Higher Percentage Of CD4 Research Articles

DHA Supplementation during Pregnancy and Lactation Affects Infants' Cellular but Not Humoral Immune Response

Background. It is currently recommended that diet of pregnant mothers contain 200–300 mg DHA/day. Aim. To determine whether DHA supplementation during pregnancy and lactation affects infants' immune response. Methods. 60 women in ≥3rd pregnancy studied; 30 randomly assigned to receive DHA 400 mg/day from 12th week gestation until 4 months postpartum. From breast-fed infants, blood obtained for anti-HBs antibodies, immunoglobulins, lymphocyte subset phenotyping, and intracellular cytokine production. Results. CD4+ lymphocytes did not differ between groups, but CD4CD45RA/CD4 (naïve cells) significantly higher in infants in DHA+ group. Proportion of CD4 and CD8 cells producing IFNγ significantly lower in DHA+ group, with no differences in proportion of IL4-producing cells. Immunoglobulins and anti-HBs levels did not differ between groups. Conclusions. In infants of mothers receiving DHA supplementation, a higher percentage of CD4 naïve cells and decreased CD4 and CD8 IFNγ production is compatible with attenuation of a proinflammatory response.

Efficient nonadhesive ex vivo expansion of early endothelial progenitor cells derived from CD34 + human cord blood fraction for effective therapeutic vascularization

Endothelial progenitor cells (EPCs) have been shown to have therapeutic potential in ischemic disease. However, the number of EPCs for cell therapy is limited. In this study, instead of the typical adherent culture method, we investigated a more efficient, clinically applicable nonadhesive expansion method for early EPCs using cord blood-derived cells to overcome rapid cellular senescence. After a suspension culture of isolated CD34(+) cells in serum-free medium containing each cytokine combination was maintained for 9 d, the number of expanded functional EPCs was assessed by an adherent culture assay. Compared to mononuclear cells, the CD34(+) fraction was superior in its expansion of functional EPCs that could differentiate into acLDL/UEA-1(+) cells without significant cellular senescence, whereas the CD34(-) fraction showed no EPC expansion. Among the cytokine combinations tested for the CD34(+) fraction, a combination (SFIb) consisting of stem cell factor (SCF), FMS-like tyrosine kinase 3 ligand, interleukin-3, and basic fibroblast growth factor resulted in a reproducible 64- to 1468-fold EPC expansion from various cord blood origins. Interestingly, the SFIb combination displayed markedly increased EPC expansion (2.43-fold), with a higher percentage of CD34(+) cells (2.17-fold), undifferentiated blasts (2.38-fold) and CXCR4(+) cells (1.68-fold) compared to another cytokine combination (SCF, thrombopoietin, and granulocyte colony-stimulating factor), although the two cytokine combinations had a similar level of total mononucleated cell expansion (∼ 10% difference). Accordingly, the cells expanded in the SFIb combination were more effective in recovery of blood flow and neovascularization in hind-limb ischemia in vivo. Taken together, these results suggest that the nonadhesive serum-free culture conditions of the CD34(+) fraction provide an effective EPC expansion method for cell therapy, and an expansion condition leading to high percentages of CD34(+) cells and blasts is likely important in EPC expansion.

Evaluation of lymphocyte levels in a random sample of 218 elderly individuals from São Paulo city

Background Age-associated changes in the immune system cause decreased protection after vaccination and increased rates of infections and tumor development.MethodsLymphocyte percentages were compared by gender and age to establish differences between subtypes. Three mL blood samples were obtained from 218 randomly selected individuals (60-101 years old) who live in São Paulo city. Blood was lysed with Tris phosphate buffer and stained for 30 minutes with monoclonal antibodies (CD3PerCP, CD4FITC, CD8Pe, CD19Pe) for analysis by flow cytometry. Statistical analysis was by ANOVA.ResultsThe percentage of CD4+ T cells (p-value = 0.005) and the CD4/CD8 ratio (p-value = 0.010) were lower in men, whereas the percentage of CD8+ T cells was lower (p-value = 0.002) in women; the percentage of B cells (CD19+ ) was similar between groups. Individuals grouped by gender and age range and compared showed a drop in CD4+ cells in 75 to 79-year-old men (female: 46.1% ± 8.1% and male: 38.8% ± 10.5%; p-value = 0.023). Also, the 80 to 84-year-old group of men had a higher percentage of CD8+ (female: 20.8% ± 8.2%, and male: 27.2% ± 8.2%; p-value = 0.032). Low percentages of B cells were detected in men in the 75 to 79-year-old (p-value = 0.003), 85 to 89-year-old (p-value = 0.020) and older than 90 year old (p-value = 0.002) age ranges.ConclusionElderly men present with more changes in lymphocyte subsets compared to elderly women. These findings could demonstrate impairment in the immune response since the lower CD4+ in men would provide less help to B cells (also lower in men) in terms of antibody production. In addition, the increase in CD8+ cells in this group could represent chronic inflammation observed during the aging process.

