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Fatty acids and lymphocyte functions.

The immune system acts to protect the host against pathogenic invaders. However, components of the immune system can become dysregulated such that their activities are directed against host tissues, so causing damage. Lymphocytes are involved in both the beneficial and detrimental effects of the immune system. Both the level of fat and the types of fatty acid present in the diet can affect lymphocyte functions. The fatty acid composition of lymphocytes, and other immune cells, is altered according to the fatty acid composition of the diet and this alters the capacity of those cells to produce eicosanoids, such as prostaglandin E2, which are involved in immunoregulation. A high fat diet can impair lymphocyte function. Cell culture and animal feeding studies indicate that oleic, linoleic, conjugated linoleic, gamma-linolenic, dihomo-gamma-linolenic, arachidonic, alpha-linolenic, eicosapentaenoic and docosahexaenoic acids can all influence lymphocyte proliferation, the production of cytokines by lymphocytes, and natural killer cell activity. High intakes of some of these fatty acids are necessary to induce these effects. Among these fatty acids the long chain n-3 fatty acids, especially eicosapentaenoic acid, appear to be the most potent when included in the human diet. Although not all studies agree, it appears that fish oil, which contains eicosapentaenoic acid, down regulates the T-helper 1-type response which is associated with chronic inflammatory disease. There is evidence for beneficial effects of fish oil in such diseases; this evidence is strongest for rheumatoid arthritis. Since n-3 fatty acids also antagonise the production of inflammatory eicosanoid mediators from arachidonic acid, there is potential for benefit in asthma and related diseases. Recent evidence indicates that fish oil may be of benefit in some asthmatics but not others.

Adult↗

[Composition of HIV-1 B-epitopes and heterologous T-cell epitopes. II. Combined analysis of immunogenicity of composite peptides in rabbits].

Rabbits were immunized with a mixture of linear cojoined peptides (composites) combining an immunodominant B-cell epitope from gp41 of HIV-1 (amino acids 593-604) and a heterogeneous T-cell epitope from tetanus toxoid or hepatitis B core antigen. HIV-1 B-epitope in the context of the composites induced HIV-1 specific antibody response, while no antibodies were produced in response to the peptide representing single HIV-1 B-epitope. Competition ELISA and ELISA of HIV-1 peptide omission analogous demonstrated that immunization induced antibodies of the correct specificity. T-Cell epitopic components also elicited potent antibody production. T-Cell epitopes appeared to be immunodominant. However, HIV-1 specific antibody response was not suppressed, on the contrary, antibody response against HIV-1 B- and heterologous T-cell epitopes developed in parallel. Cellular responses against composite peptides and/or their components were induced in all animals. The ability to develop cellular response against T-cell epitopes from the composites determined the efficacy of humoral response against the composites including HIV-1 specific one. Animals with strong cellular responses rapidly developed maximum antipeptide antibody titers. The immunization results imply that colinear fusion of B- and T-cell epitopes yielded bivalent multiepitopic products in which the immunological properties of the individual components were maintained and desired immunogenic properties were obtained. Immunogenicity of the composite peptides corresponded to that expected from the study of the composite antigenic properties.

Animals↗

Use of a 35.5 kDa cell membrane composition of Pasteurella multocida and an anti-idiotype antibody to induce protective immunity in leghorn chickens.

Using polyacrylamide gel electrophoresis and Western blot analysis, a 35.5 kDa cell membrane composition of Pasteurella multocida ATCC 11039 was identified as a dominant epitope of the bacterium. Mice inoculated with inactivated whole bacteria produce antisera primary reactive with the 35.5 kDa component (Pm35.5) in Western blot analysis. The outer membrane component was composed primarily of protein but did have lipopolysaccharide present at 133 micrograms mg-1 protein. In challenge trials (Trial 2), groups of white leghorn chickens vaccinated with Pm35.5 or an anti-idiotypic antibody (using Pm35.5 as the original antigen) had significant difference in mortality of 3.2% and 48.3%, respectively, when compared with unvaccinated controls (99%). Mortality in a group of chickens receiving a commercially available bacterin (9.2%) was higher but not significantly different from the mortality of the Pm35.5 vaccinated group (3.2%).

