The prognostic and predictive implications of the 12-chemokine score in muscle invasive bladder cancer.

466 Background: Adaptive anti-tumor immunity can be orchestrated by lymph node-like immune cell aggregates within the tumor microenvironment (TME) called tertiary lymphoid structures (TLSs). TLSs are postulated to be the gateway of lymphocyte infiltration into the TME, and are privileged sites for coordinated tumor antigen presentation and lymphocyte priming, differentiation, and proliferation, leading to a robust tumor-specific immune response. A 12-chemokine metagene grouping (12-CK score) has previously been described that correlates with the presence of TLSs in other solid tumor types. In this study, we explored the prognostic implication of the 12-CK score in bladder cancer and its correlation with the presence of TLSs. Methods: Cystectomy specimens from 132 patients with bladder cancer were arrayed on Affymetrix microarrays. 12-CK scores were normalized with > 1 denoting high scores (12-CKHi). Immunohistochemistry (IHC) antibody staining was performed for DC-LAMP, CD20, CD4, and CD8. A GU pathologist scored TLSs into Types I-III, with type III representing fully developed TLSs. The Fisher’s exact test was used to test the associations between the 12-CK scores and the type of lymphoid aggregate. Overall survival was estimated using the Kaplan Meier method. Findings were validated using 12-CK scores extracted from TCGA transcriptome sequencing data and the IMvigor210CoreBiologies package. Results: Twenty-five (n = 25) patients had 12CK scores > 1 and were classified as 12CK-High. Pathologic review of 43 bladder tumor specimens confirmed higher levels of Type III TLS patients (33% vs. 9%, p = 0.03), B cells (p = 0.002), CD8 T cells (p = 0.01), and activated DC (p = 0.01) in 12-CKHi compared to 12-CKLo. 12-CKHi was found to have a progression-free survival (PFS, HR 0.29, p = 0.003, Fig1a), disease specific survival (DSS, HR 0.29, p = 0.004, Fig1b), and overall survival (OS, HR 0.55, p = 0.03, fig1c) advantage compared to 12-CKLo in the Moffitt patient cohort. These results were validated using the publically available RNA expression data from TCGA. TCGA patients with 12-CKHi (18%,n = 72) had improved PFS ( HR 0.55, p = 0.007, fig1d), DSS (HR = 0.40, p = 0.002, fig1e), and 0S (HR = 0.59, p = 0.01, fig1f). From the IMVIGOR-210 patient who were 12-CKHi were more likely to have a complete response (p < 0.05, fig1g) and have a 11.2mo OS benefit (fig1h) after treatment using atezolizumab. Conclusions: Three important findings emerged from the current study: 12CK-High scores corresponded with formation of TLS in the TME; favorable prognosis in surgically treated MIBC patients; and CR in atezolizumab-treated patients. The findings herein suggest the 12CK gene signature to be a clinically actable biomarker for predicting response to immune checkpoint blockade. We believe the 12CK signature may serve as an important tool to refine patient selection for immune checkpoint blockade treatment.

Anti-IL-17A treatment reduces serum inflammatory, angiogenic and tissue remodeling biomarkers accompanied by less synovial high endothelial venules in peripheral spondyloarthritis

Spondyloarthritis (SpA) is characterized by inflammation and new bone formation. The exact pathophysiology underlying these processes remains elusive. We propose that the extensive neoangiogenesis in SpA could play a role both in sustaining/enhancing inflammation and in new bone formation. While ample data is available on effects of anti-TNF on angiogenesis, effects of IL-17A blockade on serum markers are largely unknown. We aimed to assess the impact of secukinumab (anti-IL-17A) on synovial neoangiogenesis in peripheral SpA, and how this related to changes in inflammatory and tissue remodeling biomarkers. Serum samples from 20 active peripheral SpA patients included in a 12 week open-label trial with secukinumab were analyzed for several markers of angiogenesis and tissue remodeling. Synovial biopsies taken before and after treatment were stained for vascular markers. Serum levels of MMP-3, osteopontin, IL-6 (all P < 0.001), IL-31, S100A8, S100A9, Vascular Endothelial Growth Factor A (VEGF-A), IL-33, TNF-α (all P < 0.05) decreased significantly upon anti-IL17A treatment. Secukinumab treatment resulted in a decrease in the number of synovial high endothelial venules and lymphoid aggregate score. These results indicate that anti-IL-17A not only diminishes inflammation, but also impacts angiogenesis and tissue remodeling/new bone formation. This may have important implications for disease progression and/or structural damage.