Role of Thymic Stromal Lymphopoietin (TSLP) in Palifermin-Mediated Immune Modulation and Protection from Acute Murine Graft-Versus-Host Disease

Using the C57BL/6→(C57BL/6 x DBA/2)F(1)-hybrid model of acute graft-versus-host disease (GVHD), we previously showed that treating the donor mice with palifermin provides protection from morbidity and a shift from Th1 to Th2 cytokine production. To determine whether thymic stromal lymphopoietin (TSLP) is involved in palifermin-mediated immune modulation, we used donors from the following groups: (1) untreated wild-type donors, (2) palifermin-treated wild-type donors, (3) untreated TSLPR(-/-) donors, and (4) palifermin-treated TSLPR(-/-) donors. Survival in the recipients was 0%, 100%, 31%, and 0%, for groups 1-4, respectively, indicating that TSLP responsiveness is required for palifermin-mediated protection from GVHD. We also found that the increases in Th2 cytokine levels that are induced by palifermin treatment are obviated in TSLPR(-/-) donors, and that protection from GVHD (group 2) is associated with a higher percentage of CD4(+)CD25(+)Foxp3(+) cells in the graft. Collectively, our findings show that when palifermin and TSLP act in concert, the predominant effect is protection in this model.

Older age of rheumatoid arthritis onset is associated with higher activation status of peripheral blood CD4+ T cells and disease activity

Rheumatoid arthritis (RA) is a chronic inflammatory disease, with a clinical manifestation both systemic and in joints. It has been suggested that age at disease onset and/or patients' age have influence on disease activity and clinical outcome. The reasons for the different course of RA in older people are not known; however, the activation status of peripheral blood lymphocytes could be responsible. Our aim was to relate expression of activation markers in peripheral blood CD4(+) T cells of RA patients with patients' age and/or onset age and disease activity measured by DAS28. Seventy RA patients were included into the immunological study. Two separation criteria were performed: based on age of RA onset and on the biological age of patients. We examined different activation markers, CD69, CD25, CD95 and human leucocyte antigen D-related (HLA-DR), on the CD4(+) T cell surface. Division of RA patients in 10-year intervals at 40, 50 and 60 years revealed that RA patients with later disease onset were characterized by higher DAS28. This phenomenon was not limited to the division at 60 years of age but, surprisingly, the major differences were found for the 40-year onset division. Analysis of all four components of DAS28 revealed that disease activity in older disease onset was dependent on all components. Older-onset RA patients had a higher percentage of CD4(+) CD25(+) and CD4(+) CD95(+) T cells. Summarizing the major differences in DAS28 and activation status of CD4(+) T cells observed for onset of disease at 40 years seems to be the most informative about the immunological status of RA patients.

A reversed CD4/CD8 ratio of tumor-infiltrating lymphocytes and a high percentage of CD4+FOXP3+ regulatory T cells are significantly associated with clinical outcome in squamous cell carcinoma of the cervix

