Activation of bovine lymphocyte subpopulations by staphylococcal enterotoxin C

Staphylococcus aureus is a major mastitis-causing pathogen in cattle. The chronic nature of bovine staphylococcal mastitis suggests that some products or components of S. aureus may interfere with the development of protective immunity. One class of molecules that could be involved are superantigens (SAgs). Although a significant number of mastitis isolates produce SAgs, the effect of these molecules on the bovine immune system is unresolved. To determine if immunosuppression caused by SAgs could play a role in pathogenesis, we monitored bovine lymphocytes exposed to staphylococcal enterotoxin C1 (SEC1). Activation of bovine lymphocytes by either SEC1 or concanavalin A (ConA) was influenced by the gammadelta/alphabeta T-cell ratio in the culture. Compared to ConA-induced stimulation, cultures stimulated with SEC1 generated small numbers of CD4+ alphabeta T cells expressing high levels of interleukin-2 receptor alpha chain (IL-2R alpha) and major histocompatibility complex class II (MHCII), suggesting that SAg exposure does not lead to full activation of these cells. This state of partial activation was most pronounced in cultures with a high gammadelta/alphabeta ratio. In contrast, significant numbers of CD8+ alphabeta T cells expressed high levels of IL-2R alpha and MHCII, regardless of the gammadelta/alphabeta ratio and the stimulant used. CD8+ blasts in cultures stimulated with SEC1 also expressed another activation marker, ACT3, previously detected predominantly on thymocytes and CD4+ T cells. Although gammadelta CD2- and CD2+ T cells expressed MHCII and IL-2R alpha following stimulation with SEC1, only a few cells increased to blast size, suggesting that they were only partially activated. The results suggest ways in which SAgs might facilitate immunosuppression that promotes the persistence of bacteria in cattle and contributes to chronic intramammary infection.     

Analysis of cytokine mRNA profiles in the lungs of Pneumocystis carinii-infected mice

Severe combined immunodeficient (scid) mice lack functional CD4+ lymphocytes, and therefore develop life-threatening Pneumocystis carinii infection. However, when scid mice are immunologically reconstituted with spleen cells, including CD4+ cells, a protective inflammatory response is mounted against the organism. To determine whether these lymphocytes induce elevated cytokine mRNA levels in response to P. carinii infection, steady-state levels of cytokine mRNAs were measured in the lungs of both reconstituted and unaltered scid mice. Despite significant numbers of organisms and the presence of functional alveolar macrophages in the lungs of 8- and 10-wk-old scid mice, there was neither evidence of pulmonary inflammation, nor increased proinflammatory cytokine expression. However, when 8-wk-old scid mice were immunologically reconstituted, signs of intense, focal pulmonary inflammation were observed, and levels of interleukin (IL)-1alpha, IL-1beta, IL-3, IL-6, interferon-gamma (IFN-gamma), tumor necrosis factor (TNF)-alpha, and TNF-beta mRNAs were all significantly elevated. Cytokine expression was increased at day 10 post-reconstitution (PR), maximal at day 12 PR, and returned to baseline by day 22 PR. In situ hybridization demonstrated that at day 12 PR, increased IL-1beta and TNF-alpha expression was localized to sites of intense inflammation and focal P. carinii colonization. Many of the cells expressing high levels of IL-1beta and TNF-alpha in these regions were in direct contact with organisms, or contained degraded organisms within their cytoplasm. Thus, even though functional macrophages are present in scid mice, CD4+ T cells are required for proinflammatory cytokine expression, which is associated with the generation of a protective inflammatory response at sites of P. carinii infection.     

