Histochemical Localization of T-Cells in Tissue Sections

1979 ◽  
Vol 23 (4) ◽  
pp. 886 ◽  
Author(s):  
S. Odend'hal ◽  
E. C. Player
Keyword(s):  
T Cells ◽  
Histochemical Localization ◽  
Tissue Sections

Compartmentalisation between Gut and Lung Mucosae in a Model of Secondary Immunodeficiency: Effect of Thymomodulin

2003 ◽  
Vol 16 (2) ◽  
pp. 151-156 ◽  
Author(s):  
M.E. Roux ◽  
M.G. Marquez ◽  
S. Olmos ◽  
C.A. Frecha ◽  
A. Florin-Christensen
Keyword(s):  
T Cells ◽  
Oral Administration ◽  
Therapeutic Agent ◽  
Intraepithelial Lymphocytes ◽  
Mucosal Immune Response ◽  
Oral Route ◽  
Cd4 T Cell ◽  
Secondary Immunodeficiency ◽  
Tissue Sections ◽  
Intestinal Intraepithelial Lymphocytes

Compartmentalisation of mucosal immune response seems to be the result mainly of the preferential migration of activated cells back to their inductive sites. The aim of this report was to demonstrate, in a model of secondary immunodeficiency in Wistar rats (severely protein deprived at weaning and refed with casein 20 %; group R21), that the oral administration of Thymomodulin (group: R21TmB) has different effects on gut and BALT (Bronchus-associated lymphoid tissue). Tissue sections (5μ) were studied by immunohistochemistry 1). The oral administration of Thymomodulin restores only in gut Lamina propria (LP) the IgA B and CD4 T cell populations to control levels. The CD8a and CD25 subpopulations do not vary in gut as they return to control levels when refed with 20% casein diet. All the populations mentioned above remained decreased even after receiving Thymomodulin by the oral route. However, the same behaviour was observed for the TCRδ T cells that were decreased and return to normal levels in both mucosae by the effect of the immunomodulator; 2) when studying the iIEL (intestinal intraepithelial lymphocytes) CD8α, CD25 and TCRγδ T cells, that were increased in R21, return to control levels in R21TmB. In BALT intraepithelium CD8α and CD25 T cells remained decreased, while only TCRγδ T cells (increased in R21) return to control values. Conclusions: 1) there exists a compartmentalisation between both mucosae, as T CD4+ and IgA B+ cells are restored by TmB only in gut; 2) only those iIEL involved in inflammation (CD8α+/CD25+ and TCRγδ+/CD25+) are normalised by means of the Thymomodulin 3) however, in BALT, only TCRγδ+ T cells are restored 4) the oral administration of the present immunomodulator may be useful as a therapeutic agent, although the preferential survival in the tissue of initial stimulation is the major factor in the preferential distribution of activated cells.

Histochemical localization of mushroom tyrosinase in whole tissue sections on nitrocellulose

1989 ◽  
Vol 90 (5) ◽  
pp. 379-381 ◽  
Author(s):  
B. M. Moore ◽  
B. Kang ◽  
W. H. Flurkey
Keyword(s):  
Histochemical Localization ◽  
Mushroom Tyrosinase ◽  
Tissue Sections

scholarly journals The Determination of Immunomodulation and Its Impact on Survival of Rectal Cancer Patients Depends on the Area Comprising a Tissue Microarray

Cancers ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 563 ◽  
Author(s):  
Elisabeth S. Gruber ◽  
Georg Oberhuber ◽  
Dietmar Pils ◽  
Theresa Stork ◽  
Katharina Sinn ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
T Cells ◽  
Survival Analysis ◽  
T Cell ◽  
Cell Density ◽  
Tumor Heterogeneity ◽  
Tissue Sections ◽  
Cell Quantification ◽  
Cell Densities ◽  
Tumor Area