Animals↗

X-linked B-lymphocyte immune defect in CBA/HN mice. I. Studies of the function and composition of spleen cells.

A study of the composition and functional properties of spleen cells from the immune deficient CBA/HN mice and their F1 progeny is reported. While abnormalities were seen in both the numbers and function of thymus-independent (B) lymphocytes, all studies involving thymus-dependent (T) lymphocytes were normal. The X-linked nature of the immune defect in these mice was therefore attributed to abnormal or absent B lymphocytes. The possible nature of this defect and the similarity of the immune defect in these mice to certain human X-linked immunodeficiency diseases are discussed.

Animals↗

Age-related changes in immune reactivity: the influence of intrinsic defects and of a changed composition of the CD4+ T cell compartment.

Aging is accompanied by a decline in immune reactivity which to a major extent can be attributed to changes at the level of regulatory CD4+ T cells. In addition to evidence pointing to intrinsic defects, resulting in improper responsiveness of lymphocytes, it is likely that many age-related phenomena can be explained by a changed composition of the T cell compartment. Most likely as a consequence of thymic involution, the fraction of naive T cells in the periphery decreases, resulting in poor responses to neoantigens in particular. Moreover, due to antigenic exposure the fraction of memory cells increases. It is likely that, regardless of their phenotype, cells from aged individuals are subject to intrinsic defects or to immunosuppression, resulting in a lower responsiveness. As far as CD4+ T cells are concerned, recent studies have demonstrated that naive and memory cells behave differently with regard to activation requirements and lymphokine production. Age-related changes in T cell reactivity will be discussed in the context of these observations.

Aging↗

Effect of high/low dietary linoleic acid levels on the function and fatty acid composition of T-lymphocytes of normal and diabetic rats.

T-lymphocytes play a central role in the initiation, regulation and effector functions of immune responses. Changes in membrane composition can alter the membrane fluidity, receptor distribution and consequently function of these cells. Altered immune response in the diabetic state may be attributed partly to diabetes induced alterations in the metabolism of essential fatty acids. We have investigated the effect of dietary linoleic acid on T cell membrane composition and T cell mediated immune responses in normal and diabetic rats. Streptozotocin induced diabetes was found to produce lower T cell proliferative responses in mixed lymphocyte reactions and upon mitogen stimulation. Feeding of a diet rich in linoleic acid did not improve these responses. Feeding a diet low in linoleic acid further lowered the T cell dependent immune responses. We also found lower levels of 18:2 omega 6 fatty acids in the membrane phospholipids of these T cells. Levels of 20:4 omega 6 fatty acids were altered as a result of diabetes and diet composition. These fatty acids are the precursors of prostaglandins which are known to influence immune responses. It is concluded that diabetes results in significant alterations in T cell membrane composition and function in a manner that can be manipulated by modifications of the fatty acid composition of the diet. The present study suggests that diet fat modification may be important in regulating T cell-mediated immunity in insulin dependent diabetes mellitus.

Animals↗

[Morphocytochemical reaction of the lymph nodes in dogs to the administration of lysozyme].

