Maya Gulubova

Prognostic significance of mast cell number and microvascular density for the survival of patients with primary colorectal cancer

Background and Aim:  The prognostic relevance of tumor‐related angiogenesis and mast cell presence in colorectal cancer remains controversial. The aim of the current study was to assess the mast cell and microvessel densities (MCD and MVD) in the invasive front of colorectal cancers and to determine their prognostic relevance for survival of the patient with colorectal carcinoma.

Mast cells and inflammatory mediators in chronic ulcerative colitis

Chronic ulcerative colitis (CUC) is an inflammatory destructive disease of the large intestine characterized by motility and secretion disorders. In the past decade, attention has been paid to the role of neuronal structures and mast cells in regulating inflammatory and immune responses in inflammatory bowel disease (IBD). The present study was performed to demonstrate neuronal fibres (NF) and cells containing substance P (SP), tryptase and serotonin (SER) in the colonic wall of patients with CUC in remission. Biopsy specimens of 6 patients with CUC were investigated with immunocytochemical methods. Normal colon tissue obtained from 6 patients with rectal carcinoma was used as a control. An increased number of SP- and SER-positive NF was found in all the layers of the intestinal wall. The number of SER-containing endocrine cells in the mucosal glands was also increased per crypt. Tryptase-, SP- and SER-immunopositive mast cells were found in higher amounts than in control specimens in close apposition to the basal lamina of the glands among the epithelial cells and in other layers of the gut wall. Two types of mast cells were found: mast cells containing both tryptase and SP, and mast cells containing tryptase only. It is concluded that interactions between neuronal elements and mast cells play a significant role in the progress and maintenance of inflammatory processes in CUC.

Role of dendritic cells in progression and clinical outcome of colon cancer.

PurposeThe dendritic cells (DCs) are key players in the initiation and regulation of immune responses including antitumor immunity. In the current study, we aimed to elucidate the role of different subtypes of DCs infiltrating the tumor stroma and invasive margin for tumor progression and survival of patients with colon cancer.MethodsThe presence of immature (CD1a- and S100 protein+) and mature (CD83- and HLA-DR+) DCs was evaluated by immunohistochemistry in tissue samples from 145 patients with colon cancer. Patients were dichotomized according to the number of DCs in the tumor stroma and invasive margin, and clinical, histological, and survival data were compared between the two groups of patients.ResultsThe number of the mature CD83+ DCs in the tumor stroma and in the invasive margin significantly correlated with the tumor stage: the lower level of infiltration was found in patients that have advanced tumor stage. The frequency of distant metastases was higher in patients who had lower numbers of immature CD1a+ DCs in tumor stroma and of CD83+ DCs in invasive margin. Patients showing a relatively high number of S100+ DCs in the tumor stroma and HLA-DR+ DCs in the invasive margin had a longer overall survival (p < 0.05). Patients with lower CD83+ DCs infiltration in invasive margin had worse prognosis after surgical therapy compared with those with higher CD83+ DCs infiltration (p = 0.0397).ConclusionsOur results demonstrate that the infiltration of colon cancer with DCs is related with tumor progression and patient prognosis, suggesting a central role for DCs in controlling local antitumor immunity.

Expression of ICAM-1, VCAM-1, E-selectin and TNF-α on the endothelium of femoral and iliac arteries in thromboangiitis obliterans

Immunohistochemical light and electron microscopical analysis of surgical biopsies obtained from femoral and iliac arteries of patients with thromboangiitis obliterans (TAO) were performed to investigate the presence of tumour necrosis factor-alpha (TNF-alpha) and expression of the endothelial cell adhesion molecules intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Expression of ICAM-1, VCAM-1 and E-selectin was increased on endothelium and some inflammatory cells in the thickened intima in all TAO patients. Ultrastructural immunohistochemistry revealed contacts between mononuclear blood cells and ICAM-1-, and E-selectin-positive endothelial cells. These endothelial cells showed morphological signs of activation. The present data indicate that endothelial cells are activated in TAO and that vascular lesions are associated with TNF-alpha secretion by tissue-infiltrating inflammatory cells, ICAM-1-, VCAM-1- and E-selectin expression on endothelial cells and leukocyte adhesion via their ligands. The preferential expression of inducible adhesion molecules in microvessels and mononuclear inflammatory cells suggests that angiogenesis contributes to the persistence of the inflammatory process in TAO.

