PKCδ deficiency inhibits fetal development and is associated with heart elastic fiber hyperplasia and lung inflammation in adult PKCδ knockout mice.

Protein kinase C-delta (PKCδ) has a caspase-3 recognition sequence in its structure, suggesting its involvement in apoptosis. In addition, PKCδ was recently reported to function as an anti-cancer factor. The generation of a PKCδ knockout mouse model indicated that PKCδ plays a role in B cell homeostasis. However, the Pkcrd gene, which is regulated through complex transcription, produces multiple proteins via alternative splicing. Since gene mutations can result in the loss of function of molecular species required for each tissue, in the present study, conditional PKCδ knockout mice lacking PKCδI, II, IV, V, VI, and VII were generated to enable tissue-specific deletion of PKCδ using a suitable Cre mouse. We generated PKCδ-null mice that lacked whole-body expression of PKCδ. PKCδ+/- parental mice gave birth to only 3.4% PKCδ-/- offsprings that deviated significantly from the expected Mendelian ratio (χ2(2) = 101.7, P < 0.001). Examination of mice on embryonic day 11.5 (E11.5) showed the proportion of PKCδ-/- mice implanted in the uterus in accordance with Mendelian rules; however, approximately 70% of the fetuses did not survive at E11.5. PKCδ-/- mice that survived until adulthood showed enlarged spleens, with some having cardiac and pulmonary abnormalities. Our findings suggest that the lack of PKCδ may have harmful effects on fetal development, and heart and lung functions after birth. Furthermore, our study provides a reference for future studies on PKCδ deficient mice that would elucidate the effects of the multiple protein variants in mice and decipher the roles of PKCδ in various diseases.

Age-associated proinflammatory elastic fiber remodeling in large arteries.

Elastic fibers are the main components of the extracellular matrix of the large arterial wall. Elastic fiber remodeling is an intricate process of synthesis and degradation of the core elastin protein and microfibrils accompanied by the assembly and disassembly of accessory proteins. Age-related morphological, structural, and functional proinflammatory remodeling within the elastic fiber has a profound effect upon the integrity, elasticity, calcification, amyloidosis, and stiffness of the large arterial wall. An age-associated increase in arterial stiffness is a major risk factor for the pathogenesis of diseases of the large arteries such as hypertensive and atherosclerotic vasculopathy. This mini review is an update on the key molecular, cellular, functional, and structural mechanisms of elastic fiber proinflammatory remodeling in large arteries with aging. Targeting structural and functional integrity of the elastic fiber may be an effective approach to impede proinflammatory arterial remodeling with advancing age.

Effects of micro-amounts of Porphyromonas gingivalis lipopolysaccharide on rabbit inflammatory immune response and development of atherosclerosis.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis (Pg) lipopolysaccharide is associated with the immune response and atherosclerosis. This study aimed to evaluate the effects of micro-amounts of Pg-lipopolysaccharide on rabbit inflammatory immune response and the development of atherosclerosis. MATERIAL AND METHODS: Twenty-four New Zealand white rabbits were randomly divided into four groups (n = 6). Group A was fed a regular diet and normal saline. Group B was supplied with a high-fat diet and normal saline. Group C was treated with a normal diet and Pg-lipopolysaccharide. Group D was given a high-fat diet and Pg-lipopolysaccharide. After 14 wk, the rabbits were killed to determine the changes in pathological indices. RESULTS: The serum lipid levels of groups B and D were significantly higher than that of group A (p < 0.01), and that of group C was higher (p < 0.05). Serum interleukin-6, monocyte chemoattractant protein-1 and tumor necrosis factor-α levels were significantly elevated by individual high-fat diets or Pg-lipopolysaccharide stimulation (p < 0.05). Groups A and C did not undergo evident aortic pathological damages, while foam cells appeared in the other two groups. Real-time polymerase chain reaction detection showed that toll-like receptor-2, interleukin-6, matrix metalloproteinase-9 and monocyte chemoattractant protein-1 were highly expressed in groups B and D (p < 0.05), and toll-like receptor-4, C-reactive protein and tumor necrosis factor-α levels were higher than those of group A (p < 0.05). Western blotting showed that transcription factor NF-κB p65 was expressed more highly in the three experimental groups than in group A (t = 9.26, p < 0.01). CONCLUSION: Micro-amounts of Pg-lipopolysaccharide induced the high expressions of inflammatory factors and mediated the inflammatory response. Pg-lipopolysaccharide elevated the blood lipid level less significantly than the high-fat diet did, but it may promote atherosclerosis.