Expression of lymphocyte subpopulations, cytokine serum levels and blood and urine trace elements in nickel sensitised women

In this study, 20 non-allergic and 20 non-atopic women sensitised only to nickel (Ni) showed similar levels of urine and serum Ni and serum chromium (Cr). On the contrary, serum copper, a marker of inflammation, was significantly higher in the Ni-sensitised group. Sensitised women also had higher values of blood B CD19+, CD5--CD19+ and B and natural killer CD3--CD25+ lymphocytes, but not alterations of some other lymphocyte subsets and serum cytokines. Urine Ni was correlated with "memory" CD4+-CD45RO+ lymphocytes in the non-allergic women and with T CD4+-CD45RO+ and CD3+-CD25+ cells in the atopic women; these subjects also showed a statistically significant correlation of serum Ni with B CD5+-CD19+ lymphocytes and serum IL-13. Moreover, serum Cr of both groups of women was positively or negatively correlated with activated HLA-DR+ cells and/or serum IL-5 and interferon gamma. These results (confirming in part those of a previous study on non-allergic men) suggest that both Ni and Cr are involved in mechanisms regulating the immune response and that allergy to these metals could be considered an alteration of their physiological role.     

Prenatal ontogeny of lymphocyte subpopulations in pigs

Although porcine lymphocytes have been classified into numerous subpopulations in postnatal animals, little is known about the ontogeny of these complex cell subsets. Using double- and triple-colour flow cytometry (FCM), we investigated the surface phenotype of fetal lymphoid cells in the thymus, cord blood, spleen and mesenteric lymph nodes at different stages of gestation. It was found that the major lymphocyte subpopulations started to appear at the beginning of the second third of the gestation period, with B cells being the earliest lymphocyte subpopulation to appear in the periphery. The T-cell receptor (TCR) gamma delta+ cells were the earliest detectable T-cell subset, developing first in the thymus and subsequently arriving in the periphery. Later in ontogeny, however, the number of TCRalpha beta+ lymphocytes rapidly increased, becoming the predominant T cells both in the thymus and in the periphery. Cells with the phenotype of adult natural killer cells were also identified in pig fetuses, though their nature and functional roles remain to be investigated. In addition, CD2 was expressed on most B cells whilst very few CD4+ TCRalpha beta+ cells or CD2+ TCRgamma delta+ cells expressed CD8, suggesting that the expression of CD2 and CD8 may reflect the functional status of the cells in postnatal animals. Taken together, this study has provided a systematic analysis of fetal porcine lymphocyte subpopulations and may provide the base for studies to establish the physiological roles of these lymphocyte subsets.     

Modulation of in vitro lymphocyte stimulation by subpopulations of transforming lymphocytes

The finding of less-than-expected lymphocyte transformation by both PHA-P and PWM acting together supports the possibility that subpopulations of lymphocytes may interact to suppress their transformation and thereby help regulate the immune response.     

Analysis of splenic and thymic lymphocyte subpopulations in chickens infected with Salmonella enteritidis

Lymphocytes expressing CD3, CD4, CD8, pan lymphocyte, IgA, IgG and IgM cell surface antigens were assessed by in the spleen and thymus of chickens following infection with Salmonella enteritidis using flow cytometric analysis. At 6 days post primary infection and 2 days post secondary infection with S. enteritidis, the percentages of IgA+ and IgM+ lymphocytes in the spleen were significantly increased (P < 0.05). At 2 days post secondary infection with S. enteritidis, the percentage of CD4+ T lymphocyte in the spleen and CD8+ T lymphocyte percentage in the thymus were significantly increased (P < 0.05). These results indicate that S. enteritidis infection induces changes in the spleen and thymus that reflect the dynamics of the host protective immune response.     

Natural killing and antibody-dependent cytotoxicity by lymphocyte subpopulations in young and aging humans

Natural killer (NK) and antibody-dependent cellular cytotoxicity (ADCC) were examined in the peripheral blood lymphocytes and their major subpopulations from young and aging subjects. Monocyte-depleted unseparated lymphocyte-mediated NK activity (against cells of K-562) and ADCC (against IgG-coated chicken erythrocytes) were comparable between young and aging subjects. Similarly no significant difference was observed in T cell-mediated NK and ADCC and non-T cell-mediated ADCC between young and aging subjects. Non-T cell-mediated NK activity, however, was significantly (P less than 0.025) greater in aging humans compared to that of young subjects. When the data were analyzed according to gender, T cell-mediated ADCC in aging males was significantly (P less than 0.05) greater than that found in young males. No significant difference was observed between T-cell ADCC among young and aging females. T cell-mediated NK was comparable among young and aging males and young and aging females. Non-T cell-mediated NK as well as ADCC activity was significantly (P[ less than 0.025 or less than 0.05) greater in aging males compared to that in young males. Both non-T-cell NK and ADCC were comparable among young and aging females. This study demonstrates an increase in NK and ADCC activity in aging subjects that is primarily shared by males and not by females. No correlation was observed between the proportion of T gamma cells and T-cell NK or ADCC activity.     

