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T and B lymphocyte subsets in spermatic granulomas in five rams.

Epididymal spermatic granulomas from five adult (greater than 4 years old) rams from a commercial flock were evaluated by quantitative histologic methods. Sections were stained for isotype specific immunoglobulin (Ig)-containing cells or specific lymphocyte subsets, as indicated by plasma membrane staining with monoclonal antibodies (MoAb). For the purpose of analysis, individual spermatic granulomas were allocated to one of three groups (early, resolving, or intermediate) on the basis of histologic examination. Ig-containing cells were most prevalent in early spermatic granulomas, and IgG-containing cells predominated in all spermatic granulomas regardless of their presumed age. IgM immunoglobulin-containing cells were the next in prevalence, and IgA-containing cells were infrequent. Assessment of lymphocyte subsets confirmed that lymphocytes present were of both T- and B-cell lineage and were well represented in all spermatic granulomas. CD4+ (MHC Class II-restricted "helper") T-cells were more prevalent in early spermatic granulomas than in intermediate or resolving types, while CD8+ (MHC Class I-restricted, "cytotoxic/suppressor") T-cells were more prevalent in spermatic granulomas of the intermediate type. These results suggest that in addition to an essentially foreign body reaction to extravasated spermatozoa, both local production of antibody and accumulation of a variety of B- and T-lymphocytes are present in ovine spermatic granulomas.

Animals↗

A correlation study between serum adenosine deaminase activities and peripheral lymphocyte subsets in Parkinson's disease.

Adenosine deaminase (ADA) and its isozyme activities in serum were measured together with peripheral lymphocyte subsets in 42 patients with idiopathic Parkinson's disease. The total and ADA 2 activities were significantly higher than normal controls (p < 0.01). As regards the peripheral lymphocyte subsets, the proportion of OKT 10+ cells (activated T lymphocytes) and the proportions of interleukin-2 receptor+ and HLA-DR+ cells (mainly activated T lymphocytes) were significantly higher than normal controls (p < 0.05, 0.01, 0.01, respectively). On the other hand, OKT 10+ cells demonstrated a significant correlation not only with total ADA but also with ADA 2 activity. These results suggest that high serum ADA activity may be involved in the pathogenesis of Parkinson's disease through peripheral T lymphocyte activation.

Adenosine Deaminase↗

Lymphocyte subsets in patients with aplastic anemia.

Lymphocyte subsets were enumerated in a group of 31 patients with aplastic anemia. Abnormal numbers of immunoregulatory T-cells were found in some patients: 26% of them showed a reversed helper/suppressor ratio. Seven of 18 patients showed significantly decreased proliferation in response to PWM; this hyporesponsiveness was present in 75% of patients with a reversed helper/suppressor ratio and in 10% of those with a normal helper/suppressor ratio (R = 0.66, P = 0.008). Eight of 18 patients showed suppressor activity over PWM-induced allogeneic cell proliferation. This suppressive activity did not correlate with T-cell phenotype. Of the patients with a low number of T-cells, 73% had responded to treatment, whereas of those patients with a normal number of T-cells, 26% had responded (P = 0.016). The results are consistent with abnormal immune response in selected patients with aplastic anemia, and suggest a possible influence of T-cells on disease process.

Adolescent↗

Stellate ganglion block modifies the distribution of lymphocyte subsets and natural-killer cell activity.

BACKGROUND: Recent investigations suggest that the sympathetic nervous system affects the immune system. This study examined whether stellate ganglion block (SGB) affects the immune response, specifically the distribution of lymphocyte subsets and natural-killer (NK) cell activity. METHODS: Ten volunteers received three different treatments in random order at 7-day intervals: (1) SGB with 7 ml of 1% lidocaine; (2) an identical volume of normal saline injected at the same site as SGB; and (3) an identical volume of 1% lidocaine injected intramuscularly. Blood samples were drawn before and 30 min after treatment. The distribution of lymphocyte subsets was analyzed, and NK cell activity was measured. Plasma concentrations of epinephrine, norepinephrine, adrenocorticotropic hormone, and cortisol were measured. RESULTS: Any value in the normal saline and intramuscular treatments did not change significantly. After SGB, the plasma concentrations of epinephrine and norepinephrine decreased significantly (P < 0.01), but adrenocorticotropic hormone and cortisol values were unchanged. Increases were observed in the proportion of B cells (from 18.4 +/- 3.0% to 20.0 +/- 3.8%; P < 0.01) and T cells (from 64.2 +/- 4.1% to 67.1 +/- 4.2%; P < 0.01). A decrease in the proportion of NK cells was observed (from 13.4 +/- 2.7% to 9.8 +/- 2.2%; P < 0.01). The proportion of CD4+ cells increased (P < 0.01), and that of CD8+ cells decreased (P < 0.01), so that the CD4+ cell/CD8+ cell ratio increased (P < 0.01). The proportion of CD29+ (helper-inducer T) cells increased (P < 0.05), but that of CD45RA+ (suppressor-inducer T) cells did not change. NK cell activity decreased significantly (from 33.6 +/- 8.3% to 29.1 +/- 7.6%; P < 0.01). CONCLUSIONS: A small but significant alteration in lymphocyte subsets and in NK cell activity by SGB indicates that local sympathetic nerve block may modulate the immune response.