By means of morphological, morphometrical and histochemical methods pelvic and tracheobronchial lymph nodes have been studied in dogs and concentration of lysozyme has been estimated in blood serum, in lymph and the lymph nodes after a single intramuscular injection of lysozyme (2 mg/kg of body mass). In the material investigated total concentration of lysozyme reaches its maximal values in 6 h after injection, then it gradually decreases and in 48 h reaches its control level. Morphometrically changes in cell composition are revealed predominantly of immune-competent cells in T- and B-dependent zones of the lymph nodes. Thus, the volumetric part of lymphoblasts in the germinative centers of the lymphoid nodules reaches its maximal indices by 48 h after lysozyme injection, while plasmatization of the paracortical zone and of medullary cords increases up to the 7th day. By the 14th day the volumetric part of lymphoblasts, immunoblasts and plasmocytes decreases gradually, and in 21 days after injection of the drug contents of the blast forms of the cells in the structural-functional zones of the lymph nodes does not differ from that in the control. The data obtained demonstrate the immunomorphological rearrangement of the lymph nodes in response to the exogenic lysozyme administration.

Animals↗

[Influence of various factors on glucocorticoid receptors of bronchoalveolar lavage (BAL) cells from rabbits].

It is supposed that the effects of glucocorticoid on target cells are mediated by glucocorticoid receptors. Recent investigations have demonstrated that changes in the composition and function of immune cells in bronchoalveolar lavage (BAL) cells of patients with immunological lung diseases are associated with the intensity of alveolar septal inflammation, or indicate steroid responsiveness. In order to clarify the physiologic changes of glucocorticoid receptor (GR) we measured the GR content of BAL cells obtained from rabbits with various conditions of the host (aging, immunological activation, malnutrition and pretreatment with prednisolone). The specific binding to GR was carried out by incubating BAL cells with 1, 2, 4, 10, 20, 40 nM 3H-prednisolone in the presence or absence of 1.0 x 10(-6) M unlabeled prednisolone. BAL cells from rabbits of various ages exhibited age-dependent reduction of GR content. There were striking differences of GR content in BAL cells from normal and BCG-vaccinated rabbits. The former yielded 3124 +/- 365 binding sites per cell, whereas activated BAL cells showed considerably higher values (6807 +/- 766). Chronological changes in GR content in BAL cells were also observed in normal rabbits pretreated with intravenous injection of prednisolone (2 mg/kg). The GR content was lowest at 3 hours after administration, reaching a peak at two days and then returned to control level. As for nutritional modulation, GR content tended to increase in the initial phase of fasting (days 4 to 8), and then decreased after day 12. In contrast, there were no significant changes in dissociation constants among BAL cells from animals in normal, immunologically activated and malnourished conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

[Effect of composite blood-activating decoction on expression of adherent molecule and cyclin in bone marrow hematopoietic cells in mice of immune-induced aplastic anemia].

OBJECTIVE: To study the effect of composite blood-activating decoction (CBAD) on expression of adherent molecule CD49 d and cyclin D2 in bone marrow hematopoietic cells in experimental immune-mediated aplastic anemia (AA) mice. METHODS: After the model of AA was established, the animals were fed with 0.2 ml of 100% CBAD, twice a day for 12 days. Te CD49 d and cyclin D2 expression level of bone marrow hematopoietic cells in model mice were measured by flow cytometor analysis system at the 13th day of experiment. RESULTS: CD49 d expression level in CBAD group was significantly higher than that in the AA group (P < 0.05), and was similar to that in the normal group (P > 0.05). The expression of cyclin D2 in CBAD group was significantly higher than that in the AA group (P < 0.01), but the cell count of G0 + G1 phase was significantly lower in the CBAD group, as compared with the AA group, P < 0.01. CONCLUSION: CBAD can increase the expression of adherent molecule CD49 d and cyclin D2 in bone marrow hematopoietic cells, and promote the growth of hematopoietic cells.

Anemia, Aplastic↗

Effects of candidate vaginally-applied microbicide compounds on innate immune cells.