The Rare First Manifestation Of Mantle Cell Lymphoma As Immune Complex Glomerulonephritis With Crescents And Membranoproliferative Features

Introduction/Objective Initial presentation of Non-Hodgkin Lymphoma (NHL) with immune complex glomerulonephritis (ICGN) is rare. Mantle cell lymphoma (MCL) is a rare aggressive lymphoma comprising 3–7 % of all NHL and ICGN as the first manifestation of MCL is very unusual. Methods We report a 73 year-old-man with increased serum creatinine (2.1 mg/dl) from normal baseline 3 months ago. Urinalysis showed the presence of dysmorphic RBCs, RBC casts and 300 mg/dl of protein. Physical examination revealed diffuse lymphadenopathy. Hepatitis C and B Ab, HIV screen, ANA, ANCA, Anti dsDNA Ab, Anti GBM Ab, and Serum protein electrophoresis were normal or negative. Results The renal biopsy showed Immune complex glomerulonephritis with (25%) fibrocellular/cellular crescents and focal membranoproliferative (MPGN) features. There were foci of dense monomorphic lymphoid aggregates of small lymphocytes and 30% interstitial fibrosis with corresponding tubular atrophy. No tubulitis was present. Direct Immunofluorescence showed 2+ to 4+ granular mesangial and capillary loop staining for Total Ig, IgG, IgA, IgM, C1q, C3 and C4. The lymphoid aggregate was monoclonal Kappa. Electron microscopy showed widespread podocyte foot process effacement, subendothelial and rare mesangial dense deposits. Excisional lymph node biopsy flow cytometry showed a monoclonal population of CD19+, CD20+, CD22+, CD5+, skappa-restricted, CD10-, CD11c-, CD23-, CD25- CD38-, CD103-, CD200- B- cells. Cyclin D1 was positive on immunostain, consistent with stage IV (renal) MCL. He received chemotherapy with steroids, bendamustine, and rituximab leading to resolution of lymphadenopathy and proteinuria and improvement in serum creatinine. Conclusion It is important to recognize that the first manifestation of MCL may present as renal involvement in the form of ICGN and, as in this case, with fibrocellular/cellular crescents and a focal MPGN pattern. This is an extremely rare, but important, cause of renal insufficiency that can be successfully treated with lymphoma chemotherapy.

Incidence of Helicobacter Pylori Infections in a group of patients in Erbil City, Kurdistan Region, Iraq and Its Association with Gastritis and Adenocarcinoma