Background: T cell density in colorectal cancer (CRC) has proven to be of high prognostic importance. Here, we evaluated the influence of a hyperfractionated preoperative short-term radiation protocol (25 Gy) on immune cell density in tumor samples of rectal cancer (RC) patients and on patient survival. In addition, we assessed spatial tumor heterogeneity by comparison of analogue T cell quantification on full tissue sections with digital T cell quantification on a virtually established tissue microarray (TMA). Methods: A total of 75 RC patients (60 irradiated, 15 treatment-naïve) were defined for retrospective analysis. RC samples were processed for immunohistochemistry (CD3, CD8, PD-1, PD-L1). Analogue (score 0–3) as well as digital quantification (TMA: 2 cores vs. 6 cores, mean T cell count) of marker expression in 2 areas (central tumor, CT; invasive margin, IM) was performed. Survival was estimated on the basis of analogue as well as digital marker densities calculated from 2 cores (Immunoscore: CD3/CD8 ratio) and 6 cores per tumor area. Results: Irradiated RC samples showed a significant decrease in CD3 and CD8 positive T cells, independent of quantification mode. T cell densities of 6 virtual cores approximated to T cell densities of full tissue sections, independent of individual core density or location. Survival analysis based on full tissue section quantification demonstrated that CD3 and CD8 positive T cells as well as PD-1 positive tumor infiltrating leucocytes (TILs) in the CT and the IM had a significant impact on disease-free survival (DFS) as well as overall survival (OS). In addition, CD3 and CD8 positive T cells as well as PD-1 positive TILs in the IM proved as independent prognostic factors for DFS and OS; in the CT, PD-1 positive TILs predicted DFS and CD3 and CD8 positive T cells as well as PD-1 positive TILs predicted OS. Survival analysis based on virtual TMA showed no impact on DFS or OS. Conclusion: Spatial tumor heterogeneity might result in inadequate quantification of immune marker expression; however, if using a TMA, 6 cores per tumor area and patient sample represent comparable amounts of T cell densities to those quantified on full tissue sections. Consistently, the tissue area used for immune marker quantification represents a crucial factor for the evaluation of prognostic and predictive biomarker potential.

scholarly journals Immunofluorescence analysis of normal and malignant lymphoid tissues with selected combination of antisera.

1980 ◽  
Vol 28 (11) ◽  
pp. 1207-1214 ◽  
Author(s):  
G Janossy ◽  
J A Thomas ◽  
J A Habeshaw
Keyword(s):  
T Cells ◽  
Lymph Nodes ◽  
Close Association ◽  
Lymphoid Tissues ◽  
Immunofluorescence Analysis ◽  
Tissue Sections ◽  
Reticular Cells ◽  
Lymphocyte Antigens ◽  
Relationship Of

Tissue sections stained with combinations of antisera labeled with different fluorochromes (i.e., conventional antisera to human immunoglobulin classes, T lymphocyte antigens, and Ia-like p28,33 antigens used in various double combinations with each other or with different mouse monoclonal antibodies) allow the identification of the different areas of lymph nodes in serial sections and provide great flexibility as well as precision in the analysis of the distribution and relationship of normal and malignant cells. Lymphoid microenvironments in the thymus and the paracortical areas of lymph nodes are described. The close association of T lymphocytes and nonlymphoid cells expressing large amounts of Ia-like antigens (such as interdigitating reticular cells and endothelium) may be relevant for the understanding of immunoregulatory disorders such as dermatopathic and angioimmunoblastic lymphadenopathies and some malignancies (e.g., mycosis fungoides) were the expression of Ia-like antigens on non-T cells seems to be abnormally abundant. The analysis of immunoglobulin and membrane marker expression of normal and malignant B cells and their relation to T cells can also be related to the histology of the disease. These studies are clinically useful for the classification of childhood lymphomas, the differential diagnosis of anaplastic carcinomas and lymphomas, and in the study of the early stages of lymphomas.

Sunitinib's effect on tumor infiltration of CD8 T cells in renal cell carcinoma (RCC) and modulation of their function by altering VEGF-induced upregulation of PD1 expression.