In this study, 40 biopsy samples collected from cervical cancer patients at the First Affiliated Hospital of Xi'an Jiaotong University, China, were retrospectively assessed using immunohistochemistry for CD4(+) and CD8(+) tumor-infiltrating lymphocytes (TILs) and were analyzed for the expression of FOXP3, OX40, granzyme B (GrB) and perforin (Prf). The proliferating index of the TILs was determined by assessing Ki67 expression. We determined the prognostic value of low and high numbers of TILs on survival by performing Kaplan-Meier analysis using median values as the cut-off points. Except for the number of CD4(+)FOXP3(+) regulatory T cells (Tregs) and the CD4/CD8 ratio, none of the CD4(+), CD8(+), OX40(+), GrB(+) or Prf(+) TILs were associated with the overall 5-year survival rate. The 5-year survival rate was significantly lower in patients who had a high percentage of Tregs as compared with the those who had a lower percentage (35.3% versus 88.9%, P=0.001), while the 5-year survival rate was significantly higher in patients with a high CD4/CD8 ratio as compared with patients who had a low CD4/CD8 ratio (82.4% versus 44.4%, P=0.029). When we considered the deaths and surviving cases as separate groups, we found that both the number of CD4(+) T cells and the CD4/CD8 ratio were significantly lower in patients who died as compared with those who survived (26.33±11.80 versus 47.79±38.18, P=0.023 and 0.60±0.25 versus 1.17±1.02, P=0.019, respectively). In conclusion, decreased proportions of tumor-infiltrating CD4(+) T cells with high percentages of Tregs and reversed CD4/CD8 ratios were significantly associated with the clinical outcome of patients with cervical carcinoma.

Novel immunomodulatory effects of adiponectin on dendritic cell functions

Adiponectin (ADN) is an adipocytokine with anti-inflammatory properties. Although it has been reported that ADN can inhibit the immunostimulatory function of monocytes and macrophages, little is known of its effect on dendritic cells (DC). Recent data suggest that ADN can regulate immune responses. DCs are uniquely specialised antigen presenting cells that play a central role in the initiation of immunity and tolerance. In this study, we have investigated the immuno- modulatory effects of ADN on DC functions. We found that ADN has only moderate effect on the differentiation of murine bone marrow (BM) derived DCs but altered the phenotype of DCs. The expression of major histocompatibilty complex class II (MHCII), CD80 and CD86 on ADN conditioned DCs (ADN-DCs) was lower than that on untreated cells. The production of IL-12p40 was also suppressed in ADN-DCs. Interestingly, ADN treated DCs showed an increase in the expression of the inhibitory molecule, programmed death-1 ligand (PDL-1) compared to untreated cells. In vitro co-culture of ADN-DCs with allogeneic T cells led to a decrease in T cell proliferation and reduction of IL-2 production. Concomitant with that, a higher percentage of CD4 +CD25 +Foxp3 + regulatory T cells (Tregs) was detected in co-cultures of T cells and ADN-DCs. Blocking PD-1/PDL-1 pathway could partially restore T cell function. These findings suggest that the immunomodulatory effect of ADN on immune responses could be at least partially be mediated by its ability to alter DC function. The PD-1/PDL-1 pathway and the enhancement of Treg expansion are implicated in the immunomodulatory mechanisms.

1400 Nosocomial Infection and Cd4+ Lymphocytes in Very Low Birth Weight (VLBW) Infants' Cord Blood

Very low birth weight infants are at high risk for nosocomial infection specially due to higher survival rates related to advances in intensive care units. Prolonged hospital stay, instrumentalization and immaturity of their immune system are risk factors for infection, being gram-negative bacteria responsible for higher mortality rates and longterm sequelae. Patients and Methods: Prospective observational study in inborn VLBW infants, describing epidemiological features, morbidity and mortality in relation to risk factors sepsis and type of pathogen isolated in blood cultures. We perfomed flowcytomety analyses in cord blood samples and compared the lymphocyte subpopulations between the group of patients with nosocomial sepsis and the asymptomatic group. Results: 40 preterm infants were enrolled, mean birth weight (1059± 197 grams) and 29,18±2,05 weeks postmenstrual age. 70% developed late-onset sepsis, with 65% coagulase-negative Stahylococcus, 31% gramnegative bacteria and 4,4% Candida spp. Compared to those without sepsis, patent ductus arteriosus and bronchopulmonary dysplasia was significantly more frequent in the sepsis group (91,7% vs 8,3%; p=0,02; 85,7% vs 14,3%; p=0,064), with longer duration of mechanical ventilation and O2 requirement (3,40 vs 15,04days; p=0,044; 16 vs 46,96 days; p=0,01). CD4+ T lymphocytes were higher in umbilical cord blood in 28 VLBW neotates with sepsis (45,52+/-; 12,13 vs 36,05+/-;10,21; p=0,01) but no differences were observed in CD8+ T cells. Conclusions: Umbilical cord blood phenotype of immune system differs in VLBW infants with nosocomial infection, with a higher percentage of CD4+ cells. The meaning of this finding and other possible differences remain to be fully interpreted.