Ideally, a vaginally-applied microbicide would be effective against a broad range of pathogens but would have minimal effects on the female genital tract. The aim of this study was to determine if representative candidate detergent-type and sulfated/sulfonated polymer-type microbicides altered the composition or function of innate immune cells normally found in the vaginal mucosa. The effect of microbicide on the composition of vaginal leukocytes was tested using a flow cytometric approach. Application of the detergent cholic acid, but not the sulfated polysaccharide lambda carrageenan, resulted in a significant increase in macrophages at the vaginal epithelial surface compared to control treatment (19.3% macrophages compared to 2.8%; p<0.0004). Phagocytosis of fluorochrome-labeled bacteria by macrophages was inhibited greater than 50% in the presence of 1.0mg/ml of the sulfonated polymer PRO 2000 but was not inhibited by the same concentration of lambda carrageenan. PRO 2000-pulsed macrophages regained phagocytic function after being washed free of the compound. Culture of macrophages with PRO 2000 also resulted in diminished detection of the surface proteins CD11b and CD18. After treated cells were washed free of PRO 2000, these proteins were detected at levels similar to control treated cells. In conclusion, application of a detergent-type microbicide, but not a sulfated polymer, resulted in the infiltration of inflammatory cells at the vaginal epithelial surface. Phagocytic function of macrophages was lost in the presence of 1mg/ml PRO 2000 which may have reflected masking of important cell surface proteins by the microbicide; however, there was no evidence of permanent loss of function upon removal of the compound.

Administration, Intravaginal↗

The induction of systemic and mucosal immune responses to antigen-adjuvant compositions administered into the skin: alterations in the migratory properties of dendritic cells appears to be important for stimulating mucosal immunity.

The properties of various vaccine-adjuvant formulations that are capable of inducing both systemic and common mucosal immunity subsequent to their intradermal administration are described. Effective mucosal adjuvants, including bacterial toxins, chemical enhancers of cyclic AMP, and the active form of vitamin D3, all shared the ability to promote dendritic cell migration from the skin to Peyer's patches subsequent to antigen induced maturation. Our data suggests that skin dendritic cells may function as effective antigen presenting cells for the induction of mucosal immune responses, if microenvironmental conditions are appropriately manipulated subsequent to their stimulation by antigen.

Administration, Topical↗

Temporal arteritis: cell composition and the possible pathogenetic role of cell-mediated immunity.

A biopsy specimen exhibiting the typical morphologic characteristics of temporal arteritis was studied by using light immunofluorescent, and electron microscopy and immunohistochemical techniques. The granulomatous lesion consisted of clusters of macrophages, epithelioid cells, giant cells, and the peripheral lymphocyte mantle, and was localized mainly in the media. Neutrophils were rare, and fibrinoid necrosis was absent. In immunofluorescent and immunohistochemical studies, no significant deposition of immunoglobulins or complement was observed. Immunohistochemical study with monoclonal antibodies to leukocyte surface antigens demonstrated that the central aggregated granulomatous infiltrate consisted of OKTM1+, Leu-M3+, HLA-DR+ epithelioid macrophages and multinucleated giant cells, whereas OKT8+, HLA-DR+ (suppressor/cytotoxic) T cells predominated in the peripheral lymphocyte mantle. These findings suggest that cell-mediated immunity, especially T cell-regulated granulomatous reaction, may play an important role in the pathogenesis of temporal arteritis. By electron microscopy, smooth muscle cells often exhibited closely attached macrophages, epithelioid cells, and giant cells, and displayed a variety of cell injuries. It therefore seems likely that smooth muscle cells are a primary target of the granulomatous reaction.

Aged↗

Xanthogranulomatous cholecystis. Cell composition and a possible pathogenetic role of cell-mediated immunity.

