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Characterization of morphologic changes and lymphocyte subset distribution in lymph nodes from cats with naturally acquired feline immunodeficiency virus infection.

Lymph nodes were collected at biopsy or necropsy from 18 cats with naturally acquired symptomatic feline immunodeficiency virus (FIV) infection and from 18 seronegative cats. Thirty-five of the cats were domestic shorthairs and one was a Persian cross. The cats ranged from 7 months to 16 years of age and were mainly obtained from California veterinary practitioners, a California cattery, and a Veterinary Teaching Hospital. Based on clinical signs present at tissue collection, ten FIV-infected cats fell into the acquired immunodeficiency syndrome (AIDS)-related complex (ARC) clinical stage and eight in the terminal (AIDS) stage of FIV disease. All cats were FeLV negative by antigen ELISA. Histologic sections of lymph nodes from each cat were examined blindly and were categorized as hyperplastic, involuting, mixed hyperplastic and involuting, depleted, or normal based upon subjective evaluation of follicles and paracortex. The relative abundance of plasma cells was evaluated in methyl green pyronin (MGP) and hematoxylin and eosin-stained sections. Similar numbers of FIV-seropositive and -seronegative cats fell into each lymph node category. The only difference evident between FIV-infected cats and control cats was in the degree of plasmacytosis present; moderate to marked plasmacytosis was present in 13/18 FIV-infected cats but in only 3/18 control cats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Lymphocyte subsets in granulomas of human tuberculosis: an in situ immunofluorescence study using monoclonal antibodies.

The immunophenotypic characteristics of lymphocyte subpopulations in tissue lesions of human tuberculosis were investigated in 18 cases. Frozen sections of affected cervical lymph nodes were stained with a panel of monoclonal antibodies using avidin-biotin immunofluorescence technique. The mean percentage of cells staining with different antibodies was as follows: CD3 44%; CD4 43%; CD8 36%; HLA-DR 86%; and IgM 29%. The mean CD4/CD8 ratio was 1.3:1. Leu7 and CD1 staining cells were very sparsely distributed in the granulomas. Thus, infection with tuberculosis is associated with a predominantly T cell infiltrate in lesional tissues. The proportion of CD4 positive (helper/inducer) cells at these sites is much lower than that seen (i) in the normal circulation, (ii) in reactive lymph nodes and (iii) at the site of purified protein derivative (PPD) injection in healthy tuberculin-responsive individuals.

Adolescent↗

[Relationship between syndrome types of TCM in lung neoplasm patients and peripheral T lymphocyte subsets, carcino-embryonic antigen].

Fifty-six cases with lung carcinoma were detected for peripheral blood T lymphocyte by direct staphylococcal protein A (SPA) bacterial rose ring for carcino-embryonic antigen (CEA) by radioimmunoassay, and the relationship between T lymphocyte. CEA and Syndrome of TCM was explored. The results were as follows:the values of OKT3, OKT4, OKT4/OKT8 were lower in the lung carcinoma than that in healthy subjects (P < 0.05-0.01), but OKT8 was higher (P < 0.05); The values of T lymphocyte lowered or increased in the order of Yin Deficiency with internal Heat, Qi Stagnation with blood stasis, Phlegm-Dampness in Lung, Qi-Yin Deficiency, but that of CEA was the highest in Qi-stagnation with blood stasis. There was a significant difference between all Syndrome-types of TCM and the healthy subjects (P < 0.01). Therefore, it is suggested that the values of peripheral T lymphocytes and CEA with the lung carcinoma were regarded as good referential parameters in reflection the condition of Deficiency in Vitality and Excess in Superficiality, which offered the objective evidence for the TCM treatment.

Adenocarcinoma↗

Hematologic values and lymphocyte subsets in fetal blood.

