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PubMed2003

The source of fatty acids incorporated into proliferating lymphoid cells in immune-stimulated lymph nodes.

To explore the hypothesis that proliferating lymphoid cells in immune-stimulated lymph nodes obtain nutrients locally from adjacent adipose tissue, adult guinea pigs were fed for 6 weeks on standard chow or on chow supplemented with 100 g suet, sunflower oil or fish oil/kg. All the guinea pigs ate standard chow for the last 5 d, during which swelling of one popliteal lymph node was stimulated by repeated local injection of lipopolysaccharide. The fatty acid compositions of phospholipids in both popliteal and in several mesenteric lymph nodes, and of triacylglycerols in eleven samples of adipose tissue defined by their anatomical relations to lymph nodes, were determined by GC. The proportions of fatty acids in the phospholipids extracted from the stimulated popliteal node correlated best with those of triacylglycerols in the surrounding adipocytes, less strongly with those of adipocytes elsewhere in depots associated with lymphoid tissue, but not with those of nodeless depots. The composition of triacylglycerols in the perinodal adipose tissue changed under local immune stimulation. We conclude that proliferating lymphoid cells in activated lymph nodes obtain fatty acids mainly from the triacylglycerols in adjacent perinodal adipose tissue. Immune stimulation prompts changes in the fatty acid composition of the triacylglycerols of adipocytes in node-containing depots that equip the adipose tissue for provisioning immune responses. Such local interactions show that specialised adipocytes can act as an interface between whole-body and cellular nutrition, and may explain why mammalian adipose tissue is partitioned into a few large and many small depots.

Adipocytes↗
PubMed1976

Idiopathic late-onset immunoglobulin deficiency with functional T-cell deficiency.

The common features in our two patients were late onset of infections that are known to complicate both T-cell and B-cell deficiencies, decreased numbers of circulating B-cells with low serum immunoglobulin levels, and normal numbers of circulating T-cells, which were, however, defective in the response to delayed hypersensitivity skin test antigens and to mitogens in vitro. The composite of clinical and immunologic aberrations is consistent with the presence of an immune deficiency involving the B-cell system quantitatively and the T-cell system qualitatively.

Adolescent↗
PubMed2003

Archaeosomes as self-adjuvanting delivery systems for cancer vaccines.

Archaeal ether glycerolipid vesicles (archaeosomes) efficiently deliver exogenous antigen for induction of humoral and cell-mediated immunity. Because induction of CD8 cytotoxic T cells is critical for protective vaccination against tumors, we compared the ability of various archaeosome lipid compositions to evoke a strong CD8 CTL response to entrapped antigen. Subcutaneous immunization of mice with ovalbumin (OVA) entrapped in all archaeosome lipid compositions evoked a primary (day 10) splenic CTL response indicating processing for MHC class I presentation. Interestingly, several polar lipid compositions from halophilic archaea were very potent to adjuvant this early CTL response. Despite this, the lytic units reduced substantially by weeks 6-7. More importantly, at >50 weeks, only Methanobrevibacter smithii and Thermoplasma acidophilum both rich in bipolar membrane-spanning caldarchaeols, demonstrated recall memory CTLs. Immunization of mice with OVA entrapped in M. smithii, Halobacterium salinarum or T. acidophilum vesicles provided prophylactic protection against challenge with OVA-expressing solid tumors at 6 weeks. Even a dose of 3 microg OVA in archaeosomes significantly delayed tumor growth. Tumor protection was also noted in a therapeutic design wherein OVA-archaeosomes were injected concurrent with the tumor challenge. Interestingly, antigen-free T. acidophilum but not antigen-free H. salinarum archaeosomes provided innate therapeutic protection. Vaccination with a CTL peptide epitope from the melanoma differentiation antigen, tyrosinase-related protein 2, in archaeosomes induced a protective CD8 response against B16OVA metastasis, indicating potential for targeting self, tumor antigens. Thus, lipid structural properties of archaea may differentially modulate primary, long-term and/or innate immunity, impacting adjuvant choice for vaccine design.

Animals↗
PubMed1995

Epidermal growth factor promotes a neural phenotype in thymic epithelial cells and enhances neuropoietic cytokine expression.

