Expression Of D2-40 Research Articles

Frequency of Expression of D240 in Dermatofibroma – A diagnostic study

Aim: To establish the diagnostic utility of D2-40 IHC stain in dermatofibroma, by determining its positivity of expression. Study design: It was a Cross-sectional study Setting: The study was conducted in Department of Histopathology, Chughtai Institute of Pathology Duration of study: Six months (1st August 2018 to 31st January 2019) Methods: A total 70 samples were included. The clinical parameters like age and gender were recorded. The histological preparation was performed by classic method for inclusion in paraffin followed by haematoxylin-eosin staining. The immunohistochemical analysis was performed on serial sections using immune-enzymatic soluble complex method. The antibody used was D2-40 polyclonal antibody from DAKO. Diffuse crisp cytoplasmic D2-40 staining was considered positive. Results: The mean age was 32.76±10.65 years and male to female ratio was 1:1.4. There were 70% cases have D2-40 positivity for dermatofibroma and 30% have no D2-40 positivity for dermatofibroma and mean duration of disease was 9.05±6.33 months. There were significant difference (P<0.05) between age and duration of disease with respect to D2-40 positivity for dermatofibroma; Conclusion: D2-40 immunoreactivity is sensitive and is useful in the differential diagnosis of dermatofibroma. Keywords: Dermatofibroma, Haematoxylin, Immunohistochemical, Positivity of D2-40

Immunohistochemical evaluation of CD31 and D2-40, expression in oral squamous cell carci-noma

Background: The present study was carried out to evaluate the relation between angiogenesis, and tumor stage of oral squamous cell carcinoma (OSCC).Materials and methods: Thirty formalin-fixed paraffin embedded blocks were used, ten of them were previously diagnosed as well differentiated OSCC, ten moderately differentiated OSCC and ten poorly differentiated OSCC. To determine the expression of CD31 and D2-40 proteins, streptavidin-biotin immunoperoxidase staining technique was used. The areas with the most vascular density (hot spots) were determined. The stained vessels were counted independently in intratumoral and peritumoral stroma. Results: CD 31 protein showed positive expression in the peritumoral and intratumoral blood vessels subjacent to the malignant invading nests. D2-40 expression was positive in lymphatic vessel in the peritumoral and intratumoral stroma subjacent to the invading nests. The highest mean values of both CD31 and D2-40 proteins expression were recorded in poor differentiated OSSC followed by moderate differentiated OSSC then well differentiated OSSC. There was statistically significant difference found between the three studied groups regarding CD31 and D2-40 levels. Also there was statistically significant positive correlation found between CD31 level and D2-40 level.Conclusion: CD31 and D2-40 are related to stage of OSCC and are consistent with angiogenesis in tumor progression.

Diagnostic significance of CyclinD1 and D2-40 expression for follicular neoplasm of the thyroid

ObjectivesTo evaluate the expression and differential diagnostic significance of CyclinD1 and D2–40 in follicular neoplasm (FN) and other thyroid adenomatoid lesions. MethodsA total of 144 cases of thyroid adenomatoid lesions were enrolled. Immunohistochemistry for CyclinD1 and D2–40 was performed. ResultsWe found two patterns of CyclinD1 expression: nuclear (N) and cytoplasmic (C). The expression of N-CyclinD1 / C-CyclinD1 in FN (77.4%, 48/62; 50.0%, 31/62) was much higher than that in multinodular goiters with dominant nodules (MNG-DN) (16.4%, 10/61; 4.9%, 3/61) (p < 0.05). In contrast, the expression of D2–40 in MNG-DN (82.0%,50/61) was much higher than that in FN (4.8%, 3/62) (p < 0.05). In addition, unique staining patterns were observed: CyclinD1 showed no immunostaining only in all 8 cases of oncocytic cell tumors (OCT); D2–40 staining showed the characteristic wide distribution of lymphatic vessels in all 8 cases of poorly differentiated thyroid carcinoma (PDTC). Finally, the expression of CyclinD1 and D2–40 did not differ among follicular thyroid adenoma and follicular thyroid carcinoma / noninvasive follicular thyroid neoplasm with papillary-like nuclear features (p > 0.05). ConclusionsCyclinD1 and D2–40 are helpful diagnostic markers of FN, which can assist to discern FN from MNG-DN / OCT / PDTC.

Immunohistochemical evaluation of D2-40, Galectin-3, Maspin and MCM7 expression in palate squamous cell carcinomas.