Thirty-three cases of xanthogranulomatous cholecystitis (XGC) exhibiting the typical morphologic features were studied by light and electron microscopy and immunohistochemical techniques. Incidence of XGC was 4.2% of the surgically resected gallbladder diseases. Histologically, the granulomatous lesion of XGC principally consisted of accumulations of foam cells and lymphocytes. Variable numbers of multinucleated giant cells, granulocytes and fibroblastic cells were also noted. With respect to the origin of foam cells, it was considered that the vast majority of foam cells were derived from monocytes/macrophages because they were invariably positive for KP1, HAM56, CD11b and CD68. Interspersed among macrophage foam cells, many T lymphocytes were identified. The subtyping of T cells indicated a heterogenous population composed of both CD4+ and CD8+ lymphocytes typically in a ratio of 1:2. Macrophages and T lymphocytes demonstrated a marked expression of HLA-DR antigen. Electron microscopic and immunohistochemical double-staining observation demonstrated intimate apposition of T lymphocytes to macrophages or macrophage foam cells. The results indicate that XGC is a granulomatous disorder characterized by accumulations of macrophage foam cells and T cells. Delayed type hypersensitivity reaction of cell-mediated immunity may be implicated in the pathogenesis of XGC.

Adult↗

Preoperative granulocyte/macrophage colony-stimulating factor (GM-CSF) increases hepatic dendritic cell numbers and clustering with lymphocytes in colorectal cancer patients.

Despite surgery with curative intent, approximately 30% of colorectal carcinoma patients will develop liver metastases during follow-up. Synchronous occult micrometastases, tumor cell shedding into the portal circulation and postoperative immune impairment have all been suggested to facilitate outgrowth of liver metastases. In experimental models, increases in both number of resident macrophages of the liver, the so-called Kupffer cells (KC), and tumoricidal capacity of KC were observed after pretreatment with granulocyte/macrophage colony-stimulating factor (GM-CSF), a potent immuno-stimulatory agent. Following perioperative recombinant human GM-CSF (rhGM-CSF), we previously showed activation of the systemic immune response in the postoperative period, which is normally transiently down-modulated after surgery. Therefore, in this pilot study, effects of preoperative rhGM-CSF administration on the composition of human liver immune cell population were evaluated in patients undergoing surgery for colorectal cancer. No difference in KC numbers of rhGM-CSF-treated patients was observed. Importantly, however, a 6-fold increase in dendritic cell (DC) numbers was observed compared to control patients, as quantified by immunohistochemistry of liver biopsies, taken during laparotomy. Furthermore, direct contact between liver CD8+ cells and DC was significantly enhanced in rhGM-CSF-treated patients. Both increases in DC numbers and DC interaction with CD8+ T cells suggest enhanced immunological activation, which may reduce liver metastases formation and ultimately improve survival after initial colorectal surgery.

Cell Adhesion↗

Factors affecting engraftment of allogeneic hematopoietic stem cells after reduced-intensity conditioning.

Several factors influence the engraftment of allogeneic hematopoietic stem cells (HSC). Recently, there has been increased utilization of transplant-conditioning regimens that use reduced doses of chemotherapy and radiation that are considered to be non-myeloablative. These non-myeloablative (or reduced-intensity) allogeneic HSC transplants (RIST) decrease early post-transplant complications, but they are associated with higher incidences of mixed chimerism and graft rejection compared with transplantation after myeloablative condition-ing. RIST provides a unique opportunity to study allogeneic HSC engraftment. In particular, host immune status and stem cell graft composition have emerged as important factors affecting engraftment after RIST Based on these observations, it has been hypothesized that conditioning regimens and allograft composition can be tailored to an individual patients immune and disease status prior to transplant.

Animals↗

Distribution of immune cells in the epididymis of the aging Brown Norway rat is segment-specific and related to the luminal content.