Neural crest-derived cells populate the thymus, and their coexistence with epithelial cells is required for proper organ development and T cell education function. We show here that epidermal growth factor (EGF), a major epithelial cell growth-enhancing agent, has a morphogenetic action to promote the expression of a neuronal phenotype (e.g., neurofilament expression) in cultured thymic epithelial cells that are characterized by a cytokeratin-positive epithelial cell background. The proliferation of such neurodifferentiated cells is also enhanced by EGF. Furthermore, the growth factor enhances cells that express the genes encoding the preprotachykinin A-generated neuropeptides and bipotential neuropoietic and lymphopoietic cytokines ciliary neurotrophic factor and interleukin-6. These cytokines also enhance the neuronal phenotype of thymic epithelial cells. Therefore, EGF appears to be a composite autocrine/paracrine neuromodulator in thymic stroma. This suggests that EGF may regulate thymus-dependent immune functions by promoting neuronal gene expression in neural crest-derived cells.

Animals↗
PubMed2001

Spatial organization of signal transduction molecules in the NK cell immune synapses during MHC class I-regulated noncytolytic and cytolytic interactions.

The cytolytic activity of NK cells is tightly regulated by inhibitory receptors specific for MHC class I Ags. We have investigated the composition of signal transduction molecules in the supramolecular activation clusters in the MHC class I-regulated cytolytic and noncytolytic NK cell immune synapses. KIR2DL3-positive NK clones that are specifically inhibited in their cytotoxicity by HLA-Cw*0304 and polyclonal human NK cells were used for conjugate formation with target cells that are either protected or are susceptible to NK cell-mediated cytotoxicity. Polarization of talin, microtubule-organizing center, and lysosomes occurred only during cytolytic interactions. The NK immune synapses were analyzed by three-dimensional immunofluorescence microscopy, which showed two distinctly different synaptic organizations in NK cells during cytolytic and noncytolytic interactions. The center of a cytolytic synapse with MHC class I-deficient target is comprised of a complex of signaling molecules including Src homology (SH)2-containing protein tyrosine phosphatase-1 (SHP-1). Closely related molecules with overlapping functions, such as the Syk kinases, SYK, and ZAP-70, and adaptor molecules, SH2 domain-containing leukocyte protein of 76 kDa and B cell linker protein, are expressed in activated NK cells and are all recruited to the center of the cytolytic synapse. In contrast, the noncytolytic synapse contains SHP-1, but is lacking other components of the central supramolecular activation cluster. These findings indicate a functional role for SHP-1 in both the cytolytic and noncytolytic interactions. We also demonstrate, in three-cell conjugates, that a single NK cell forms a cytolytic synapse with a susceptible target cell in the presence of both susceptible and nonsusceptible target cells.

Cell Polarity↗
PubMed1993

Association of the lactococcin A immunity factor with the cell membrane: purification and characterization of the immunity factor.

The physicochemical characteristics of the lactococcin A immunity protein, as deduced from its gene sequence, were used to devise a procedure for its purification. The protein was purified from cell extracts by cation-exchange and reverse-phase chromatography. As judged from the amino acid composition and amino acid sequencing, the immunity protein is not post-translationally processed by cleavage at its N- or C-terminus. Consequently, the absorption coefficient at 280 nm, the isoelectric point, and the molecular mass of the immunity protein may be calculated to be, respectively, 8.2 x 10(3) M-1 cm-1, 10.2 and 11,163 Da from the amino acid sequence predicted from the nucleotide sequence. The immunity protein is a major cell protein component--one cell may contain (to an order of magnitude) 10(5) molecules--and it is in part associated with the cell membrane, as judged by immunoblot analysis of membrane vesicle-associated proteins. Exposing lactococcin-A-sensitive cells to an excess of the immunity protein did not affect the lactococcin-A-induced killing of the cells, indicating that the immunity protein does not protect cells by simply binding to lactococcin A, nor to externally exposed domains on the cell surface. Exposing immune-positive cells to antiserum against the immune protein did not sensitize the cells to lactococcin A, suggesting that the immunity protein in fact does not act extracellularly.

