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PubMed2005

Lymphatic organ development in dogs: major lymphocyte subsets and activity.

In the present study, we have characterized lymphocyte subsets and activity in peripheral blood, spleen, mesenteric and popliteal lymph nodes in pups from birth till the age of one month and compared the results with the situation in the group of three adult dogs. In neonatal pups, lower numbers of CD3(+) T-cells were detected in both the spleen and peripheral blood than in lymph nodes. In contrast to the other compartments, CD21(+) B-cells prevailed in the spleen, which resulted in low values (<1) of the CD3(+)/CD21(+) ratio. Low numbers of CD8(+) lymphocytes were characteristic in all compartments immediately after birth; consequently a high CD4(+)/CD8(+) ratio has been calculated. Postnatal development was characterized by an increasing frequency of CD8(+) lymphocytes in all organs studied. Another typical feature of the early period of life was a relative decrease of B-cell numbers, which was compensated by an increasing proportion of T-lymphocytes, particularly in the peripheral blood and spleen. DNA synthesis in newborn pups' cells as measured by in vitro thymidine incorporation was surprisingly high in non-stimulated control samples, notably in the spleen. Further development of lymphocyte activity was characterized by the decline in spontaneous activity in all organs. Stimulation indices upon mitogen-induced proliferation increased proportionally to the decrease in spontaneous activity. Based on our experimental data, we have concluded that pups are born with a relatively competent immune system the structure of which, however, markedly develops during a few postnatal weeks.

Age Factors↗
PubMed1991

Characterisation of the in vitro responsiveness of lymphocyte subsets to locomotor stimuli by immunocytochemical methods.

The in vitro locomotion of lymphocyte subsets has previously been determined by use of highly purified cell populations. A method is now described in which mixed peripheral blood lymphocytes (PBL) migrate to the undersurface of polycarbonate filters in a 48 well microassay, the responding cells being characterised by alkaline phosphatase-anti-alkaline phosphatase immunocytochemistry. Recombinant interleukin (rIL)-1 alpha, zymosan activated plasma (ZAP) and rIL-8 were shown to induce concentration-related migration of mixed PBL in the 48 well assay and were therefore used as reference agonists. Total T cells, B cells, T helper/inducer and T suppressor/cytotoxic cells, as well as lymphocytes stained with the monoclonal antibodies UCHL1 and SN130, have now been quantified after migration to the undersurface of 8 microns pore size polycarbonate filters, in response to optimal concentrations of rIL-1 alpha, ZAP and rIL-8. The value of the analytical method was demonstrated by the selective responses seen. rIL-1 alpha selectively stimulated the migration of T helper/inducer and a small number of B cells without affecting T suppressor/cytotoxic cell locomotion. ZAP and rIL-8 significantly stimulated the migration of T helper/inducer, T suppressor/cytotoxic and B cells. ZAP and rIL-1 alpha also stimulated the migration of both UCHL1 and SN130 positively stained cells. This method may therefore be used to investigate the selective actions of lymphocyte locomotor stimuli on PBL sub-populations without the need to purify specific cell subsets, and to study the specificity of certain inhibitors or drugs on lymphocyte responses.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1995

[Immunological study of the postoperative adjuvant treatment with interferon gamma in patients with renal cell cancer--measurement of peripheral blood lymphocyte subsets with three color flow cytometry].

The antitumoral effects of interferon (IFN) against renal cell carcinomas (RCC) are considered to consist of a direct tumoricidal activity and an indirect action related to the host immune system. Monitoring to T lymphocytes is an immunological parameter to evaluate the antitumoral effects of IFN. To examine in detail the change of CD8 positive cells reported previously, we analyzed the change of the peripheral blood T lymphocyte subsets using three color flowcytometry during the period of IFN-gamma administration. We administered recombinant interferon gamma (rIFN-gamma) to the patients with renal cell cancer. Eighteen patients received a prophylactic injection of rIFN-gamma (300 x 10(4) units per week) after nephrectomy. Among them three patients developed distant metastases and one had local recurrence after the start of rIF-gamma therapy. We evaluated the immunological change by measurement of the peripheral blood lymphocyte subsets consisting of activated cytotoxic T lymphocytes (ACTL), activated suppressor T lymphocytes (ASTL), activated suppressor-inducer T lymphocytes (AITL), cytotoxic T lymphocytes (CTL), helper T lymphocytes (HTL), and suppressor-inducer T lymphocytes (SITL), from the start of rIFN-gamma injection. We also estimated the natural killer (NK) activity by a cytolytic test at the same points. NK activity consecutively increased throughout the period of administration of rIFN-gamma. We found significant enhancement of ACTL at the sixth month (P < 0.05), ASTL at the third month (P < 0.01) and NK activity at the second week, the first month, the third month and the sixth month (P < 0.01) after the start of rIFN-gamma injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1984

Lysis of tumor biopsy cells by blood lymphocyte subsets of various densities. Autologous and allogeneic studies.