Remarkable changes occur during aging in the testis and epididymis of the Brown Norway rat. A dramatic increase in the number of halo cells, which are present in the epididymal epithelium and originate from the immune system, is found in animals of increasing age. Halo cells have been postulated to be either lymphocytes or monocytes. We hypothesized that halo cells are a mixture of different immune cells and that their relative composition changes with age. To verify this hypothesis, markers for helper T lymphocytes, cytotoxic T lymphocytes, B lymphocytes, and monocytes-macrophages were used to identify the major categories of immune cells in the epididymides of Brown Norway rats ranging in age from 3 to 24 mo. The numbers of immunocompetent cells in the epididymis were determined in relation to age, epididymal segment, and luminal content. We found that monocytes, helper T lymphocytes, and cytotoxic T lymphocytes belong to the population of halo cells. In addition, a segment-specific increase with age in the number of these immune cells was noted. Finally, we report a segment-specific recruitment of cytotoxic T lymphocytes and monocytes-macrophages in the epididymal epithelium of aged rats whose epididymal lumen contained few spermatozoa. We postulate that accumulation of damaged epithelial cells and antigens of germ cell origin, leaking through a dysfunctional blood-epididymis barrier, may contribute to the active recruitment of immune cells with age.

Aging↗

Dose-dependent effects of dietary gamma-linolenic acid on rat spleen lymphocyte functions.

Feeding rodents a diet rich in evening primrose oil (EPO), which contains 5-10 g gamma-linolenic acid (GLA)/100 g total fatty acids, has been shown to decrease lymphocyte proliferation and natural killer cell activity. However, EPO contains a very high level of linoleic acid which itself can affect lymphocyte functions and it is not clear to what extent the effects of EPO can be attributed to GLA. The current study investigated the effect of two levels of GLA in the rat diet upon immune cell functions; the level of linoleic acid was maintained below 30 g/100 g total fatty acids. Weanling rats were fed on high fat (178 g/kg) diets which contained 4.4 g or 10 g GLA/100 g total fatty acids in place of a proportion of linoleic acid. The total polyunsaturated fatty acid content and the n-6 to n-3 polyunsaturated fatty acid ratio of the diet were maintained at 35 g/100 g total fatty acids and 7, respectively. The fatty acid compositions of the serum and of spleen leukocytes were markedly influenced by that of the diet, with an increase in the proportions of GLA and dihomo-gamma-linolenic acid when the diets containing GLA were fed; these diets also increased the proportion of arachidonic acid in spleen leukocytes. Spleen lymphocyte proliferation in response to concanavalin A was significantly reduced (by 60%) by feeding the diet containing the higher level of GLA, but not by the diet containing the lower level of GLA. Spleen natural killer cell activity and prostaglandin E (PGE) production by spleen leukocytes were not significantly affected by inclusion of GLA in the diet, although there was a tendency towards decreased natural killer cell activity by cells from rats fed the high GLA diet. Thus, this study shows that dietary GLA is capable of altering the fatty acid composition of cells of the immune system and of exerting some immunomodulatory effects, but that the level of GLA in the diet must exceed 4.4 g/100 g total fatty acids for these effects to become apparent.

Animals↗

HEPC-based liposomes trigger cytokine release from peripheral blood cells: effects of liposomal size, dose and lipid composition.

The immune response caused by liposome stimulation was studied by assessing the level of several cytokines released from human peripheral blood cells. Liposome stimulation resulted in the release of IL-6, IL-10, IL-1beta, TNF-alpha and IFN-gamma. The size of the liposomes affected the degree of the cytokine releases with larger sized liposomes causing higher levels of cytokine induction. In addition, it appears that the lipid composition of liposomes had no effect on the degree of cytokine release. The release of cytokines occurred even in the absence of serum, suggesting that serum proteins did not contribute to liposome stimulation in peripheral blood cells. The release of cytokines induced by liposome stimulation was inhibited by the presence of either protein kinase-C (PKC) or protein tyrosine kinase (PTK) inhibitor, but not by the presence of an endocytosis inhibitor. This indicates that signal transduction via PKC or PTK is necessary, in order for human peripheral blood cells to release cytokines (IL-6, IL-10, IL-1beta, TNF-alpha and IFN-gamma) as the result of liposome stimulation. These quantitative data on the release of cytokines by liposomal stimulation provide useful information for the development of rational drug delivery systems and the safety of cytokine induction via the use of liposomes.

Cytokines↗