Background: Adenocarcinoma is one of the most common causes of Gastric cancer related deaths worldwide. Helicobacter pylori is the causative agent of most cases of gastritis, it can cause chronic active gastritis and known as a risk factor for the development of gastric cancer. This study aimed to assess the prevalence of H. pylori among patients with symptoms of dyspepsia and other gastritis related symptoms and its association with adenocarcinoma.Methods: This study was carried out during the period of January 2018 to October 2019 with a total of 227 patients with gastritis related symptoms. The presence of H. pylori was detected by Rapid Urease Test (RUT) and histo-pathological tests using biopsy specimens. Statistical Analysis was done by using Chi-square test. P < 0.05 was considered to be statistically significant.Results: From the total of 227 patients with gastritis related symptoms, 26 cases (13.61%) were diagnosed with adenocarcinoma. Their ages were between 13 and 90 years with mean of 47.81± 18.23. The result showed that low severity prevalence of H. pylori was highest (111 cases) compared to 17 and 63 cases for high and moderate severity, respectively. Comparison between positive low, moderate, and high H. pylori cases for rapid urease test was highly significant (P<0.000). The results showed no association between H. pylori severity across various age groups and gender. Moreover, goodness of fit test for metaplasia, activity, glandular atrophy, and endoscopic finding across severity status of H. pylori showed highly significant. Four composite categorized groups were initiated based on positive/negative prevalance of H. pylori and adenocarcinoma status. Results revealed statistical significance between combination of H. pylori and adenocarcinoma with inflammation, lymphoid aggregate, metaplasia, activity of neutrophils, glandular atrophy, rapid urease test, and endoscopic findings.Conclusion: Histopathology tests are reliable diagnostic tools for the detection of H. pylori. Data showed that H. pylori was seen more in middle age patients with mucosal lymphoid follicle formation and more than one third of patients with adenocarcinoma. Therefore, screening of these infections is an important strategy for preventing gastric adenocarcinoma.

Sex differences in lymphoid follicles in COPD airways

Background Female smokers have increased risk for chronic obstructive pulmonary disease (COPD) compared with male smokers who have a similar history of cigarette smoke exposure. Tertiary lymphoid follicles are often found in the lungs of patients with severe COPD but sex-related differences have not been previously investigated. We determined the impact of female sex hormones on chronic cigarette smoke-induced expression of lymphoid aggregates in mice with COPD-like pathologies. Methods Lymphoid aggregate counts, total aggregate cross-sectional area and foamy macrophage counts were determined morphometrically in male, female, and ovariectomized mice exposed to air or cigarette smoke for 6 months. B-cell activating factor (BAFF) protein expression and markers of oxidative stress were evaluated in mouse lung tissues by immunofluorescence staining and gene expression analyses. Quantitative histology was performed on lung tissue sections of human COPD lungs to evaluate follicle formation. Results Lymphoid follicle and foamy macrophage counts as well as the total follicle cross-sectional area were differentially increased in lung tissues of female mice compared to male mice, and these differences were abolished by ovariectomy. These lymphoid aggregates were positive for CD45, CD20, CD21 and BAFF expression. Differential increases in Mmp12 and Cxcl2 gene expression correlated with an increase in foamy macrophages in parenchymal tissues of female but not male mice after smoke exposure. Parenchymal tissues from female mice failed to induce antioxidant-related genes in response to smoke exposure, and this effect was restored by ovariectomy. 3-nitrotyrosine, a stable marker of oxidative stress, positively correlated with Mmp12 and Cxcl2 gene expression. Hydrogen peroxide induced BAFF protein in mouse macrophage cell line. In human lung tissues, female smokers with severe COPD demonstrated increased numbers of lymphoid follicles compared with males. Conclusions Chronic smoke exposure increases the risk of lymphoid aggregate formation in female mice compared with male mice, which is mediated female sex hormones and BAFF expression in an oxidative environment.

Circulating LOXL2 Levels Reflect Severity of Intestinal Fibrosis and GALT CD4+ T Lymphocyte Depletion in Treated HIV Infection