Adult↗

The distribution of lymphocyte subsets in normal ovine skin.

The distribution of lymphocyte subsets in the skin of clinically normal sheep was studied using monoclonal antibodies to OvCD5, OvCD4, OvCD8, WC1, and CD45RA. Four different anatomical sites were examined in each of 38 sheep. Four different age groups ranging from 7 to 10-day-old lambs to 12 to 14-month-old adults were represented. The majority of lymphocytes in all age groups and at all sites were angiocentrically located within the superficial dermis. Total lymphocyte numbers at each site increased with age. The predominant cell type identified at all sites was WC1+ and the proportion of lymphocytes of this phenotype was significantly higher at wooled sites. Only occasional CD45RA +/- cells were present in any section.

Aging↗

Lymphocyte subsets in patients with acute myopericarditis, arrhythmias and dilated cardiomyopathy.

To investigate the role of immunoregulatory function in determining the clinical course of acute myopericarditis, lymphocyte subsets were analysed by laser flow cytometry in 20 patients with acute myopericarditis, 30 with various arrhythmias or atrio-ventricular block and 31 with dilated cardiomyopathy. During the healing stage of acute myopericarditis, patients with residual electrocardiographic or left ventricular wall motion abnormalities presented altered frequencies of lymphocyte subsets, increased B 1 and reduced OKT 8 positive cells with an elevated OKT 4/8 ratio. The abnormal pattern was not evident in patients with acute pericarditis nor in those with acute myocarditis who recovered completely without residual abnormalities. This observation suggested that an imbalance of helper/suppressor T cells could modulate the clinical course of acute myopericarditis, either by producing extensive and irreversible myocardial damage during acute illness or by inducing chronic smoulding myocardial inflammation. Patients with ventricular arrhythmias and left ventricular wall motion abnormalities also presented reduced suppressor/cytotoxic T cells, implying that they had been suffering from chronic smoulding myocarditis mediated by immunoregulatory dysfunction. However, we could not determine whether the imbalance of helper/suppressor T cells could mediate the progression from myocarditis to dilated cardiomyopathy, since no association was demonstrated between the abnormal lymphocyte subsets and mononuclear cell infiltration in endomyocardial biopsy sample from patients with dilated cardiomyopathy.

Acute Disease↗

Lymphocyte subsets in jejunal and ileal Peyer's patches of normal and gnotobiotic minipigs.

The size and location of Peyer's patches (PP) in the jejunum and ileum and their composition of lymphocyte subsets (B, CD2+, CD4+, CD8+) have been studied in conventional and gnotobiotic Göttingen minipigs. Each PP in the small intestine remained at the same site and was of comparable length between 2 and 12 months of age. In 1.5-month-old conventional minipigs the histology of the compartments differed between the continuous PP in the terminal ileum (ilPP) and the discrete PP in the jejunum (jejPP). No such difference was seen in gnotobiotic or in 12-month-old animals. The composition of lymphocyte subsets showed striking differences with significantly more B and less T, CD4+ and CD8+ cells in ilPP in 1.5-month-old minipigs in comparison with 12-month-olds. Mesenteric lymph nodes and jejPP displayed a typical pattern of lymphocyte subsets. The size of the lymphocyte compartments in PP and their cellular composition depends largely on age and microbial influences from the gut lumen, which might be of major importance for studies on the function of the gut-associated immune system in the pig.

Aging↗

[The T-lymphocytes subsets expression in the human recurrent nasal polyps].

