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PubMed2006

T-lymphocyte subset distribution in human spleen.

BACKGROUND: When analyzing human cellular immune responses, most focus is placed on the peripheral blood (PB) and, to a lesser extent, the lymph nodes. To date the spleen has not been analyzed with regard to its role in adaptive cellular immunity and more notably not with respect to T-cell immune responses. MATERIALS AND METHODS: We analyzed the splenic lymphocyte compartment in comparison with the PB lymphocyte compartment regarding the number of NK cells, B cells, CD4(+), CD8(+) T cells and CMV-specific CD8(+) T cells. Furthermore, we analyzed the distribution of naive, memory and effector subsets of CD4(+) and CD8(+) T cells in these compartments. RESULTS: The spleen contains proportionally more B cells and less CD4(+) and CD8(+) T cells than PB. The percentage of CD8(+) T cells is greater in the spleen, leading to an inverse CD4/CD8 ratio. Both splenic CD4(+) and CD8(+) T-cell populations show a greater number of activated cells, and splenic CD8(+) T cells show a more differentiated cytotoxic CD27(-)CD45RA(+) memory phenotype. CONCLUSIONS: Our findings show that the distribution of the different lymphocyte subsets is markedly different between the spleen and the PB, thus inferring an important and distinct role for the spleen in CD4(+) and CD8(+) T-cell activation.

Adult↗
PubMed1992

Changes in lymphocyte subsets and mitogen responsiveness following open-heart surgery and possible therapeutic approaches.

Septic multi-organ failure represents a common cause for operative mortality following open-heart surgery. One major reason might be the depression of cell-mediated immunity. The purpose of this prospective randomized trial was to quantify and specify the effects of open-heart surgery on cell-mediated immune mechanisms. In addition, the immunorestorative potential of a combined immunomodulatory therapy with the cyclooxygenase inhibitor indomethacin and the thymomimetic substance Thymopentin versus single drug administration of indomethacin was investigated. Twenty patients were given indomethacin for the first 5 postoperative days (group A). Another 20 patients also received Thymopentin perioperatively and on the second and fourth postoperative days (group B), while 20 patients underwent conventional therapy (group C). Cell-mediated immune response was quantified in vitro by measuring CD3+ T-lymphocytes and their subsets, CD4+ T-helper and CD8+ T-suppressor cells. Lymphocyte responsiveness to a specific (Antigen-Cocktail) and non-specific mitogen (phytohemagglutinin) provided information about the quality of cell-mediated immune response. On the first postoperative day CD3+ T-lymphocyte counts and Antigen-Cocktail-induced lymphocyte proliferation decreased significantly in all groups. The number of CD4+ T-Helper cells fell significantly only in groups A and C, while the decrease in group B was not statistically significant; the same applied to phytohemagglutinin-induced lymphocyte response. The CD4+/CD8+ ratio was significantly depressed only in group C, decreased slightly in group A and did not change as compared to baseline values in group B. All investigated parameters remained significantly depressed until the seventh postoperative day in group C.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1995

Increased chemiluminescence of whole blood and normal T-lymphocyte subsets in severe nodular acne and acne fulminans.

To investigate the inflammatory and immunological aspects of severe acne, we examined the luminol-enhanced chemiluminescence of whole blood, T-cell subsets and natural killer cell functions in 11 patients with severe nodular acne and 4 patients with acne fulminans. In patients with severe nodular acne, the active phase of the disease, compared to the values in remission (means 47 mV, SD 24.8 and 32 mV, SD 8.3, p < 0.05). The patients with acne fulminans also showed high values in the active phase of the disease (mean mV 68.3, SD3.5) compared to remission (mean 30.5 mV, SD 15.3). No marked alterations were seen in the percentages of T-helper cells, T-suppressor cells or DR-positive lymphocytes or in the levels of soluble interleukin 2 receptor. The percentages and activities of natural killer cells did not show any significant changes either. Five patients (4 with severe nodular acne and one with acne fulminans, accounting for 33% of all patients) carried HLA Cw6 antigen, which is a significantly increased frequency compared to health controls (pc = 0.015). The present chemiluminescence results suggest that peripheral blood neutrophils are activated in patients with severe acne.

Acne Vulgaris↗
PubMed2002

Reference values of lymphocyte subsets in the normal healthy adult Kuwaiti Arab population.

