Age-related changes in blood lymphocyte subsets of Saudi Arabian healthy children

The age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T-lymphocyte population, the CD8(+) subset increased but the CD4(+) subset decreased, resulting in a declining CD4(+)/CD8(+) ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA-DR+ T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease-related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups.     

Lipid peroxidation and changes in T lymphocyte subsets and lymphocyte proliferative response in experimental iron overload

This study was undertaken to investigate the hypothesis that lipid peroxidation might be associated with immunological abnormalities in experimental hemosiderosis. The correlation between the degree of plasma and spleen lipid peroxidation with lymphocyte proliferative response and with the proportion of T lymphocyte subsets was studied in normal and iron overloaded male Sprague Dawley rats. The iron-loading protocol consisted of a total dose of iron-dextran (1.5 mg/Kg body weight) divided in daily i.m. injections over twenty consecutive days. Lipid peroxidation was measured by the thiobarbituric acid assay in plasma and in homogenates of spleen. Plasma lipid peroxide level increased rapidly after i.m. administration of iron-dextran and decreased sharply at 48 h after the last injection. Conversely, a progressive increase of lipid peroxidation in homogenates of spleen was observed in the course of the iron overload protocol, remaining high even at 50 days after initiation of iron-dextran injections. The increase of spleen lipid peroxide levels was associated with decreased lymphocyte proliferative response to Con A in iron overloaded rats. The addition of superoxide dismutase and catalase to lymphocyte cultures reversed the inhibition of the proliferative response, implicating reactive species of oxygen as the causative agents of these alterations. These effects may be related with the enhanced membrane and DNA damage occurring during intracellular and extracellular peroxidation. Negative correlations between helper/cytotoxic ratio and malondialdehyde levels were obtained in blood and spleen during iron administration. These results supports the hypothesis that lipid peroxidation plays a role in the immunological abnormalities observed in experimental hemosiderosis.     

Identification of lymphocyte subsets in pregnancy gingivitis

The percentage of gingival tissue and peripheral T, T Subsets and B lymphocytes were assessed in ten pregnant women during the first, second and third trimesters. Clinically the gingival index (GI) was found to be higher during pregnancy than in the control group. The percentage of the T3, T4 and B cells appeared to decrease in peripheral blood and gingival tissues during pregnancy as compared to control group.     

Alterations of lymphocyte subsets in patients with recurrent fetal wastage positive for antiphospholipid antibodies treated with Chinese herbal medicine

The delay technique is an established method of enhancing flap survival. This investigation attempts to determine which of two delay techniques results in the best delay effect by measuring their relative abilities to capture adjacent vascular territories in a rat model. A dorsal flap based on the iliac branch of the iliolumbar artery with a captured random zone corresponding to the axial territory of the lateral thoracic artery was used in the evaluation. Sprague-Dawley rats (350-400 g) were randomly assigned into three groups. Group I was the control group. In group II, the circumferential borders of the animal's dorsum were incised without undermining and the dominant pedicle of the lateral thoracic flap was divided. In group III, the medial and lateral borders of the flap were incised and undermined as a bipedicled flap, violating the musculocutaneous perforators. The dominant pedicle of the lateral thoracic artery was also divided. Group III had the greatest survival with only 9% of flap area necrosis compared to 28% and 21% for groups I and II, respectively. These differences were statistically significant. The results suggest that musculocutaneous perforators provide a substantial vascular source to the tissue at risk and should be considered in selecting a delay technique.     