Hematologic values and lymphocyte subpopulations were determined in normal fetal blood during the second trimester of gestation. In these samples the platelet, erythrocyte, and leukocyte counts were significantly lower than in adults. Large red blood cells with a high hemoglobin content were present. Before the twentieth week of gestation, erythroblasts made up about half of the nucleated elements. Lymphocytes formed most of the leukocytes, and their absolute numbers were comparable to those in adults. Most of the fetal blood lymphocytes expressed T- or B-cell surface differentiation antigens. The percentage of T-cells was lower and that of B-cells was higher than in the adult. A high OKT4/OKT8 ratio was present. It was due to a low percentage of OKT8-positive cells. Lymphocytes with a natural killer cell phenotype were rare. Most lymphocytes were OKT10 positive, but almost none reacted with the antithymocyte antibody OKT6. These results give additional information about the development of blood cells in early human life. They can be used as reference values for the prenatal diagnosis of hereditary or acquired anomalies of the hematologic and immunologic systems.

Fetal Blood↗

Cytochemical analysis of human peripheral blood lymphocyte subsets defined by monoclonal antibodies.

Human blood lymphocyte subpopulations, revealed by a panel of commercially available monoclonal antibodies by means of a rosetting technique, were submitted to direct cytochemical analysis and were shown to have distinguishing characteristics. T cells reactive with OKT3 antibody (T3+) displayed higher beta-glucuronidase and dot-like alpha-naphthyl acetate acid esterase (ANAE) activity than T3- cells. Helper/inducer cells (T4+) were characterized by a high level of dot-like ANAE activity, whereas cytotoxic/suppressor cells (T8+) displayed selective naphthol-ASD-chloroacetate esterase activity. These results provide evidence for association of some cytoplasmic enzyme activities with the expression of membrane differentiation markers defined by monoclonal antibodies.

Acid Phosphatase↗

Carbohydrate-labelled glycoproteins as markers of human lymphocyte subsets stimulated with mitogen and alloantigen.

Human lymphocytes from thymus, spleen, and blood were stimulated by concanavalin A, phytohemagglutinin or pokeweed mitogen to incorporate radioactive thymidine and various carbohydrates. Peripheral-blood lymphocytes were also activated by alloantigen in a mixed lymphocyte reaction. Incorporation of thymidine and carbohydrates was parallel in stimulated cells. Carbohydrate-labelled cells were extracted with Triton X-100 buffer and the extracts subjected to sodium dodecylsulfate/polyacrylamide gel electrophoresis and subsequent autoradiography. From a comparison of the resulting glycoprotein patterns the following conclusions could be drawn. (a) The same population of thymocytes responds to all three mitogens. (b) Pokeweed-mitogen-stimulated blood lymphocytes are similar to this mitogen-responsive thymocyte population. (c) The glycoprotein pattern of pokeweed-mitogen-responsive spleen cells shows two characteristic glycoproteins, GP 50 and GP 78, and resembles that of murine B cells. (d) Whereas concanavalin A and phytohemagglutinin activate identical subpopulations of T cells in the spleen, this is not the case in the blood. (e) Concanavalin-A-activated and alloantigen-activated blood lymphocytes express a glycoprotein GP 185 which is not found on phytohemagglutinin-responsive cells. (f) The kinetic relationship during alloantigen stimulation in a mixed lymphocyte reaction between the appearance of glycoprotein GP 185 and subsequent maximal killing activity in an assay using release of 51Cr makes it likely that glycoprotein GP 185 is a marker for cytotoxic T lymphocytes.

Carbohydrate Metabolism↗

Effects of repeated exposure to 4 ppm nitrogen dioxide on bronchoalveolar lymphocyte subsets and macrophages in healthy men.