Fluorescent monoclonal antibody labelling followed by a lysed whole blood method and flow cytometry was used to determine the lymphocyte subpopulations in 127 (64 males and 63 females) normal healthy individuals in the adult (age 18-59 years) Kuwaiti population. Relative percentages and absolute values of CD2+, CD3+, CD19+, CD4+, CD8+, HLADR+, CD56+, CD45RO+, and CD45RA+ cells were determined. The reference ranges were CD2+, 73-92% (0.95-2.99 x 10(9) per l), CD3+, 64-85% (0.83-2.71 x 10(9) per l), CD19+, 6-22% (0.05-0.61 x 10(9) per l), CD4+, 34-54% (0.45-1.65 x 10(9) per l), CD8+, 20-42% (0.29-1.17 x 10(9) per l), HLADR+, 4-23% (0.02-0.62 x 10(9) per l), CD56+, 4-22% (0.06-0.58 x 10(9) per l), CD45RO+, 16-53% (0.26-1.42 x 10(9) per l) and CD45RA+, 35-72% (0.34-2.05 x 10(9) per l). The mean CD4/CD8 ratio was 1.50+/-0.35. CD3+ cells were positively correlated to both CD4+ and CD8+ cells (P<0.001), and CD4+ cells showed a significant positive correlation with CD8+ cells (P<0.001).

Adolescent↗
PubMed2003

Expression of CD30 and Ox40 on T lymphocyte subsets is controlled by distinct regulatory mechanisms.

Members of the TNF receptor (TNFR) superfamily are cell-surface proteins that can be found on most cell types including lymphocytes. Although some TNFR-related molecules are constitutively expressed, others, such as CD30 and Ox40, are induced upon activation of lymphocytes. CD30 and Ox40 are predominantly expressed on activated T helper (T(h))2 cells. Both receptors can activate c-Jun N-terminal kinase (JNK) and nuclear factor-kappaB (NF-kappaB) and have been suggested to play costimulatory roles in lymphocyte activation. To gain further insight into events triggered by both TNFR-related molecules, a detailed analysis of their expression patterns has been performed. We found that CD30 and Ox40 were coexpressed on T(h)2 cells. However, in contrast to CD30, Ox40 was also expressed on T(h)1 cells. Although expression of both receptors is augmented by interleukin-4, only CD30 expression is dependent on signal transducer and activator of transcription (STAT)-6-mediated signaling. Differences in the regulatory pathways controlling expression of CD30 and Ox40 suggest distinct, functional effects triggered by the two TNFR-related molecules during lymphocyte activation.

Animals↗
PubMed2003

Lymphocyte subsets and specific IgG antibody levels in clindamycin-treated and untreated dogs experimentally infected with Babesia gibsoni.

This study was carried out to clarify the role of lymphocyte subpopulations and Babesia-specific antibody on the treatment of clindamycin in dogs infected with B. gibsoni. Ten beagle dogs were divided into two groups: an untreated group (5 dogs) and a clindamycin-treated group (5 dogs), which was administered clindamycin at 25 mg/ kg body weight, per os, q 12 hr from 7 days to 21 days post-infection (PI). On the acute stage of infection, clindamycin treatment resolved anaemia and other clinical findings. There were no significant differences between treated and untreated dogs either in parasitemia levels or Babesial IgG antibody levels. However, morphological changes that indicated degeneration in the majority of parasites were observed. The numbers of CD4(+) showed a significant increase in treated dogs, especially after treatment. On the chronic stage, CD4(+) cells maintained high level both of the treated and untreated dogs. Although parasitemia maintained low level, their relapses were occurred on the 49th day PI in treated dogs and on the 42nd and 63rd PI in untreated dogs. A rapid humoral antibody response was observed in treated dogs, however, lower humoral antibody responses in untreated dogs after relapses. The antibody levels of treated dogs were significantly higher than those of untreated dogs. These results suggested that clindamycin might not eliminate rapidly parasites from peripheral blood, but damage parasites, which might stimulate efficiently humoral and cellular immunity against Babesia infection, and result in an improvement of clinical conditions.

Animals↗
PubMed1991

Peripheral blood lymphocyte subsets in urinary bladder carcinoma patients.

The percentages of pan T (CD3+), T helper (CD4+), T cytotoxic/suppressor (CD8+), B (CD22+) and natural killer (CD57+) cells in peripheral blood lymphocytes of 15 urinary bladder carcinoma patients and in parallel, 10 healthy donors were estimated, using monoclonal antibodies in indirect membrane immunofluorescence. A significant decrease in the percentage of CD3+ lymphocytes and a highly significant decrease in the proportion of CD8+ cells was revealed in urinary bladder cancer patients. This change was accompanied by a significant increase in the CD4/CD8 ratio and in the frequency of CD57+ (HNK-1+) cells. Our data document, for the first time, the complete lymphocyte profile of patients with advanced (T3) urinary bladder carcinoma. The reason and significance of the decline in CD8+ lymphocyte percentage and the increase of CD57+ cells are discussed.

