Immunohistochemical Phenotyping Research Articles

Adenoid cystic carcinoma of the thymus gland

BackgroundThymic carcinomas are rare and aggressive tumours. They constitute a heterogeneous group of tumours with various histological patterns and subtypes resembling epithelial tumours arising from other organs.Case presentation.We hereby represent a case of primary thymic carcinoma with adenoid cystic carcinoma-like features (TCACC) which is an extremely rare variant of thymic adenocarcinoma. To date and to the best of our knowledge, there are nine reported cases in literature and ours is the tenth. Our case was treated surgically but the implementation of adjuvant chemoradiotherapy has been reported in few of the published cases.ConclusionsTCACC constitutes a rare entity of thymic adenocarcinoma with limited available literature. The current data is derived from few case reports and case series. The histological overlap of these tumours and primary ACC of salivary glands poses a diagnostic challenge. Radiological investigations, immunohistochemical phenotyping and genetic analysis are crucial in establishing the diagnosis.

CELLULAR PHENOTYPES OF GLIOBLASTOMA AND PROGNOSIS OF TUMOR OUTCOME

In solid malignant tumors, as well as in hematological tumors, cancer stem cells (CSCs), quiescent, pluripotent, self-renewing neoplastic, also known as tumor initiating cells, have been identified. CSC tumors are resistant not only to chemotherapy, but also to radiation therapy, as well as they play their role in the occurrence of relapses after surgical treatment, An unfavorable prognosis due to an aggressive course and complications after treatment of the most common primary brain tumor - glioblastoma multiforme (GBM), is also associated with CSC. One of the mechanisms that affect the physiological regeneration of tissues, but do not work under conditions of malignancy, is the environment and composition of the cellular tumor ensemble, represented by the phenotypes of cellular and adaptive immunity. Using immunohistochemical phenotyping methods, the authors identified immunocytes/phagocytes and established the features of their localization in the malignant tissue and at the border around the tumor. It was concluded that due to apoptosis around the tumor, intercellular signaling and migration of immunocytes/phagocytes to the focus of the tumor process are disrupted. The primary tumor process is not associated with damage to the tissue cambium genome, but with the migration of immature stem cells of the hematogenous pool, capable of proliferating and partially entering into incomplete differentiation under conditions of hypoxia and the absence of blood vessels. At the second stage, connective tissue is formed, which grows into vessels, the wall of which is represented by immature endothelium, with impaired transport function. According to the authors, metastasis of cancer cells does not occur by hematogenous and lymphogenous pathways, but is associated with the development of additional foci of hematopoiesis as a result of the adaptation of the human body to the loss of stem and semi-stem cells, precursors for normal regeneration of the nervous tissue.

The antiatherosclerotic action of 1G244 – An inhibitor of dipeptidyl peptidases 8/9 – is mediated by the induction of macrophage death

BackgroundTargeting cell death to induce favorable functional and morphological changes within atherosclerotic plaques has long been postulated as a promising anti-atherosclerotic strategy. In this regard, inhibition of dipeptidyl peptidases 8/9 has received special attention in the context of chronic inflammatory diseases due to its regulatory role in macrophage death in vivo. MethodsThe present study investigates the influence of prolonged treatment with 1G244 – an inhibitor of dipeptidyl peptidases 8/9 – on the development of the advanced atherosclerosis plaque in apoE-knockout mice, using morphometric and molecular methods. Results1G244 administration has led to a reduction in atherosclerotic plaque size in an apoE-knockout mice model. Moreover, it reduced the content of in-plaque macrophages, attributed by immunohistochemical phenotyping to the pro-inflammatory M1-like activation state of these cells. Inhibition of dipeptidyl peptidases 8/9 augmented the lytic form of death response of activated macrophages in-vitro. ConclusionsIn summary, inhibition of DPP 8/9 elicited an anti-atherosclerotic effect in apoE−/− mice, which can be attributed to the lytic form of death induction in activated macrophages, as assessed by the in vitro BMDM model. This, in turn, results in a reduction of the plaque area without its transformation towards a rupture-prone morphology.

Cover Image: Volume 82 Issue 6

Cover Caption: The cover image is based on the Original Article Multiplex immunohistochemical phenotyping of T cells in primary prostate cancer by Busra Ozbek et al., https://doi.org/10.1002/pros.24315.

Multiplex immunohistochemical phenotyping of T cells in primary prostate cancer.