Following the basal descriptive studies of the bronchoalveolar inflammatory cell response induced by single exposure with nitrogen dioxide (NO2) in man, it was considered important to clarify the cell response to repeated exposure with NO2. This investigation was, therefore, undertaken with bronchoalveolar lavage (BAL) 3 weeks before and 24 h after six repeated exposures with 4 ppm NO2 (7 mg.m-3) in ten healthy volunteers. The exposures were performed during 20 min and every second day. Analysis of the recovered BAL fluid demonstrated that repeated exposures to NO2 caused a lung cell response different from that reported after a single exposure. Amounts of lavaged alveolar macrophages, B-cells, and natural killer (NK)-cells were decreased and the T-helper-inducer/cytotoxic-suppressor cell ratio was altered, but there was no lymphocytosis or mastocytosis as after single exposure. Lymphocyte numbers in peripheral blood were reduced after exposure. These results suggest that repeated exposure with NO2 adversely affects the immune defence. This could contribute to the increased susceptibility to airway infections reported to be associated with NO2 exposure.

Adult↗

T-lymphocyte subsets and their cytotoxic capacity in patients with type I and type II diabetes mellitus.

Distribution and cytotoxic function of lymphocyte subpopulations were studied in 20 patients with type I, in 20 patients with type II diabetes mellitus and in 40 control subjects. The percentage, the absolute number of (EA), (E Thr), (TM) subsets and the rate of (E Thr) (E Ths) and (TM/) (TG) cells were found to be higher in type I and lower in type II diabetic patients than in controls. This opposite tendency in the distribution of T-lymphocyte subsets may be related to the duration of diabetes. Simultaneously a significant cytotoxic capacity of U, null, T, (TG)-lymphocytes was observed against human pancreas extract-coated targets in almost all 18 out of 20 of type I, but only in a few cases (5 out of 20) of type II diabetic patients. These five patients needed insulin therapy six months later. The T and (TG)-lymphocytes had the largest killer activity in both diabetic groups. Lymphocyte-mediated cytotoxicity seems to be a specific method which is suitable for the study of in vitro sensitization against pancreas tissue, and it might predict insulin dependency in type II diabetic patients.

Adolescent↗

Blood and alveolar lymphocyte subsets in pulmonary cytomegalovirus infection after lung transplantation.

BACKGROUND: Cytomegalovirus (CMV) pneumonitis has been shown to be associated with lymphocytic alveolitis after lung transplantation. In the present study, we investigated a series of bronchoalveolar (BAL) and blood samples, collected in the absence of rejection or acute infectious episodes. in order -1: to evaluate intra-alveolar cell population changes concomitant with CMV replication and -2: to reappraise the value of cell population analysis in the management of patients after lung transplantation. METHODS: We used flow cytometry to investigate modifications of lymphocyte subpopulations related to pulmonary cytomegalovirus infections in blood and BAL samples from a series of 13 lung transplant recipients. After exclusion of samples obtained during pulmonary rejection, bronchiolitis obliterans or acute bacterial infection, 48 blood and BAL samples were retained for analysis: 17 were CMV positive by shell-vial assay and 31 were CMV negative in blood and BAL. RESULTS: Our results demonstrate that pulmonary CMV infection is associated with a significant increase in the total lymphocyte population in BAL samples, but with minor modifications of the various lymphocyte subpopulations and a significantly higher absolute number of B lymphocytes in blood samples. CONCLUSIONS: Cytomegalovirus pulmonary infection is accompanied by only minor changes in BAL lymphocyte subpopulations. The study of BAL lymphocyte subpopulations therefore appears to be of limited clinical value in the diagnosis of pulmonary CMV infection. However, increased blood B-lymphocytes seems to be a clinical feature associated with CMV infection.

Bronchoalveolar Lavage↗

Functional and phenotypic characterization of human B lymphocyte subsets isolated by unit gravity sedimentation.

We report the use of unit gravity sedimentation with a CelSep apparatus to generate two volumetrically similar but functionally and phenotypically distinct subsets of human peripheral blood B cells. One subset, comprised of small B lymphocytes, underwent a significant size change in response to anti-mu, proliferated synergistically to low concentrations of anti-mu plus B cell growth factor (BCGF) or phorbol myristate acetate plus BCGF, and could be induced to produce immunoglobulin in response to pokeweed-mitogen-derived T-lymphocyte-replacing factors. These cells were primarily sIg+, B1+, B2+, and were virtually free of monocytes (less than 0.01%). Unlike these resting B lymphocytes, the large cells proliferated directly to BCGF, without displaying synergy with anti-mu. These cells displayed very little B2 (less than 7%), did not increase in volume in the presence of anti-mu, and made more immunoglobulin in response to TRF than the small resting B lymphocytes. However, neither population synthesized immunoglobulin spontaneously. This technique, which is highly reproducible, not equipment intensive, and produces high cell recovery (greater than 90%), allows for a precise analysis of the steps involved in the maturation of a resting B lymphocyte to an immunoglobulin-secreting cell.