Telomere length dynamics in human lymphocyte subpopulations measured by flow cytometry

To measure the average length of telomere repeats at chromosome ends in individual cells we developed a flow cytometry method using fluorescence in situ hybridization (flow FISH) with labeled peptide nucleic acid (PNA) probes. Results of flow FISH measurements correlated with results of conventional telomere length measurements by Southern blot analysis (R = 0.9). Consistent differences in telomere length in CD8+ T-cell subsets were identified. Naive and memory CD4+ T lymphocytes in normal adults differed by around 2.5 kb in telomere length, in agreement with known replicative shortening of telomeres in lymphocytes in vivo. T-cell clones grown in vitro showed stabilization of telomere length after an initial decline and rare clones capable of growing beyond 100 population doublings showed variable telomere length. These results show that flow FISH can be used to measure specific nucleotide repeat sequences in single cells and indicate that the very large replicative potential of lymphocytes is only indirectly related to telomere length.     

Vinorelbine: cell cycle kinetics and differential sensitivity of human lymphocyte subpopulations

Antiproliferative activity induced by vinorelbine (VRB) was studied in whole blood cultures (WBC) and in lymphocyte subpopulations evaluated by the MAC (morphology-antibody-chromosome) method. In WBC a significant delay in cell cycle kinetics in VRB-treated cultures compared with controls (P < 0.001) was observed. The highest dose (1.0 microg/ml) arrested all cells at the first metaphase. Both WBC and isolated lymphocyte cultures showed a significant increase in the mitotic index (MI) in VRB-treated cultures compared with controls (P < 0.001). Moreover, a significant increase in the MI was found in VRB-treated CD4 cells compared with VRB-treated CD8 (P < 0.01) and B-lymphocytes (P < 0.001).     

Studies on lymphocyte subpopulations and the effect of age on immune competence in turkeys

We characterized the lymphocyte subpopulations and investigated the effect of age on cellular and humoral immunity, development of lymphoid organs, and the relative proportions of CD4+ and CD8+ cells in turkeys. The mitogenic responses of peripheral T cells were poorly developed at hatch but developed rapidly after hatch and reached adult levels by 2 weeks-of-age. The average percentage of CD4+ cells was 45, 29.8, and 26.3 in the thymi, peripheral blood, and spleens, respectively, in turkeys. The mean percentage of CD8+ cells in the thymi, peripheral blood, and spleens of turkeys was 53.8, 13.6, and 15.5, respectively. Age did not influence the relative proportions of CD4+ and CD8+ T cells in the spleens and peripheral blood of turkeys. The mean percentages of IgM+ cells in the bursae and spleens were 78.5 and 26.8, respectively. Day-old turkeys did not develop detectable antibodies to either thymus dependent or independent antigens. However, 2 week or older turkeys showed good humoral responses. Inoculation of BSA at hatch induced tolerance, whereas injection of SRBC did not. Analysis of relative organ weights of turkey lymphoid organs showed that spleens and thymi developed rapidly during the first week-of-age.     

Lymphocyte phenotyping in infants: maturation of lymphocyte subpopulations and the effects of HIV infection

