Lymph Node Tissues Of Patients Research Articles

Оценка метилирования генов MIR-143 и MIR-145 в лимфоузлах больных диффузной В-крупноклеточной лимфомой

Introduction. The expression of miR-143 and miR-145 oncosuppressive microRNAs is observed in all types of normal tissues studied and, according to numerous studies, it is lost in malignant neoplasms. A decrease in the expression of these microRNAs in diffuse large B-cell lymphoma (DLBCL) is described, the mechanisms of which require additional study. Aim. To evaluate the methylation of MIR-143 and MIR-145 genes in the lymph node tissue of patients with DLBCL and lymph nodes of patients with reactive follicular hyperplasia. Materials and methods. The study included biopsies of 14 formalin-fixed and paraffinized tumor lymph nodes of patients with DLBCL and 11 lymph nodes of patients with reactive lymphadenopathy. The methylation status of the MIR-145 gene in samples was determined by the method of methyl-specific polymerase chain reaction. Direct bisulfite sequencing based on the Sanger method was used to quantify the methylation of the MIR-143 gene. Results. It was found that all the studied samples, both reactive and tumor, had methylation of the MIR-145 gene. A similar level of methylation of CpG dinucleotides of the MIR-143 gene was registered in all samples of reactive lymph node tissue, whereas tumor samples showed greater heterogeneity. In the samples of DLBCL, namely a non-germinal B cell phenotype, the average level of methylation of the studied fragment of the MIR-143 gene sequence was significantly lower, than in the samples of reactive lymph nodes (p = 0.026). Conclusion. The methylation of MIR-143 and MIR-145 genes detected in the lymph nodes of patients with DLBCL is not tumor-specific. The complex cellular composition of the analyzed samples, as well as the different density of microvessels, may explain the differences in the level of MIR-143 methylation in the tissue of reactive and tumor lymph nodes.

Macroelement and microelement composition of mediastinal lymph nodes of patients with sarcoidosis

This study was aimed at measuring concentrations and investigation of relationships between macro- and microelements in mediastinal lymph nodes of patients with sarcoidosis. Methods. The study involved 28 patients (50% were males; median age, 42.5 (23; 53) years) with confirmed diagnosis of sarcoidosis. Autopsy samples from 13 individuals died from occasional causes (41.2% were males; median age, 43 (38.7; 61.3) years) were used for comparison. The groups did not differ in age and gender. Neutron activation analysis was used to investigate chemical elements in mediastinal lymph node tissue samples. Results. Concentrations of Ca, Cr, Fe, Co, Zn, Rb, Sb, Cs, Ba, Eu, Lu, Yb, Hf, Th, and U in the lymph node tissue of patients with sarcoidosis were increased, and concentration of zinc was decreased compared to control samples. Concentrations of Sm, Au, Sr, Nd, As, Br, Ag, Tb, Sc, Ta, Na, and La did not differ between the groups. Direct relationships were found in element groups Fe-Cr-Sb-Ca, Fe-U-Th, Co-U-Th, U-Cr, Th-Br, Sc-Zn, Eu-La, and Ce-Lu.

Human Herpesvirus Type 8-positive Multicentric Castleman Disease.