CD34-Subpopulations and Clonogenic Progenitors in Mobilized Peripheral Blood Cells from Patients with Acute Myeloid Leukemia

BACKGROUND: The use of autologous peripheral blood progenitor/stem cells as part of dose-intensified treatment protocols for patients with acute myeloid leukemia (AML) is under current clinical investigation. METHODS: We analyzed the frequency of CD 34 + 38 - and CD 34 + DR - subpopulations, clonogenic cells and long-term culture-derived clonogenic cells (LTC-DC) in peripheral blood (PB) samples from patients with AML after chemotherapy and G-CSF given for mobilization in comparison to a control population of patients with non-hematological malignancies. RESULTS: We observed a lower number of CD 34 + cells (31.2 ± 14.8 vs. 114 ± 36 /μl) and a reduction to less than 15 % for clonogenic progenitors (CFU) and LTC-DC in peripheral blood mononuclear cells from AML patients after consolidation therapy. In contrast, the quality of these CD 34 + cells was not significantly impaired after consolidation therapy determined by a moderately reduced frequency of CFU/CD34 + cells (5.0 ± 1.5 % vs. 10.0 ± 2.8 %), a nearly identical frequency of LTC-DC/CD34 + cells (0.5 ± 0.1 % vs. 0.6 ± 0.2 %), and a higher percentage of CD 34 + 38 - cells/CD34 + cells (8.4 ± 1.8 % vs. 3.8 ± 1.1 %) in comparison to the control population. CONCLUSION: Our data confirm that it is possible to mobilize CD 34 + cells into the peripheral blood of AML patients using chemotherapy and G-CSF. Nevertheless, the low absolute number of CFU and LTC-DC after high-dose ara-C treatment for AML has to be taken into consideration for the design of treatment protocols using autologous progenitor/stem cell transplantation.

Analysis of peripheral blood T‐cell subsets, natural killer cells and serum levels of cytokines in children with Down syndrome

The objective of this study was to evaluate the relationship between humoral and cell-mediated immune response parameters and impairment of immune functions in children with Down syndrome (DS). The patient group was consisted of cytogenetically documented 32 children with DS. Lymphocyte subsets and natural killer cells were counted by flow-cytometry system. Levels of interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10 and tumour necrosis factor-alpha (TNF-alpha) were detected by enzyme-linked immunosorbent assay method. Serum IgG, IgM, IgA levels were measured by turbidimetric methods. The percentage of CD8+ lymphocytes and CD56+ cells of patients with DS were significantly higher, whereas CD20+ lymphocytes were lower than that of controls (P < 0.05). The percentage of CD2 and CD4 levels and CD4/CD8 ratio of patients with DS and normal controls were similar (P > 0.05). Levels of IL-4 and IL-10 were significantly increased, but IL-6 and TNF-alpha levels were decreased in children with DS (P < 0.05). Levels of other studied cytokines between patients with DS and controls were not statistically different (P > 0.05, for all). Serum IgG, IgM and IgA levels were found to be similar between the groups (P > 0.05). It has been known that IL-4 and IL-10 are anti-inflammatory molecules which inhibit the synthesis of proinflammatory cytokines such as IL-6 and TNF-alpha. In this study, levels of IL-4 and IL-10 were significantly increased, but IL-6 and TNF-alpha levels were decreased in children with DS. These results may suggest that continuing anti-inflammatory state in DS and this process may explain the cause of recurrent infection of the disease. On the other hand, in contrast to the low percentage of CD20+ cells, high percentage of CD8+ and CD56+ cells were found. Our findings may demonstrate that the cell-mediated and humoral immune system parameters in children with DS were altered according to healthy children.

