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T-lymphocyte subsets modifications in multiple sclerosis: correlation with clinical disease activity.

Serial analyses of peripheral blood T-lymphocyte subsets were performed in 280 samples collected over a 27 months period from 14 multiple sclerosis patients. A significant decrease of T8+ cells was found in 47.1% (47/87) of blood samples collected within four weeks of onset of a relapse opposed to 2.1% (1/47) of samples collected during the four weeks before a relapse and to 3.4% (5/146) of those collected over four weeks after a relapse. Changes of T-cells subsets appear to correlate with disease activity, but normal findings observed before relapses suggest that the decrease of T8+ cells cannot be used as valid predictor of relapse.

Antigens, Differentiation, T-Lymphocyte↗

IFN-gamma influences the migration of thoracic duct B and T lymphocyte subsets in vivo. Random increase in disappearance from the blood and differential decrease in reappearance in the lymph.

Thoracic duct lymphocytes (TDL) continuously patrol through the body, facilitating immune responses at most sites. IFN-gamma might regulate immune responses by influencing the migration of TDL. Therefore, it was investigated in vivo whether IFN-gamma affects the migration of thoracic duct B, T, CD4+, and CD8+ lymphocytes from blood to lymph. Labeled TDL were injected i.v. into rats continuously receiving IFN-gamma via a central venous catheter. The numbers of B, T, CD4+, and CD8+ lymphocytes were determined in blood and thoracic duct lymph for 120 h. IFN-gamma increased the disappearance of TDL from the blood to a similar extent in all subsets. In contrast, the reappearance of B and T lymphocyte subsets in the lymph was decreased: B lymphocytes were affected significantly more than T lymphocytes, whereas CD4+ and CD8+ lymphocytes were affected to a similar extent. Our study suggests that differential retention within the tissue rather than preferential immigration into the tissue creates a microenvironment with a distinct composition of lymphocyte subsets.

Animals↗

Role of T-lymphocyte subsets in recovery from herpes simplex virus infection.

Our investigations probed the nature of different T-lymphocyte subsets effecting clearance of herpes simplex virus after infection of the pinna. Cell populations from animals recently infected subcutaneously or intraperitoneally (acute population) or from animals infected 6 weeks previously (primed population) or the latter cells reimmunized in vitro with virus (memory population) were studied. Viral clearance was a function of the Lyt 1+2- subset in the acute population, but with the memory population both Lyt 1+ and Lyt 2+ cells affected clearance. In primed populations, viral clearance was effected only by the Lyt 2+ subset. The ability of the various cell populations to adoptively transfer delayed-type hypersensitivity was also studied. Only acute population cells from animals infected subcutaneously and memory population cells transferred delayed-type hypersensitivity. In both cases, the cell subtype was Lyt 1+2-. Our results demonstrated that the delayed-type hypersensitivity response does not always correlate with immunity to herpes simplex virus. Multiple subsets of T cells participate in viral clearance, and their respective importances vary according to the stage of the virus-host interaction.

Animals↗

[Humoral immunity and lymphocyte subsets in patients with dilated cardiomyopathy].

We examined immunological abnormalities in patients with dilated cardiomyopathy (DCM) and reviewed their medical histories, NYHA classifications, electrocardiogram findings and chest X-Ray findings with regard to severity of DCM. Elevated CD4/CD8 ratio as well as decreased number of CD8 cells in lymphocyte subsets was recognized. These abnormalities were particularly noted in patients with past history of myocarditis. NYHA IV class increases in CTR and left ventricular dimension, and a reduced ejection fraction. These findings suggest that lasting of myocardial damage causes immunological abnormalities and the examination of DCM patients' immunological status is useful in anticipating disease process and determining medical treatment.

Autoantibodies↗

Alterations induced by chronic stress in lymphocyte subsets of blood and primary and secondary immune organs of mice.