Expression of cell adhesion molecules, their ligands and tumour necrosis factor α in the liver of patients with metastatic gastrointestinal carcinomas

The expression of the following cell adhesion molecules, their β1 and β2 integrin ligands and the cytokine tumour necrosis factor-α (TNF-α) was investigated by light and electron microscope immunohistochemistry in the liver tissue in 20 patients with colorectal and gastric cancer also presenting with liver metastases: intercellular adhesion molecule-1 (ICAM-1), vascular endothelial adhesion molecule-1 (VCAM-1), E-selectin, leucocyte function-associated antigen-1 (LFA-1), macrophage antigen-1 (Mac-1), and very late antigen-4 (VLA-4). We have found a parallel enhancement of the adhesion molecules and of TNF-α in liver sinusoids surrounding metastases. The expression of ICAM-1 was enhanced on sinusoidal cells in all zones of the acinus. VCAM-1 immune reactivity was diffuse but less intensive in the lobule. E-selectin expression was observed in sinusoidal cells attached to metastases. In tumour metastases the expression of ICAM-1, VCAM-1, and E-selectin was visible on the tumour vascular endothelium. Tumour infiltrating host cells sowing positive immunoreactivity for ICAM-1, VCAM-1, LFA-1, Mac-1, and VLA-4 were located mainly at the boundary between liver parenchyma and the metastasis. At the ultrastructural level, ICAM-1-positive immune deposits were observed on the cellular membrane and in some transport vesicles of gastric metastatic cells. Further, the expression of all adhesion molecules was confirmed to sinusoidal endothelial cells and tumour vessels. It is concluded that the enhanced expression of adhesion molecules in liver sinusoids could be a marker for the assessment of the ability of sinusoidal endothelial cells to control the recruitment of leukocytes and monocytes to the metastatic site. They could also direct the adhesion of new circulating tumour cells to sinusoidal endothelium.The expression of the following cell adhesion molecules, their β1 and β2 integrin ligands and the cytokine tumour necrosis factor-α (TNF-α) was investigated by light and electron microscope immunohistochemistry in the liver tissue in 20 patients with colorectal and gastric cancer also presenting with liver metastases: intercellular adhesion molecule-1 (ICAM-1), vascular endothelial adhesion molecule-1 (VCAM-1), E-selectin, leucocyte function-associated antigen-1 (LFA-1), macrophage antigen-1 (Mac-1), and very late antigen-4 (VLA-4). We have found a parallel enhancement of the adhesion molecules and of TNF-α in liver sinusoids surrounding metastases. The expression of ICAM-1 was enhanced on sinusoidal cells in all zones of the acinus. VCAM-1 immune reactivity was diffuse but less intensive in the lobule. E-selectin expression was observed in sinusoidal cells attached to metastases. In tumour metastases the expression of ICAM-1, VCAM-1, and E-selectin was visible on the tumour vascular endothelium. Tumour infiltrating host cells sowing positive immunoreactivity for ICAM-1, VCAM-1, LFA-1, Mac-1, and VLA-4 were located mainly at the boundary between liver parenchyma and the metastasis. At the ultrastructural level, ICAM-1-positive immune deposits were observed on the cellular membrane and in some transport vesicles of gastric metastatic cells. Further, the expression of all adhesion molecules was confirmed to sinusoidal endothelial cells and tumour vessels. It is concluded that the enhanced expression of adhesion molecules in liver sinusoids could be a marker for the assessment of the ability of sinusoidal endothelial cells to control the recruitment of leukocytes and monocytes to the metastatic site. They could also direct the adhesion of new circulating tumour cells to sinusoidal endothelium.

