinct lung compartments show a different ability to produce interferon-gamma (IFN-gamma) during a pulmonary immune response.

Lymphocytes play an important immunoregulatory role in pulmonary immune responses. By releasing cytokines they can control the cell-cell communication of other participating cells. Although it is well established that the lung lymphocytes, localized in distinct compartments, differ in their subset composition, little is known about cytokine production in these compartments during immune responses. Lewis rats were immunized by intravenous administration of sheep erythrocytes on day 0 and day 7 and challenged intratracheally with sheep erythrocytes on day 10. Four days after intratracheal (i.t.) challenge the composition of lymphocyte subsets (CD2+, CD4+, CD8+, B cells, natural killer (NK) cells) in the spleen, blood, lung perfusate, lung tissue and bronchoalveolar lavage fluid (BALF) was characterized, and intracellular IFN-gamma was detected in these subsets by flow cytometry. Comparing control and immunized animals, no changes were found in lymphocyte numbers, subsets or the percentage of IFN-gamma-producing lymphocytes in the spleen, blood and lung perfusate. In lung tissue and BALF, however, the absolute number of all lymphocyte subsets and the percentage of IFN-gamma-producing lymphocytes were increased. When the lymphocyte subsets were analysed an increased percentage of IFN-gamma-producing T cells was found in lung tissue (4.5 +/- 0.6% versus 12.8 +/- 1.1%) and in BALF (7.8 +/- 1.4% versus 14.8 +/- 1.9%) of immunized animals opposed to controls, this increase being seen in both CD4+ and CD8+ cells. Thus, there is an accumulation of T cells with an increased potential to produce IFN-gamma in the lung interstitium and the bronchoalveolar space during pulmonary immune responses.

Animals↗

Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys.

Rhesus macaques are invaluable experimental animals in biomedical research. Using three color flow cytometry, we screened anti-human antibodies for crossreactivity with macaque cells in order to determine the distribution of functionally important lymphocyte subsets in blood, lymph nodes (LN), and spleen. NK-cells are almost completely absent in LN. The percentage of B-cells expressing CD80, CD86, and the level of expression of CD20 is higher in blood than in LN. In contrast, a higher proportion of B-cells in LN stains positive for CD21 and CD35. Whereas the number of CD29hi expressing T-cells is lower, CD69 is expressed on more T-cells in LN than in blood. About one-third of CD8+ T-cells in blood are CD28-, a subset with a unique pattern of antigen expression which cannot be found in LN. In contrast to humans, a relatively high proportion of T-cells in blood also express the co-stimulatory molecules CD80 and CD86. With increasing age, the proportion of B-cells in blood declines, whereas the percentage of T-cells rises. In addition, the proportion of CD29hi expressing T-cells increases among both the CD4+ and CD8+ subsets.

Animals↗

Influence of pretransplant dialysis modality on the change of lymphocyte subset populations and acute rejection rates after renal transplantation.

BACKGROUND: The influence of dialysis modality on the acute rejection (AR) rate after renal transplantation is controversial. We investigated whether the pretransplant dialysis modality correlated with the lymphocyte subset populations and the incidence of AR after renal transplantation. METHODS: Thirty-eight first living renal transplant recipients, consisting of 22 patients on pretransplant hemodialysis (HD) and 16 patients on pretransplant peritoneal dialysis (PD), were studied. Peripheral blood samples were taken on days -1 through 28 after transplantation, and the lymphocyte fractions were exposed to the monoclonal antibodies anti-CD3, CD19, CD4, CD8 and CD28 for a flow cytometer analysis. Biopsy specimens were obtained at the time of presumed AR episodes and on day 28 after transplantation. RESULTS: The PD patients had a higher frequency of AR (37.5% in PD vs 9.1% in HD patients, P = 0.034). In contrast, two HD patients showed graft loss at 18 and 30 months after transplantation. The increases of CD3, CD19, CD4 and CD4+ CD28+ cells in the PD patients occurred earlier than in the HD patients and the numbers of these cells in the PD group were higher than those in the HD between days 3-28 after transplantation, most significantly on day 7. CONCLUSIONS: These findings suggest that the PD patients with similar clinical characteristics could potentially have a higher immunocompetence and immune responsiveness associated with a higher rate of AR in the early stage of renal transplantation when compared with the HD patients.

Acute Disease↗

Lymphocytapheresis in combination with immunosuppressive drugs for refractory myasthenia gravis: two-color flow cytometric analysis of changes in peripheral blood lymphocyte subsets.

We carried out lymphocytapheresis (LCP) in combination with the administration of immunosuppressive drugs in patients with myasthenia gravis (MG), who were resistant to conventional immunosuppressive therapy, and examined its efficacy and effects on peripheral blood lymphocyte subsets. LCP was carried out once a week for 1 month (one course, 4 times) using a continuous-flow blood cell separator. Immunosuppressive medication (prednisolone or prednisolone and azathioprine) was continued during the course of treatment. After LCP, clinical improvement was noted in 5 of 6 patients. Anti-AChR antibody titers and the number of lymphocytes were significantly reduced in all patients. A significant decrease in CD4+CD45RA- (memory) T cell level and significant increase in CD4+CD45RA+ (naive) T cell level were also observed. In the patients having good response to LCP, follow-up evaluation showed long-term clinical improvements, as well as the memory T cell level staying at the decreased level. Our study suggests that LCP in combination with the administration of immunosuppressive drugs can suppress the disease activity of MG.