Castleman disease (CD) is rare lymphoproliferative disorder with local lesionsor with multiple lessions (multicentric CD [MCD]-usually with plasma cell or mix cell morphology). Patients with human herpesvirus (HHV) type 8-positive MCD were included in a separate group owing to its extremely aggressive course and the high risk of transformation into HHV8(+) plasmablastic lymphoma. At our hematologic center, from 1996 to the present, the clinical and morphologic features of 87 patients with CD were analyzed. Immunohistochemical examination revealed DNA HHV8(+) lymph node tissue in patients with plasma cell and mixed cell morphology. In 45 patients, plasma cell or mixed cell variant CD was diagnosed. In 21 patients (8%), the manifestation of CD was local and in 29 (9%), it was multicentric. HHV8 was identified in only 6 cases (23.1%) of MCD (5 men and 1 woman, with a median age of 48.2 years; range, 36-77 years). The median follow-up point was 39.2 months. In 4 patients, the mixed cell variant was diagnosed and in 2, the plasma cell variant was diagnosed. In all the patients, constitutional symptoms, generalized lymphadenopathy, and hepatosplenomegaly were detected. Various laboratory changes were observed, but the most significant were anemia, thrombocytopenia, hypergammaglobulinemia, M-component, increased erythrocyte sedimentation rate, and circulating immune complexes. In 2 cases of HHV8(+) CD, MCD was combined with autoimmune hemolytic anemia and in 2 cases with non-Hodgkin lymphoma. At the last follow-up point, 2 patients were still alive after CHOP (cyclophosphamide, prednisone, Adriamycin, vincristine) and R-CHOP (rituximab, cyclophosphamide, doxorubicin hydrochloride, vincristine sulfate [Oncovin], prednisone) therapy with rituximab maintenance. HHV8(+) MCD results in aggressive multiorgan lesions and pronounced changes in laboratory test results. It is characterized by an unfavorable prognosis with a high risk of transformation to plasmablastic lymphoma and a lethal outcome. Timely chemotherapy for patients with HHV8(+) MCD can result in remission and prolong life.

Lymph node micrometastases in initial stage cervical cancer and tumoral recurrence.

To evaluate the prevalence of micrometastases in lymph node tissue of patients with stage Ib1-IIA cervical cancer, the correlation of micrometastases with tumor recurrence and survival, and the expression of D2-40 in the primary tumor of patients with recurrence and/or micrometastases and its correlation with histopathologic findings. In a retrospective study, the medical records of all patients with cervical cancer treated at a hospital in São Paulo, Brazil, between 2001 and 2007 were reviewed. Patients with no lymph node metastases and treated with radical hysterectomy without adjuvant treatment were included. Tumor sections were reviewed and lymph nodes were analyzed with AE1/AE3. Patients with and without recurrence were compared. The presence of lymph node micrometastasis or isolated tumor cells was also evaluated. Of the 83 patients evaluated, 15 (18%) had recurrence. Significant differences between patients with and without recurrence were observed with regard to tumor greatest axis, clinical stage, number of micrometastases, and negative lymph nodes (P≤0.04). Lymph node micrometastases and isolated tumor cells were significantly different for a stromal invasion depth greater than 2/3 (P=0.046). The presence of lymph node micrometastases is an important risk factor for tumor recurrence. These patients should be considered eligible for adjuvant radiochemotherapy treatment.

External validation of FXYD3 and KRT20 as predictive biomarkers for the presence of micrometastasis in muscle invasive bladder cancer lymph nodes

IntroductionRecent studies have proposed that FXYD3 and KRT20 mRNA quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in paraffin could be biomarkers to detect lymph nodes with micrometastases that avoid detection by conventional analysis with hematoxylin–eosin (HE). A validation study was conducted on the lymph nodes of patients who underwent radical cystectomy. ObjectiveTo classify the adenopathic state of a sample of patients who underwent cystectomy, based on the lymph node expression of FXYD3 and KRT20. The secondary objective was to assess whether there is a differential oncologic evolution for the patients, depending on the lymph node expression of these proteins. Material and methodThe study included lymph nodes from 64 patients who underwent cystectomy for infiltrating bladder tumor: The model was developed using metastatic lymph nodes from 15 patients and lymph nodes from four patients with no known tumor. Genetic expression was measured using real-time qRT-PCR. We calculated (using qRT-PCR) the median expression of FXYD3 and KRT20 mRNA in the lymph node tissue. We then analyzed the receiver operating characteristic (ROC) curves, according to the function y=0.1400+0.250FXYD3−2.532. The cutoff was established using an ROC curve. The formula was applied to the remaining lymph node tissue, based on the previously established cutoff. The sample was classified into four subgroups: HE− qRT-PCR−, HE− qRT-PCR+, HE+ qRT-PCR+ and HE+ qRT-PCR−. A descriptive, comparative analysis was performed, as well as a metastatic progression-free survival analysis, calculating the Kaplan and Meyer curves for the three established subgroups. The test results were considered statistically significant at p<0.05. ResultsUsing qRT-PCR, we verified that there were differences in the median expression of FXYD3 (p=0.05) and KRT20 (p=0.009) between the lymph node tissues of patients with benign prostate hyperplasia and those of patients with lymph node metastasis. A cutoff was assigned to 0.377. The sample was classified as follows: 37.5% of the patients were pN0 by HE and pN0 by qRT-PCR (−HE −qRT-PCR), 39.1% were pN0 by HE but metastatic by qRT-PCR (−HE +qRT-PCR), and 15 patients (23.4%) were metastatic by both techniques (+HE +qRT-PCR). The Kaplan and Meyer curves showed poorer metastatic progression-free survival for the patients who were +HE and +qRT-PCR than for the other subgroups, with no significant differences between −HE +qRT-PCR and −HE −qRT-PCR. ConclusionsAccording to our results, 39.1% of the patients with infiltrating vesical tumors overexpressed the FXYD3 and KRT20 biomarkers and were N0 by HE. We observed no differential clinical behavior among the patients who underwent cystectomy according to their expression of FXYD3 and KRT20 when they were N0 by HE.