Effect of Feeding a Formula Supplemented With Long‐chain Polyunsaturated Fatty Acids for 14 Weeks Improves the Ex Vivo Response to a Mitogen and Reduces the Response to a Soy Protein in Infants at Low Risk for Allergy

Feeding long-chain polyunsaturated fatty acids (LCP) influences immunity in adults; however, less is known about their effect during development. The aim of the study was to determine the effect of feeding LCP on immunity in healthy infants during the first 4 months of life. Formula-fed infants were randomized at <or=14 days of age to standard term formula (Formula) or formula containing LCP (Formula+LCP). Infants exclusively fed human milk (HM) were included for comparison. At 16 weeks of age, blood was collected and phenotypes, the ability to proliferate and produce cytokines (interleukin [IL]-2, IL-4, IL-6, IL-10, IL-12, interferon [IFN]-gamma, tumor necrosis factor [TNF]-alpha, TGF-beta) after incubation with phytohemaglutinin (PHA), beta-lactoglobulin, or soy protein were measured. Feeding LCP resulted in a higher than and more similar proliferation rate to PHA in HM-fed infants, possibly because of a greater TH1 type cytokine response and a higher percentage of antigen mature (CD45RO+) cells (P < 0.05). The response to beta-lactoglobulin did not differ among groups. After incubation with soy protein Formula+LCP, compared with Formula produced less IL-2 and more TNF-alpha and had a higher percentage of CD8+ and a lower percentage of CD20+ (CD20+CD54+) cells poststimulation (P < 0.05). Both formula groups produced less IL-2 after PHA, had a lower percentage of CD80+ cells, and a higher percentage of CD54+ cells after incubation with food proteins (P < 0.05). Formula-fed infants, at low risk for allergy, respond differently to mitogen and food proteins ex vivo than those fed HM. Feeding LCP altered some of these differences in the direction that is hypothesized to confer immune benefits.

W1799 Defective In Vitro Macrophage Maturation in Crohn's Disease

Introduction Whist the cause of Crohn9s disease (CD) remains unclear, an innate defect in the handling of intracellular bacteria appears to be involved. Evidence for this comes from observations of a defect in phagocyte recruitment and the presence of viable E coli within lamina propria (LP) macrophages in these patients. LP macrophages are derived from blood monocytes and develop into pro inflammatory cells under the influence of Th1 cytokine (M1) or anti inflammatory regulatory macrophages (M2) by Th2 cytokines. It is known that macrophage turnover in CD is higher and maturation incomplete compared to those in healthy individuals so these cells are a good candidate for the observed differences. Methods To study in vitro the cytokine-driven differentiation of macrophages from patients with IBD (5 CD, 5 UC) and five healthy controls and follow their behaviour when challenged with CD derived strains of E coli . Monocytes were stimulated for 6 days with INF-γ, IL-4 + IL-13 or IL-10 to generate M1, M2a or M2c macrophage populations, respectively. Specific maturation markers (CD14, CD16, CD33, CD163, CD206, CD209, CCR2 and CCR7) were then measured using flow cytometry. These cells were then challenged with E coli and phagocytic function measured using the gentamicin protection assay. Results Monocytes stimulated with INFγ, from both the control and IBD groups gave a similar M1 type profile. IL-4+IL-13 stimulation gave rise to M2a cells, however the cells isolated from the CD group expressed a higher percentage of CD14 and a greater proportion of these cells expressed low levels of CD163 and CD206. In the CD group, IL-10 gave rise to a population that expressed CD14+, CD16+ and were CD163++, CD206+ and CD209+. When these different cell populations were challenged with E coli the number of recovered viable bacteria at 30 min gave similar numbers of internalised bacteria. However when we recovered bacteria 3 h post infection, significantly higher survival of bacteria in the IL-10, M2c population in the CD Group (M1 cells, 14±9; M2a cells, 23±8; M2c cells, 135±41) was found. No bacteria survived in the controls or UC. Conclusion We have demonstrated a population of IL-10 induced M2c cells that are unable to complete the process of bacterial killing effectively in patients with CD. These CD163+ cells are similar to a subpopulation of lamina propria macrophages that have been shown harbour viable bacteria. This may indicate that a defect in the migration and maturation of monocytes, unable to clear bacteria efficiently, may be the underlying innate defect in CD.

