Cells In Lymph Node Tissue Research Articles

Pathogenicity of Porcine Circovirus Type 2d (PCV2d) in Pigs Infected with PCV2d or Co-infected with Mycoplasma hyopneumoniae and PCV2d or with Porcine Reproductive and Respiratory Syndrome Virus and PCV2d

The objective of this study was to determine the pathogenicity of porcine circovirus type 2d (PCV2d) in pigs inoculated intranasally with PCV2d alone, PCV2d in combination with Mycoplasma hyopneumoniae or PCV2d in combination with porcine reproductive and respiratory syndrome virus (PRRSV). Pigs infected with PCV2d alone were asymptomatic. All pigs inoculated with either M. hyopneumoniae and PCV2d or with PCV2d and PRRSV developed porcine circovirus-associated disease (PCVAD), as characterized by a sudden onset of clinical signs and disseminated granulomatous inflammation. Inflammation was mainly present in lymph nodes and spleen, and occasionally in liver and kidney. Pigs in both of these dually infected groups also had significantly higher (P<0.05) microscopic lymphoid lesion scores and a significantly higher (P<0.05) number of PCV2-positive cells in lymph node tissue than did pigs inoculated with PCV2d alone. The M. hyopneumoniae and PRRSV combination potentiated the PCV2d load in the blood. Co-infection with PRRSV and PCV2d resulted in a significantly higher blood load of PCV2d compared with the M. hyopneumoniae and PCV2d combination. Successful reproduction of PCVAD in pigs appears to require PCV2d with at least one additional infectious agent, such as M. hyopneumoniae or PRRSV, for the full manifestation of disease.

Prolonged Posttreatment Virologic Control and Complete Seroreversion After Advanced Human Immunodeficiency Virus-1 Infection

Possible human immunodeficiency virus (HIV)-1 clearance has rarely been reported. In this study, we describe a unique case of an HIV-positive, combination antiretroviral therapy (cART)-experienced woman with prior acquired immunodeficiency syndrome (AIDS) who has not experienced viral rebound for over 12 years since discontinuing cART. Leukapheresis, colonoscopy, and lymph node excision were performed for detailed examination of virologic (including HIV reservoir) and immunologic features. Comparisons were made with chronically infected patients and healthy controls. No HIV-specific antibodies were detected in serum. Plasma HIV ribonucleic acid (RNA) levels were <0.2 copies/mL, and, except for low-frequency HIV deoxyribonucleic acid (DNA)+ cells in lymph node tissue (1 copy/3 × 106 cells), HIV antigen could not be detected by quantitative virus outgrowth (<0.0025 infectious units/106 CD4+ T cells) or by most measurements of HIV RNA or DNA in blood, lymph node, or gut-associated mononuclear cells. Human immunodeficiency virus-specific T-cell responses were detectable but low. Brain imaging revealed a prior biopsy site and persistent white matter disease since 1996. Human immunodeficiency virus DNA+ cells in the 1996 brain biopsy specimen confirmed her identity and initial HIV diagnosis. This represents the first report of complete seroreversion, prolonged posttreatment virus suppression, a profoundly small HIV reservoir, and persistent HIV-specific T cells in an adult with prior AIDS.

In Situ Staining and Laser Capture Microdissection of Lymph Node Residing SIV Gag-Specific CD8+ T cells--A Tool to Interrogate a Functional Immune Response Ex Vivo.

While a plethora of data describes the essential role of systemic CD8+ T cells in the control of SIV replication little is known about the local in situ CD8+ T cell immune responses against SIV at the intact tissue level, due to technical limitations. In situ staining, using GagCM9 Qdot 655 multimers, were here combined with laser capture microdissection to detect and collect SIV Gag CM9 specific CD8+ T cells in lymph node tissue from SIV infected rhesus macaques. CD8+ T cells from SIV infected and uninfected rhesus macaques were also collected and compared to the SIV GagCM9 specific CD8+ T cells. Illumina bead array and transcriptional analyses were used to assess the transcriptional profiles and the three different CD8+ T cell populations displayed unique transcriptional patterns. This pilot study demonstrates that rapid and specific immunostaining combined with laser capture microdissection in concert with transcriptional profiling may be used to elucidate phenotypic differences between CD8+ T cells in SIV infection. Such technologies may be useful to determine differences in functional activities of HIV/SIV specific T cells.

Bioactivity of Combination Elephantopus scaber and Sauropus androgynus on the Level of B220 cells of Lymph Node in Pregnant Typhoid BALB/c Mice

Elephantopus scaber and Sauropus androgynus are herbal remedies that contain flavonoids as immunomodulatory agents. This study was aimed to observe the changes in the relative number of B cells in lymph node tissue of pregnant typhoid mice. Mice were divided into seven groups K-, K+, E. scaber 200 mg.kg -1 BW, E. scaber 150 mg.kg -1 BW + S. androgynus 37.5 mg.kg -1 BW, E. scaber 100 mg.kg -1 BW + S. androgynus 75 mg.kg -1 BW, E. scaber 50 mg.kg -1 BW + S. androgynus 112.5 mg.kg -1 BW, dan S. androgynus 150 mg.kg -1 BW. Mice were dissected on the 12 th and 18 th day after herbal treatment. Data was analyzed using one way ANOVA (p < 0.05) and Duncan. The result show that E. scaber combination of 200 mg.kg -1 BW and E. scaber 150 mg.kg -1 BW + S. androgynus 37.5 mg.kg -1 BW can increase the number of B220 cells (p < 0,05) on pregnant mice typoid model. Keywords: B220, Elephantopus scaber , pregnant, Sauropus androgynus , typhoid fever