OBJECTIVE: To investigate the expression of T-lymphocyte subsets (CD4+, CD8+ and CD45RO+ cells) in human recurrent nasal polyps. METHOD: Nasal polyps tissue samples and peripheral blood were obtained from 17 patients, normal human inferior turbinate mucosa and peripheral blood were obtained as well. Flow cytometry was adopted to detect the expression of surface markers of the T-lymphocytes. All data were analyzed with t-test. RESULT: There were significantly large number of CD4+, and CD45RO+ cells in the tissue of recurrent nasal polyps. The expression percentage of CD3+ CD4+ cells was significantly higher than that of CD3+ CD8+ cells. Ratio of CD3+ CD4+/CD3+ CD8+ was 1.956 +/- 0.093. CONCLUSION: There were generous of T-lymphocytes expression in human recurrent nasal polyps. The ratio of the T-lymphocytes subsets was abnormal high than the usual and this indicated the immunological function were in disorder in the local area. The abnormally higher expression of CD3+ CD4+ cells in the local area may play an important role in the recurrence of human nasal polyps.

Adolescent↗

[Studies on immunological parameters in patients with gastric carcinoma--with special reference to analyses with monoclonal antibodies of T-lymphocyte subsets and interleukin-2].

We have investigated the distributions of T-lymphocyte subsets and interleukin-2 in peripheral blood of 62 patients with gastric carcinoma. The antibody (OKT3, OKT4, OKT8, OKT4/8, Leu11, and Leu7) treated cells were analyzed on a Spectrum III automated flow cytometer. Interleukin-2 was measured by the tritium-labelled thymidine CTLL assay on the supernatants of peripheral blood lymphocytes of gastric cancer patients after 24 hours stimulation with phytohemagglutinin. The number of OKT3, OKT4 cells and the ratio of OKT4/8 were decreased in patients with advanced gastric carcinoma compared with early gastric carcinoma. The numbers of Leu11 and Leu7 cells in patients with stage II and III were in higher levels than those with stage IV. Interleukin-2 production was depressed in patients in advanced stage compared with those of early stage. We concluded that the T-lymphocyte subsets and the interleukin-2 production were useful immunological parameters for gastric cancer patients.

Adult↗

Cytofluorometric analysis of lymphocyte subsets in thyroid aspirates from patients with autonomously functioning nodules.

A microscale method, based on two-colour dye immunolabelling and flow sorting cytofluorometry, was used to characterize lymphocyte subsets in thyroid tissue specimens obtained by fine-needle aspiration (FNA) in 21 patients with autonomously functioning thyroid nodule (AFTN) and five patients with cold thyroid nodule (CTN). Inversion of the ratio between CD4+ and CD8+ lymphocytes, due to a relative increase of CD8+ cells, was found among intrathyroidal lymphocytes in AFTN patients. The abnormal lymphocyte subset distribution was not observed in the peripheral blood of the same group of patients. In patients with CTN the lymphocyte subset distribution was normal, both in the thyroid and in the peripheral blood. In AFTN patients, a significant correlation was observed between the decrease of intrathyroidal CD4+/CD8+ ratio and the increase of plasma thyroid hormone levels. Whether the immunological abnormalities found in AFTN could play a pathogenetic role in the clinical presentation and outcome of the disease remains to be established. The FNA-applied micromethod used in this study could be extended as a routine investigation to characterize the immunogenic substrate of thyroid disorders.

Adult↗

Alterations in T-lymphocyte subsets in the airways of asthmatic patients with active hepatitis C virus infection.

BACKGROUND: The interaction between hepatitis C virus (HCV) infection and certain pulmonary disorders is of considerable interest. We have previously found that active HCV infection in asthmatic patients is associated with impaired responses to inhaled corticosteroid therapy. OBJECTIVES: To examine differences in T-lymphocyte subsets in asthmatic airways with or without HCV infection. METHODS: 12 HCV-negative and 29-HCV positive asthmatics were included in this study. All HCV-positive asthmatics received interferon (IFN) therapy (11 IFN responders and 18 IFN non-responders). T-lymphocyte subsets in induced sputum were analyzed before and 1 year after IFN therapy. RESULTS: Before IFN therapy, percentages of lymphocytes and CD8+ T lymphocytes in induced sputum were higher in HCV-positive asthmatics than in HCV-negative asthmatics. After IFN therapy, there was no significant change in percentages of lymphocytes and T-lymphocyte subsets in IFN non-responders. However, percentage of lymphocytes and CD8+ T lymphocytes in IFN responders significantly decreased, while CD4+ T lymphocytes increased. Therefore, the CD4/CD8 ratio significantly increased following IFN therapy in IFN responders. In IFN responders, forced expiratory volume in 1 s (FEV(1)) after salbutamol administration significantly increased following IFN therapy, while it did not change in non-responders. In addition, in IFN responders, the change in the CD4/CD8 ratio after IFN therapy was significantly correlated with the increase in FEV(1) after salbutamol administration. CONCLUSIONS: The number of CD8+ T lymphocytes increases in the airways of asthmatic patients with active HCV infection, and this may be associated with airway dysfunction in these patients.