Amino Acid Sequence↗
PubMed1983

Comparison of highly NK active human lymphocyte subsets separated by various procedures involving E, EA rosetting, density gradients and adherence to immune complexes.

Lymphocyte subsets from human blood obtained by different procedures were analyzed for cytotoxic potential and phenotypic characteristics. Nylon wool column passed lymphocytes were fractionated on the basis of: (1) E and Fc gamma receptor expression, (2) cell density and Fc gamma receptor expression, (3) Fc gamma receptor expression. The cytotoxic subsets obtained by separation on the basis of E and EA rosetting differed in their phenotypic composition from those separated on the basis of density or on immune complex monolayers. The E- Fc gamma- population contained few LGL and OKM1 positive cells. The E- Fc gamma+ population was made up almost entirely of LGL and OKM1 positive cells. The low density population was highly enriched in LGLs; among these the Fc gamma- cells were OKT3 positive. In contrast to the E- population the low dense Fc gamma+ cells were mainly LGLs and were OKM1 positive. Fc gamma+ subsets had less killer activity against Daudi cells. The choice of procedure for obtaining a strongly cytotoxic population depends on the needs of particular experiments. Separation on the basis of E rosetting gave lower cell (62%) and cytotoxic (43%) recovery and required about twice the amount of blood and twice the time, as compared with the other 2 procedures. The cell fractions obtained this way allowed characterization of several phenotypically different active populations and showed a difference in cytotoxic potential against K562 and Daudi cells. Density fractionation isolated a highly cytotoxic subset with LGL morphology but this population was still heterogeneous phenotypically. With regard to enrichment of NK activity, the immune complex monolayer attachment method was the most efficient for total cell recovery and for the time taken to perform it.

Antigen-Antibody Complex↗
PubMed2006

PCL-PU composite vascular scaffold production for vascular tissue engineering: attachment, proliferation and bioactivity of human vascular endothelial cells.

A new compliant scaffold suitable for small-diameter vascular grafts has been developed that promotes strong attachment of endothelial cells. Composite scaffolds were produced by wet spinning polycaprolactone (PCL) fibres which form the luminal surface, then electrospinning porous polyurethane (PU) onto the back of the PCL fibres to form the vessel wall substitute. Human endothelial cells demonstrated strong attachment to the composite PCL-PU scaffold, and proliferated to form a monolayer with strong PECAM-1 expression and cobblestone morphology. Attached cells demonstrated abundant release of von Willebrand factor, nitric oxide and ICAM-1 under physiological stimuli, and exhibited an immune response to lipopolysaccharide. The composite scaffold may also deliver bioactive molecules. Active trypsin, used as a test molecule, had a defined 48 h pattern of release from luminal PCL fibres. These data confirm the potential of this novel composite scaffold in vascular tissue engineering.

Biocompatible Materials↗
PubMed1974

Local antibody production against the murine toxin of Yersinia pestis in a golf ball-induced granuloma.

The method of Garra and Baygorria, for localized antibody production, has been adapted for obtaining high-titered and monospecific antibodies against the murine toxin of Yersinia pestis. A hollow, perforated plastic golf ball surgically implanted under the skin of rabbits induced the formation of a granuloma. When the murine toxin of Y. pestis was injected directly into the granulomatous cavity, an increased amount of antibody was found in the granuloma fluid as compared with serum or with serum antibody obtained by conventional immunization. The granuloma antibody consisted mainly of immunoglobulin G, probably produced locally by lymphoid cells of the granuloma. The immune granuloma fluid and the granuloma tissue were rich in plasma cells and lymphocytes. The chemical composition of the granuloma fluid indicated that it was a transudate.

Animals↗
PubMed1995

The in vivo effects of neutralizing antibodies against IFN-gamma, IL-4, or IL-10 on the humoral immune response in young and aged mice.