BACKGROUND: The immune system is particularly sensitive to stress. Although acute stress generally has positive effects, chronic stress typically provokes immunosuppression. The elucidation of the mechanisms involved in immunosuppression are of interest for the design of therapeutic approaches to avoid the appearance of stress disorders. This study aimed to investigate chronic stress-induced alterations on lymphocyte subset distribution and percentages. The experiments were performed with C57BL/6 mice subjected to chronic immobilization stress. RESULTS: Stress caused a marked increase in apoptosis inside the thymus, and a reduction in the total number of thymocytes. Furthermore, the proportion of immature thymocytes declined significantly suggesting that the increased apoptosis mainly affected cells of immature phenotype. In blood, the total number of lymphocytes diminished but not all lymphocyte populations showed the same tendency: while the relative proportion of B cells declined slightly, the relative proportion of circulating CD3+ cells, and particularly some T cell subsets showing an immature phenotype (CD3+PNA+), increased under stress. The spleen and lymph nodes show a marked reduction in cellularity, but the relative proportion of T cells increased, while no change or only a slight reduction was observed in the relative proportion of B cells. Similarly, the relative proportion of T cells increased in bone marrow. CONCLUSIONS: Detailed data on the alterations of lymphoid cell subsets occurring under immobilization stress, both in the bloodstream and in different lymphoid tissues, are obtained. In general, T cells are more affected than B cells and, in particular, a marked increase in the percentage of a subset of circulating PNA+CD3+ T cells is observed.

Animals↗

Twenty-four-hour rhythms of mitogenic responses, lymphocyte subset populations and amino acid content in submaxillary lymph nodes of growing male rats subjected to calorie restriction.

Peripubertal male Wistar rats were submitted to a calorie restriction diet enriched in proteins and low in fat and carbohydrates for 4 weeks, and starting on day 35 of life. Mitogenic responses, lymphocyte subset populations, interferon (IFN)- gamma release and amino acid content were determined in submaxillary lymph nodes at six time intervals during the 24-h span. The diet employed completely arrested growth in growing rats. After caloric restriction, mean values of Con A response, lymph node T and CD4+ cell number and CD4+/CD8+ ratio augmented, whereas those of B cell number, IFN-gamma release and glutamine and glutamate concentration decreased. Calorie restriction modified 24-h rhythmicity of lymph node mitogenic responses to Con A and LPS, and of T, T-B, CD4+ and CD4+ -CD8+ lymph node cell subsets. It also changed the 24-h pattern of lymph node IFN-gamma release and glutamine, aspartate, glutamate and taurine lymph node content. Availability of nutrients presumably affects the mechanisms that modulate the circadian variation of immune responsiveness in growing rats.

Aging↗

Immunoperoxidase localization of lymphocyte subsets in the host response to melanoma and nevi.

Using an avidin:biotin immunoperoxidase system with monoclonal antibodies to lymphocyte subsets, we have investigated the host response to malignant melanoma and melanocytic nevi in frozen sections. Eight primary melanomas, eight metastases, three dysplastic nevi, and two dermal nevi were studied with antibodies T11, T4, T8, and B1. Sections were read in a semiquantitative manner by two observers. Virtually all lymphocytes in these lesions were T-cells (T11 positive). In all primary melanomas, in the majority of metastases, and in all dysplastic nevi, both T4- and T8-positive cells were present. In two of eight metastases, tumor cells stained with T4, and in one case, melanoma cells stained with B1 antibody. The host response to melanoma involves primarily T-cells and includes both the helper:inducer (T4) and suppressor:cytotoxic (T8) subsets.

Horseradish Peroxidase↗

A comparative study of alteration in lymphocyte subsets among varicella, hand-foot-and-mouth disease, scarlet fever, measles, and Kawasaki disease.

Changes in the lymphocyte subsets of 13 patients with varicella, 5 with hand-foot-and-mouth disease, 4 with scarlet fever, 10 with measles and 20 with Kawasaki disease were examined by immunofluorescent flow cytometric analysis using monoclonal antibodies against lymphocyte cell surface antigens. The results were compared with those of age-matched normal controls. A significant increase in the percentage of Leu-2a positive (Leu-2a+) cells was shown during the early convalescence of varicella, scarlet fever and measles. A significant decrease in the percentage of Leu-3a+ cells during the acute phase was common to all the diseases examined, and a significant decrease of Leu-4+ cells was observed except in measles. As a result, a significant decrease in the Leu-3a+/Leu-2a+ ratio was common to all the diseases examined during the acute and/or early convalescent phases. Leu-M3+ cells increased significantly in varicella, scarlet fever, and Kawasaki disease. HLA-DR+ cells increased significantly in varicella and Kawasaki disease. No significant changes in the proportions of Leu-7+, Leu-10+, and 2H7+ cells were found throughout the course of all the diseases examined.

Adolescent↗

Antigen-specific in vitro activation of T-lymphocyte subsets of cattle immunized with a modified live bovine herpesvirus 1 vaccine.