Decrease in intrahepatic CD56+ lymphocytes in gastric and colorectal cancer patients with liver metastases

Gulubova M, Manolova I, Kyurkchiev D, Julianov A, Altunkova I. Decrease in intrahepatic CD56+ lymphocytes in gastric and colorectal cancer patients with liver metastases. APMIS 2009; 117: 870–9.

Sarcoidosis and molecular mimicry--important etiopathogenetic aspects: current state and future directions.

SummarySarcoidosis is a disease of uncertainty in terms of its cause, presentation, and clinical course. The disease has a worldwide distribution and affects all ages, races, and both sex. Sarcoidosis of the skin may have an extremely heterogeneous clinical presentation, so that the definitions of ‘great imitator’ and ‘clinical chameleon’ have long been used.The factors that influence clinical picture and severity of the disease are probably linked to the etiopathogenesis of sarcoidosis, which continues to be shrouded in mystery.The current state of the art on the pathogenesis of sarcoidosis is that it is an immunological response in a genetically susceptible individual to an as-yet undefined antigenic stimulus. How exposure occurs in genetically predisposed patients is not completely clear, but the most likely explanation is that these agents or antigens are either inhaled into the lungs or enter through contact with the skin, as these are the common target organs that are constantly in contact with the environment. An autoimmune etiology of sarcoidosis could possibly occur through a process of molecular mimicry of infectious or other environmental antigens to host antigens. This could lead to a cross-mediated immune response and induction of autoimmune disease. This molecular mimicry may probably be responsible for the heterogeneous clinical presentations of the disease.Several investigations and studies have provided valuable evidence on the etiopathogenesis of sarcoidosis, which may lead to the future development of targeted and innovative treatment strategies. Nevertheless, we are still a long way from unravelling the underlying cause of this mysterious disease.ZusammenfassungDie Sarkoidose ist eine Erkrankung voll der Unsicherheiten in Bezug auf ihre Ursache, Präsentation und den klinischen Verlauf. Die Erkrankung kommt überall auf der Welt vor. Sie tritt in jedem Alter, bei allen Rassen bei Mann und Frau auf. Die Sarkoidose der Haut kann sich klinisch extrem unterschiedlich präsentieren – seit Langem wird sie daher auch als „Großer Nachahmer“ und „klinisches Chamäleon“ bezeichnet.Die Faktoren, die das klinische Bild und die Schwere der Erkrankung beeinflussen, sind wahrscheinlich mit der Ätiopathogenese der Erkrankung verbunden. Diese sind aber auch heute noch immer ungeklärt. Die aktuelle Lehrmeinung besagt, dass die Sarkoidose eine immunologische Antwort auf einen bis jetzt undefinierten antigenen Reiz in einem – genetisch bedingt – empfindlichen Individuum darstellt. Wie es zur Exposition der genetisch prädisponierten Patienten kommt ist nicht ganz klar. Die wahrscheinlichste Erklärung ist, dass diese Substanzen, beziehungsweise Antigene entweder über die Lungen inhaliert werden oder durch Kontakt mit der Haut in den Körper eintreten. Haut und Lunge sind die am häufigsten befallenen Zielorgane, die dauernd mit der Umwelt in Kontakt stehen. Eine autoimmune Ätiologie der Sarkoidose könnte möglicherweise durch einen Prozess der molekularen Mimikrie von infektiösen oder anderen Antigenen aus der Umwelt auf Wirt Antigene erklärt werden. Dadurch könnte es zu einer „cross mediated“ Immunantwort und Auslösung einer autoimmunen Erkrankung kommen. Diese molekulare Mimikrie kann möglicherweise für die unterschiedliche klinische Präsentation der Erkrankung verantwortlich sein.Verschiedene Untersuchungen und Studien haben wertvolle Evidenz in Bezug auf die Ätiopathogenese der Sarkoidose geliefert. Dadurch könnte es in der Zukunft zur Entwicklung von zielgerichteten neuen Behandlungsstrategien kommen. Heute sind wir allerdings trotz allem noch weit entfernt von der Entwirrung der zugrunde liegenden Ursache dieser geheimnisvollen Krankheit.