Analysis of the scalp of women with AIDS subjected to autopsy: epithelial, follicular, and immunologic aspects.

Hair keeps the scalp warmer and slightly moister than the rest of the skin, which contributes to a favorable environment for mycotic, bacterial, and parasitic infections. It is well established that AIDS makes the patient more susceptible to opportunistic infections and cutaneous manifestations. Because of this, the aim of this study was to analyze scalp fragments of autopsied women with AIDS. Twenty-eight scalp samples of women aged between 18 and 46 years were observed. These women were divided into 2 groups: with AIDS (n = 14) and without AIDS (n = 14). We conducted histochemical (hematoxylin-eosin, Picrosirius, and Verhoeff), morphometric (Image J; National Institutes of Health, Hamilton, ON, Canada and KS-300 Kontron-Zeiss; Kontron Elektronik, Carl-Zeiss, Germany), and immunohistochemical (S-100) analyses of the scalp. In patients with AIDS, epithelial thickness, number of epithelial cell layers, number of immature Langerhans cells in the epidermis, and percentages of elastic fibers in the dermis were significantly lower, whereas telogen hair follicles were significantly higher. The percentage of collagen fibers in the dermis and the diameter of the epithelial cells were smaller in patients with AIDS, without significant difference. AIDS possibly causes immunologic and morphologic alterations in the scalp. This study may establish parameters for better clinical and morphologic diagnostic in patients with AIDS.

Immunophenotyping and extracellular matrix remodeling in pulmonary and extrapulmonary sarcoidosis.

OBJECTIVE: To investigate the significance of cellular immune markers, as well as that of collagen and elastic components of the extracellular matrix, within granulomatous structures in biopsies of patients with pulmonary or extrapulmonary sarcoidosis. METHODS: We carried out qualitative and quantitative evaluations of inflammatory cells, collagen fibers, and elastic fibers in granulomatous structures in surgical biopsies of 40 patients with pulmonary and extrapulmonary sarcoidosis using histomorphometry, immunohistochemistry, picrosirius red staining, and Weigert's resorcin-fuchsin staining. RESULTS: The extrapulmonary tissue biopsies presented significantly higher densities of lymphocytes, macrophages, and neutrophils than did the lung tissue biopsies. Pulmonary granulomas showed a significantly higher number of collagen fibers and a lower density of elastic fibers than did extrapulmonary granulomas. The amount of macrophages in the lung samples correlated with FVC (p < 0.05), whereas the amount of CD3+, CD4+, and CD8+ lymphocytes correlated with the FEV1/FVC ratio and VC. There were inverse correlations between TLC and the CD1a+ cell count (p < 0.05), as well as between DLCO and collagen/elastic fiber density (r = -0.90; p = 0.04). CONCLUSIONS: Immunophenotyping and remodeling both showed differences between pulmonary and extrapulmonary sarcoidosis in terms of the characteristics of the biopsy samples. These differences correlated with the clinical and spirometric data obtained for the patients, suggesting that two different pathways are involved in the mechanism of antigen clearance, which was more effective in the lungs and lymph nodes.

Imunofenotipagem e remodelamento da matriz extracelular na sarcoidose pulmonar e extrapulmonar / Immunophenotyping and extracellular matrix remodeling in pulmonary and extrapulmonary sarcoidosis

OBJETIVO: Investigar o significado de marcadores de imunidade celular e de componentes elásticos/colágeno da matriz extracelular em estruturas granulomatosas em biópsias de pacientes com sarcoidose pulmonar ou extrapulmonar. MÉTODOS: Determinações qualitativas e quantitativas de células inflamatórias, de fibras de colágeno e de fibras elásticas em estruturas granulomatosas em biópsias cirúrgicas de 40 pacientes com sarcoidose pulmonar e extrapulmonar foram realizadas por histomorfometria, imuno-histoquímica, e técnicas de coloração com picrosirius e resorcina-fucsina de Weigert. RESULTADOS: A densidade de linfócitos, macrófagos e neutrófilos nas biópsias extrapulmonares foi significativamente maior do que nas biópsias pulmonares. Os granulomas pulmonares apresentaram uma quantidade significativamente maior de fibras de colágeno e menor densidade de fibras elásticas que os granulomas extrapulmonares. A quantidade de macrófagos nos granulomas pulmonares correlacionou-se com CVF (p < 0,05), ao passo que as quantidades de linfócitos CD3+, CD4+ e CD8+ correlacionaram-se com a relação VEF1/CVF e com CV. Houve correlações negativas entre CPT e contagem de células CD1a+ (p < 0,05) e entre DLCO e densidade de fibras colágenas/elásticas (r = -0,90; p = 0,04). CONCLUSÕES: A imunofenotipagem e o remodelamento apresentaram características diferentes nas biópsias dos pacientes com sarcoidose pulmonar e extrapulmonar. Essas diferenças correlacionaram-se com os dados clínicos e espirométricos dos pacientes, sugerindo que há duas vias envolvidas no mecanismo de depuração de antígenos, que foi mais eficaz nos pulmões e linfonodos.