B-Lymphocytes↗

CD45 (leucocyte common antigen) expression in T and B lymphocyte subsets.

CD45 is the dominant tyrosine phosphatase in haematopoietic cells and can modulate the effects of many other signaling molecules by dephosphorylation. The extracellular portion of CD45 has considerable variability due to differential splicing and glycosylation. This may allow for interactions with a variety of ligands expressed on interacting cells or on the same cell surface. Monoclonal anti CD45 antibodies that are reactive with epitopes that result from differential splicing and glycosylation can distinguish between cell populations that differ in maturation and function. These reagents can be used in the immunophenotyping of hematopoietic malignancies as well as in immunodeficiencies and autoimmune diseases. Several studies have shown that different anti CD45 reagents have different activating or inhibiting effects in vitro on a variety of T and B cell activation events. There are some indications that anti CD45 reagents can also selectively modify lymphocyte function in vivo. Such applications could potentially allow for the selective upregulation and down regulation of lymphocyte functions in a variety of immunologically mediated diseases.

B-Lymphocyte Subsets↗

B and T lymphocyte subsets in fetal and cord blood: age-related modulation of CD1c expression.

Peripheral blood lymphocytes from fetal, term and adult subjects were analyzed with a panel of B- and T-lymphoid differentiation markers. CD1c +ve and CD5 +ve B cell (IgM +ve) percentage in fetal, cord and adult blood decreased significantly. Fetal and cord blood contained less CD2+ and CD3+ T lymphocytes and a higher percentage of CD38 +ve cells than adult blood. The phenotypic changes occurring during perinatal age in humans relate to the changes in the functional activities of the B and T lymphoid subsets from fetal to postnatal age.

Adult↗

A longitudinal study of circulating lymphocyte subsets in the peripheral blood during the acute stage of Guillain-Barré syndrome.

Activated T cells are implicated in the pathogenesis of Guillain-Barré syndrome (GBS). Blood samples from 16 patients with GBS were studied with flow cytometry during the acute stage of their disease to define circulating lymphocyte populations. During the progressive phase of GBS the T-suppressor/inducer (CD4/CD45RA) subset was decreased (10.3 +/- 4.4%, P < 0.05) and the T-helper/ inducer (CD4/CD29) subset was increased (34.9 +/- 9.2%, P < 0.05) compared to sex and age matched patients with other neurological diseases (OND, 15.5 +/- 5.7% and 27.8 +/- 8.6%, respectively) and healthy controls (16.5 +/- 6% and 28.1 +/- 8.5%, resp.). Within the CD8 population, the activated T-cytotoxic/suppressor (CD8/CD38) subset was increased during the progressive (13 +/- 10.1%, P < 0.05) and plateau phase (16.4 +/- 16.9%, P < 0.01) of GBS compared to OND patients (6.2 +/- 2.3%) and healthy controls (5.8 +/- 2.5%). The proportion of activated T cells (CD3/CD25) was increased during the progressive (9.3 +/- 3.8%, P < 0.05) and plateau phase (11.5 +/- 5.5%, P < 0.01) compared to OND patients (6.6 +/- 2.7%) and healthy controls (5.5 +/- 2.5%). The changes of T cell subsets normalized during the early recovery phase of GBS. 2 patients with serological evidence of antecedent cytomegalovirus (CMV) infection had abnormal high proportions (mean +/- 2 (SD) of healthy controls) of CD8 lymphocytes and correspondingly abnormal low proportions of CD4 lymphocytes during all phases of GBS. In contrast, the CD8 proportions were abnormal low in 3 patients with serological evidence of recent Campylobacter jejuni infection. There was no correlation between the proportions of lymphocyte subsets and the disability score during the maximum of the disease and after half a year. In conclusion, we found further evidence of T cell activation during the acute stage of GBS by the demonstration of an increased proportion of activated CD8+ T cells, which may be directly cytotoxic to Schwann cells. The abnormalities of the CD8 subset in some GBS patients seem to depend on the nature of the preceding infection.