Biological characteristics and antitumor activity of CIK cells activated by recombinant human fibronectin for human lung cancer cell lines in vitro

背景与目的CIK细胞是过继免疫治疗的重要手段之一，简化体外培养过程从而提高其增殖率和杀瘤活性仍是目前研究的一个热点课题。本研究观察重组人纤维连接蛋白（recombinant human fibronectin, RN）诱导CIK细胞的生物学特性，建立一种高效、简便的体外CIK细胞扩增方法。方法抽取10名健康人外周静脉血各50 mL，用淋巴细胞分离液分离单个核细胞，分别采用RN诱导法和传统方法培养CIK细胞，记录细胞增殖数；用流式细胞术测定免疫细胞表型和分泌IFN-γ、IL-4、穿孔素和颗粒酶B细胞的百分比；用MTT法测定CIK细胞对4种人肺癌细胞株的体外杀伤率。结果RN诱导的CIK细胞扩增倍数为传统方法的2.0倍-3.5倍，具有统计学差异（P < 0.05）；RN诱导组和传统方法组CD3+CD16+CD56+细胞绝对数分别增加了3 778倍和2 069倍；RN诱导组细胞中CD3+CD8+细胞比例明显高于传统方法组（P < 0.05）；但CD3+CD4+细胞比例无统计学差异（P > 0.05）；对4种肺癌细胞株的体外杀伤活性无统计学差异（P > 0.05）。RN诱导的CIK较诱导前：分泌IFN-γ的细胞比例明显增加；分泌IL-4的细胞比例略有降低；释放穿孔素、颗粒酶B的阳性细胞比例较诱导前增加。结论RN诱导法是一种高效、简便的体外扩增CIK方法，可以替代传统方法。

Could a loss of memory T cells limit responses to hepatitis C virus (HCV) antigens in blood leucocytes from patients chronically infected with HCV before and during pegylated interferon-α and ribavirin therapy?

The proportions and activation status of T cells may influence responses to hepatitis C virus (HCV) and treatment outcome in patients receiving pegylated interferon (IFN)-alpha/ribavirin therapy. We confirmed that IFN-gamma enzyme-linked immunospot (ELISPOT) responses to HCV are poor in HCV patients and showed that responses to HCV and cytomegalovirus (CMV) antigens decrease during therapy. This was most apparent in patients with sustained virological response (SVR). Baseline frequencies of CD4+ effector memory (TEM) T cells were lower in SVR than non-SVR. Proportions of CD4+ and CD8+ TEM and terminally differentiated effector memory (TEMRA) T cells declined on therapy in SVR, as did proportions of Fas+ CD8+ TEMRA T cells. Baseline frequencies of programmed death (PD)-1-expressing CD4+ TEM and TEMRA T-cells were higher in SVR. Therapy increased percentages of PD-1+ CD4+ central memory (TCM) T cells and PD-1+ CD8+ TEM and TEMRA T cells in SVR. We conclude that successful therapy depletes circulating antigen-specific CD4+ T cell responses. This paralleled decreases in proportions of effector memory T cells and higher percentages of CD4+ TCM T cells expressing PD-1.

The role of CD8+ T cells in cutaneous leishmaniasis caused by Leishmania braziliensis (40.9)

Abstract The leishmaniases are a group of diseases that affect millions of people worldwide. In the present study, we investigated the participation of CD8+ T cells during infection with L. braziliensis, the main etiolological agent of mucocutaneous leishmaniasis. Following inoculation of L. braziliensis into the ear dermis of BALB/c mice, we observed a high proportion of CD8+CD25+, at the time of lesion appearance, whereas CD8+CD69+ T cells were found at a greater proportion in the draining lymph nodes, at this same time point. Regarding cytokine production, a high percentage of CD8+IFN-g+ cells was detected as early as two weeks post infection, at the ear dermis. The same kinetics was observed for expression of CD107b. Interestingly, presence of IL-4+ and IL-10+ T cells was also detected during the course of infection in the ear dermis. Parasite load in β2-m deficient mice was smaller than in wild type (WT) mice. However, transfer of CD8+ cells obtained from L. braziliensis-primed mice induced the control of lesion development upon transfer to infected mice. These data indicate that CD8+ T cells may play a protective role in cutaneous leishmaniasis caused by L. braziliensis.