Chromogranin A-, serotonin-, synaptophysin- and vascular endothelial growth factor-positive endocrine cells and the prognosis of colorectal cancer: An immunohistochemical and ultrastructural study

Background and Aim:  Endocrine differentiation in colorectal adenocarcinoma has been reported but its significance as a prognostic marker remains uncertain. The aim of the present study was to analyze the prognostic significance of endocrine differentiation in colorectal cancer.

Ile105Val GSTP1 polymorphism and susceptibility to colorectal carcinoma in Bulgarian population.

Background and aimsEtiologically, the sporadic colorectal cancer (CRC) is a complex and multifactorial disease that is linked to both exogenic and endogenic factors. Accumulating evidence indicates that susceptibility to cancers, including CRC, is mediated by genetically determined differences in the effectiveness of detoxification of potential carcinogens. A member of the glutathione-S-transferase (GST) family, GSTP1, is an important candidate for involvement in susceptibility to carcinogen-associated CRC. An A→G transition in exon 5 of the GSTP1 gene resulting in Ile105Val amino acid substitution has been identified. This change leads to alteration in catalytic efficiency of variant enzyme. The aim of the current study was to evaluate the influence of Ile105Val GSTP1 polymorphism on susceptibility to CRC.Materials and methodsThe GSTP1 genotyping was conducted in a case-control study of 80 ethnic Bulgarian CRC patients and 126 unaffected controls using polymerase chain reaction restriction fragment length polymorphism method.ResultsA statistically significant case-control difference in genotype frequencies was observed: 0.69 vs 0.54 for Ile/Ile, 0.22 vs 0.39 for Ile/Val, and 0.09 vs 0.07 for Val/Val (p = 0.049). The odds ratio (OR) for Val/Val was close to 1 (0.96, 95%CI: 0.35–2.66, p = 0.942), whereas the OR for Ile/Val was significantly lower, 0.45 (95%CI: 0.24–0.86, p = 0.016), compared to the referent Ile/Ile genotype. Although a prevalence of the GSTP1 variant allele-containing genotypes (Ile/Val or Val/Val) was found in controls than in patients (OR = 0.53, 95%CI: 0.30–0.96, p = 0.035), the allele frequencies did not show significant difference between cases and controls (p = 0.127).ConclusionsBased on the obtained protective effect of Ile/Val GSTP1 genotype, we could suggest that Ile105Val GSTP1 polymorphism may play some role in susceptibility to CRC.

Pathophysiological mechanisms of acute pancreatitis define inflammatory markers of clinical prognosis.

Development of acute pancreatitis illustrates the need to understand the basic mechanisms of disease progression to drive the exploration of therapeutic options. Cytokines play a major role in the pathogenesis of acute pancreatitis as underlying systemic inflammatory response, tissue damage, and organ dysfunction. However, little is known about circulating concentrations of these inflammatory markers and their real impact on clinical practice. Experimental studies have suggested that the prognosis for acute pancreatitis depends on the degree of pancreatic necrosis and the intensity of multisystem organ failure generated by the systemic inflammatory response. This suggests an intricate balance between localized tissue damage with proinflammatory cytokine production and a systemic anti-inflammatory response that restricts the inappropriate movement of proinflammatory agents into the circulation. Implication of such mediators suggests that interruption or blunting of an inappropriate immune response has the potential to improve outcome. A detailed understanding of pathophysiological processes and immunological aspects in patients with acute pancreatitis is the basis for the development of therapeutic strategies that will provide significant reductions in morbidity and mortality.