Validación externa de los ARNm: FXYD3 y KRT20 como biomarcadores predictivos de la presencia de micrometástasis en ganglios de pacientes con tumor vesical infiltrante

IntroductionRecent studies have proposed that FXYD3 and KRT20 mRNA quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in paraffin could be biomarkers to detect lymph nodes with micrometastases that avoid detection by conventional analysis with hematoxylin-eosin (HE). A validation study was conducted on the lymph nodes of patients who underwent radical cystectomy. ObjectiveTo classify the adenopathic state of a sample of patients who underwent cystectomy, based on the lymph node expression of FXYD3 and KRT20. The secondary objective was to assess whether there is a differential oncologic evolution for the patients, depending on the lymph node expression of these proteins. Material and methodThe study included lymph nodes from 64 patients who underwent cystectomy for infiltrating bladder tumor: The model was developed using metastatic lymph nodes from 15 patients and lymph nodes from 4 patients with no known tumor. Genetic expression was measured using real-time qRT-PCR. We calculated (using qRT-PCR) the median expression of FXYD3 and KRT20 mRNA in the lymph node tissue. We then analyzed the receiver operating characteristic (ROC) curves, according to the function y=0.1400+0.250FXYD3-2.532. The cutoff was established using an ROC curve. The formula was applied to the remaining lymph node tissue, based on the previously established cutoff. The sample was classified into 4 subgroups: HE- qRT-PCR-, HE- qRT-PCR+, HE+ qRT-PCR+ y HE+, qRT-PCR-. A descriptive, comparative analysis was performed, as well as a metastatic progression-free survival analysis, calculating the Kaplan and Meyer curves for the 3 established subgroups. The test results were considered statistically significant at P<.05. ResultsUsing qRT-PCR, we verified that there were differences in the median expression of FXYD3 (P=.05) and KRT20 (P=.009) between the lymph node tissues of patients with benign prostate hyperplasia and those of patients with lymph node metastasis. A cutoff was assigned to 0.377. The sample was classified as follows: 37.5% of the patients were pN0 by HE and pN0 by qRT-PCR (-HE -qRT-PCR), 39.1% were pN0 by HE but metastatic by qRT-PCR (-HE +qRT-PCR), and 15 patients (23.4%) were metastatic by both techniques (+HE +qRT-PCR). The Kaplan and Meyer curves showed poorer metastatic progression-free survival for the patients who were +HE and +qRT-PCR than for the other subgroups, with no significant differences between -HE +qRT-PCR and -HE -qRT-PCR. ConclusionsAccording to our results, 39.1% of the patients with infiltrating vesical tumors overexpressed the FXYD3 and KRT20 biomarkers and were N0 by HE. We observed no differential clinical behavior among the patients who underwent cystectomy according to their expression of FXYD3 and KRT20 when they were N0 by HE.