Squamous cell carcinoma (SCC) is the most frequent cancer in oral cavity and its prognosis has exhibited little improvement in the last decades. Although much less common palate SCCs manifests a higher local aggression invading very quickly the adjacent muscles and jawbones, thus being able frequently to lead to dysfunctions in chewing, swallowing, and speech. To elucidate what underlies such local aggression, we investigated the immunohistochemical expression in palate SCCs of Podoplanin (D2-40), Galectin-3 (Gal-3), mammary serine protease inhibitor (Maspin) and minichromosome maintenance complex component 7 (MCM7), markers that are known to be involved in tumor invasiveness. We found a progressive increase in reactivity for D2-40 and MCM7 from the normal epithelium toward dysplastic epithelium and respectively to SCC, which suggests the intervention of these markers in the early stages of squamous cell carcinogenesis in the palate. The highest D2-40, Gal-3 and MCM7 reactivity was observed in basaloid and in poorly differentiated (G3) palate SCCs, while for Maspin the well-differentiated (G1) palate SCCs were the most reactive. The first three markers mentioned above were most intensely expressed at the invasion front, while the Maspin reactivity was low or absent at this level. Statistically, we found significant stratification on localization, grading, muscle invasion, and survival for all investigated markers, but with very high direct correlations between D2-40, Gal-3, and MCM7 immunoreactive score (IRS) values, while between the Maspin and each of the previous markers there were very high inverse correlations. Overall, all these investigate markers proved to be responsible for the local invasiveness and regional lymph node metastasis, thus allowing a prognostic and therapeutic stratification of patients with palate SCCs.

Significance of D2-40 and CXCL5 Expression in Urinary Bladder Transitional Cell Carcinoma: An Immunohistochemical Study

Background: Transitional cell carcinoma remains a challenge in the oncology field, representing an ideal candidate for research on biomarkers that could identify patient’s progression and prognosis. D2-40 and CXCL5 have been implicated in progression of many cancers, but their significance in urinary bladder TCC remains unclear. The aim of this study is to assess their possible significance in urinary bladder transitional cell carcinoma.Methods: Immunohistochemistry was performed to examine the expression of D2-40 and CXCL5 in 50 cases of urinary bladder TCC and 10 cases of normal urothelium taken as a normal control. Statistical analysis methods were used to evaluate the relationship between D2-40 and CXCL5 and various clinicopathological parameters.Results: D2-40 was expressed in the lymphatic endothelial cytoplasm of all cases highlighting the tumor emboli in cancer lymphatic vessels. CXCL5 was found to be expressed in all urinary bladder TCC cases but not in normal control. The two markers significantly correlated with associated necrosis, tumor grade, T, N stage and presence of H&E detected LVI. CXCL5 was also positively correlated with presence of associated necrosis and CIS while D2-40 was insignificantly correlated with both. D2-40 detected intra-lymphatic tumor emboli are associated with lymph node metastasis in a highly significant correlation. Conclusions: The results suggested that CXCL5 might be involved in urinary bladder TCC carcinogenesis. It could be concluded that D2-40 and CXCL5 may have a possible role in urinary bladder TCC progression and prognosis. D2-40 might be considered as more reliable method in predicting tumor spread and lymph-node metastasis.

Peri-tumoral Lymphovascular density by antipodoplanin antibody D2-40, as a predictor of nodal metastasis in oral squamous cell carcinoma

BACKGROUND:&#x0D; The involvement of regional lymph nodes by cancer is a major determinant of prognosis in oral squamous cell carcinoma (OSCC) and is a significant predictor of patient survival. Peri-tumoral lymphangiogenesis is being studied as an important tool in predicting neck lymph node metastasis. Podoplanin, a transmembrane glycoprotein is expressed on lymphatic vessel endothelium and considered as a marker of lymphangiogenesis. The high podoplanin expression has been associated with invasion, progression, and metastasis of oral cancer. D2-40, an anti-podoplanin monoclonal antibody, is a specific marker of lymphatic endothelial cells. The present study was designed to investigate the peritumoral lymphovascular density (PT-LVD) by evaluating D2-40 expression and its potential utility in predicting lymph node metastasis in OSCC.&#x0D; METHODS: Seventy-two patients with OSCC were enrolled (36 with early and 36 with advanced-stage tumors). Differential expression of peritumoral lymphovascular density (PT-LVD) was evaluated using D2-40 expression by immunohistochemistry (IHC). Expression was categorized as low and high, derived from the mean of a number of vessels per 10 high power fields in hot-spots. Statistical methods were employed to correlate expression with age, gender, tumor site, stage, and neck lymph node involvement. &#x0D; RESULTS: We observed a significant association between peritumoral lymphovascular density (PT-LVD) with clinical neck lymph node status (p=0.001) and tumor stage (p=0.033). There was no significant correlation of D2-40 expression with age, gender, tumor site, T stage, and tumor differentiation.&#x0D; CONCLUSION: PT-LVD is a positive predictor of neck lymph node metastasis. It has translational potential, along with other clinical parameters for decision making in the treatment of the neck in oral cancer.