Most prostate cancers are "immune cold" and poorly responsive to immune checkpoint inhibitors. However, the mechanisms responsible for the lack of a robust antitumor adaptive immune response in the prostate are poorly understood, which hinders the development of novel immunotherapeutic approaches. Most inflammatory infiltrates in the prostate are centered around benign glands and stroma, which can confound the molecular characterization of the antitumor immune response. We sought to analytically validate a chromogenic-based multiplex immunohistochemistry (IHC) approach applicable to whole slide digital image analysis to quantify T cell subsets from the tumor microenvironment of primary prostatic adenocarcinomas. As an initial application, we tested the hypothesis that PTEN loss leads to an altered antitumor immune response by comparing matched regions of tumors within the same individual with and without PTEN loss. Using the HALO Image Analysis Platform (Indica Labs), we trained a classifier to quantify the densities of eight T cell phenotypes separately in the tumor epithelial and stromal subcompartments. The iterative chromogenic approach using 7 different antibodies on the same slide provides highly similar findings to results using individually stained slides with single antibodies. Our main findings in carcinomas (benign removed) include the following: i) CD4+ T cells are present at higher density than CD8+ T cells; ii) all T cell subsets are present at higher densities in the stromal compartment compared to the epithelial tumor compartment; iii) most CD4+ and CD8+ T cells are PD1+; iv) cancer foci with PTEN loss harbored increased numbers of T cells compared to regions without PTEN loss, in both stromal and epithelial compartments; and v) the increases in T cells in PTEN loss regions were associated with ERG gene fusion status. This modular approach can apply to any IHC-validated antibody combination and sets the groundwork for more detailed spatial analyses. Iterative chromogenic IHC can be used for whole slide analysis of prostate tissue samples and can complement transcriptomic results including those using single cell and spatial genomic approaches.

The Anti-Atherosclerotic Action of FFAR4 Agonist TUG-891 in ApoE-Knockout Mice Is Associated with Increased Macrophage Polarization towards M2 Phenotype.

Background: Over the past few years, a better understanding of the biology of G-protein coupled receptors (GPRs) has led to the identification of several receptors as novel targets for free fatty acids (FFAs). FFAR4 has received special attention in the context of chronic inflammatory diseases, including atherosclerosis, obesity and NAFLD, through to its anti-inflammatory effect. Methods: The present study investigates the influence of prolonged treatment with TUG-891-FFAR4 agonist on the development of atherosclerosis plaque in apoE-knockout mice, using morphometric and molecular methods. Results: TUG-891 administration has led to the reduction of atherosclerotic plaque size and necrotic cores in an apoE-knockout mice model. TUG-891-treated mice were administered subcutaneously at a dose of 20 mg/kg three times a week for 4 months. The FFAR4 agonist reduced the content of pro-inflammatory M1-like macrophages content in atherosclerotic plaques, as evidenced by immunohistochemical phenotyping and molecular methods. In atherosclerotic plaque, the population of smooth muscle cells increased as evidenced by α-SMA staining. We observed changes in G-CSF and eotaxin markers in the plasma of mice; changes in the levels of these markers in the blood may be related to macrophage differentiation. Importantly, we observed a significant increase in M2-like macrophage cells in atherosclerotic plaque and peritoneum. Conclusions: Prolonged administration of TUG-891 resulted in significant amelioration of atherogenesis, providing evidence that the strategy based on macrophage phenotype switching toward an M2-like activation state via stimulation of FFAR4 receptor holds promise for a new approach in the prevention or treatment of atherosclerosis.

Предиктивный потенциал фенотипирования макрофагальной популяции в малигнизации H. pylori-ассоциированного хронического гастрита