In the present study we investigated whether age-related changes in the composition and functional properties of murine CD4+ T cells are reflected in vivo by a changed humoral response to influenza vaccine in aged mice. After the primary immunization, the titers of influenza-specific IgM, IgG1, IgG2a, and IgG2b, but not of IgG3 and IgE, were significantly reduced in aged mice compared to young mice. Treatment of aged mice with anti-IFN-gamma, anti-IL-4, or anti-IL-10 resulted in levels of IgM and IgG1 comparable to those found in young mice, whereas IgG2a and IgG2b were further decreased. After the booster immunization IgE was significantly enhanced in aged mice, whereas no differences were observed with regard to the other isotypes. During the primary response in young mice, anti-IFN-gamma stimulated IgG1 and IgE, whereas an inhibition of IgG2a, IgG2b, and IgG3 was observed. Anti-IL-4 caused a decrease only in IgG3 while anti-IL-10 increased IgM and IgG1 and decreased IgG2b and IgG3. During the primary response in aged mice, all anti-cytokine antibodies enhanced IgM and IgG1 while IgE was only enhanced by anti-IL-10. By contrast, IgG3 was inhibited by anti-IFN-gamma and anti-IL-10. Anti-cytokine treatment of young mice increased all isotypes, except IgG3, in the secondary response, whereas the secondary response in aged mice was largely insensitive to anti-cytokine treatment. These data therefore support the idea that the in vivo effects of cytokines on isotype switching are dependent on the differentiation stage of B cells which may be different in young and aged mice.

Aging↗
PubMed2026

Plasmacytoid dendritic cell-mediated L-glutamate catabolism links gut microbiota to male infertility.

Emerging evidence suggests that gut microbiota composition influences male reproductive health; however, the immunometabolic mechanisms underlying this association remain insufficiently characterized. We investigated whether specific immune cell-mediated metabolic pathways, particularly plasmacytoid dendritic cell (pDC)-driven L-glutamate catabolism via the hydroxyglutarate pathway, contribute to the causal link between gut microbiota and male infertility. We conducted a 2-sample, 2-step Mendelian randomization (MR) analysis using inverse-variance weighting as the primary estimator and Bayesian weighted MR for robustness. Exposure data comprised 412 gut microbial taxa/metabolic pathways and 731 immune cell phenotypes from large European-ancestry genome-wide association studies. Male infertility genome-wide association studies data (1429 cases; 128,710 controls) were obtained from FinnGen R10. Only exposure-mediator-outcome pairs meeting stringent pleiotropy, heterogeneity, and reverse-causality criteria were retained for mediation analysis. Nine microbial taxa/metabolic pathways and 18 immune traits exhibited putative causal associations with male infertility. The L-glutamate degradation V pathway via hydroxyglutarate was linked to reduced infertility risk (inverse-variance weighting odds ratio [OR] = 0.68; 95% confidence interval, 0.52-0.89; P = .005). Two-step MR suggested that forward scatter area on pDCs may mediate this association, although the mediation effect was imprecise (effect = 0.0277; 95% confidence interval, -0.0348 to 0.0903). This study provides suggestive genetic evidence that pDC-mediated glutamate catabolism may connect gut microbial metabolic activity to male infertility. These findings highlight immunometabolic pathways as testable targets for mechanistic validation and microbiota-directed interventions.

Male↗
PubMed1994

[Significance of fatty acid composition in plasma and in food for cellular immune function in elderly men].

The relation between fatty acid composition in plasma and natural killer (NK) cell activity and the relation between fatty acid composition of diet and NK cell activity was evaluated in healthy elderly men. The correlations between basal NK activity and the fraction of plasma fatty acids consisting of total polyunsaturated fatty acids (PUFA), total n-6 fatty acids and linoleic acid were r = -0.68, p = 0.006, r = 0.62, p = 0.014 and r = 0.52, p = 0.048, respectively. Significant negative correlations were also found between alpha-interferon stimulated NK cell activity and the three groups of fatty acids and between interleukin-2 stimulated NK cell activity and PUFA. Likewise, negative correlations between grammes of PUFA in diet, determined from two four-day registration-periods, and basal NK and alpha-interferon stimulated NK cell activity were found. No significant negative correlation between percentage intake of n-3 fatty acids and NK cell activity was found. It is concluded that the type of dietary fatty acids influence NK cell activity in elderly men. A high intake of n-6 polyunsaturated fatty acids may be detrimental to cellular immune defence mechanisms in the elderly.