Differential activity of P-glycoprotein in normal blood lymphocyte subsets

To better understand the phenomenon of P-glycoprotein (P-170) expression we investigated lymphocyte subpopulations for P-170 function in healthy volunteers. Studies were based on three-colour flow cytometry including the fluorescent probe rhodamine 123 (Rh123), which is transported by P-170. Marked Rh123 efflux was detected in CD8+ T lymphocytes with CD8+/CD45RA+ T cells (naive cells) showing significantly higher P-170 activity as compared with CD8+/CD45RA- cells (P<0.04). Vice versa, CD8+/CD45RO+ T cells (memory cells) demonstrated less P-170 activity than CD8+/CD45RO- cells (P<0.04). P-170 function was less prominent in CD4+ T cells, however, Rh123 efflux was higher in the CD4+/CD45RA+ and CD4+/CD45RO- subpopulations (P<0.025) corresponding to the CD8+ results. Dye efflux differed significantly between activated and non-activated CD8+ and CD4+ as well as CD8+/CD11b+ and CD8+/CD11b- T lymphocytes. Since CD16+ natural killer cells (NK) expressed the highest level of P-170, the NK cytotoxicity against 51Cr-labelled K562 target cells was assayed in the presence or absence of P-170 inhibitors. NK related cytotoxicity was significantly reduced in the presence of R-verapamil and dexnigaldipine-HCP in a dose-dependent manner. The differential expression of P-170 activity in naive and memory T cells together with the reduced NK related cytotoxicity in the presence of MDR-modulators suggest a physiological role of P-170 in immunological functions of these lymphocyte subsets. Consequently, the addition of MDR modulators to conventional chemotherapy as a strategy to overcome drug resistance should consider possible adverse immunosuppressive effects.     

Neutralization of HIV-1 primary isolates by polyclonal and monoclonal human antibodies

To examine antibody-mediated neutralization of HIV-1 primary isolates in vitro, we tested sera and plasma from infected individuals against four clade B primary isolates. These isolates were analyzed further for neutralization by a panel of several human anti-HIV-1 mAb in order to identify the neutralizing epitopes of these viruses. Each of the HIV-1+ serum and plasma specimens tested had neutralizing activities against one or more of the four primary isolates. Of the three individual sera, one (FDA-2) neutralized all of the four isolates, while the other two sera were effective against only one virus. The pooled plasma and serum samples reacted broadly with these isolates. Based on the neutralizing activities of the mAb panel, each virus isolate exhibited a distinct pattern of reactivity, suggesting antigenic diversity among clade B viruses. Neutralizing epitopes were found in the V3 loop and CD4-binding domain of gp120, as well as near the transmembrane region (cluster II epitope) of gp41. A mAb directed to the cluster I epitope of gp41 near the immunodominant disulfide loop weakly neutralized one primary isolate. None of the mAb in the panel affected one primary isolate, US4, although this virus was sensitive to neutralization by some of the polyclonal antibody specimens. This isolate was also resistant to neutralization by a cocktail of 10 mAb, most of which individually inhibited at least one of the other three viruses tested. These results suggest that neutralizing activity for this latter virus is present in certain HIV-1+ sera/plasma, but is not exhibited by the mAb in the panel. Thus, effective neutralizing antibodies against primary isolates can be generated by humans upon exposure to HIV-1, but not all of these antigenic specificities are represented in a large panel of human anti-HIV-1 mAb.     

FIV vaccine studies. II. Clinical findings, hematological changes and kinetics of blood lymphocyte subsets

Five groups of cats were vaccinated with different recombinant feline immunodeficiency virus (FIV) SU vaccines expressed either in Escherichia coli or in the Baculovirus system. In Part I of this series, we described the humoral immune response and outcome of intraperitoneal FIV challenge exposure. Additionally, all cats were monitored for clinical and hematological changes and the course of blood lymphocyte subsets. These results are described in this present paper. A great increase of antibodies was found after vaccination with different recombinant FIV antigens, which did not protect the cats from intraperitoneal FIV challenge infection. This observation was paralleled by an increase of eosinophils during vaccination which was even more pronounced after challenge infection. After FIV challenge, infection lymphadenopathy, gingivitis, pharyngitis, changes in total leukocytes and neutrophils and a decrease in the CD4+:CD8+ ratio were found in cats of all groups and were considered as a sign of the FIV infection taking place, independent of vaccination. The following observations suggest that in these cats a TH2-like immune response was elicited: the high counts of eosinophils, the nature of antigen and adjuvant (aluminium hydroxide) and the high amounts of antigens used for immunization. Clearly, this type of immune response did not protect the animals from intraperitoneal FIV challenge infection.     