Tumor-associated macrophages are able to regulate the tumor cell proliferation and to affect the tumor cell dissemination. The study was aimed to assess the predictive potential of the macrophage population immunohistochemical phenotyping in early malignization of H. pylori-associated chronic gastritis. Gastic biopsy samples of male and female patients aged 48 ± 7.2 infected with Helicobacter pylori were used as the research material. The patients were divided into three groups: non-atrophic chronic gastritis (NACG, n = 10), atrophic chronic gastritis (ACG, n = 10), G1/G2 gastric adenocarcinoma (GAC, n = 10). The macrophage population was visualized using the CD68 pan-macrophage marker and the type 2 monocyte/macrophage marker CD163. Intensity of neoangiogenesis was defined using the CD31 endothelial marker by assessing the total cross sectional area of blood vessels. It was found that chronic gastritis was accompanied by the dynamic increase in the size of the general macrophage population with the progression of atrophic and metaplastic processes. According to immunohistochemical study of biopsies obtained from patients with NCG, the CD163 : CD68 ratio was 0.67 ± 0.02, and the total cross sectional area of blood vessels was 3590.92 ± 356.27 µm2. Atrophic gastritis and adenocarcinoma were characterized by vector redistribution of monocytes/macrophages into the 2nd functional phenotype. The CD163 : CD68 expression index in the group with ACG was 0.81 ± 0.04, and in the group with GAC it was 0.88 ± 0.03. Microvascular area was significantly increased in the groups with ACG and GAC, which reflected tumor neoangiogenesis intensification under the influence of М2 monocytes/macrophages. The increased expression of CD163 can serve as a predictor of chronic gastritis malignization together with evaluation of the glandular epithelium atrophy and metaplasia degree.

Molecular heterogeneity of pancreatic intraductal papillary mucinous neoplasms and implications for novel endoscopic tissue sampling strategies

AimsIntraductal papillary mucinous neoplasms (IPMNs) may be precursor lesions of pancreatic cancer. The path towards malignancy is associated with mutations in tumour suppressor—and oncogenes that may serve as biomarkers during...

The Exostosin Immunohistochemical Status Differentiates Lupus Membranous Nephropathy Subsets With Different Outcomes

The Exostosin Immunohistochemical Status Differentiates Lupus Membranous Nephropathy Subsets With Different Outcomes

Immunohistochemical phenotyping of macrophages and T lymphocytes infiltrating in peripheral nerve lesions of dourine-affected horses.

Dourine is a deadly protozoan disease in equids caused by infection with Trypanosoma equiperdum. Neurological signs in the later stage of infection may be caused by peripheral polyneuritis and related axonal degeneration. This neuritis involves T lymphocytes, B lymphocytes, and macrophages, and is observed in cases without obvious neurological signs. However, the pathogenesis of neuritis remains unclear. We identified M2 macrophages and CD8 T cells as the predominant phenotypes in neuritis of dourine-affected horses with or without neurological signs. In contrast, the populations of M1 macrophages and CD4 T cells were small. This result indicates that inflammation was chronic and suggests that dourine-associated neuritis occurs at the early stage of infection.

Evidence for a role of autoinflammation in early-phase psoriasis.

Psoriasis is a common, inflammatory immune-mediated skin disease mainly presenting with plaques whose pathogenesis is based on the central role of the interleukin (IL)-23/IL-17 axis. However, the mechanisms acting in papular lesions of early-phase psoriasis are not fully understood. The aim of this study was to assess the involvement of autoinflammation, a state of sterile inflammation mainly driven by IL-1 over-production that has been recently hypothesized to act in the early phase of disease. Lesional skin of 10 patients with recent onset, untreated psoriasis has been investigated for expression of IL-1β, IL-17, IL-23 and other cytokines involved in the disease in comparison with normal skin of 10 healthy controls using a protein array method. Immunohistochemical phenotyping of inflammatory infiltrate and co-localization experiments with immunofluorescence confocal microscopy were conducted. IL-1β was significantly more expressed in psoriasis than in normal skin (P<0·0001). The chemokine IL-8 was also over-expressed in psoriasis (P=0·03) while IL-12, IL-17, IL-23, tumour necrosis factor-α and interferon-γ were only slightly more expressed in psoriasis than in normal skin, without reaching statistical significance. The inflammatory infiltrate consisted mainly of neutrophils with a relevant number of macrophages and dendritic cells and only scattered, predominantly T helper 1 lymphocytes. IL-1β co-localized mainly with CD66b, a neutrophil marker, suggesting that neutrophils were the major source of this cytokine. IL-1β over-expression in combination with low expression of cytokines that are predominant in late-phase plaque psoriasis may support the role of autoinflammation in early-phase disease, possibly paving the way to randomized trials with IL-1 antagonists.