Blood lymphocyte subsets of various densities from 14 healthy donors and 25 patients with solid tumors were used in tests for cytotoxicity against allogeneic or autologous tumor cells. The allogeneic combinations comprised 36 tests on 28 tumors. In these the unmanipulated total lymphocyte population was cytotoxic in one experiment. In tests with density-separated subsets, 16 experiments were positive. The lytic effect resided in the light subsets. Auto-tumor lysis was detected in 7 of 25 cases, when the total lymphocyte population was used. Eleven additional tests showed cytotoxicity with the separated subsets. The autologous tumor cells were damaged by the light lymphocytes of 15 patients. Of these patients, 6 had cytotoxic cells also in the high-density subset. In seven cases auto-tumor lysis was exerted only by the dense lymphocytes. Thus, in 13 cases the profile of auto-tumor lysis differed from that observed with the allogeneic combinations and against K562. Pretreatment of the effectors with Hu-IFN-alpha enhanced or induced cytotoxicity in the light lymphocytes, against both allogeneic and autologous tumors, while the dense subset was rarely potentiated. The autologous system may represent an immune situation and therefore it is not unexpected that the active cells should have different phenotypic characteristics from those of the operationally natural killer system.

Biopsy↗
PubMed1991

Determination of T-lymphocyte subsets in bronchoalveolar lavage of patients with pulmonary sarcoidosis.

Bronchoalveolar T-lymphocyte subsets were evaluated in seven patients with pulmonary sarcoidosis and eight patients with other interstitial lung disorders. Both disease groups considerably differed in the distribution of bronchoalveolar CD4+ and CD8+ cells. The lymphocytic alveolitis in sarcoidosis was characterized by excess CD4+ (helper-inducer) cells; mean CD4/CD8 ratio was 6.8 +/- 3.8. Inversely, in the lungs of patients with other interstitial pulmonary disorders a less pronounced but clear predominance of CD8+ (suppressor-cytotoxic) lymphocytes was observed with CD4/CD8 ratio 0.7 +/- 0.4. The results of the study suggest that the determination of the major immunoregulatory subsets of bronchoalveolar lymphocytes contributes to the reliable differentiation of sarcoidosis from other interstitial lung disorders. The recent advances improving our understanding of the pathogenesis of sarcoidosis are briefly reviewed in the discussion.

Adult↗
PubMed2005

Lymphocyte subsets' reference ranges in an age- and gender-balanced population of 100 healthy adults--a monocentric German study.

Region-specific reference ranges for adult peripheral blood lymphocyte subsets have been established in few countries around the world, but most studies were restricted to younger adults for monitoring of HIV patients. The aim of this investigation was to establish age- and gender-specific reference ranges for healthy adults. Lymphocyte subsets were examined in 100 healthy volunteers (50 males, 50 females) aged 19-85 years by two-color flow cytometric analysis with a FACSCalibur. A statistically significant decline in the mean numbers of CD3+/CD8+ T cells and CD19+ B cells was observed beyond an age of 50 years, whereas the mean counts of NK cells and CD4+/CD8+ ratio significantly increased beyond the age of 50. Females < or = 50 years had significantly higher mean CD4+ T cell counts and lower NK cell counts than males < or = 50 years. Based on these results, we established reference values for three subgroups: males < or = 50 years, females < or = 50 years, and males/females > 50 years.

Adult↗
PubMed1986

Tumor histology and the immunoregulatory T lymphocyte subsets in lung cancer patients.

The immunoregulatory T lymphocyte subsets (CD4+, CD8+) in 39 patients with primary lung cancer were analyzed as for the possible correlation with tumor histology. Compared to the healthy controls (n = 36), cancer lymphocytes were generally characterized by a higher proportion of CD8+ subset (p less than 0.01) with no difference in either CD4+ subset or CD3+ cells. There was, however, a significant difference (p less than 0.05) between one extreme of epidermoid carcinoma with the highest CD8+ and the lowest ratio (CD4+/8+) and the other end of adenocarcinoma with the lowest CD8+ and the highest ratio. Large cell and small cell carcinomas were of the intermediate values. A spectrum of the subset marker profiles was thus found in associated with tumor histology. A diversity in the host immune status was suggested in primary lung cancer, although clinical implication remains to be clarified.