Polysaccharides from Agaricus blazei stimulate lymphocyte T-cell subsets in mice

Subset analysis of splenic lymphocytes using flow cytometry showed that the percentages of Thy1.2-(pan T-cells), L3T4-(CD4, helper T-cells), and Lyt2-(CD8, cytotoxic T-cells) positive cell populations were significantly increased in mice orally administered a hot water-soluble fraction from Agaricus blazei as compared with mice treated only with saline. 13C-NMR data indicates that the main component in the active polysaccharide is the complex of alpha-1,6- and alpha-1,4-glucan, which had already been shown to have anti-tumor activity against Sarcoma 180. It seems that the polysaccharide from Agaricus blazei may be an effective prophylactic, protecting humans against cancer by stimulating lymphocytes such as cytotoxic T-cells.     

Requirement for multiple lymphocyte subsets in protection by a live attenuated vaccine against retroviral infection

STUDY DESIGN: A case report and literature review of thoracic hyperkyphosis deformity secondary to glucocorticoid-induced osteoporosis in Cushing's disease. OBJECTIVES: To identify the pathophysiology of glucocorticoid-induced osteoporosis and to outline the diagnosis and treatment options for a patient with severe spinal deformity secondary to unrecognized excess glucocorticoid activity. SUMMARY OF BACKGROUND DATA: Glucocorticoid-induced osteoporosis is seen in patients exposed to supraphysiologic levels of endogenous or exogenously administered glucocorticoids. In these patients, glucocorticoids act to suppress bone formation and increase bone resorption by indirect and direct effects. These patients have a high prevalence of trabecular bone loss, resulting in much higher rates of vertebral body collapse and pathologic fracture and thus causing an increased propensity toward kyphotic spinal malalignment. METHODS: The literature was reviewed and case reports studied. This case report highlights the pathophysiology of the disease process that caused the spinal deformity and the surgical intervention used to correct the kyphotic deformity after the metabolic problem was resolved. RESULTS: This patient has responded well to treatment and surgical intervention to correct a thoracic hyperkyphotic deformity without complication. CONCLUSIONS: Unrecognized endogenous production of glucocorticoids in Cushing's disease should be considered in young adult patients with progressive osteoporotic spinal deformities.     

Increased insulin binding by lymphocyte receptors induced by beta-OH butyrate

The serum concentration of ketones increases in both fasting and diabetic ketoacidosis. Whether ketones alter, independent of changes in pH, in number and affinity of insulin receptors is not known. The binding of insulin 125I by 24 X 10(6)IM-9 lymphocyte cells was examined in both the presence and absence of 8 mM beta-OH butyrate. Studies were conducted at multiple concentrations of insulin. A consistent increase of insulin binding was induced by beta-OH butyrate. The mean maximum increase of insulin binding was 35%, which achieved significance at the 1% level.     

T lymphocyte subsets and activation status in patients following liver transplantation