Oxygen administration is one of the most important therapeutic interventions for a child with severe acute lower respiratory tract infection (ALRI). Inexpensive and efficient methods of oxygen administration are highly desirable in hospitals in developing countries. The objectives of this study were to compare the frequency and nature of complications when nasopharyngeal catheters or nasal prongs are used to deliver oxygen. One hundred and twenty-one children between the ages of 2 weeks and 5 years with hypoxia due to ALRI were randomized to receive oxygen via a catheter (61 children) or via nasal prongs (60 children). The two groups were similar in terms of diagnoses, clinical severity, oxygen saturation on admission and case fatality rates. There was no difference in the incidence of hypoxaemic episodes between the two groups. The oxygen flow rates required on the day of admission for adequate oxygenation (SaO2 > 90%) ranged from 0.8 litres per minute to 1.2 litres per minute. The required oxygen flow rate decreased during the course of treatment. Mucus production was more of a problem in the catheter group, and nasal blockage, intolerance of the method of oxygen administration and nursing effort were generally higher amongst the catheter group, but none of these differences was significant. Ulceration or bleeding of the nose was significantly more common in the catheter group (19.7% vs 6.7%, p < 0.05). Abdominal distension and nasal perforation were not seen in either group. This study suggests that nasal prongs are safer, more comfortable and require less nursing expertise than nasopharyngeal catheters for administration of oxygen to children.     

Studies of lymphocyte subpopulations of bronchoalveolar lavage fluid in HTLV-I seropositive patients with uveitis

To determine whether HTLV-I infection is associated with uveitis, we investigated the bronchoalveolar lavage fluid (BALF) cells in 23 patients with uveitis. Of these, sarcoidosis was diagnosed in 13 patients, but could not be confirmed in the remaining 10 patients (non-sarcoidosis). Three of 13 patients (23.1%) with sarcoidosis and 7 of 10 non-sarcoidosis patients (70.0%) were HTLV-I seropositive. In BALF, no significant differences were observed between the HTLV-I seropositive and seronegative patients. However, in non-sarcoidosis patients, total cell counts, CD3+ and CD+ HLA-DR+ cells in BALF were significantly higher in seropositive patients than in seronegative patients. CD3+ CD25+ cells in BALF were markedly increased in 3 of 7 non-sarcoidosis seronegative patients. These findings indicated that BALF lymphocytes in HTLV-I seropositive non-sarcoidosis patients were more activated than those in seronegative non-sarcoidosis patients, supporting the hypothesis that HTLV-I infection may be associated with uveitis.     

Effects of GnRH antagonist on lymphocyte subpopulations in primary and secondary lymphoid tissues of female mice

PROBLEM: GnRH analogs are playing an increasing role in the treatment of many clinical disorders. Recent studies have indicated that GnRH agonists suppress immune function in mice in vivo. The present study investigated the effects of GnRH antagonist of functional lymphocyte subsets of mice in vivo. METHOD: Three- and 10-wk old female mice received 10 micrograms of Nal-Glu daily for 15 and 30 days; changes in the immunophenotypic expression of lymphocytes from thymus, bone marrow, spleen and blood were analyzed by flow cytometry. RESULTS: The administration of GnRH antagonist to pre- and postpubertal female mice induced slight increases in lymphocyte subpopulations in primary and secondary lymphoid tissues. These effects are opposite those obtained with GnRH agonist in our earlier studies in mice. CONCLUSIONS: Assuming similar effects in humans and rodents, the gonadal steroid suppression achieved by GnRH antagonist treatment has no apparent suppressive effects on the immune system.     

Longitudinal follow-up of blood lymphocyte subpopulations from birth to 1 year of age

The features of the rare interstitial hernia, as seen in four children, are described. All were boys ranging in age from 7 days to 7 years. The presenting sign was swelling in the right side of the abdomen associated with undescended testis. The hernias arose at the deep inguinal ring and expanded superiorly between the internal and external oblique muscles. Repair involved a standard herniotomy and orchidopexy through an oblique inguinal incision-except in one patient with a vanishing testis. The importance of being aware of this rare type of hernia is stressed.     

Seed extract of Aeginetia indica L induces cytokine production and lymphocyte proliferation in vitro

The development of the human medial superior olivary nucleus was studied in serial sections of 10 fetuses at 12-35 weeks of gestation (WG), an infant at 2 months of age and an adult of 63 years using an electronic planimeter with a computer. Morphometric analysis suggested that the development of the human medial superior olivary nucleus accelerates between 16 and 21 WG in terms of columnar lengths and volumes, neuronal sizes and circularity ratios, while it matures gradually in terms of the amount of Nissl bodies.