Distinct Abnormalities in the Innate Immune System of Children with Down Syndrome

To analyze the frequency and phenotype of cells of the innate immune system in the peripheral blood of children with Down syndrome (DS). Flow cytometric analysis of expression of cell surface markers was performed in children with DS (n = 41) and healthy age-matched controls (n = 41). Compared with controls, children with DS had significantly lower absolute total leukocyte counts, lymphocytes, monocytes, and granulocytes, but 1.5-times higher absolute numbers of CD14(dim)CD16(+) monocytes (147 x 10(6)/L vs 93 x 10(6)/L; P = .02). This difference is fully explained by a higher percentage of CD14(dim)CD16(+) monocytes within the monocyte compartment (28.7% vs 13.4%; P <.001). The absolute numbers of myeloid dendritic cells were lower in DS (13.8 x 10(6)/L vs 22.7 x 10(6)/L; P <.001). The numbers of plasmacytoid dendritic cells and natural killer cells were normal. Absolute numbers of invariant natural killer T cells were very low overall, but significantly lower in children with DS than in controls (1.2 x 10(6)/L vs 3.7 x 10(6)/L; P = .01). Children with DS exhibited distinct abnormalities in cells of the innate immune system. Most strikingly, they had a high number of proinflammatory CD14(dim)CD16(+) monocytes. This elevated level of CD14(dim)CD16(+) monocytes may play an important role in the onset and maintenance of chronic inflammatory disease in DS.

Forkhead Box P3+ Regulatory T Cell Underlies Male Resistance to Experimental Type 2 Autoimmune Hepatitis

Autoimmune hepatitis (AIH), like many autoimmune diseases, is most prevalent in young women. The immunological basis of this age and sex susceptibility bias was investigated in a murine model of AIH. Xenoimmunization of 7-week-old female C57BL/6 mice resulted in more severe AIH with higher levels of liver inflammation, serum alanine aminotransferase, specific T-cell cytotoxicity, and autoantibody than younger and older females. Vaccinated males developed minimal liver inflammation and higher percentages of CD4(+)CD25(+)FoxP3(+) regulatory T cell in peripheral blood mononuclear cells, spleen, and liver than females. Regulatory T cells (Tregs) were virtually absent in liver-lymphocytes infiltrates of females. Castration of C57BL/6 mice, with or without 17beta-estradiol supplementation, did not modify susceptibility in males, nor Treg numbers, suggesting minimal contribution of testosterone and estradiol to autoimmune hepatitis (AIH) susceptibility. Xenoimmunized Aire(+/0) mouse displayed similar AIH susceptibility, sex bias, and Tregs numbers as C57BL/6 mice, suggesting that susceptibility in females is not the result of less stringent thymic central tolerance. Autoreactive B cell response against formiminotransferase-cyclodeaminase correlated with disease activity, possibly linking B-cell autoreactivity and AIH pathogenesis. Peripheral tolerance and development of regulatory T cells after self-mimicking antigen exposure, and not sexual hormone nor central tolerance, are the main factors for susceptibility to AIH in females.

Short-term administrations of a combination of anti-LFA-1 and anti-CD154 monoclonal antibodies induce tolerance to neonatal porcine islet xenografts in mice.

OBJECTIVEThe objective of this study was to determine whether tolerance to neonatal porcine islet (NPI) xenografts could be achieved by short-term administrations of anti–LFA-1 and anti-CD154 monoclonal antibodies (mAbs).RESEARCH DESIGN AND METHODSDiabetic B6 mice received NPI transplants and short-term injections of combined anti–LFA-1 and anti-CD154 mAbs. Mice with long-term islet graft function were treated with depleting anti-CD25 mAb or re-transplanted with a second-party NPI. At the end of the study, grafts from mice with long-term islet function were examined. Their spleen cells were characterized and used for in vitro proliferation and adoptive transfer studies.RESULTSAll mAb-treated NPI recipients maintained normoglycemia for >100 days post-transplantation. Only 5 of 50 mice rejected their grafts before 300 days post-transplantation. Intact islets, foxp3+ immune cells, as well as interleukin (IL)-10 and transforming growth factor (TGF)-β regulatory cytokine transcripts were detected in the NPI xenografts from tolerant mice. A higher percentage of CD4+ T-cell population from these mice expressed regulatory markers, suggesting that tolerance to NPI xenografts may be mediated by T regulatory cells. This was confirmed when tolerant mice treated with depleting anti-CD25 mAb became diabetic. Lymphocytes from tolerant mice inhibited the proliferation of lymphocytes from B6 mice immunized with porcine cells and they displayed limited proliferation when adoptively transferred. All protected B6 mice transplanted with a second-party NPI xenograft maintained long-term normoglycemia even after removal of the first NPI graft-bearing kidney.CONCLUSIONSThese results demonstrate that tolerance to NPI xenografts can be achieved by transient administrations of combined anti–LFA-1 and anti-CD154 mAb therapy.