Autoantibodies↗

Experimental Actinobacillus pleuropneumoniae infection in piglets with different types and levels of specific protection: immunophenotypic analysis of lymphocyte subsets in the circulation and respiratory mucosal lymphoid tissue.

Actinobacillus pleuropneumoniae (APP) infection in piglets results in severe and fatal fibrinous hemorrhagic necrotizing pneumoniae. The aim of our study was to analyze changes in lymphocyte subset distribution in peripheral blood, bronchoalveolar lavage fluid (BALF) and tracheobronchal lymph nodes (TLN) in non-immune piglets upon a challenge with a high dose of APP and to compare the quality of such changes in unprotected piglets with counterparts exhibiting specific immunity mediated by high titers of colostrum-derived APP-specific antibodies and/or a low dose APP infection in the early postnatal period. Challenge with APP resulted in a massive increase in CD8-negative gammadelta T-cells in parallel with a reduction in numbers of CD3-CD8low cells in BALF independent of the type and level of immunity and this seems to be a general phenomenon associated with experimental infection. An increase in B-lymphocyte numbers in TLN was another characteristic feature accompanying APP infection in all experimental groups. In piglets with colostrum-derived APP-specific antibodies, this was associated with higher relative numbers of IgM+CD2+ lymphocytes in TLN, while B-cells with the CD2- surface phenotype apparently expanded in the absence of passive humoral immunity.

Actinobacillus Infections↗

Expression of lymphocytes and lymphocyte subsets in patients with severe acute respiratory syndrome.

In a cohort of 38 patients with severe acute respiratory syndrome (SARS), we observed leukopenia in 47% of patients, lymphopenia in 84%, and T lymphopenia in 95%. CD4(+) T lymphocyte levels were reduced in 100% of patients, CD8(+) T lymphocyte levels were reduced in 87%, B lymphocyte levels were reduced in 76%, and natural killer cell levels were reduced in 55%. Our data suggested that these patients' immune systems were impaired during the course of SARS. The absolute counts of lymphocyte subsets demonstrated a clinical significance for patients with SARS.

Adolescent↗

Changes in T-lymphocyte subsets during acute rheumatic fever.

Relative proportions and numbers of helper [OKT4(+)] and suppressor [OKT8(+)] peripheral blood lymphocytes were examined in 32 young patients during the acute phase of rheumatic fever. No significant decrease in percentage T cells during acute rheumatic fever was noted in comparison to normal children controls from the same population. A decrease in absolute numbers of OKT4(+) T cells was also noted in acute rheumatic subjects not receiving corticosteroids (P less than 0.05). In addition, a significant decrease was documented in both proportions and total numbers of OKT8(+) putative suppressor cells during acute rheumatic attacks. C-reactive protein binding to T-lymphocyte subsets showed no preferential reactivity in vivo for suppressor or helper T cells. Antigen-reactive T lymphocytes identified with the T29 mouse hybridoma reagent showed similar proportions and numbers in rheumatic children as were noted in controls. The present data indicate profound alterations in both helper- and suppressor-cell types in the peripheral blood profile of children with acute rheumatic fever.

Acute Disease↗

Lymphocyte subsets and activation in prediabetes.

Peripheral blood lymphocytes were obtained from 65 individuals: 34 nondiabetic patients with islet cell autoantibodies (ICA) (prediabetic phase), 9 patients with newly diagnosed insulin-dependent diabetes mellitus (IDDM), 6 ICA-negative siblings or offsprings of IDDM patients, and 16 ICA-negative controls. The presence of lymphocyte abnormalities and/or activation was examined with dual fluorescence flow cytometry. The percentages of B cells, total T-lymphocyte, and helper T-lymphocyte (Th) and cytotoxic/suppressor T-lymphocyte (Tc/s) subsets and their ratio were not significantly different among the patient groups. No increased expression of interleukin 2 receptor on T-lymphocyte was found in newly diagnosed IDDM or prediabetic individuals. Sixteen of 49 patients had significantly increased number of T-lymphocyte expressing HLA-DR. A significant increase in the number of both Th and Tc/s subsets expressing HLA-DR was found in only 3 of 16 patients. This increase was unrelated to the patients's relative ICA titer or HLA-DR phenotype. On the other hand, the relative density of the DR antigen (RAD-DR) was significantly increased on both Th (886 +/- 120) and Tc/s (1250 +/- 273) in 13 of 38 patients compared with control patients (Th 484 +/- 129 and Tc/s 460 +/- 166). The RAD-DR on Tc/s correlated with the relative ICA titer and was greatest on DR3/4-phenotyped T-lymphocytes. In addition, significantly increased RAD-DR was found in noncontrol patients with impaired insulin release responses at 1 and 3 min to intravenous glucose.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