Adult↗
PubMed1994

The effect of dietary lipid manipulation on rat lymphocyte subsets and proliferation.

Polyunsaturated fatty acids (PUFA) have been shown to suppress immune cell functions in vitro. Dietary studies investigating the effects of PUFA-containing oils on lymphocyte functions have yielded contradictory findings: such studies are difficult to compare as there are many variations in protocols. The present study investigated the effects of diets containing oils rich in saturated fatty acids, monounsaturated fatty acids, n-6 PUFA or n-3 PUFA on rat lymphocyte proliferation and on receptor and surface marker expression. Rats were fed for 10 weeks on a low-fat (LF) diet (approximately 2% fat by weight) or on one of five high-fat diets, which contained 20% (by weight) hydrogenated coconut oil (HCO), olive oil (OO), safflower oil (SO), evening primrose oil (EPO) or menhaden (fish) oil (MO). Compared with feeding the LF diet, all of the high-fat diets suppressed the proliferation of lymphocytes from the spleen: although there was no significant effect of diet on the proliferation of lymphocytes from the thymus, there was a trend towards decreased proliferation with high-fat feeding. Feeding the OO, EPO or MO diets significantly suppressed proliferation of mesenteric lymph node lymphocytes compared with feeding the LF, HCO or SO diets. Dietary lipid manipulation had no effect on the proportion of T cells, B cells or monocytes/macrophages in the spleen, thymus or lymph nodes. Dietary lipid manipulation also had no significant effect on the proportions of CD4+ or CD8+ lymphocytes in spleen, thymus or lymph nodes, either in freshly prepared cells or in cells cultured in the presence of mitogen. There were no significant effects of dietary lipid manipulation on the expression of IL-2 receptors or transferrin receptors by concanavalin A (Con A)-stimulated lymphocytes. However, there was a trend towards a decrease in transferrin receptor expression by Con A-stimulated lymphocytes from the thymus and lymph nodes of the MO-fed rats and towards a decrease in the expression of IL-2 receptors by lymphocytes from the spleens and thymi of the MO-fed rats. These observations provide evidence that some dietary oils, particularly OO, EPO and MO, possess immunosuppressive properties and so may be useful in the therapy of diseases involving inappropriate lymphocyte activation.

Animals↗
PubMed1991

Flow cytometric analysis of lymphocytic subsets in haemorrhagic fever with renal syndrome virus infection.

Percentage counts of T-lymphocytes (helper and suppressor subpopulation) and B-lymphocytes were performed in the peripheral blood of two confirmed cases of HFRS infection using Coulter Clone monoclonal antibodies and Becton Dickinson flow cytometry. The individual values were compared with those from healthy subjects. HFRS patients had lower total T-cell (CD2) and T-helper (CD4 subset) counts. This seems linked with the role of the CD4 determinant as viral receptor. The measuring of specific lymphocyte helper and suppressor subpopulations might be useful in predicting the course of the disease and in following the effects of therapy.

B-Lymphocytes↗
PubMed2001

Distribution of lymphocyte subsets in healthy human immunodeficiency virus-negative adult Ethiopians from two geographic locales.

Immunological values for 562 factory workers from Wonji, Ethiopia, a sugar estate 114 km southeast of the capital city, Addis Ababa, Ethiopia, were compared to values for 218 subjects from Akaki, Ethiopia, a suburb of Addis Ababa, for whom partial data were previously published. The following markers were measured: lymphocytes, T cells, B cells, NK cells, CD4(+) T cells, and CD8(+) T cells. A more in depth comparison was also made between Akaki and Wonji subjects. For this purpose, various differentiation and activation marker (CD45RA, CD27, HLA-DR, and CD38) expressions on CD4(+) and CD8(+) T cells were studied in 60 male, human immunodeficiency virus-negative subjects (30 from each site). Data were also compared with Dutch blood donor control values. The results confirmed that Ethiopians have significantly decreased CD4(+) T-cell counts and highly activated immune status, independent of the geographic locale studied. They also showed that male subjects from Akaki have significantly higher CD8(+) T-cell counts, resulting in a proportional increase in each of the CD8(+) T-cell compartments studied: naïve (CD45RA(+)CD27(+)), memory (CD45RA(-)CD27(+)), cytotoxic effector (CD45RA(+)CD27(-)), memory/effector (CD45RA(-)CD27(-)), activated (HLA-DR(+)CD38(+)), and resting (HLA-DR(-)CD38(-)). No expansion of a specific functional subset was observed. Endemic infection or higher immune activation is thus not a likely cause of the higher CD8 counts in the Akaki subjects. The data confirm and extend earlier observations and suggest that, although most lymphocyte subsets are comparable between the two geographical locales, there are also differences. Thus, care should be taken in extrapolating immunological reference values from one population group to another.