Effect of Different Dialysis Modalities on Microinflammatory Status and Endothelial Damage

We studied the relationship between microinflammation and endothelial damage in chronic kidney disease (CKD) patients on different dialysis modalities. Four groups of CKD stage 5 patients were studied: 1) 14 nondialysis CKD patients (CKD-NonD); 2) 15 hemodialysis patients (HD); 3) 12 peritoneal dialysis patients with residual renal function >1 ml/min (PD-RRF >1); and 4) 13 peritoneal dialysis patients with residual renal function <or=1 ml/min (PD-RRF <or=1). Ten healthy subjects served as controls. CD14(+)CD16(+) cells and apoptotic endothelial microparticles (EMPs) were measured by flow cytometry. Serum vascular endothelial growth factor (VEGF) was measured by ELISA. CKD-NonD and HD patients had a higher percentage of CD14(+)CD16(+) monocytes than PD groups and controls. CD14(+)CD16(+) was similar in the PD groups, regardless of their RRF, and controls. The four uremic groups displayed a marked increase in apoptotic EMPs and VEGF compared with controls. Apoptotic EMPs and VEGF were significantly higher in HD patients than in CKD-NonD and both PD groups. However, there were no significant differences between CKD-NonD and the two PD groups. There was a correlation between CD14(+)CD16(+) and endothelial damage in CKD-NonD and HD patients, but not in PD and controls. There was an increase in CD14(+)CD16(+) only in CKD-NonD and HD patients. In these patients, there was a relationship between increased CD14(+)CD16(+) and endothelial damage. These results strongly suggest that other factors unrelated to the microinflammatory status mediated by CD14(+)CD16(+) are promoting the endothelial damage in PD, regardless of their RRF.

Differential Cell Analysis and Lymphocyte Subset Analysis in Bronchoalveolar Lavage Fluid from Patients with Miliary Tuberculosis

Background: Bronchoalveolar lavage (BAL) is a useful technique to recover lower airway fluid and cells involved in many respiratory diseases. Miliary tuberculosis is potentially lethal, but the clinical manifestations are nonspecific and typical radiologic findings may not be seen until late in the course of disease. In addition, invasive procedures are often needed to confirm disease diagnosis. This study analyzed the cells and the T-lymphocyte subset in BAL fluid from patients with miliary tuberculosis to determine specific characteristics of BAL fluid that may help in the diagnosis of miliary tuberculosis, using a less invasive procedure. Methods: On a retrospective basis, we enrolled 20 miliary tuberculosis patients; 12 patients were male and the mean patient age was years. We analyzed differential cell counts of BAL fluid and the T-lymphocyte subset of BAL fluid. Results: Total cells and lymphocytes were increased in number in the BAL fluid. The percentage of CD4+ Tlymphocytes and the CD4/CD8 ratio in BAL fluid were significantly decreased and the percentage of CD8+ T-lymphocytes was relatively higher. These findings were more prominent in patients infected with the human immunodeficiency virus (HIV). In the HIV-infected patients, the proportion of lymphocytes was significantly higher in BAL fluid than in peripheral blood. There were no significant differences between the BAL fluid and the peripheral blood T-lymphocytes subpopulation. Conclusion: BAL fluid in patients with miliary tuberculosis demonstrated lymphocytosis, a lower percentage of CD4+ T-lymphocytes, a higher percentage of CD8+ T-lymphocytes, and a decreased CD4/CD8 ratio. These findings were more significant in HIV-infected subjects.