Albuterol↗

The normalization of guinea pig leukocyte fractions and lymphocyte subsets in blood and lymphoid tissues using a flow cytometric procedure.

Many hematological and immunological parameters remain unclear in the study of the guinea pig. In this study, we established the mean values of blood counts, the percentage of leukocyte fractions and lymphocyte subsets in blood and various lymphoid tissues of the guinea pig with a flow cytometric procedure using MIL4/SSC. The mean counts of WBC and RBC in the blood were lower, and MCV and MCH were higher than those of other rodents, resembling those of humans. Furthermore, the mean percentages of blood lymphocytes were smaller and that of granulocyte was larger than those of other rodents, resembling those of humans. We further established a flow cytometric procedure for lymphocyte subsets and clarified the mean percentages of T- and B-cells, CD4(+)-, CD8(+)- and MHC Class II(+)- T-cells, and CD4(-)CD8 (-) T-cells. The latter were morphologically larger in cell size and cytoplasm than CD4(+)- plus CD8(+) T-cells, and this subset had a significantly higher percentage in newborn animals. Furthermore, the appearance of the MHC Class II(+) T-cell subset was suggested to be a marker of hyper-activation of T-cells in BCG-immunized animals. Thus, both the novel flow cytometric procedure for leukocyte fractions and lymphocyte subsets, and the established normal values will be useful tools in studying guinea pigs as models of various diseases and biological phenomena.

Animals↗

[Study on abnormality and regulation of T-lymphocyte subsets in peripheral blood of rheumatoid arthritis patients].

OBJECTIVE: To investigate the change of T-lymphocyte subsets in peripheral blood of rheumatoid arthritis (RA) patients and analyze the effects of Fuzheng Qubi Decoction (FZQBD) on T-cell subsets. METHODS: Thirty RA patients were randomly divided into two groups, and treated with FZQBD or western medicine combination therapy respectively for one month, the percentage of peripheral CD3+, CD4+, CD8+ as well as the serum IgG and IgA levels were determined. RESULTS: The percentage of peripheral CD4+, CD4+/CD8+ ratio, IgG and IgA levels increased significantly (P < 0.001) in RA patients before treatment. After 1 month of FZQBD treatment, the CD4+/CD8+ ratio decreased obviously (P < 0.05). CONCLUSION: Abnormal cellular immunity exists in RA patients, FZQBD could adjust the abnormal T-lymphocyte subsets to normalize it.

Adolescent↗

[Lymphocytic subsets of tumor tissue, non tumorous kidney, and the peripheral blood in primary renal cell carcinoma].

Lymphocyte subsets were examined in renal cell carcinoma (TILs), adjacent non-tumor renal tissue and peripheral blood (PBLs) by flow cytometry and histochemistry in eighteen patients with renal cell carcinoma. CD5-positive cells were predominant in the TILs in 14 patients. In the renal cell carcinoma tissue, CD8-positive cells were predominant over CD4-positive cells, resulting in a less than unity ratio of CD4/CF8-positive cells. The lymphocyte number was significantly in adjacent normal renal tissue than in renal cell carcinoma. However, lymphocyte subsets ratios were not significantly different between these two tissues. PBLs showed the same proportions (CD4/CD8 mean 1.9 +/- 0.8) as the previously published healthy controlled data. The proportions of CD8-positive cells were significantly increased (p less than 0.05) and those of CD4-positive cells were also significantly decreased (p less than 0.01) in the TILs. The infiltrating pattern of TILs in 17 patients was divided histochemically into cluster (N = 7), single (N = 4), and mixed types (N = 6). The cluster and mixed types were significantly more common in grade 1 tumors and the single type was more common in the grade 2 tumors (p less than 0.05). The pT3 tumors showed the single type of TIL infiltration pattern, but showed no significant difference. In the cluster pattern of TILs, CD8-positive cells were surrounded by CD4-positive cells. Non-tumorous kidneys showed no infiltration of lymphocytes, except in 2 patients of pyelonephritis. These results suggest that cytotoxic T-cells stained as CD8 play an immunoreactive role against renal cell carcinoma.

CD4-CD8 Ratio↗

High-dose continuous venous infusion of interleukin-2: influence of dose and infusion rate on tumoricidal function and lymphocyte subsets.