Background: Incomplete immune reconstitution may occur despite successful antiretroviral therapy (ART). Gut-associated lymphoid tissue (GALT) fibrosis may contribute via local CD4+ T lymphocyte depletion, intestinal barrier disruption, microbial translocation, and immune activation.Methods: In a cross-sectional analysis, we measured circulating fibrosis biomarker levels on cryopreserved plasma from adult HIV-infected (HIV+) SCOPE study participants on suppressive ART who also had fibrosis quantification on recto-sigmoid biopsies. Relationships among biomarker levels, clinical and demographic variables, GALT lymphoid aggregate (LA) collagen deposition, and LA CD4+ T lymphocyte density were analyzed using simple regression. Biomarker levels were also compared to levels in HIV+ viremic SCOPE participants and a convenience sample of HIV-uninfected (HIV-) samples. Results: HIV+ aviremic participants (n=39) were 92% male and 41% non-white, with median age 48 years, CD4+ T lymphocyte count 277 cells/mm3, and 17 years since HIV diagnosis. Most biomarkers were lower in HIV− (n=36) vs HIV+ aviremic individuals, although CXCL4 levels were higher. HIV+ viremic individuals (N=18) had higher median TGF-ß3, CIC-C1Q, and TIMP-1 (P<0.05) and lower LOXL2 levels (P=0.08) than HIV+ aviremic individuals. Only higher LOXL2 levels correlated with more GALT collagen deposition (R=0.44, P=0.007) and lower LA CD4+ T lymphocyte density (R=−0.32, P=0.05) among aviremic individuals.Conclusions: Circulating LOXL2 levels may be a noninvasive measure of intestinal fibrosis and GALT CD4+T lymphocyte depletion in treated HIV infection. LOXL2 crosslinks elastin and collagen, and elevated LOXL2 levels occur in pathologic states, making LOXL2 inhibition a potential interventional target for intestinal fibrosis and its sequelae.

Bacteria-driven peribronchial lymphoid neogenesis in bronchiectasis and cystic fibrosis

We aimed to characterise lymphoid neogenesis in bronchiectasis and cystic fibrosis (CF) lungs and to examine the role of bacterial infection.Lymphoid aggregates were examined using immunohistochemical staining and morphometric analysis in surgical lung sections obtained from nonsmokers and patients with bronchiectasis or CF. Sterile, Pseudomonas aeruginosa- or Staphylococcus aureus-coated agarose beads were instilled intratracheally in mice. Kinetics of lymphoid neogenesis and chemokine expression were examined over 14 days.Lymphoid aggregates were scarce in human lungs of nonsmokers, but numerous peribronchial lymphoid aggregates containing B-lymphocytes, T-lymphocytes, germinal centres and high endothelial venules were found in bronchiectasis and CF. Mouse lungs contained no lymphoid aggregate at baseline. During persistent P. aeruginosa or S. aureus airway infection peribronchial lymphoid neogenesis occurred. At day 14 after instillation, lymphoid aggregates expressed markers of tertiary lymphoid organs and the chemokines CXCL12 and CXCL13. The airway epithelium was an important site of CXCL12, CXCL13 and interleukin-17A expression, which began at day 1 after instillation.Peribronchial tertiary lymphoid organs are present in bronchiectasis and in CF, and persistent bacterial infection triggered peribronchial lymphoid neogenesis in mice. Peribronchial localisation of tertiary lymphoid organs and epithelial expression of chemokines suggest roles for airway epithelium in lymphoid neogenesis.

Bone marrow trephine biopsy in Haematological diseases: a study of 53 cases

Bone marrow aspiration (BMA) and biopsy (BMTB) are important investigations for diagnosis of haematolgical malignancies and non-malignant diseases both in adults and children. BMA and BMTB are complementary and if both are done a comprehensive analysis of bone marrow involvement is possible. 53 cases of BMTB were studied in order to underscore the indications and importance of BMTB. BMTB was done to determine cellularity in aplastic anaemia (AA) (33.96%, n=18) and in cases of failure of aspiration (32.08%, n=17). Failure of aspiration was attributable to bone marrow (BM) fibrosis (76%, n=13) due to acute leukaemia (35.30%, n=6) and myelofibrosis (43.17%, n=7). BMTB upstaged non Hodgkin’s lymphoma (NHL) from IIIB to IVB in 22.22% cases. 1 case of AA showed focal lymphoid aggregate which later evolved into acute lymphoblastic leukaemia (ALL). BMTB is a safe procedure and increased bleeding was noted only in a case of polycythaemia vera. DOI: http://dx.doi.org/10.3329/jdmc.v22i2.21544 J Dhaka Medical College, Vol. 22, No.2, October, 2013, Page 207-210