Bovine peripheral blood mononuclear cells from cattle immunized with a modified live virus bovine herpesvirus 1 (BHV1) vaccine and from BHV1-negative cattle were incubated in vitro with inactivated BHV1 for 6 days. Activation of T-lymphocyte subsets was measured by two-color flow cytometric analysis of T-cell phenotype and surface expression of CD25, the alpha-subunit of the high-affinity interleukin-2 receptor. Vaccinated animals, but not unvaccinated animals, had CD3+, CD4+, and gamma-delta T cells that significantly (p < 0.05) increased expression of CD25 when incubated with BHV1. CD8+ T cells from vaccinated animals did not consistently increase CD25 expression when incubated with inactivated BHV1.

Animals↗

Perforin expression can define CD8 positive lymphocyte subsets in pigs allowing phenotypic and functional analysis of natural killer, cytotoxic T, natural killer T and MHC un-restricted cytotoxic T-cells.

In this study we have used the expression of perforin to characterize subsets of porcine cytotoxic lymphocytes. Perforin positive lymphocytes expressed both CD2 and CD8alpha, most were small dense lymphocytes (SDL) and up to 90% were CD3 negative. However, the numbers of perforin positive T-cells increased with the age of the animal and their populations increased after specific antigen stimulation in vitro. The remaining perforin positive lymphocytes were large and granular and contained more CD3+CD5+CD6+ T-cells (-40%) of which a substantial proportion also co-expressed CD4. Perforin was expressed in subpopulations of both CD8alphaalpha and CD8alphabeta lymphocytes, but was not expressed in gammadelta T-cells or monocyte/macrophages. The perforin positive CD3- subset was phenotypically homogeneous and defined as CD5-CD6-CD8beta-CD16+CD11b+. This population had NK activity and expressed mRNA for the NK receptor NKG2D, and adaptors DAP10 and DAP12. Perforin positive T-cells (CD3+) could be divided into at least three subsets. The first subset was CD4-CD5+CD6+CD11b-CD16- most were small dense lymphocytes with cytotoxic T-cell activity but not all expressed CD8beta. The second subset was mainly observed in the large granular lymphocytes. Their phenotype was CD4+CD5+CD6+CD8beta+CD16-CD11b- and also showed functional CTL activity. Thus not all of double positive T-cells are memory helper T-cells. The third subset did not express the T-cell co-receptor CD6, but up to half of them expressed another T-cell co-receptor CD5. The majority of this subset expressed CD11b and CD16, thus the third perforin positive T-cell subset was CD3+CD4-CD5+CD6-CD8beta+/-CD11b+CD16+, and possessed MHC-unrestricted cytotoxicity and LAK activity.

Animals↗

Determination of co-expression of activation antigens on proliferating CD4+, CD4+ CD8+ and CD8+ lymphocyte subsets by dual parameter flow cytometry.

The incidence of activation markers on proliferating CD4+, CD4+ CD8+ and CD8+ lymphocyte subsets was determined in a single laser Epics-C fluorescence-activated cell sorter system, using a series of double staining combinations. Experiments were performed after 3 days of culture with PHA on cell fractions enriched for CD4+ or CD8+ lymphocytes before initiation of culture. The percentage of CD4+, CD4+ CD8+ and CD8+ lymphocytes in the total population was determined using double staining with Leu3 PE for the detection of CD4+ cells, and Leu2 FITC for the detection of CD8+ cells. Next, double stainings with Leu3 and Leu2 antibodies conjugated with PE and antibodies directed against activation markers (M) IL-2 receptor, transferrin receptor, HLA-DR antigen and CALLA conjugated with FITC were performed, using the following combinations: Leu3 and Leu2/M, Leu3/M and Leu2/M. The expression of activation markers on CD4+ CD8+ lymphocytes was calculated from the results. Our findings indicate that CALLA is expressed on most CD4+ and all CD4+ CD8+ cells, and on a small percentage of CD8+ lymphocytes; the IL-2 receptor was expressed on most CD4+ cells, on approximately three-quarters of CD4+ CD8+ cells and half the CD8+ cells; HLA-DR was expressed on a small percentage of CD4+ cells, all CD4+ CD8+ cells and half of CD8+ cells. The transferrin receptor was almost exclusively expressed on CD4+ CD8+ cells. The standard deviation of the calculated values did not exceed 13% and this analysis can generally be applied to determine the co-expression of a third marker in a mixture of single and double stained cells using conventional methods.