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 591-591 ◽  
Author(s):  
Samuel Haywood ◽  
Roy Chen ◽  
Paul Pavicic ◽  
Lin Lin ◽  
C. Marcela Diaz-Montero ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Renal Cell ◽  
Checkpoint Inhibitors ◽  
Vegf Inhibitors ◽  
Effector Cells ◽  
Immune Infiltrate ◽  
Multiple Tumor ◽  
Sunitinib Treatment ◽  
Tissue Sections

591 Background: The effect of VEGF inhibitors such as sunitinib to modulate RCC tumor microenvironment and the effect on immunotherapy using checkpoint inhibitors is unknown and of clinical interest. Methods: Immune infiltrate in tissue sections from clear cell renal cell carcinomas (ccRCC) (n = 13) treated with sunitinib in the neoadjuvant setting were compared to that of untreated RCC patients with localized disease (n = 15). Immune infiltrate was scored in H&E stained tissue sections. Infiltrates were further characterized by IHC using antibodies against CD4 (Novocastra NCL-CD4-1F6 clone 1F6), CD8 (BiogeneX MU422-UC clone 1A5) or PD1 (Abcam ab52587 clone EPR4877). Intensity of staining was quantified using a Leica microscope equipped with a Retiga SRV camera. Scanned images of multiple tumor fields were analyzed using ImagePro Plus software. For pre-clinical studies, Balb/c mice bearing Renca tumors (RCC) were treated with sunitinib (40mg/kg) daily for 14 days, and tumors were dissociated and analyzed for CD8 and CD4 infiltrate by FACS analysis. Results: Qualitative analysis showed a more prominent accumulation of lymphocytes in tumor sections of patients receiving sunitinib. Quantitative analysis of these infiltrates revealed higher levels of CD8+ T cells in sunitinib treated patients (p = 0.04). By contrast, sunitinib treatment was associated with a reduction of intratumoral expression of PD1. Preclinical studies in the Renca model also showed an increase in the frequency of tumor infiltrating CD8+ T cells following sunitinib treatment. Remarkably, a higher fraction of these tumor infiltrating CD8 T cells were found to co-express the activation marker of cytotoxic effector cells (CD107a) when compared to untreated mice. Conclusions: These findings suggest that sunitinib not only promotes the accumulation of CD8+ T cells within the tumors, but also affects their activity by modulating PD1 expression and enhancing cytotoxic function.

scholarly journals Localization of Peroxidase in Grapes using Nitrocellulose Blotting of Freezing/Thawing Fruits

HortScience ◽  
1993 ◽  
Vol 28 (1) ◽  
pp. 38-40 ◽  
Author(s):  
Antonio A. Calderón ◽  
Jose M. Zapata ◽  
Romualdo Muñoz ◽  
A. Ros Barceló
Keyword(s):  
Vitis Vinifera ◽  
Vascular Bundle ◽  
Peroxidase Activity ◽  
Histochemical Localization ◽  
Vascular Bundles ◽  
Reaction Products ◽  
Tissue Sections ◽  
Bundle Structure

A technique has been developed to study the histochemical localization of peroxidase in Vitis vinifera by blotting freezing/thawing tissue sections on nitrocellulose membranes. After being stained with 4-methoxy- α -naphthol and H2O2, peroxidase-mediated reaction products in mature `Gamay' grapes were seen principally in the skin and, to a lesser extent, the pericarp, where discrete areas of reaction products were located in the vascular bundles. However, for immature `Gamay' and `Grenache' grapes, peroxidase activity in the skin was low and similar to that found in the pericarp. With this technique, fruit vascular bundle structure was preserved. The reliability of the technique in the histochemical localization of peroxidase in grapes was confirmed by fractionation and determining the peroxidase activity in the various tissues.

Densities of decidual high endothelial venules correlate with T-cell influx in healthy pregnancies and idiopathic recurrent pregnancy losses

2020 ◽  
Vol 35 (11) ◽  
pp. 2467-2477
Author(s):  
Karin Windsperger ◽  
Sigrid Vondra ◽  
Andreas Ian Lackner ◽  
Victoria Kunihs ◽  
Peter Haslinger ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Cell ◽  
First Trimester ◽  
High Endothelial Venules ◽  
Tissue Sections ◽  
Decidual Tissue ◽  
Vascular Phenotype ◽  
Depth Analysis ◽  
Recurrent Pregnancy Losses