Changes in T-lymphocyte subsets have previously been shown to relate to clinical events following liver transplantation and be of prognostic significance following renal transplantation. The aim of this study was to examine T lymphocyte subsets, their activation status and the mean fluorescence intensity of cell surface markers by flow cytometric analysis, in peripheral blood of patients following liver transplantation. Stable transplant patients (n=11) had a significantly higher level of activation (HLA-DR expression ) of all T cell subsets: CD3, CD4 and CD8 compared to healthy controls: 17.5% +/- 14.0 (mean +/- SD) vs 4.7 +/- 1.8 (p=0.04), 13.7% +/- 10.3 vs 4.3 +/- 1.7 (p=0.03) and 23.8% +/- 19.9 vs 3.6 +/- 2.4 (p=0.02) respectively. A further increase in activation status occurred in all T cell subsets in association with acute cellular rejection, reaching significance for the CD4+ population: 13.7% +/- 10.2 vs 23.3% +/- 20.6 (p=0.04). The mean fluorescence intensity of the CD3+DR- and CD3+ DR+ populations were increased to 1397 +/- 869 and 1282 +/- 810 following liver transplantation compared to values of 425 +/- 204 and 376 +/- 166 respectively for controls (p<0.05). T-lymphocytes maintain a high level of activation following liver transplantation and continue to express high levels of the surface marker CD3, which may account for the occurrence of acute cellular rejection despite immunosuppression in these patients.     

Role of T lymphocyte subsets in immunity to spotted fever group Rickettsiae

To evaluate the roles of CD4 and CD8 T lymphocytes in immunity to disseminated endothelial infection with Rickettsia conorii (Malish 7 strain), these T cell subsets were depleted or adoptively transferred into subsequently infected C3H/HeN mice. CD4 T lymphocyte-depleted and sham-depleted mice underwent a similar course of illness with a sublethal rickettsial dose, cleared the infection by day 10, and recovered on days 10 to 11. In contrast, mice depleted of CD8 lymphocytes or CD4 and CD8 lymphocytes died or remained persistently infected through day 15 with the ordinarily sublethal dose. Endothelium was the major site of rickettsial persistence, including sites in the vital organs, brain, and lungs of CD8 lymphocyte-depleted mice. In nondepleted animals, CD8 T lymphocytes were observed in apposition to endothelial cells on day 10 at the time of rickettsial clearance. Adoptive transfer of immune CD4 or CD8 T lymphocytes protected mice against a lethal dose of R. conorii in the disseminated endothelial target model. Nonimmune CD4 or CD8 lymphocytes and immune lymphocytes that had passed through columns that depleted both CD4 and CD8 lymphocytes failed to protect mice against R. conorii. These studies represent the first analysis of the role of T lymphocyte subsets in immunity to spotted fever group rickettsiae and the first demonstration that clearance of spotted fever group rickettsiae from endothelial cells requires immune CD8 T lymphocytes.     

Comparative analysis of equine lymphocyte subsets in whole blood and gradient-purified samples

In the present study, two methods of lymphocyte preparation, whole blood lysis and Ficoll-Paque separation, prior to FACS analysis were compared. The comparison was done with single and dual-colour staining techniques. Monoclonal antibodies (mAb) against eCD4, eCD5, eCD8 and eMHC class II were used. There was no significant difference in the results obtained by these two methods.     

Analysis of the peripheral blood lymphocyte subsets in patients with bladder carcinoma

OBJECTIVES: To evaluate the immune system of patients with bladder transitional cell carcinoma (TCC) by using peripheral blood lymphocyte subsets and to further compare the relationship between these subsets with respect to tumor stage and grade (superficial versus invasive and low versus high grade). METHODS: Thirty patients with superficial TCC of the bladder, 30 patients with invasive TCC of the bladder, and 30 age- and sex-matched control subjects without any malignancy or immunologic abnormality were included in this study. The peripheral blood lymphocyte subset analysis was performed in all patients before any treatment was performed. RESULTS: All lymphocyte subset values of patients with invasive bladder cancer, except B cell value, were significantly lower (P < 0.01) than the values of the control group. There were no significant differences between the lymphocyte subset values of patients with superficial bladder cancer and those of control subjects. The comparison of the lymphocyte subset values of the patients with superficial versus invasive bladder carcinoma revealed that in patients with invasive bladder carcinoma, the numbers of T and natural killer (NK) cells were significantly lower (P < 0.05) than those of patients with superficial bladder carcinoma. Patients with high-grade tumors had significantly fewer (P < 0.05) T and NK cells than patients with low-grade tumors. CONCLUSIONS: Our results indicate that analysis of mean NK and T cell values and the mean ratio of CD4+/CD8+ cells in peripheral blood might be a useful adjunct for the clinical evaluation of patients with bladder cancer.     