Adult↗

[Effect of jiawei dachaihu decoction on blood T lymphocyte subsets and tumor necrosis factor in patients with obstructive jaundice].

OBJECTIVE: To observe the effect of the preoperative Jiawei Dachaihu decoction in improving cellular immune functions in patients with obstructive jaundice. METHODS: Forty patients were randomly divided into two groups with (group B) and without (group A) the herbal therapy. And their preoperative T lymphocyte subsets, TNF and the postoperative dynamic changes were measured. RESULTS: CD3, CD4 and CD4/CD8 decreased and TNF increased in the patients. CD3, CD4 and CD4/CD8 decreased much less and were lower than preoperative levels 2 days after the operation. CD3, CD4 and CD4/CD8 were higher and TNF lower than preoperative levels in group A after 20 days of the operation (all of them P < 0.05). CONCLUSIONS: The operation could be helpful to recover immune functions of T lymphocyte and decrease TNF levels. The perioperative herbal therapy is an important adjuvant measure.

Adult↗

[Peripheral blood lymphocyte subsets in acute relapsing multiple sclerosis].

Anti-Leu series of monoclonal antibodies were used to evaluate lymphocyte subsets with ABC staining method. The peripheral blood total T cells (Tt), helper T cells (Th), suppressor T cells (Ts) and B cells were counted in 10 cases of acute relapsing MS, in which 7 cases had been investigated consecutively. Controls included 12 cases of remitting MS, 18 patients with other neurological diseases and 56 healthy subjects. The results showed that the peripheral blood Ts decreased in number and the Th/Ts ratio elevated during acute relapsing stage of MS, and they tended to return to normal when the disease showed a gradual recovery (The patients with longer periods of high Th/Ts ratio had more serious manifestations of the illness more severe disability) and the relapsing stage would be longer too.

Adolescent↗

Immunization with Trichinella spiralis Korean isolate larval excretory-secretory antigen induces protection and lymphocyte subset changes in rats.

Protection and immune responses were studied in rats immunized with Trichinella spiralis muscle stage larval excretory-secretory antigen (ES Ag) without adjuvant. Protection was assessed by the degree of adult worm burden and the yield of muscle (diaphragmatic) larvae after challenge infection with live larvae. Lymphocyte subsets were identified by flow cytometry in the spleen and peripheral blood. Cytokine production and specific IgG, IgG1 and IgG2a antibody responses were measured. Immunization with ES Ag produced highly significant protection against adult stages (98.4%) and muscle larvae (82.9%). Th2 type cytokines (IL-10, IL-4) were predominant. Anti-muscle stage larval ES Ag antibody was significantly elevated in the order IgG2a > IgG1 > IgG on the 2nd day after final immunization and on the 7th day after challenge infection. Expression of CD4+ and the CD4+/CD8+ ratio from spleen and blood were significantly increased compared to the control. These results demonstrate that immunization with T. spiralis antigen can elicit robust immune response, resulting in complete protection against infective larvae, and this protection can be achieved without the use of any adjuvant.

Animals↗

[Peripheral lymphocyte subset of patients with pancreatic diabetes mellitus--about a decreased ratio of T-LGL and ability of host defense].

Patients with diabetes mellitus (DM) often suffer from various and severe infection. Patients with pancreatic DM have malnutrition due to chronic pancreatitis. Therefore it is believed that patients with pancreatic DM have a lower ability of host defence to infectious diseases than normal subjects. We examined the primpheral lymphocyte subsets (T cell, B cell, NK cell, CD3+ 56+ T) of eighteen patients (males, age = 52.8 +/- 12.7 years old, mean +/- SD) with pancreatic DM by two color flow-cytometry to evaluate the lymphocyte function. Ratios of T cell (T%, 66.9 +/- 9.3%, mean +/- SD). B cell (B%, 13.1 +/- 5.8%) and NK cell (Nk%, 19.3 +/- 8.7%) to total peripheral lymphocytes of patients with pancreatic DM were not significantly different from those (T% = 66.4 +/- 7.8%, B% = 13.5 +/- 6.7%, NK% = 19.9 +/- 9.1%) of thirteen normal subjects (males, age = 51.2 +/- 13.1). CD3+56+ T cell % (4.1 +/- 1.9%) of patients with pancreatic DM was lower than that (6.0 +/- 3.0%) of normal subjects (p < 0.05). CD3+56+ T cells have cytotoxic activity and it is likely that this activity is similar to that of NK cells. These results suggested that a decrease in peripheral CD3+56+ T cell % is a factor showing a weak host defense mechanism to infectious diseases in patients with pancreatic DM.