Abstract STUDY QUESTION Do high endothelial venules (HEVs) appear in the uterus of healthy and pathological pregnancies? SUMMARY ANSWER Our study reveals that HEVs are present in the non-pregnant endometrium and decidua parietalis (decP) but decline upon placentation in decidua basalis (decB) and are less abundant in decidual tissues from idiopathic, recurrent pregnancy losses (RPLs). WHAT IS KNOWN ALREADY RPL is associated with a compromised decidual vascular phenotype. STUDY DESIGN, SIZE, DURATION Endometrial (n = 29) and first trimester decidual (n = 86, 6–12th week of gestation) tissue samples obtained from endometrial biopsies or elective pregnancy terminations were used to determine the number of HEVs and T cells. In addition, quantification of HEVs and immune cells was performed in a cohort of decidual tissues from RPL (n = 25). PARTICIPANTS/MATERIALS, SETTING, METHODS Position and frequency of HEVs were determined in non-pregnant endometrial as well as decidual tissue sections using immunofluorescence (IF) staining with antibodies against E-selectin, intercellular adhesion molecule, von Willebrand factor, ephrin receptor B4, CD34 and a carbohydrate epitope specific to HEVs (MECA-79). Immune cell distribution and characterization was determined by antibodies recognizing CD45 and CD3 by IF staining- and flow cytometry-based analyses. Antibodies against c-c motif chemokine ligand 21 (CCL21) and lymphotoxin-beta were used in IF staining and Western blot analyses of decidual tissues. MAIN RESULTS AND THE ROLE OF CHANCE Functional HEVs are found in high numbers in the secretory endometrium and decP but decline in numbers upon placentation in decB (P ≤ 0.001). Decidua parietalis tissues contain higher levels of the HEV-maintaining factor lymphotoxin beta and decP-associated HEVs also express CCL21 (P ≤ 0.05), a potent T-cell chemoattractant. Moreover, there is a positive correlation between the numbers of decidual HEVs and the abundance of CD3+ cells in decidual tissue sections (P ≤ 0.001). In-depth analysis of a RPL tissue collection revealed a decreased decB (P ≤ 0.01) and decP (P ≤ 0.01) HEV density as well as reduced numbers of T cells in decB (P ≤ 0.05) and decP (P ≤ .001) sections when compared with age-matched healthy control samples. Using receiver-operating characteristics analyses, we found significant predictive values for the ratios of CD3/CD45 (P < 0.001) and HEVs/total vessels (P < 0.001) for the occurrence of RPL. LIMITATIONS, REASONS FOR CAUTION Analyses were performed in first trimester decidual tissues from elective terminations of pregnancy or non-pregnant endometrium samples from patients diagnosed with non-endometrial pathologies including cervical polyps, ovarian cysts and myomas. First trimester decidual tissues may include pregnancies which potentially would have developed placental disorders later in gestation. In addition, our cohort of non-pregnant endometrium may not reflect the endometrial vascular phenotype of healthy women. Finally, determination of immune cell distributions in the patient cohorts studied may be influenced by the different modes of tissue derivation. Pregnancy terminations were performed by surgical aspiration, endometrial tissues were obtained by biopsies and RPL tissues were collected after spontaneous loss of pregnancy. WIDER IMPLICATIONS OF THE FINDINGS In this study, we propose an inherent mechanism by which the endometrium and in particular the decidua control T-cell recruitment. By demonstrating reduced HEV densities and numbers of T cells in decB and decP tissues of RPL samples we further support previous findings reporting an altered vascular phenotype in early pregnancy loss. Altogether, the findings provide important information to further decipher the etiologies of unexplained RPL. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Austrian Science Fund (P31470 B30 to M.K.) and by the Austrian National Bank (17613ONB to J.P.). There are no competing interests to declare. TRIAL REGISTRATION NUMBER N/A.

T cell infiltration in matched renal biopsy (bx) and nephrectomy (nx) samples in renal cell carcinoma (RCC).