Expression of microRNA-1234 related signal transducer and activator of transcription 3 in patients with diffuse large B-cell lymphoma of activated B-cell like type from high and low infectious disease areas

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease, and infectious agents are suspected to be involved in the tumorigenesis of DLBCL. MicroRNAs (miRNAs) are non-coding RNAs modulating protein expression. We compared miRNA expression profiles in lymph node tissues of patients with DLBCL of the activated B-cell like (ABC) type from two geographical areas with different background exposures, Sweden and Egypt. We showed previously that DLBCL tissues of the ABC-type in Swedish patients had a higher expression of signal transducer and activator of transcription 3 (STAT3) compared to Egyptian patients. Here, we analyzed the involvement of miRNAs in STAT3 regulation. miR-1234 was significantly up-regulated in Egyptian patients with DLBCL compared to Swedish patients (p < 0.03). The miR-1234 expression level correlated inversely with the expression of STAT3. The Stat3 protein was down-regulated in cells transfected with miR-1234, suggesting that STAT3 might be a potential target for miR-1234. miR-1234 and STAT3 might be involved in the tumorigenesis of DLBCL of ABC type and possibly associated with environmental background exposure.

Detection and preliminary characterization of CD8+T lymphocytes specific for Wilms’ tumor antigen in patients with non-Hodgkin lymphoma

Wilms’ tumor antigen (WT1) is overexpressed in many different solid tumors and hematologic malignancies. However, little is known about WT1 expression or WT1-specific immune responses in patients with non-Hodgkin lymphoma (NHL). In a cross-sectional survey study, we investigated the immune recognition of WT1 by patients with NHL. Utilizing a WT1 overlapping peptide library, we discovered that a large percentage of patients with NHL of all grades maintain WT1-specific T cells. Ex vivo frequencies of these T cells measured from unfractionated samples by the CD137 activation marker assay were high in many patients (some > 1% CD8+). Using standard in vitro techniques we discovered that they were cytotoxic to WT1 peptide library-loaded T2 cells and WT1 antigen-primed autologous Epstein–Barr virus-transformed B cell lines (EBV-LCLs) and expressed interferon gamma (IFN-γ). In addition, we detected WT1 mRNA transcripts in diseased lymph node tissues of patients with NHL utilizing real-time quantitative polymerase chain reaction (RT-qPCR) technology. These results are the first example of strong T cell reactivity against WT1 in patients with NHL which also demonstrate strong cytotoxicity against peptide-loaded tumor cells. The potential for developing WT1 as a target for immunotherapy in NHL deserves further exploration.

Abstract 405: The role of Stat3 in pre-metastatic niche formation: Examination in murine models and benign lymph nodes from prostate cancer patients

Abstract Objectives: High-risk prostate cancer (PC) represents a heterogeneous disease, and metastatic potential varies within this subset of patients. The occurrence of metastasis involves both tumor cells and tumor-associated stromal cells. In the present study, murine models were used to examine the association between Stat3 activation and tumor-associated stromal cells. The latter may confer metastatic potential through the formation of pre-metastatic niches. These parameters were subsequently assessed in the benign lymph nodes of patients with high-risk PC. Methods: To induce pre-metastatic conditions, we injected tumor-conditioned media from murine tumor cell lines over-expressing the Stat3 activator, S1PR1, into C57BL/6 mice. Subsequently, mice were challenged with parental tumor cells via tail vein injection. For assessing a potential role of Stat3 in formation of the pre-metastatic niche, we analyzed mouse lung tissue by immunofluorescent staining and Western blot. We also increased and decreased myeloid cell Stat3 activity by retrovirus transduction and genetic ablation, respectively. For clinical correlation, paraffin-embedded lymph node tissue derived from 50 patients with high-risk PC was acquired. High-risk PC was defined by standard criteria as follows: (1) baseline prostate specific antigen (PSA) ≫ 20, (2) clinical stage T3a disease, or (3) Gleason score 8-10. Immunohistochemistry (IHC) staining was performed to assess pStat3 levels and CD68+ myeloid cell clustering. Results: In the murine models examined, increasing Stat3 activity in tumor cells by its newly identified activator, S1PR1, produced factors that activated Stat3 in lung tissue. This led to a dramatic increase in myeloid clustering and metastasis. Up-regulating Stat3 activity within myeloid cells enhanced the interaction between myeloid cells and endothelial cells in vivo, increased angiogenesis, and promoted subsequent formation of metastasis. In contrast, ablation of Stat3 in myeloid cells prevented the formation of pre-metastatic niches and metastasis. IHC staining of benign lymph node tissue in patients with high-risk PC yielded a spectrum of pStat3 levels. Higher levels of pStat3 were qualitatively associated with infiltration of CD68+ macrophages, appearing in distinct clusters. Conclusion: The current study indicates a critical role of Stat3 in promoting metastatic potential, possibly through mediating myeloid cell infiltration and formation of pre-metastatic niches. In the context of high-risk PC, differential levels of pStat3 and a potential association with myeloid cell clustering was observed. While correlation between these parameters and clinical outcome is ongoing, these preliminary findings suggest a putative role for Stat3 as a therapeutic target to prevent and/or reduce pre-metastatic niche formation and metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 405. doi:10.1158/1538-7445.AM2011-405