Acute Disease↗

Circulating lymphocyte subsets in obsessive compulsive disorder, major depression and normal controls.

BACKGROUND: Obsessive compulsive disorder (OCD) shares several features with depressive illness (e.g., comorbidity, early escape from dexamethasone suppression, effectiveness of serotonergic pharmacotherapy). It was of interest to establish whether OCD, like major depression, was also associated with immune alterations, notably elevations of circulating natural killer (NK) cells. METHOD: Circulating lymphocytes were determined from morning blood samples taken from OCD and major depressive patients, as well as from age- and sex-matched controls. Stress perception and coping styles were evaluated in order to assess whether such variables accompanied the NK alterations. Finally, in a subset of patients, symptoms of the illness, stress/coping, and circulating lymphocytes, were also evaluated following 12 weeks of antidepressant medication (serotonergic reuptake inhibitor). RESULTS: The major depressive and OCD patients reported increased perception of day-to-day stresses, coupled with reliance on emotion focused coping styles. Moreover, circulating NK cells were elevated among male OCD and major depressive patients, whereas only a modest increase of NK cells was seen in female major depressives. Twelve weeks of medication alleviated depressive and OCD symptoms, and resulted in normalization of NK cells in the major depressives. However, in OCD patients the reduction of symptoms was not accompanied by significant variations of circulating NK cells. CONCLUSIONS: Although major depression and OCD are both accompanied by elevated circulating NK cells, at least in males, normalization of NK cells following treatment was only evident in depression. The persistent elevations of NK cells among male OCD patients may reflect either a trait characteristic of the illness, or symptom reduction and not true remission. LIMITATIONS: Although elevations of lymphocyte subsets in major depressive and OCD patients were observed, conclusions concerning immune status in OCD ought to be held in abeyance pending assessment of other indices of immune and cytokine functioning.

Adaptation, Psychological↗

Changes in peripheral blood lymphocyte subsets in patients undergoing radiotherapy.

PURPOSE: To investigate the changes in peripheral blood lymphocyte subpopulations in patients undergoing radiotherapy. MATERIALS AND METHODS: In 8 patients undergoing external beam radiotherapy to the pelvis, the different lymphocyte subpopulations were followed during treatment. The lymphocyte populations were determined using two-colour flow cytometry. The study comprises the T-helper, T-suppressor/cytotoxic cells, the B-lymphocytes and natural killer (NK) cells. RESULTS: The B-cells were characterized by a steep decrease at the beginning of the radiotherapy. They reached their lowest level at an equivalent total body dose of approximately 1.5 Gy and remained constant during the rest of the therapy (10% of the initial level). In T-cells (both T-helper and T-suppressor subsets) the steep decrease was less pronounced. T-lymphocytes reached a base level at 2.5 Gy equivalent total body dose (20% of the initial level). No significant differences between the T-helper and the T-suppressor/cytotoxic cells were observed. NK cells were characterized by a weak decline during the first weeks of therapy, being less pronounced than in the other populations. Near the end of therapy, the NK cells reached the level of the T-lymphocytes. CONCLUSION: In vivo, NK cells were the most radioresistant and B-cells the most radiosensitive lymphocytes. No significant differences between T-helper and T-suppressor/cytotoxic cells were observed. These data are in agreement with the differences in apoptosis induction in peripheral blood lymphocyte subpopulations after in vitro gamma-irradiation of whole blood lymphocytes.

Adult↗