Adult↗

Rapid flow cytometric method for measuring lymphocyte subset activation.

Standard in vitro methods for assessing T-cell activation have typically measured either the proliferative responses of peripheral blood mononuclear cell (PBMC) cultures to various provocative stimuli employing tritiated thymidine incorporation or the secretion of specific cytokines from activated cells. These bulk assay methods suffer the drawback of being lengthy assays, and, in addition, they do not provide information about functional responses of individual lymphocyte subsets. This report describes a three-color flow cytometric method for the rapid analysis (4 hours) of individual T-cell subsets in whole blood responding to various provocative stimuli, including pokeweed mitogen, the comitogenic monoclonal antibodies (mAbs) CD2/CD2R, the superantigen staphylococcal enterotoxin B (SEB), and the specific antigen Candida albicans. After 4 hours, CD69 expression in response to CD2/CD2R paralleled thymidine incorporation measured after 72 hours. Variations in the proportions of CD4+ and CD4- T cells expressing CD69 were observed with different stimuli. These observations demonstrate the potential of multiparameter flow cytometry for the investigation of functional responses of individual T-cell subsets to a variety of stimuli in whole blood.

Antigens, CD↗

Immunodepression secondary to malnutrition: assay by lymphocyte subset analysis using monoclonal antibodies and the cell sorter.

It has been well documented that the hospitalized child frequently is malnourished, and the considerably greater morbidity and mortality of such children, in large part, is due to the associated secondary immunodepression. To assess the mechanism of such immunodepression in acute and chronically malnourished subjects, we chose an experimental murine model of malnutrition. Immune function was assayed by lymphocyte subset population and mixed lymphocyte culture (MLC) assessment, determining whether an alteration of the T helper (TH) to T suppressor-cytotoxic (TS-C) lymphocyte ratio is the mechanism that produces immunoincompetence in malnutrition. Ten-week-old A/J mice were rendered acutely or chronically malnourished by graded protein restriction using a 2.5% protein diet. These animals were studied before and after protein depletion with the fluorescent activated cell sorter (FACS) and the MLC. FACS cell populations were defined by the monoclonal antibodies to THY 1.2, LYT 1 (TH), and LYT 2 (TS-C) antigens. MLC reactivity was assessed with A/J v C 57 BL/6 mouse lymphocytes and expressed as a stimulation index (S.I.). Results are expressed in Table 1, and suggest that these malnourished animals were significantly immunodepressed as measured in the MLC reaction. However, the mechanism of this depression is not mediated by an inversion of the TH/TS-C ratio. These data contrast with those previously reported for the immunodepression of the burned or traumatized patient in which the TH/TS-C ratios are reversed; these data suggest that a different, as yet to be elucidated, mechanism exists for the immunodepression of malnutrition.

Animals↗

Analysis of mitogen-stimulated lymphocyte subset proliferation and nitric oxide production by peripheral blood mononuclear cells of captive elk (Cervus elaphus).

Elk (Cervus claphus) are reservoirs for Brucella abortus, Mycobacterium bovis, and Mycobacterium avium subsp. paratuberculosis, each a serious pathogen of domestic livestock. An understanding of the basic immune responsiveness of elk would aid efforts to develop methods to diagnose and prevent these diseases of elk. Peripheral blood mononuclear cells (PBMC) isolated from captive elk were examined for phenotype, lymphocyte subset proliferative capacity, and ability to produce nitric oxide (NO) upon pokeweed mitogen (PWM) stimulation. Although gamma delta TCR+ cells represented a high percentage of the peripheral blood lymphocyte pool, these cells responded poorly to PWM stimulation. B cells (i.e., sIgM+ cells), conversely, were responsive to PWM stimulation. Addition of PWM to PBMC cultures also resulted in a significant production of nitrite, the stable oxidation product of NO. Similar to other ruminant species, the majority of elk peripheral blood sIgM+ cells co-expressed MHC class II and B-B4, a B cell lineage marker that varies in expression during B cell development. Findings from the present study provide basic information on several parameters of cellular immunity of elk.

Animals↗