Adenocarcinoma↗
PubMedDate not supplied

In situ localization of lymphocyte subsets in synovial membranes of patients with rheumatoid arthritis with monoclonal antibodies.

Immunological analysis of the mononuclear infiltrates in 12 rheumatoid synovial membranes was performed by means of modified peroxidase antiperoxidase technique using a panel of monoclonal antibodies, directed against T cell differentiation antigens and HLA-DR (Ia-like) antigens. Helper/inducer T lymphocytes (OKT4 +, Leu3a+) were found in large numbers in nodular lymphoplasmocellular infiltrates, whereas the number of cytotoxic/suppressor T lymphocytes (OKT8 +m Leu2a+) was very low, resulting in a high T4/T8 or Leu3a/Leu2a ratio (6á14:1). In diffusely localized lymphoplasmocellular infiltrates this ratio was only slightly increased as compared with the peripheral blood of patients with rheumatoid arthritis (2á4:1). Moreover, most of these T lymphocytes appeared to have Ia-like antigens and seemed to have contact with HLA-DR+, sometimes weakly OKT6+ dendritic nonlymphoid cells. The results showed a constant basic localization pattern of T lymphocyte subsets and suggest that interactions between dendritic nonlymphoid cells, T lymphocyte subsets and B lymphocytes determine the ultimate architecture of the inflammatory infiltrates in the rheumatoid synovial membrane.

Antibodies, Monoclonal↗
PubMed1987

Antibodies anti-HTLV III and lymphocyte subsets of high risk subjects.

208 assay for the research into anti-HTLV III antibodies and lymphocytes subsets were carried out on the same number of patients at risk. 11 homosexual man, 143 intravenous drug users (i.d.u.) and 3 children of drug addicts from hospitals in the Marche and Abruzzo and 51 haemophiliacs from hospital in Florence were examined. 3 determination of anti-HTLV III antibodies were taken from each subject using 3 different commercial Kits. The results concur with and confirm similar epidemiological studies that have been done. The haemophiliac group had the highest positive percentage (39.2%), then came i.d.u. (11.9%) and the homosexuals (10.0%). Furthermore, of the 38 positive totals, there were 22 with only one kit, 18 with two, and 15 with all three. The evaluation of the lymphocyte subsets did not strictly correlate with the presence of the antiretrovirus antibodies.

Acquired Immunodeficiency Syndrome↗
PubMed1989

[Natural killer (NK) activity of peripheral blood and regional lymph nodes in patients with genitourinary malignant tumors. 2. NK activity and T-lymphocyte subsets in peripheral blood and regional lymph nodes].

We investigated natural killer (NK) activity and T-lymphocyte subsets in peripheral blood and regional lymph nodes in order to examine the host's defence mechanisms against cancer. The materials were obtained from 26 patients with genitourinary cancer and 9 patients with benign diseases, who were used as controls. NK activity was measured by ATP-chemiluminescence (ATP) assay, a new method which is well correlated to the 51Cr-release assay, and T-lymphocyte subsets were stained with anti-Leu-2, -3, -4, -7, and -11 monoclonal antibody. The NK activity of regional lymph nodes was lower than that of the peripheral blood in all 35 cases, and the percentage of Leu 7+ and Leu 11+ cells was in agreement with that of the NK activity detected by ATP assay. In the peripheral blood, the percentage of Leu 3+ cells was lower in high stage patients than it was in low stage patients and controls, but in the regional lymph nodes it was paradoxically higher. The NK activity of regional lymph nodes against autologous tumor cells was low in 8 patients with bladder cancer. It appears from this study that the regional lymph nodes have no function as immunological barriers for metastasis and we therefore conclude that they should be dissected during surgery.

Adult↗
PubMed2002

Longterm decrease in the CD57 lymphocyte subset in a patient with chronic Lyme disease.

Lyme disease is a tickborne illness caused by the spirochete Borrelia burgdorferi. In a previous report we described a decrease in the CD57 lymphocyte subset in patients with chronic Lyme disease. We have now identified a patient with chronic relapsing and remitting symptoms of Lyme disease who had decreased levels of CD57 lymphocytes over 10 years. This observation represents the longest duration of an immunologic abnormality ever documented in chronic Lyme disease. The CD57 lymphocyte subset appears to be a useful marker of longterm infection with the Lyme disease spirochete.