OBJECTIVE: To investigate the significance of cellular immune markers, as well as that of collagen and elastic components of the extracellular matrix, within granulomatous structures in biopsies of patients with pulmonary or extrapulmonary sarcoidosis. METHODS: We carried out qualitative and quantitative evaluations of inflammatory cells, collagen fibers, and elastic fibers in granulomatous structures in surgical biopsies of 40 patients with pulmonary and extrapulmonary sarcoidosis using histomorphometry, immunohistochemistry, picrosirius red staining, and Weigert's resorcin-fuchsin staining. RESULTS: The extrapulmonary tissue biopsies presented significantly higher densities of lymphocytes, macrophages, and neutrophils than did the lung tissue biopsies. Pulmonary granulomas showed a significantly higher number of collagen fibers and a lower density of elastic fibers than did extrapulmonary granulomas. The amount of macrophages in the lung samples correlated with FVC (p < 0.05), whereas the amount of CD3+, CD4+, and CD8+ lymphocytes correlated with the FEV1/FVC ratio and VC. There were inverse correlations between TLC and the CD1a+ cell count (p < 0.05), as well as between DLCO and collagen/elastic fiber density (r = -0.90; p = 0.04). CONCLUSIONS: Immunophenotyping and remodeling both showed differences between pulmonary and extrapulmonary sarcoidosis in terms of the characteristics of the biopsy samples. These differences correlated with the clinical and spirometric data obtained for the patients, suggesting that two different pathways are involved in the mechanism of antigen clearance, which was more effective in the lungs and lymph nodes.

The pathology of large-vessel vasculitides.

Vasculitis affecting large elastic arteries, including the aorta and major proximal branches, encompasses various diseases including Takayasu arteritis, giant cell (or temporal) arteritis, and tertiary syphilis, but also may occur as a rare complication of Behçet's disease, rheumatoid arthritis, sarcoidosis, Cogan syndrome, Kawasaki disease, ankylosing spondylitis, systemic lupus erythematosus and Wegener's granulomatosis. Recent reports have also established a link between inflammatory abdominal aortic aneurysm as well as lymphoplasmacytic thoracic aortitis with an overabundance of IgG4-producing plasma cells and the burgeoning constellation of 'Hyper-IgG4' syndromes. This review focuses on morphologic aspects of large-vessel vasculitis pathology associated with giant cell arteritis, Takayasu arteritis, idiopathic or isolated aortitis, lymphoplasmacytic thoracic and ascending aortitis, and the inflammatory aneurysm/retroperitoneal fibrosis syndrome.

Blocking TNF-alpha attenuates aneurysm formation in a murine model.

Abdominal aortic aneurysm (AAA) is one of a number of diseases associated with a prominent inflammatory cell infiltrate and local destruction of structural matrix macromolecules. This chronic infiltrate is predominately composed of macrophages and T lymphocytes. Activated macrophages produce a variety of cytokines, including TNF-alpha. Elevated levels of TNF-alpha were observed in patients with AAA, suggesting that TNF-alpha may play a role in the pathogenic mechanisms of AAA. In the present study, we investigated the role of TNF-alpha in AAA formation. By studying a murine aneurysm model, we found that both mRNA and protein levels of TNF-alpha were increased in aneurysm tissue compared with normal aortic tissues. Therefore, we tested the response of mice lacking expression of TNF-alpha. These mice were resistant to aneurysm formation. Our results show that TNF-alpha deficiency attenuates matrix metalloproteinase (MMP) 2 and MMP-9 expression and macrophage infiltration into the aortic tissue. These data suggest that TNF-alpha plays a central role in regulating matrix remodeling and inflammation in the aortic wall leading to AAA. In addition, we investigated the pharmacological inhibition of AAA. A Food and Drug Administration-approved TNF-alpha antagonist, infliximab, inhibited aneurysm growth. Our data also show that infliximab treatment attenuated elastic fiber disruption, macrophage infiltration, and MMP-2 and MMP-9 expression in aortic tissue. This study confirms that a strategy of TNF-alpha antagonism may be an important therapeutic strategy for treating AAA.