Aged↗
PubMed1981

[Effect of inducers of mixed-function microsomal oxidases on the immune response of mice induced by heterologous erythrocytes].

In experiments on male white mice immunized with sheep red blood cells the influence of various substances, differing in their chemical composition and biological activity, on the antibody titers and the number of rosette-forming cells in the spleen has been studied. These substances, acting as inductors of microsomal oxidases with mixed function (cytochrome P-450 and P-448), are phenobarbital, 3-methylcholanthrene, DDT, diphenin, rifampicin, benzodiazepin derivatives (diazepam and phenazepam). All monoxygenase activity inductors have been shown to exert a more or less pronounced immunosuppressive effect. Thus, reciprocal relations between the induction of monoxygenases and the induction of immune response have been established. The results obtained in this research are discussed from the viewpoint of the new hypothesis stating that immunity is one of the functions of the general system metabolizing chemical compounds in the body.

Animals↗
PubMed1987

Exogenous growth hormone treatment alters body composition and increases natural killer cell activity in women with impaired endogenous growth hormone secretion.

In order to assess the potential relationship between human growth hormone (GH) and body composition (BC) and natural immunity (NI), we measured the effects of exogenous GH on fat weight (FW), fat-free weight (FFW), and the cytotoxic activity of natural killer (NK) cells in women with impaired GH secretion. Mean peak serum concentrations of GH in response to L-dopa/arginine stimulation were 6.2 +/- 1.1 (SEM) ng/mL in 6 untreated subjects (US) and 5.4 +/- 1.5 ng/mL in 6 GH-treated subjects (TS). Moreover, the pretreatment circulating levels of IGF-I were low in both groups (US 684 +/- 121 mU/mL and TS 583 +/- 83 mU/mL), and they correlated with pretest levels of NK cell activity (r = .59, P less than .05) when both groups were combined. The TS were given 700 micrograms of human GH IM for an average of 14 days while the US were studied in parallel without GH treatment. As measured by hydrodensitometry or skinfold anthropometry, FW decreased (26.1 +/- 6.8 kg to 23.8 +/- 6.3 kg, P less than .05) and FFW increased (44.9 +/- 3.3 kg to 46.2 +/- 3.8 kg, P less than .05) in the TS. In the US, there were no significant (P less than .05) changes in either FW or FFW. Using a standard 51Cr release assay to measure the specific lytic (SL) activity of NK cells, mean SL activity increased from 24.4 +/- 7.0% to 44.1 +/- 8.9% (P less than .05) in the TS, whereas levels in the US were not altered significantly (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1997

Immunosuppressive phenotype of corticotropin-releasing factor transgenic mice is reversed by adrenalectomy.

Stress elicits a wide range of physiological changes involving the nervous, endocrine, and immune systems. Corticotropin-releasing factor (CRF) plays a key role in orchestrating this response, activating both the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis, resulting in release of corticosteroids. The present study examines the immunological phenotype and responsiveness of CRF-transgenic (CRF-Tg) mice. The immune system of the CRF-Tg animals has profound changes compared to littermate controls, including a marked reduction in both cell number and immune responsiveness. There were also phenotypic changes in the lymphocytic composition of the various lymphoid organs, most notably in the spleen, where CRF-Tg mice had a greater percentage of T lymphocytes compared to littermate controls. Adrenalectomy of CRF-Tg reversed the immunological phenotype observed and restored immune responsiveness. These results demonstrate that CRF overexpression leads to profound impairment on lymphocyte development and function mediated via corticosteroids.

Adrenalectomy↗
PubMed2000

Essential fatty acids in the nutrition of severely neurologically disabled children.