Acute and long-term changes in T-lymphocyte subsets in response to clinical and subclinical measles. A community study from rural Senegal

Experiments carried out on 30 cats using retrograde axonal transport of markers (horseradish peroxidase and luminophores) were used to study the organization of the afferent projection system of the pallidal complex (the globus pallidum, the entopeduncular nucleus, the ventral pallidum) formed by fibers from functionally different cortical and subcortical structures (the ventral tegmental area, the substantia nigra, the amygdaloid body). The distribution of afferent projection fibers in this complex of nuclei led to identification of the following zones: a "limbic" zone, corresponding to the ventral pallidum, and a "motor" zone, corresponding to the caudal part of the globus pallidum. On the one hand, the features of the afferent organization as demonstrated here can be regarded as a structural basis for the functional heterogeneity of the pallidal complex; on the other, significant regions within these structures (the rostral part of the globus pallidum and the entopeduncular nucleus) were found to receive projection fibers from functionally different structures, suggesting the existence of convergence and integration processes in these regions.     

Effect of zinc supplementation on cell-mediated immunity and lymphocyte subsets in preschool children

The pattern and distribution of periodontitis were investigated in 162 randomly selected dogs available for necropsy in veterinary practice. There were 82 males and 80 females of 50 different breeds (150 dogs were pure-bred and 12 were mongrels, aged between 7 months and 14 yr. Presence of periodontitis was determined by assessment of alveolar bone loss on radiographs of the skulls and jaws. Periodontitis occurred frequently with increasing age, although the prevalence varied markedly among and within different breeds. Of the breeds most represented in the sample, periodontitis was most frequently seen in poodles and dachshunds but was rarely recognized in German shepherd dogs. Regardless of age, the vast majority of the dogs displayed either one or both of two different radiographic patterns of alveolar bone loss. One pattern was characterized by slight, horizontal alveolar bone loss involving interradicular and interdental areas. The other pattern was one of predominantly crater-like, or narrow, vertical bone defects which, when advanced, often extended around a single root or tooth to surround the root apices. The two types of patterns did not seem to be breed-dependent. The posterior maxillary and mandibular premolars and molars were the most frequently affected teeth. Alveolar bone loss was most severe in the maxilla, while corresponding bone loss in the mandible was more often related to increasing age.     

Changes in lymphocyte subsets in the intestine and mesenteric lymph nodes in caprine paratuberculosis

In five normally hearing subjects and seven subjects with damaged cochleas, detection thresholds for sinusoidal frequency modulation (FM) and amplitude modulation (AM) were measured using 1 s stimuli with a 500 Hz carrier frequency (Fc) at a 'comfortable' loudness (given by subject-dependent SPLs and SLs). The modulation frequency (Fmod) was 2 Hz or 10 Hz. FM (but not AM) detection was poorer in the hearing-impaired group, especially when the hearing loss at Fc exceeded 50 dB. Fmod had a different effect on FM and AM detection. The corresponding interaction was essentially identical for the two groups of subjects. Previous studies strongly suggested that normal listeners use mainly neural phase-locking cues for the detection of FM when Fmod = 2 Hz, but mainly tonotopic cues when Fmod = 10 Hz. The present results suggest that cochlear damages reduce the usefulness of these two types of cues to an approximately equal degree.     

Female premenopausal tophaceous gout induced by long-term diuretic abuse

We describe 3 cases of tophaceous gout affecting premenopausal women. The only precipitating factor to be found was the chronic and unnecessary overuse of furosemide, apparently resulting from a psychological profile of anorexia nervosa. Our cases emphasize the need for physicians to look for hidden abuse of diuretics in the presence of gouty arthritis in menstruating women, especially if tophi are noted.