Adult↗
PubMed1994

New medical technologies and clinical practice: a survey of lymphocyte subset monitoring.

BACKGROUND: Since its inception in the early 1980s, lymphocyte subset (LS) monitoring of transplant patients has been a controversial technique. The clinical literature is replete with contradictory claims concerning its usefulness. No systematic information is however available on clinicians' attitudes towards the new technology. METHOD: We carried out a mail survey of the members of The Transplantation Society concerning the availability, use, value, and critical assessment of LS monitoring. RESULTS: Results show that LS monitoring technology is available in most clinical settings surveyed and is regularly used by about half of the respondents associated with a clinical transplant program. About half of the users obtain diagnostically relevant measurements (T4/T8 ratios), as opposed to measurements related to OKT3 anti-rejection therapy. While claiming that LS measurement is a useful tool, respondents attribute a low average score to the diagnostic value of the technique; about 2/3 believe that its absence would not affect their clinical judgment. Finally, 25% of the respondents send blood for LS measurements although they do not believe the technique is useful. CONCLUSIONS: The results suggest that research instruments which generate clinical interest may develop clinical-diagnostic routines despite a lack of consensus concerning their usefulness. More importantly, these routines avoid a critical assessment of key notions such as "immune monitoring" which tend to blur the distinction between research and therapy.

Attitude of Health Personnel↗
PubMed1998

[Circadian variation of lymphocyte subsets in health subjects].

In our study cortisol and interleukin 2 (IL-2) levels were measured and lymphocyte sub-population analyses were performed on blood samples collected every four hours, for 24 hours from 10 healthy subjects aged 38-65 years. A clear circadian rhythm was validated for cortisol serum levels, for CD8, CD8 dim, CD16 and delta TcS1 with acrophase in the morning, and for CD2, CD4, CD/CD8 ratio, HLA-DR, CD20 and CD25 with acrophase at night. CD8 bright and TcR delta 1 presented higher levels in the morning without validation of the circadian rhythm. Changes of serum levels of IL-2 did not show circadian rhythmicity. The results show that specific lymphocyte subsets present different profiles of nyctohemeral changes and this may explain time related variations of immune responses.

Adult↗
PubMed1991

Lymphocyte subset distribution and natural killer cell activity in men with idiopathic hypogonadotropic hypogonadism.

Sex-related differences in immune responsiveness are mediated at least in part by sex steroid hormones. Lymphocyte subset distribution in peripheral blood and natural killer cell function both have been reported to be under hormonal control. In order to gain more insight into sex steroid hormone action on the immune system, we have measured the lymphocyte subset distribution and natural killer cell activity in 18 men with idiopathic hypogonadotropic hypogonadism before treatment, and after hormonal treatment had normalized plasma testosterone levels. In untreated patients, the mean plasma testosterone concentrations were significantly lower than those in the treated men (3.0 +/- 0.5 nmol/l vs 16 +/- 1.7 nmol/l, p less than 0.001). The percentage of peripheral CD3+ lymphocytes, CD8+ cells, the CD4+/CD8+ ratio, and the natural killer cell activity of peripheral mononuclear cells measured in a 51Cr release assay against target K 562 cells did not differ between patients with idiopathic hypogonadotropic hypogonadism and healthy adults, and most importantly, did not change during hormonal treatment which normalized plasma testosterone levels in the patients. In contrast, the percentage of peripheral CD4+ cells was significantly higher in untreated patients compared with normal adult subjects or patients with idiopathic hypogonadotropic hypogonadism after hormonal treatment that resulted in normal plasma testosterone levels (53 +/- 2 vs 47 +/ 2, p less than 0.05). It should be noted that the percentage of peripheral CD16+ cells was significantly lower in untreated men with low plasma testosterone levels than in normal controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2002

Evaluation of peripheral blood CD4 and CD8 lymphocyte subsets, CD69 expression and histologic rejection grade as diagnostic markers for the presence of cardiac allograft rejection.