Immunohistochemical phenotyping of T cells, granulocytes, and phagocytes in the muscle of cancer patients: association with radiologically defined muscle mass and gene expression

BackgroundInflammation is a recognized contributor to muscle wasting. Research in injury and myopathy suggests that interactions between the skeletal muscle and immune cells confer a pro-inflammatory environment that influences muscle loss through several mechanisms; however, this has not been explored in the cancer setting. This study investigated the local immune environment of the muscle by identifying the phenotype of immune cell populations in the muscle and their relationship to muscle mass in cancer patients.MethodsIntraoperative muscle biopsies were collected from cancer patients (n = 30, 91% gastrointestinal malignancies). Muscle mass was assessed histologically (muscle fiber cross-sectional area, CSA; μm2) and radiologically (lumbar skeletal muscle index, SMI; cm2/m2 by computed tomography, CT). T cells (CD4 and CD8) and granulocytes/phagocytes (CD11b, CD14, and CD15) were assessed by immunohistochemistry. Microarray analysis was conducted in the muscle of a second cancer patient cohort.ResultsT cells (CD3+), granulocytes/phagocytes (CD11b+), and CD3−CD4+ cells were identified. Muscle fiber CSA (μm2) was positively correlated (Spearman’s r = > 0.45; p = < 0.05) with the total number of T cells, CD4, and CD8 T cells and granulocytes/phagocytes. In addition, patients with the smallest SMI exhibited fewer CD8 T cells within their muscle. Consistent with this, further exploration with gene correlation analyses suggests that the presence of CD8 T cells is negatively associated (Pearson’s r = ≥ 0.5; p = <0.0001) with key genes within muscle catabolic pathways for signaling (ACVR2B), ubiquitin proteasome (FOXO4, TRIM63, FBXO32, MUL1, UBC, UBB, UBE2L3), and apoptosis/autophagy (CASP8, BECN1, ATG13, SIVA1).ConclusionThe skeletal muscle immune environment of cancer patients is comprised of immune cell populations from the adaptive and innate immunity. Correlations of T cells, granulocyte/phagocytes, and CD3−CD4+ cells with muscle mass measurements indicate a positive relationship between immune cell numbers and muscle mass status in cancer patients. Further exploration with gene correlation analyses suggests that the presence of CD8 T cells is negatively correlated with components of muscle catabolism.

Primary pulmonary lymphoma from bronchus-associated lymphoid tissue (clinical observation)

Purpose. To describe the clinical case of rare indolent extranodal non-Hodgkin’s lymphoma, which originates from bronchus-associated lymphoid tissue, and to analyze the main current trends in the diagnosis and treatment of this pathology.Using bibliosemantic and analytical methods of research in modern scientific literature, we analyzed the main historical data and clinical-radiological and immunomorphological criteria for the diagnosis of casuistically rare neoplasm – the primary pulmonary non-Hodgkin's lymphoma – the indolent extranodal B-cell lymphoma of the marginal zone, arising from the bronchus-associated lymphoid tissue (BALT). The prevalence of BALT-lymphoma is less than 0.5 % of all pulmonary malignant tumors and less than 1 % of all non-Hodgkin’s lymphomas. Nowadays, diagnostic approaches and standard systemic treatment of this disease are still widely discussed. The recommendations of the retrospective international study of primary extranodal lymphoma of lungs from the marginal zone conducted by the international extranodal lymphoma study group (IELSG) are presented and the main recommendations of the NCCN for the diagnosis and treatment of mucosa-associated lymphoma (2019) are analyzed. Treatment options for BALT-lymphomas vary in the range from antibiotic therapy and clinical observation to surgical resection or combination with chemotherapy and radiotherapy, but optimal therapy for the disease needs further study. It is noted that precise morphological definition and immunohistochemical phenotyping allow choosing an adequate protocol for chemo-therapeutic treatment (immunochemotherapy). The main options for radical surgical, chemo-therapeutic and radiological stages of BALT-lymphoma treatment and indications for their combined use within the framework of the multidisciplinary approach to treatment with lymphomas are discussed and their effectiveness is determined. The actual 4-year clinical observation of 48 year old patient with BALT-lymphoma is presented and the main radiological diagnostic criteria and difficulties of morphological diagnostics and immunohistochemical phenotyping are discussed. The above clinical observation is aimed at information of a wide range of practicing doctors – therapists, pulmonologists, phthisiologists, thoracic surgeons, oncologists, radiologists.Conclusions. BALT-lymphoma is a rare form of extranodal non-Hodgkin’s lymphoma, that does not have clear clinical guidelines and specific radiological signs and requires well-balanced histological diagnostics and immunohistochemical verification, which, in combination with computed tomography (PET-CT) methods, determines the stage of the disease and, respectively, tactics for the management of patients.