Previous clinical studies have demonstrated a dose-response relationship between enhancement of certain immune parameters and interleukin-2 (IL-2) dose in trials with low dosages of the cytokine. This has not been demonstrated for high-dose (greater than 18 x 10(6) IU/m2 per day) IL-2. We completed phase II trials of sustained administration of indomethacin and ranitidine with IL-2 given as a continuous infusion over 5 days for three courses. Peripheral blood mononuclear cells, both fresh and cultured in vitro with IL-2 or IL-2 and indomethacin, were tested for tumoricidal function against K562 and Daudi targets; these results were then correlated with actual delivered dose and mean infusion rate per course. Similar correlations were calculated between delivered dose or infusion rate and absolute and proportional counts of lymphocyte subsets as determined by flow cytometry. No enhancement of in vitro tumoricidal function with either increasing delivered dose or increasing infusion rate was seen. No consistent pattern of correlation was found between the absolute counts of lymphocyte subsets after each course of IL-2 with delivered dose or infusion rate. The percent rise in absolute counts of selected T- and NK-cell subsets at the end of course 1 compared with baseline values correlated positively with infusion rate; however, a similar correlated between the infusion rate and an increase in lymphocyte tumoricidal function was lacking. Little evidence was found for improved tumoricidal function of mononuclear cells or consistent enhancement of lymphocyte subset counts in patients able to tolerate doses of IL-2 beyond 18 x 10(6) IU/m2 per day in a 5-day continuous infusion schedule.

Carcinoma, Renal Cell↗

Lymphocyte subsets in the endometrium of patients with endometriosis throughout the menstrual cycle.

PROBLEM: Comparison and characterisation of different lymphocyte subsets in the endometrium of endometriosis patients and in healthy women on every day of the menstrual cycle with special emphasis on the CD4:CD8 ratio in the endometrium. METHOD: Immunohistochemical staining of 253 endometrial biopsies of infertile women with and without endometriosis with Anti-Leu4 (CD3), Anti-Leu3a (CD4), Anti-Leu2a (CD8), Anti-Leu7 and Anti-Human-B-cell (CD22) using the immune peroxidase reaction. Identification and counting of positive lymphocyte were performed on cryostat sections. RESULTS: Endometrial lymphocyte subsets show equal quantity and distribution in endometriosis patients and in the control group. After a peak in the early proliferative phase the absolute number of T lymphocytes decreases while a predominance of T-suppressor/cytotoxic T lymphocytes (CD8) compared to T-helper/inducer lymphocytes (CD4) occurs towards the end of the menstrual cycle. CONCLUSION: Endometrium as the potential parent epithelia of endometriosis lesions seems not to be altered in its lymphatic cell content compared to healthy women. Furthermore, endometrium is clearly characterised as part of the mucosa associated lymphatic tissue (MALT). T lymphocytes show specific quantitative changes due to different phases of the menstrual cycle.

Adult↗

Double fluorescence technique for measurement of complement-fixing antibody to lymphocyte subsets.

A double fluorescent antibody method for quantitating human complement-fixing antibody to lymphocyte subclasses has been developed. The indicators in this system are a C6-deficient serum as a non-lytic source of complement, rhodamine-labeled anti-C3 and fluorescein-labeled murine monoclonal antibodies to human lymphocyte subsets. The basic procedure is to incubate lymphocytes with the unknown serum and then to add C6-deficient serum. The binding of C3 is indicated by staining with rhodamine-labeled anti-C3 and the subset class of the lymphocyte so stained is determined by binding of fluorescein-tagged anti-OKT4 or -OKT8 antibodies. The occurrence of both red and green cell surface fluorescence denotes the presence of a complement-binding antibody to the lymphocyte subset defined by the monoclonal antibody. In addition to defining the specificity of complement-fixing anti-lymphocyte antibodies, this technique is more sensitive than the microcytotoxicity assay.

Acquired Immunodeficiency Syndrome↗

Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry.

Determination of the number of viable cells or quantification of lymphocyte subsets in heterogeneous cell populations is critically important for cytotoxicity assays, apoptosis assays, or the analysis of differential activation of T-cell subsets by distinct stimuli. In this report, we describe a rapid flow cytometry method termed Standard Cell Dilution Analysis (SCDA) specifically to quantify any subset of phenotypically definable, viable cells in heterogeneous populations using a FACScan flow cytometer. This method combines: (1) specific detection of lymphocyte subsets by phycoerythrin-conjugated monoclonal antibodies, (2) electronic exclusion of dead cells or cell debris by propidium-iodide staining and gating on forward vs. sidescatter, respectively, and (3) admixture of a known amount of fixed, fluorescein isothiocyanate stained cells immediately before analysis as a constant parameter to allow for calculation of cell quantity. We have used SCDA to analyze the in vitro growth characteristics of various human T-lymphocyte subpopulations in response to different activation stimuli.

Antibodies, Monoclonal↗