Essential fatty acids (EFA) are important for the normal development and functioning of the brain, retina and immune competent cells. Severely neurologically handicapped children often have feeding difficulties, and the composition of the diet may be critical with respect to an optimal nutrient content. The aim of the present investigation was to evaluate if the dietary intakes and serum phospholipid concentrations of EFA were adequate in a group of severely neurologically impaired children in an institution. To achieve this, a prospective study was done. The investigation showed low dietary intakes of both n-6 fatty acids (FA) and n-3 FA. The serum concentrations of total n-6 FA, linoleic acid and 22:6n-3 (docosahexaenoic acid) as proportions of the total serum phospholipid FA concentration were initially low. The serum concentrations of 20:3n-9 and 22:5n-6 cholesterol, triacylglycerol, total saturated FA, total monounsaturated FA and apolipoproteins A-I and B were high compared with levels in a reference group of healthy children. Following supplementation with fish oil and soyabean oil, the serum lipid profile approached normal. We conclude that the study children had suboptimal intakes of EFA and that elevated serum concentrations of 20:3n-9 and 22:5n6 were useful serological markers of suboptimal EFA status. Recommended dietary allowances for EFA given as a percentage of energy underestimate EFA requirements in children with a low energy intake. Severely disabled children with feeding difficulties should probably be monitored with serum phospholipid FA measurements or calculation of dietary absolute intakes of EFA.

Adolescent↗
PubMed1994

Proliferating dendritic cell progenitors in human blood.

CD34+ cells in human cord blood and marrow are known to give rise to dendritic cells (DC), as well as to other myeloid lineages. CD34+ cells are rare in adult blood, however, making it difficult to use CD34+ cells to ascertain if DC progenitors are present in the circulation and if blood can be a starting point to obtain large numbers of these immunostimulatory antigen-presenting cells for clinical studies. A systematic search for DC progenitors was therefore carried out in several contexts. In each case, we looked initially for the distinctive proliferating aggregates that were described previously in mice. In cord blood, it was only necessary to deplete erythroid progenitors, and add granulocyte/macrophage colony-stimulating factor (GM-CSF) together with tumor necrosis factor (TNF), to observe many aggregates and the production of typical DC progeny. In adult blood from patients receiving CSFs after chemotherapy for malignancy, GM-CSF and TNF likewise generated characteristic DCs from HLA-DR negative precursors. However, in adult blood from healthy donors, the above approaches only generated small DC aggregates which then seemed to become monocytes. When interleukin 4 was used to suppress monocyte development (Jansen, J. H., G.-J. H. M. Wientjens, W. E. Fibbe, R. Willemze, and H. C. Kluin-Nelemans. 1989. J. Exp. Med. 170:577.), the addition of GM-CSF led to the formation of large proliferating DC aggregates and within 5-7 d, many nonproliferating progeny, about 3-8 million cells per 40 ml of blood. The progeny had a characteristic morphology and surface composition (e.g., abundant HLA-DR and accessory molecules for cell-mediated immunity) and were potent stimulators of quiescent T cells. Therefore, large numbers of DCs can be mobilized by specific cytokines from progenitors in the blood stream. These relatively large numbers of DC progeny should facilitate future studies of their Fc epsilon RI and CD4 receptors, and their use in stimulating T cell-mediated resistance to viruses and tumors.

Animals↗
PubMed1983

Monoclonal antibodies to three widely distributed human cell surface antigens.

Four mouse monoclonal antibodies to widely distributed human cell surface antigens are described. The AJ2 antigen is one of the immunodominant antigens in mice immunized with human cells, as demonstrated by testing conventional mouse antisera. This antigen has four subunits having molecular weights (MW) of 170 kD, 140 kD, 140 kD, and 28 kD when immunoprecipitated from the immunizing cell line, an astrocytoma. With other cell lines the molecular weights and subunit composition varies, but the major component is always of MW 125-140 kD. The antigen was detected on all human tissue culture cells and on sections of all normal human tissues tested. Antibody MA99 detects the same antigenic complex but shows a slightly different pattern of reactivity with cell lines and tissues. The MA103 antigen has a similar ubiquitous distribution. It is a glycoprotein of MW 50-55 kD and the determinant detected is not denatured by treatment at 100 degrees C. MH99, the third common antigen detected, has two glycosylated subunits with MW of 29 kD and 38 kD. It is present on a subpopulation of tumor cell lines of most histologic types except that it is absent or weakly expressed on melanomas and astrocytomas. MH99 is present on some T lymphomas and leukemias and myeloid leukemias but was not detected on B lymphomas, null cell lymphomas and myelomas. It was detected preferentially on epithelial cells in tissue sections.

Animals↗