Prediction of response to pemetrexed in non-small-cell lung cancer with immunohistochemical phenotyping based on gene expression profiles

BackgroundPalliative pemetrexed-based chemotherapy remains a standard of care treatment for the majority of patients with advanced non-squamous non-small-cell lung cancer (NSCLC). Currently, no predictive markers for pemetrexed treatment are available.MethodsResected tumour samples from pemetrexed-naïve NSCLC patients were collected. Gene expression profiling with respect to predicted sensitivity to pemetrexed classified predicted responders (60%) and non-responders (40%) based on differentially expressed genes encoding for pemetrexed target enzymes. Genes showing a strong correlation with these target genes were selected for measurement of corresponding protein expressions by immunohistochemical (IHC) staining. A semi-quantitative IHC scoring method was applied to construct a prediction model for response to pemetrexed. A retrospective cohort of patients with advanced NSCLC treated with first-line pemetrexed-based chemotherapy was used for external validation.ResultsFrom ninety-one patients resected tumour samples were collected. The majority of patients had early or locally advanced NSCLC (96.3%). Gene expression profiling revealed five markers, which mRNA levels strongly correlated to pemetrexed target genes mRNA levels: TPX2, CPA3, EZH2, MCM2 and TOP2A. Of 63 (69%) patients IHC staining scores of these markers were obtained, which significantly differed between predicted non-responders and responders (P < 0.05). The optimized prediction model included EZH2 (OR = 0.56, 95% CI 0.35–0.90) and TPX2 (OR = 0.55, 95% CI 0.30–1.01). The model had a sensitivity of 86.8%, specificity of 63.6% and showed a good ability to distinct between responders and non-responders (C-index 0.86).In the external study population (N = 23) the majority of patients had metastatic NSCLC (95.7%). Partial response (PR) was established in 26.1%. The sensitivity decreased drastically to 33.3%, with a specificity of 82.4% and a C-index of 0.73.ConclusionsUsing external validation this prediction model with IHC staining of target enzyme correlated markers showed a good discrimination, but lacked sensitivity. The role of IHC markers as response predictors for pemetrexed in clinical practice remains questionable.

Macrophages of the subcutaneous and omental fatty tissue in obese patients: Immunohistochemical phenotyping of M2 subtypes in relation to type 2 diabetes.

Macrophages are linked to the initiation of the chronic inflammation believed to underlie the changes taking place in the white fatty tissue of obese people. Both the number of macrophages, but their functional status, play an important role in the development of inflammation. Classically, macrophages are divided into two types: pro-inflammatory (M1) and anti-inflammatory (M2) types, and based on current immunological studies, further views on the functional distribution of macrophages are suggested. In this study, we evaluated the M1 and M2 macrophages ratio in obese subjects with, or without diabetes. To identify all macrophages, we used CD68 expression, while CD204 expression is typically used for the M2 macrophage. During bariatric surgery, carried out in obese people with and without type 2 diabetes (T2D), we obtained subcutaneous adipose tissue from the navel and omental adipose tissue. We also obtained the same tissue from people with a physiological range of BMI from a judicial autopsy. Applying immunohistochemical staining anti-CD68 and anti-CD204, we carried out a quantitative evaluation of the number of macrophages. We found CD68+ and CD204+ positive macrophages in perivascular spaces and between fat cells, both isolated and in larger infiltrates. They were also present in so-called "crown-like structures" (CLS) around dying adipocytes. Quantitative analysis showed an increased number of macrophages in all obese patients compared to the control group of non-obese, individuals without T2D. The most striking observation was the macrophage increase in the visceral fatty tissue of diabetics. The number of CD68 and CD204 positive macrophages was statistically significantly smaller in patients without T2D. We demonstrated a significantly greater number of macrophages in visceral adipose tissue, especially in patients with T2D. Our results also show a positive correlation between the presence of T2D and the total number of macrophages; a significantly greater number of macrophages were found in visceral adipose tissue, especially in patients with T2D.

Anti-atherosclerotic action of GW9508 - Free fatty acid receptors activator - In apoE-knockout mice.