2017 ◽  
Vol 35 (6_suppl) ◽  
pp. 472-472
Author(s):  
Haris Zahoor ◽  
Paul G Pavicic ◽  
Christopher Przybycin ◽  
Paul Elson ◽  
C. Marcela Diaz-Montero ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Checkpoint Inhibitors ◽  
Locally Advanced ◽  
Cleveland Clinic ◽  
Cell Infiltration ◽  
Tissue Sections ◽  
T Cell Infiltration ◽  
Potential Biomarker ◽  
Pre Treatment

472 Background: T cell infiltration in tumors has been investigated as a biomarker of response to checkpoint inhibitors. A neo-adjuvant trial of checkpoint inhibition in locally-advanced RCC is ongoing at Cleveland Clinic, where T cell infiltration in pre-treatment renal mass bx will be compared to post-treatment nx specimens. However, there are no data regarding the association of T cell infiltration in matched bx and nx samples without intervening treatment. Understanding this association will enable further study of this potential biomarker in future neo-adjuvant studies. Methods: Matched bx and nx samples (without intervening systemic therapy) were identified from patients with non-metastatic RCC. Demographic and pathological data were collected from chart review. Selected tissue sections from bx and nx samples of each patient were reviewed, and marked for tumor and intra-tumoral lymphocytes by the pathologist. Immunohistochemistry (IHC) was utilized to stain these selected tissue sections for T cell markers (CD3, CD4 and CD8). Intra-tumoral T cells were then quantified in the pre-marked tissue sections as counts per total tumor area surveyed, using Image-Pro Plus (Media Cybernetics, Inc.). Spearman correlation (ρ) was used to measure the strength of association of T cell infiltration between matched samples. Results: 30 matched pairs were investigated. The median interval between bx and nx was 2.8 (0.2-87.7) months. Clear cell was the most common histology (29/30; 97%). 15/30 (50%) had grade 3-4 tumors, 2/19 (11%) patients had sarcomatoid features, 7/25 (29%) had necrosis, and 8/28 (29%) had lymphovascular invasion. We found a positive correlation between the frequencies of CD8+ T cells between matched bx and nx samples (ρ= 0.39; p=0.03). CD3+ and CD4+T cells did not show significant correlation. (Table) Conclusions: Bx material can be used to accurately assess the degree of CD8+T cell infiltration in RCC. [Table: see text]

FOXP3 Expression in B and T Cell Lymphomas.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4503-4503
Author(s):  
Giovanna Roncador ◽  
Juan Fernando Garcia ◽  
Jose Francisco Garcia ◽  
Lorena Maestre ◽  
Elena Lucas ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Regulatory Gene ◽  
Immune Surveillance ◽  
Foxp3 Expression ◽  
Treg Cells ◽  
Cell Leukaemia ◽  
Tissue Sections ◽  
T Cell Lymphomas ◽  
And Function

Abstract Foxp3, which encodes a forkhead/winged helix transcription factor designated Scurfin, is a key regulatory gene required for the development and function of regulatory CD4+CD25+ T cells (Treg), a subpopulation of T-cells specialized in maintaining the balance between immunity and tolerance. Humans with defects in the FOXP3 gene, develop strong activation of the immune system, leading to multiorgan autoimmune disease, allergies, inflammatory bowel disease and severe infections, collectively known as the IPEX syndrome (immune deregulation, polyendocrinopathy, enteropathy, X-linked inheritance syndrome) Because of the importance of FOXP3 in the development and function of Treg cells, and its potential use as a specific Treg marker, we have developed several monoclonal antibodies against FOXP3, for use on paraffin-embedded tissue sections and evaluated its expression in a large series (150 cases) of B- and T-cell lymphomas. In reactive lymphoid tissue, strong nuclear FOXP3 expression was observed in approximately 5% of interfollicular T-cells. FOXP3 expression in tumour cells was confined to most of Adult T-cell Leukaemia/Lymphoma (ATLL) cases (68%), with some variability in protein expression. In other lymphoma types, FOXP3 expression was only detected in the reactive T-cell background, and the number of FOXP3-positive reactive T-cells was variable, ranging from almost a complete absence (Burkitt’s lymphoma) to abundant infiltrate (common in follicular lymphoma). In conclusion, the availability of a FOXP3 monoclonal antibody, not only provides an important tool for the study of the development and function of Treg cells, but also represents a useful marker for the identification of ATLL cases in formalin-fixed paraffin-embedded tissue sections. The presence or absence of Treg cells in the tumour environment could also play a role in the immune surveillance of tumours, thus implying a potential additional value for the detection of this cell population in tumour samples.

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