Adult↗
PubMed1995

T lymphocytes from normal human peritoneum are phenotypically different from their counterparts in peripheral blood and CD3- lymphocyte subsets contain mRNA for the recombination activating gene RAG-1.

The surface antigens of peritoneal lymphocytes of healthy human individuals were studied. B lineage cells comprised 2.3% of the total peritoneal lymphocyte population. Although the majority of peritoneal cavity lymphocyte (PCL) T cells expressed alpha beta T cell receptor (TcR), up to 17% expressed gamma delta TcR. The majority of PCL exhibited markers of the thymus-dependent lineage (CD2+ CD3+ TcR alpha beta + CD4+ or CD8 alpha + beta +) and surface antigens associated with memory and activation (CD45RO+ CD11a+ CD18+ CD49d+ HLA-DR). Up to 92% of both CD4+ and CD8+ T cells bore CDw60, thus characterizing the T cell subset containing helper activity for mitogen-driven B cell differentiation. The majority of PCL T cells were CD8+ and, in addition, up to 60% of this population expressed the homodimeric CD8 alpha + beta -. Messenger RNA for the recombination activating gene RAG-1 was examined in CD3- PCL depleted of CD19+ lineage cells. The PCL population which comprised cells containing RAG-1 mRNA transcripts was CD19-, surface IgM-, cytoplasmic IgM- and CD2- CD3- CD4- CD8- CD56-. However, this population was CD7+ (approx. 75%), and contained both CD7- CD34+ (up to 3%) and CD7- CD34+ (up to 3%) cells. These findings are compatible with the hypothesis that the adult human peritoneum provides a microenvironment capable of supporting a thymus-indenpendent differentiation of T lymphocytes.

Adult↗
PubMed1991

The in vitro and in vivo effects of 1,1-dimethylhydrazine (UDMH) on murine lymphocyte subsets and Ia antigen expression.

1,1-Dimethylhydrazine or unsymmetrical dimethylhydrazine (UDMH) is a highly volatile and reactive compound used primarily as a liquid rocket propellant. Previous studies found UDMH to possess immunomodulatory activity similar to other hydrazine derivatives. Modulation of T lymphocyte subpopulations and Major Histocompatibility Complex Class II or Ia antigen were evaluated as possible mechanisms for this UDMH-induced immunomodulation. Murine lymphoid cell populations were examined by flow cytometry for changes in their cell surface marker percentages or relative number upon exposure to UDMH either in vitro or in vivo. The results show UDMH caused significant suppression of the T helper cell population derived from the thymus at the 75 mg/kg dose in vivo, but did not affect other lymphocyte subpopulations isolated from mesenteric lymph node, spleen or thymus at this or any other dose. In vivo exposure of mice at all doses of UDMH did not significantly alter expression of Ia antigens on adherent cell populations and expression of the Ia antigen following in vitro UDMH exposure was not affected as well. Results indicate that the immunomodulatory effects of UDMH are not mediated by phenotypic alteration of T lymphocyte subpopulations or Ia antigen.

Adjuvants, Immunologic↗
PubMed2000

Double staining of intracellular cytokine proteins and T-lymphocyte subsets. Evaluation of the method in blood and bronchoalveolar lavage fluid.