In the past two decades, enhanced understanding of the biology of G-protein-coupled receptors (GPRs) has led to the identification of several such receptors as novel targets for free fatty acids (FFAs). Two GPRs, FFAR1 and FFAR4, have received special attention in the context of chronic inflammatory diseases, thanks to their anti-inflammatory activities. The present study investigates the influence of prolonged treatment with GW9508 - agonist of FFAR1 and FFAR4 - on the development of atherosclerosis plaque in apoE-knockout mice, using morphometric and molecular methods. GW9508 administration has led to the reduction of atheroscletoric plaque size in an apoE-knockout mice model. Moreover, a FFAR1/FFAR4 agonist reduced the content of macrophages by almost 20%, attributed by immunohistochemical phenotyping to the pro-inflammatory M1-like activation state macrophages. Prolonged administration of GW9508 resulted in significant amelioration of atherogenesis, providing evidence that the strategy based on macrophage phenotype switching toward an M2-like activation state via stimulation of FFAR1/FFAR4 receptors holds promise for a new approach to the prevention or treatment of atherosclerosis.

Perspectives and Challenges in Molecular-Based Diagnostics and Personalized Treatment for Recurrent High-Grade Gliomas

Glioblastoma is the most common and most malignant type of intrinsic brain tumor in adults. The standard of care for glioblastoma consists of surgical debulking followed by combined radiochemotherapy. The clinical efficacy of standard therapies for newly diagnosed glioblastomas is rather modest with the highest survival rate at 5-years being less than 10%. Inevitable recurrence after cytotoxic therapies poses the major challenge in the clinical management of high grade gliomas. For recurrent glioblastomas, there is no standard therapy with lack of level one evidence for treatment efficacy. Recent evidence indicates that post-therapy recurrence in gliomas is a consequence of a plethora of molecular and cellular factors including intratumoural heterogeneity, functional hierarchy of distinct types of glioma cells, dynamic changes in the molecular landscapes and cellular composition of the tumour during therapy and the impact of particular treatment modalities. There is an emerging consensus that molecular distinctions within and between individual tumours is an important factor determining clinical outcomes. Consequently, integrated approaches based on the combination of molecular profiling with traditional methods such as immunohistochemical phenotyping, karyotyping and/or non-quantitative methylation-specific PCR have emerged as a promising venue towards increasing the predictive value of diagnostics for malignant brain tumors. The high level of inter-and intra-tumoural molecular diversity in gliomas underscores the need of integrating high throughput molecular profiling and pharmacogenomics into a diagnostic paradigm for gliomas and raises the possibility that molecular-instructed personalized treatments may provide clinical benefit to patients with glioblastoma, particularly in the setting of post-treatment recurrence. Here we discuss potential prospects and challenges of patient-tailored diagnostics and personalized treatment strategies for recurrent glioblastomas.

Treatment with a human recombinant monoclonal IgG antibody against oxidized LDL in atherosclerosis-prone pigs reduces cathepsin S in coronary lesions

BackgroundImmunization with oxidized LDL (oxLDL) reduces atherosclerosis in rodents. We tested the hypothesis that treatment with a human recombinant monoclonal antibody against oxLDL will reduce the burden or composition of atherosclerotic lesions in hypercholesterolemic minipigs. Methods and resultsThirty-eight hypercholesterolemic minipigs with defective LDL receptors were injected with an oxLDL antibody or placebo weekly for 12weeks. An 18F-fluorodeoxyglucose positron emission tomography (FDG PET) scan (n=9) was performed before inclusion and after 3months of treatment. Blood samples were obtained prior to each injection. Following the last injection all animals were sacrificed, and the heart, aorta, and iliac arteries were removed. The left anterior descending coronary artery was sectioned at 5mm intervals for quantitative and qualitative assessments of atherosclerosis, including immunohistochemical phenotyping of macrophages using a pan-macrophage marker (CD68) and markers for putative pro-atherogenic (cathepsin S) and atheroprotective (CD163) macrophages. Aorta, right coronary artery, and left iliac artery were stained en face with Sudan IV and the amount of atherosclerosis quantified. There was no effect of treatment on plasma lipid profile, vascular FDG-PET signal or the amount of atherosclerosis in any of the examined arteries. However, immunostaining of coronary lesions revealed reduced cathepsin S positivity in the treated group compared with placebo (4.8% versus 8.2% of intima area, p=0.03) with no difference in CD68 or CD163 positivity. ConclusionsIn hypercholesterolemic minipigs, treatment with a human recombinant monoclonal antibody against oxLDL reduced cathepsin S in coronary lesions without any effect on the burden of atherosclerosis or aortic FDG-PET signal.