Single Cell Analysis of Lymph Node Tissue from HIV-1 Infected Patients Reveals that the Majority of CD4+ T-cells Contain One HIV-1 DNA Molecule

Genetic recombination contributes to the diversity of human immunodeficiency virus (HIV-1). Productive HIV-1 recombination is, however, dependent on both the number of HIV-1 genomes per infected cell and the genetic relationship between these viral genomes. A detailed analysis of the number of proviruses and their genetic relationship in infected cells isolated from peripheral blood and tissue compartments is therefore important for understanding HIV-1 recombination, genetic diversity and the dynamics of HIV-1 infection. To address these issues, we used a previously developed single-cell sequencing technique to quantify and genetically characterize individual HIV-1 DNA molecules from single cells in lymph node tissue and peripheral blood. Analysis of memory and naïve CD4+ T cells from paired lymph node and peripheral blood samples from five untreated chronically infected patients revealed that the majority of these HIV-1-infected cells (>90%) contain only one copy of HIV-1 DNA, implying a limited potential for productive recombination in virus produced by these cells in these two compartments. Phylogenetic analysis revealed genetic similarity of HIV-1 DNA in memory and naïve CD4+ T-cells from lymph node, peripheral blood and HIV-1 RNA from plasma, implying exchange of virus and/or infected cells between these compartments in untreated chronic infection.

Spectral Detection of Micro-Metastases and Individual Metastatic Cells in Lymph Node Histology

The detection of micro-metastases and individual metastatic cells in lymph node tissue by spectral methods is summarized. These methods are based on instrument-based acquisition of thousands of infrared spectra of individual tissue pixels from the tissue section, and analysis of the resulting spectral hypercube by multivariate algorithms. The method of infrared image acquisition, followed by multivariate analysis, is henceforth referred to as Spectral Histopathology (SHP). SHP produces pseudo-color images of tissue sections which reveal details that compare very favorably with images collected from hematoxylin/eosin (H and E) stained tissues in that the same tissue structures are detected. However, the infrared results are based on objective and reproducible measurements and do not depend on subjective interpretation. One of the major topics of this paper is the comparison of spectral patterns observed for the same cancer type from different patients. While this is easy in some tissue types, we found it to be difficult in tissues of very different cellularity, or tissue sections that exhibit high levels of inflammatory response. In both cases, spectral quality will be compromised due to confounding effects resulting from scattering effects. The correction of these effects now permits the direct comparison of different patient samples, and paves the way for diagnostic algorithms for cancer detection to be developed.

One-step biotinylation procedure for carbohydrates to study carbohydrate–protein interactions

Protein–carbohydrate interactions play crucial roles in numerous biological processes. To study these interactions, we developed a simple and fast procedure for the biotinylation of carbohydrates based on reductive amination. The method allows complete and stable biotinylation of small quantities of oligosaccharides and includes a rapid and simple procedure to remove excess labeling reagent. After biotinylation, the structural and biological integrity of the glycans was intact as determined by HPLC, mass spectrometry, and a plant lectin assay. By using the human C-type lectin DC–SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin), we demonstrate that the biotinylated glycans can be used in a glycan array to determine binding specificities of lectins. Moreover, we show that fluorescent beads coated with selected biotinylated glycans bind to DC–SIGN-expressing dendritic cells in vitro. Finally, by using biotinylated high-mannose N-glycans, we could visualize DC–SIGN-expressing cells in lymph node tissue. The availability of easy biotinylation methods for oligosaccharides such as those described here greatly facilitates the functional analysis of lectins. In addition, the biotinylated glycans will be great tools for investigating functional lectin receptors in situ.

Biomarkers for melanoma

Given the capricious nature of melanoma, biomarkers that provide significant insight into the behavior of melanoma would greatly aid in identifying patients at risk for disease progression, those whose disease has progressed subclinically, and those who would benefit from currently available systemic therapies. This review focuses on molecular prognostic markers in primary melanoma, markers that aid in the detection of metastatic melanoma, and markers predictive of systemic therapy. Significant advances have been made in the field of melanoma biomarkers. Utilization of paraffin-embedded tissue and multiple markers have improved the RT-PCR assays for detection of melanoma cells in lymph node tissue as well as peripheral blood. Lymphangiogenesis has been identified as a novel mechanism for melanoma progression, and candidate markers in the NF-kappaB signaling pathway have been identified to play a key role in melanoma: tumor vasculature interactions. Loss of heterozygosity has been used to identify potential candidates for biochemotherapy. Furthermore, serum S100B protein has been shown to be superior to lactate dehydrogenase in predicting prognosis and response to treatment for patients with advanced melanoma. Although recent studies have contributed greatly to the development of melanoma markers, it is anticipated that the application of gene expression profiling and proteomics techniques to melanocytic neoplasms will result in the identification of even more effective biomarkers for melanoma than those currently in clinical use.

Detection of gene deletion in single metastatic tumour cells in lymphnode tissue by fluorescent in-situ hybridisation

Detection of gene deletion in single metastatic tumour cells in lymphnode tissue by fluorescent in-situ hybridisation