Anetoderma occurring after hepatitis B vaccination.

Anetoderma is an elastolytic disorder of unknown origin. To our knowledge, anetoderma secondary to hepatitis B immunization has been described only once in the literature, in two siblings vaccinated at the same time. We describe, what we believe to be an additional case of such a rare disorder in a 21-year-old man. He presented with white spots and papules on his neck, upper limbs and trunk, that had developed gradually within the last 6 years without any symptoms. The initial lesions were red macules, which gradually enlarged in size and number, becoming pale. Biopsy of a sack-like lesion revealed normal epidermis and a discrete mononuclear infiltrate throughout the dermis. Association of anetoderma with hepatitis B vaccination is speculated here, as suggested by history of vaccination two weeks prior to the onset of skin eruption and ruling out other possible causes of anetoderma.

Immunohistochemical localization of cerebrovascular cannabinoid CB1 receptor protein.

Cannabinoids are powerful hypotensives and vasodilators. However, their mode of action is controversial. This study is the first to investigate the distribution of vascular CB1 receptor protein expression in situ. We used double-fluorescence and chromogenic immunohistochemistry to investigate patterns of CB1 protein expression in cerebrovascular tissue in rat brain sections. We found a layer of intense CB1 labeling immediately adjacent to the internal elastic lamina, consistent with myointimal and vascular smooth muscle cells, and diffuse labeling adventitial to this layer. We concluded that CB1 receptor are most intensely expressed in the vascular smooth muscle layer in cerebral arteries, and are likely to be chiefly responsible for the potent vasodilatory effect of cannabinoids.

CD55 expression patterns on intestinal neuronal tissue are divergent from the brain.

BACKGROUND: and objectives: In this study, we investigated how enteric plexuses protect themselves from complement mediated attack. For this purpose, the expression patterns of membrane bound complement regulatory proteins (mCRP) and their association with C3 deposition was determined. In addition, mCRP expression patterns of enteric plexuses were compared with those in the central nervous system (CNS). METHODS: Immunohistochemical stainings were performed to discriminate neuronal cells from glial cells and to detect the presence of CD46, CD55, CD59, and C3d. RNA in situ hybridisation (RISH) was used to determine the cell types that produce CD55 mRNA. RESULTS: Enteric plexuses minimally expressed CD46 whereas CD55 and CD59 were highly expressed. CD55 expression was also observed in a ring around Auerbach's plexuses which was not observed for CD46 and CD59. C3d was deposited around the plexuses but plexus cells themselves did not stain for C3d. In contrast with CNS neurones, enteric neurones were shown to express CD55 whereas enteric glial cells did not. This was confirmed with CD55 RISH. Phospholipase C mediated cleavage of CD55 demonstrated that CD55 was most likely attached to elastic fibres surrounding the plexus. Attached CD55 might protect CD55 negative glial cells from complement mediated injury during inflammatory reactions. CD55 on elastic fibres surrounding the plexuses most likely originated from enteric neuronal cells. CONCLUSION: In contrast with the CNS, enteric neurones express CD55 and enteric glial cells lack CD55 expression. CD55, produced by neuronal cells, attached to elastic fibres surrounding the plexuses is proposed to protect the CD55 negative glial cells within plexuses.

[Unilateral blepharochalasis with IgA-deposits]. / Unilaterale Blepharochalasis mit läsionalen IgA-Ablagerungen.

A 36-year-old male patient presented with unilateral periocular skin atrophy. The blepharochalasis developed without any obvious inflammation of the eyelids over the past 10 years. Interestingly, elongated blood vessels and microaneurysmatic vessel changes were found in the tarsal conjunctiva. A punch biopsy revealed a nearly complete loss of elastic fibres in the papillary and superficial reticular dermis. The contralateral side was histopathologically normal. On immunohistology IgA-deposits could be observed especially on perifollicular elastic fibres. Immunoelectronmicroscopy confirmed the diagnosis and suggested fibulin and fibronectin as potential binding sites for the autoantibodies. This further report of elastolysis in association with IgA-autoantibodies defines the autoantibody binding site in more detail and suggests that the immune mechanisms may also play a role in vessel changes of the conjunctiva.

Middermal elastolysis in two patients with lupus erythematosus.