Mycobacterium tuberculosis Complex and Mycobacterial Heat Shock Proteins in Lymph Node Tissue from Patients with Pulmonary Sarcoidosis

We demonstrated that not whole Mycobacterium tuberculosis but its particular antigens, hsp70(Mtb), hsp65(Mtb), and hsp16(Mtb), are present in lymph node tissues of patients with sarcoidosis (SA). hsp16(Mtb) occurs in the early stage of SA, whereas hsp70(Mtb) occurs in stage II of SA. hsp65(Mtb) is highly expressed in the capillary vessels in lymph node tissues in patients with SA.

Cat scratch disease. An overview for the ENT physician

Cat scratch disease (CSD) is a relatively common cause of chronic lymphadenopathy in the USA. In the present paper the authors describe recent advances in the understanding of this disorder focusing on etiology, clinical aspects, diagnostic management, and therapy. Rochalimaea henselae and Afipia felis, two gram-negative bacteria, have recently been isolated from lymph node tissue of patients suffering from CSD. The current literature reveals that Rochalimaea henselae seems to be the most probable agent responsible for CSD. Serum samples from CSD patients' cats have shown titers of 1:64 or higher for antibodies to Rochallmaea henselae. The incidence of CSD in the USA is between 0.77 and 9.3 per 100,000 per year. The incidence of CSD in Europe is unknown, but the prevalence of antibodies to Rochalimaea henselae among cats is comparable. Most common symptoms of CSD are regional lymphadenopathy, fever, and malaise. Other manifestations occur in about 5% of patients and include encephalitis, granulomatous hepatitis, and Parinaud's oculoglandular syndrome. Fatal complications and irreversible sequelae have not been reported. To establish the diagnosis of CSD, the presence of regional lymphadenopathy, cat contact, and papula are required. Additional procedures include indirect fluorescent antibody assays and PCR. Up to now there is no standard therapy for CSD. Antibiotic treatment, however, might be considered for CSD patients with severe symptoms. Rifampicin, ciprofloxacin, trimethoprim sulfamethoxazole, and gentamicin are known to be effective antibiotic agents.

Detection of Silicone in Lymph Node Biopsy Specimens by Near-Infrared Raman Spectroscopy

Near-infrared Raman spectroscopy with a 782-nm cw laser was used to examine lymph node biopsy specimens from women with ruptured breast implants containing silicone gel. For reduction of fluorescence and sample radiation damage, a low-power (30 mW) Ti: sapphire laser, single-stage spectrograph, and CCD detector were employed. Silicone Raman features were clearly visible in lymph node tissue of patients with leaking implants, and the spectra were easily distinguished from those of normal lymph node tissue. The technique has promise for medical diagnostic purposes, and may be amenable to in vivo analysis with adaptation to a fiber-optic probe.

Cutaneous Reaction to Pine Pollen in Sarcoidosis: Study of Forty Cases

Three interesting and possibly relevant findings applicable to the etiology of sarcoidosis have been reported in the past few years. Epidemiologic surveys have related the incidence of sarcoidosis to the concentration of forest distributions in the country (1, 2). Pine pollen has been found to contain acid-fast lipids and alpha epsilon diami-nopimelic acid similar to the tubercle bacillus (3), and to stain with acid-fast technics. Further, my colic acid and diaminopimelic acid have been isolated from the lung and lymph node tissues of patients with sarcoidosis (4). Since these findings suggest a possible etiologic clue to this disease, it was felt that an intradermal test utilizing pine pollen as the antigen might contribute additional information, particularly if a positive reaction paralleling the Kveim test could be elicited.