We investigated the dynamics of the CD4+ and CD8+ lymphocyte subsets, and the expression of activation markers in cardiac transplant recipients. We tested 132 peripheral blood samples from 62 cardiac transplant recipients using fluorescent staining and flow cytometry analysis. The results were correlated with histological rejection grade of concurrently taken biopsies, and 5-year survival of the recipients. A decrease in the total T lymphocyte subset, and in CD4+ lymphocytes was associated with higher rejection grade and lesser survival. An increase (5-11%) of double positive CD4+ CD8+ lymphocytes was observed; these were mostly CD4brightCD8dim. The CD4/CD8 ratio was significantly (P < 0.00) lower in the transplant recipients than in normal individuals. CD69 expression was higher than CD54 and CD154 expression on CD4 and CD8 lymphocytes of cardiac transplant recipients; correlation between these activation markers was excellent (P < 0.001). Fluorescent staining for CD69 was often of low intensity. Multiple regression for % CD8+ CD69+ cells and survival, and for % CD69+ T cells and rejection grade yielded a significant correlation (P < 0.050). Both % CD8+ CD69+ and % CD69+ T cells were significantly higher in samples with severe and moderate rejection grade (grades 3A, 3B and 4) than in samples which showed no, minimal or mild rejection (grades < or = 2); P-values were 0.052 and 0.003, respectively. Preliminary results indicated that false negative results could be contributed to increased immunosuppression. We conclude that CD69 expression on circulating CD4 and CD8 lymphocytes is a useful parameter for the diagnosis of moderate and severe rejection.

Antigens, CD↗
PubMed2004

Blood lymphocyte subsets in canine idiopathic pericardial effusion.

The immunophenotype of peripheral blood lymphocytes was investigated in 23 dogs diagnosed with idiopathic pericardial effusion in order to provide information about a possible role of the immune system in this pathology. Flow cytometric analysis showed a significant reduction in nearly all lymphocyte subsets examined and a strong, significant (P < 0.001) reduction of the CD4 subset, which gave rise to a significantly lower CD4/CD8 ratio. Our data suggest that an imbalance in the immune system is present during the course of the disease, preferentially affecting the T helper cell response.

Animals↗
PubMed1997

T lymphocytes subsets in experimental iron overload.

Several abnormalities of the immune system have been reported in association with clinical and experimental iron overload. To dissect further such abnormalities, changes in lymphocyte subsets were evaluated in iron-loaded male Sprague-Dawley rats. The iron-loading protocol consisted of a total dose of irondextran (1.5 mg/Kg body weight) divided in daily intramuscular injections over twenty consecutive days. At days 0, 20, and 50 after initiation of iron injections lymphocyte subsets in blood, spleen and mesenteric lymph nodes were estimated by indirect immunofluorescence using monoclonal antibodies recognizing T cells (W3.13), the subset of helper T cells in (W3.25), and the subset of cytotoxic T cells (OX.8). By day 20, there was no change in the number of W3.25+ T cells in the blood of iron-loaded animals as compared to the controls, but the OX.8+ T cells were significantly elevated. At this time, the ratio W3.25 +/OX.8+ cells was significantly decreased (0.5 in experimental rats vs 2.0 in controls). Similar results were obtained at day 50. In the spleen, there was a decrease in the proportion of W3.25 +T cells and an increase in OX.8+ T cells at day 20. However, these values returned to normal by day 50. A negative correlation between W3.25 +/OX.8+ ratio and serum ferritin was observed in blood and spleen during iron administration. These changes were associated with abnormalities in lymphocyte proliferative response. No changes in W3.25 +/OX.8+ ratio were observed in mesenteric lymph nodes. These results demonstrate that iron overload alters the distribution of T lymphocytes in various compartments of the immune system.

Animals↗
PubMedDate not supplied

Lymphocyte subsets associated with major depression and dysthymia: modification by antidepressant treatment.

Major depression and dysthymia (chronic, low grade depression) were associated with an increase in the number of CD16/56 (natural killer; NK) cells in blood, whereas other lymphocyte subsets (CD3, CD4, CD8, CD19, and the CD4/CD8 ratio) did not differ from control subjects. After treatment with a specific serotonin reuptake inhibitor, the symptoms of depression were alleviated in both the major depressive and dysthymic patients. Likewise, NK cell numbers declined to control values in these treated groups. Among the major depressive patients, the NK cell number reached control values within 4 weeks, whereas 6 months of treatment was required for such an effect to be achieved in the dysthymic patients. Although plasma levels of epinephrine, norepinephrine, cortisol, and ACTH were not different between groups, among the major depressive patients ACTH was inversely correlated with total lymphocytes, CD3, and CD19, and epinephrine was directly related to the CD4 and CD4/CD8 ratio. Among dysthymics, ACTH was unrelated to any of the lymphocyte subsets, but norepinephrine was directly related to total lymphocytes, CD3, CD4, and NK cells. The data are interpreted in terms of stress perception among major depressive and dysthymic patients and the potential impact of stressor experiences on immune processes.

1-Naphthylamine↗