Despite lupus erythematosus (LE) being considered a "connective tissue disease," little has been written about the elastic fiber changes in the skin of affected patients. We report our histologic findings in two patients with unusual cutaneous lesions. Elastic fiber loss was noted, and scattered giant cells with elastic fiber phagocytosis were prominent in one patient. The findings are similar to those described for middermal elastolysis. Other authors have reported patients with LE and elastic fiber loss resembling anetoderma. We believe that a spectrum of elastic fiber changes can occur in patients with LE and may be induced by infiltrating lymphocytes and/or circulating antibodies.

Morphologic change of the internal elastic lamina in Buerger's disease.

Morphologic features and pathogenesis of arterial changes occurring in Buerger's disease (thromboangiitis obliterans) are still controversial. This study describes histopathologic features of medium sized arteries from patients with Buerger's disease, particularly of the internal elastic lamina in relation to the immunologic mechanism of the injury. Seventeen segments of occluded arteries (femoral or popliteal arteries) from 17 patients with Buerger's disease were analyzed by histopathological and immunohistochemical methods. The most characteristic features were total luminal obliteration, together with a varying degree of recanalization and deposition of hemosiderin pigments. Detailed analysis, however, showed marked undulation and multiplication of the internal elastic lamina (100%) associated with basophilic degeneration and delicate linear calcification (47%). Lymphocytic infiltration along the internal elastic lamina was seen in 71% and was associated with localized edema. Lymphocytes along the lamina were consistently positive for T cell marker. Mild to moderate fibrosis was present at the media in 24%. Adventitial changes included mild, nonspecific and irregular fibrosis seen in 53%. Immunologic injury to the internal elastic lamina associated with T-lymphocytic infiltration might be the initial morphogenetic mechanism of the thrombotic occlusion and organization of medium-sized arteries in Buerger's disease.

Actinically degenerate elastic tissue: the prime antigen in the giant cell (temporal) arteritis syndrome? New data from the posterior ciliary arteries.

OBJECTIVE: To search for evidence of actinic elastotic degeneration (actinic arteriopathy) and giant cell arteritis (GCA) in the posterior ciliary arteries of eyes from aged white Australians. METHODS: Three hundred donor eyes were given to us by the Lions Eye Bank of New South Wales at Sydney Hospital. Of these, 146 formed the basis of this study. Portions of the posterior ciliary arteries located in relation to the optic nerve heads were processed in paraffin and were then stained by a sensitive haematoxylin and eosin stain that had been especially developed to display actinic elastotic degeneration of elastic tissue. RESULTS: Among 60 "aged" subjects (70-90 yrs.), a total of 41 (approximately 68%) showed definite changes of actinic elastotic degeneration in their laminae, a condition called actinic arteriopathy. One of these subjects revealed giant cells on degenerate lamina, giving a picture regarded as early (pre-clinical) GCA. A young "control" group of 60 subjects 17-59 years of age revealed only one subject with a similar degree of actinic arteriopathy. CONCLUSIONS: Actinic arteriopathy of the posterior ciliary arteries was more frequent and advanced in the "aged", over-70 group as compared with changes in the "young" group < 60 years of age. One aged subject without a history of eye disease showed giant cells associated with elastotically degenerate internal elastic lamina. Her fortuitous lesions are regarded as indicative of how GCA is likely to begin in the damaged arteries.

Human mast cell basic fibroblast growth factor in pulmonary fibrotic disorders.

Mast cells (MCs) are abundant in fibrotic tissue, although their role in fibrogenesis remains obscure. Recent studies suggest MCs may produce basic fibroblast growth factor (bFGF). To evaluate the hypothesis that MC bFGF contributes to the fibrotic response in human interstitial lung disease, we studied lung tissue, bronchoalveolar lavage fluid and serum in 1) idiopathic pulmonary fibrosis, 2) chronic beryllium disease and sarcoidosis, 3) control subjects with no disease or who were beryllium sensitized with normal lung histology. Diseased subjects underwent clinical assessments to stage disease severity. We determined that most bFGF+ cells in lung interstitium are MCs and are most abundant in idiopathic pulmonary fibrosis. Distribution of bFGF+ MCs matched that of extracellular matrix deposition and correlated with the extent of fibrosis morphometrically. Only one bFGF isoform (17.8 kd) was found in idiopathic pulmonary fibrosis and chronic beryllium disease lung tissues and interacted with heparin-like molecules in the lung. Using a human MC line, we verified that MCs express bFGF mRNA and protein that localizes to cytoplasmic granules. Clinically, bFGF concentrations in bronchoalveolar lavage fluid and serum were highest in disease states and correlated with bronchoalveolar lavage cellularity and severity of gas exchange abnormalities, supporting a role for MC bFGF in the pulmonary fibrotic response and its clinical consequence.