An immunocytochemical staining method has been developed for simultaneous staining of both cell surface markers (CD4 and CD8) and intracellular cytokine proteins IFN-gamma, IL-4 and IL-5. Cell surface molecules were visualized with alkaline phosphatase, which was developed by Fast Blue BB. Intracellular cytokine proteins were detected by amino-ethyl carbazole. We applied this technique to T cells from T-cell lines and T-cell clones, peripheral blood mononuclear cells and broncho-alveolar lavage fluid cells. Cells were used either unstimulated or stimulated for 4 h with 1 ng/ml PMA and 1 microg/ml ionomycin, which proved to be an optimal stimulus taking cytokine staining, cell recovery and cell viability into account. We studied peripheral blood mononuclear cells from healthy subjects and found that without in vitro stimulation on average 0.4% of the cells were IFN-gamma positive cells. In unstimulated broncho-alveolar lavage fluid cells of the 2 allergic asthmatic subjects studied so far we found higher numbers of cytokine-positive cells (up to 22% of the lymphocytes being IL-4+ cells). By in vitro stimulation, the numbers of cytokine-positive peripheral blood mononuclear cells from the healthy subjects were increased to maximally 5% IFN-gamma+ cells. In stimulated lavage fluid cells from allergic asthmatic subjects maximally 34% of the lymphocytes became IFN-gamma+. We conclude that this method allows detection of intracellular cytokine proteins in both CD4+ and CD8+ T cells without the need for stimulating the cells in vitro. In vitro stimulation may change the cytokine profile detected.

Adult↗
PubMedDate not supplied

Postnatal exposure to chlorinated dioxins and related chemicals on lymphocyte subsets in Japanese breast-fed infants.

Effects of postnatal exposure to polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and coplanar polychlorinated biphenyls (Co-PCBs) on lymphocyte subpopulations were investigated in the peripheral blood of 36 breast-fed Japanese babies. As a result, estimated total intakes of these chemicals in toxic equivalent quantity (TEQ) converted into 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) equivalents from the breast milk positively and negatively correlated with the respective percentages of CD4+ and CD8+ lymphocytes in the blood of breast-fed babies. Consequently, the ratios of CD4+ to CD8+ T cells showed significant increasing tendency with the estimated total TEQ intakes. Therefore, our study suggests that exposure to background levels of the highly toxic organochlorine compounds through the breast milk influences the human neonatal immune system.

Adult↗
PubMed1997

[Characterization of leukocytes, lymphocytes and lymphocyte subsets in eating disorders].

Mental anorexia patients have severe denutrition and high cortisol levels. Nervous bulimia presents a particular psychic profile. The aim of this study was to evaluate lymphocyte population in eating disorders in order to understand the effect of these disturbances on immune system. We have found that patients with mental anorexia or bulimia present a significant decrease of leucocytes, lymphocytes, CD2 and CD4. Patients with post-anorexia bulimia have no significant variants. There is no difference between mental anorexia and nervous bulimia. There is a tendential variation between mental anorexia and controls in relation to CD45Ra. Considering the similar patterns of immune disturbance in anorexia and bulimia it looks like a dominance of neuroimmune network over malnutrition. Naive cells are particularly sensitive to malnutrition.

Adolescent↗
PubMed1998

[6-mercaptopurine levels and study of blood lymphocyte subsets during azathioprine treatment of Crohn's disease].

OBJECTIVES: Our aim was to study the relationships between clinical efficacy of azathioprine, 6-mercaptopurine pharmacokinetics and changes in peripheral blood lymphocyte subpopulations induced by azathioprine treatment in Crohn's disease. METHODS: Twenty-three patients were prospectively followed up for 1 year. Peripheral blood counts, total lymphocytes, CD3+, CD4+, CD8+, CD25+, CD16+CD56+, CD57+ and CD19+ lymphocyte subpopulations were carried out, using flow cytometry, during azathioprine treatment. Pharmacokinetic studies were performed at day 8 and month 3 by measuring 6-mercaptopurine plasma concentration after an oral dose of azathioprine (2 mg/kg). Results were compared in responders (no activity and no steroids) and non-responders. RESULTS: The decrease in peripheral blood leukocytes and neutrophils was significant after 1 month, reaching 49% and 48% of the pre-treatment values at 1 year; the one of lymphocytes was significant after 6 months and reached 41% at 1 year. Percentages of CD3+, CD4+, CD8+, CD57+, CD16+CD56+ and CD19+ lymphocytes remained unchanged whereas percentage of CD25+ lymphocytes increased from 10% to 28% (P < 0.01). There was a high inter and intraindividual variability of 6-mercaptopurine peak plasma concentration and area under the curve. No significant difference was found between responders (n = 14) and non responders (n = 7) for pharmacokinetic parameters and lymphocyte subpopulations; there was no correlation between lymphocyte subpopulation changes and 6-mercaptopurine pharmacokinetics. CONCLUSION: Monitoring of 6-mercaptopurine plasma concentration and blood lymphocyte subpopulations is of little value in Crohn's disease patients treated with azathioprine.

Adult↗