Morphometric and immunohistochemical study of regional lymph nodes in gastric cancer

Aim. To perform quantitative analysis and immunohistochemical phenotyping of regional lymph nodes structural and cellular elements in gastric cancer in the absence and the presence of metastases.&#x0D; Methods. The regional lymph nodes obtained during surgery for gastric cancer from 48 patients (31 men and 17 women, mean age 60.9 years) were studied. Histological sections were stained with haematoxylin and eosin, van Gieson stain, azure-II-eosin, pyronin by Brashe. Then morphometric analysis of lymph nodes structural and cellular elements was performed. A set of monoclonal antibodies against CD45, CD3, CD4, CD8, CD10, CD20, CD30, BLA-36, immunoglobulin (Ig) λ- and κ-chains, CD56, myeloperoxidase, CD68, lysozyme, α1-antichymotrypsin, CD35, S100, Ki-67, CD31, collagen type IV, vimentin, desmin was used for immunohistochemical reactions. Comparative study was performed in the following groups: (1) control group; (2) lymph nodes without metastases; (3) lymph nodes with metastases of different size.&#x0D; Results. Paracortical and follicular hyperplasia, sinus histiocytosis, microcirculatory disorders and fibroplastic processes were found in regional lymph nodes in gastric cancer development. Paracortical hyperplasia was characterized by following immunohistochemical phenotype: CD 45, CD 3, CD 4, CD 8, CD 10, CD 30, CD 56, S 100, CD 31, Ki-67. CD45, CD20, BLA-36, CD10, CD30, CD35, Ig λ- and κ-chains, Ki-67 were mainly expressed in follicular hyperplastic reaction. The sinus reaction was accompanied by total cellularity increase with a predominance of CD68(+), α1-antichymotrypsin(+) and lysozyme(+) cells. Expression of CD3, CD4, CD8, CD10, CD56, Ki-67 decreased amid the paracortical zone area reduce in metastases development. Sinuses were devastated and poor with cellular elements, often with lymphostasis or sclerosis signs. Imunohistochemically there was moderate reaction with monoclonal antibodies against α1-antichymotrypsin, low - against lysozyme, myeloperoxidase, and high - against vimentin. Wherein follicular hyperplasia with high levels of CD20(+) B-cells, BLA-36(+) activated lymphocytes and Ig(+) antibody producing plasma cells persisted. At the same time microcirculatory disorders and fibroplastic processes progressed.&#x0D; Conclusion. Structural components area ratio, cellular composition and immunohistochemical phenotype change in the regional lymph nodes in gastric cancer development, with significant differences between intact and affected lymph nodes; paracortical zone and sinuses area reduction with T-, NK-cells and macrophage markers expression decrease amid the preservation of follicular hyperplasia with high level of activated B lymphocytes and antibody producing plasma cells can promote metastases engrafment in lymph nodes and further cancer generalization.

Lymphocytic Infiltration as a Prognostic Factor in Patients With Colon Cancer.

Host-defense mechanisms may have an important role in predicting the outcome of colorectal cancer patients. We designed our study to evaluate the possible prognostic significance of the presence of lymphocytic infiltration (LI) and subgroups of lymphocytes (CD3 and CD20) in the primary tumors. We randomly selected 195 patients operated for colorectal carcinoma from a larger cohort of 1527 patients with colorectal cancer. Histological slides were blindly reevaluated for the presence of LI that was graded 0 to 3. Immunohistochemical phenotyping of the lymphocytes was performed only for tumors with LI score 3 and included antibodies CD3 and CD20. CD3 and CD20 immunostaining were graded in the same manner as LI. The mean duration of follow-up was 63.8 months. The distribution of patients with colorectal cancer according to LI scores was as follows: score 0, 20/195 (10.2%); score 1, 61/195 (31.3%); score 2, 78/195 (40%); and score 3, 36/195 (18.5%). There was no correlation between any clinicopathological pattern and LI. Score 3 staining for CD3 was more common than for CD20 (64.7% vs 8.8%, P < .0001). Prominent lymphocytic infiltration (score 3) was associated with better disease-free survival (P = .062). Recurrence was diagnosed among 2/22 (9.1%) patients with prominent CD3 staining versus 62/171 (36.2%) of all other patient groups (P = .054) and they correspondingly had better disease-free survival (P = .018). It seems we can identify a group of patients with colorectal cancer who have an excellent prognosis according to a single immunological test unrelated to other known prognostic factors.