Investigation of Targeting Relationship between Micro-Rna-22 and Vegfr3 in Lung Squamous Cell Carcinoma.

The present study explored the clinical significance of microRNA-22 (miR-22) expression in lung squamous cell carcinoma and to explore the targeting relationship with vascular endothelial growth factor receptor 3 (VEGFR3). A total of 49 patients with lung squamous cell carcinoma who underwent surgical treatment were selected. The expression of miR-22 was detected by fluorescence quantitative realtime PCR (qPCR), the expression of VEGFR3 was detected by Western blotting assays, and D240 labeled microlymphatic vessels density (MLVD) was detected by immunohistochemistry (IHC). Lung squamous cell carcinoma cell line SK-MES-1 was selected and the targeting relationship between miR-22 and VEGFR3 was analyzed by double luciferase reporter gene assay. Western blotting assays were used to detect the expression of vascular endothelial growth factor-D (VEGFD) and D240 in the blank control group, empty vector transfection group, miR-22 transfection group, miR-22 and VEGFR3 co-transfection group. The expression range of miR-22 in lung squamous cell carcinoma was 0.8-3.5. The expression of miR-22 in lung squamous cell carcinoma was significantly different by tumor maximum diameter, lymph node metastasis, vascular invasion and TNM stage. The expression of miR-22 was linked to survival time. There was a negative correlation between miR-22 and VEGFR3, miR-22 and MLVD. Double luciferase reporter gene assays showed that miR-22 reduced the luciferase activity of pGL3-VEGFR3-WT transfected cells. Compared with the control group, the expression of VEGF-D and D2-40 in the miR-22 transfection group was significantly decreased. However, VEGF-D and D240 in the miR-22 and VEGFR3 co-transfection group reversed the changes. We assumed that the abnormal expression of miR-22 in lung squamous cell carcinoma may be involved in the development and progression of lung squamous cell carcinoma. MiR-22 negatively regulated the target gene VEGFR3 to mediate lymphangiogenesis. The expression of miR-22 may also be linked to the prognosis of the disease.

Transient Expression of Lymphatic Markers in Retrobulbar Intraconal Orbital Vasculature During Fetal Development.

PurposeThe aim of this study is to investigate the presence of orbital lymphatic vessels during fetal and neonatal development and in adults using a panel of lymphatic markers.MethodsThis was a retrospective observational case series. For analyzing lymphatic vessels, we used formalin-fixed paraffin-embedded enucleated eyes from 25 human fetuses between 13 and 24 weeks of gestation and postnatal eyes from 15 children and 5 adults. Immunohistochemical analysis of lymphatic vessels was performed for the markers: lymphatic vessel endothelial hyaluronic acid receptor-1 (LYVE-1), podoplanin (D2-40), Prospero-related homeobox gene-1 (Prox-1), pan-endothelial marker CD31, and blood vessel endothelium specific CD34.ResultsVasculature showing endothelial expression of LYVE-1, D2-40, Prox-1, and CD31 in combination with absence or weak expression of CD34, as would be expected for lymphatic vessels, was seen in 11 of 25 fetuses in an age range from 14 weeks to 23 weeks of gestation (44%). This lymphatic vascular staining pattern was also observed in 4 of 15 liveborn children (27%), all within 1 month of age, of which two were born prematurely at 32 and 34 weeks of gestation. Interestingly, an incomplete lymphatic staining pattern was observed in another 4 fetuses and two liveborn children of 4 months and 7 years old. No expression of lymphatic markers was observed in adult orbital vasculature.ConclusionsNo retrobulbar intraorbital lymphatic vessels were observed in adults, however, we did observe transient expression of lymphatic markers in retrobulbar intraconal orbital vasculature during fetal and early neonatal development. The orbit may, therefore, be proposed to possess a full range of lymphatic plasticity.

Maspin suppresses cell invasion and migration in gastric cancer through inhibiting EMT and angiogenesis via ITGB1/FAK pathway.

This study aims to investigate how Maspin affects the EMT and angiogenesis of gastric cancer (GC) cells via ITGB1/FAK pathway. Immunohistochemistry was used to evaluate the expressions of Maspin, ITGB1, FAK, E-cadherin, Vimentin, D2-40, and CD34 in GC and adjacent normal tissues from 160 patients. Then, the human GC cells with different degree of differentiation were transfected with Maspin CRISPR activation plasmid, ITGB1 siRNA and/or Maspin siRNA, followed by the following experiments, including qRT-PCR, western blotting, tube formation assay, Transwell assay and wound healing. GC tumor tissues manifested decreased Maspin with the activated ITGB1/FAK pathway. In tumor tissues, Maspin was negatively correlated with the expressions of ITGB1 and FAK, as well as Lauren's classification, differentiation degree, and TNM stage. Besides, Maspin was negatively related with lymphatic vessel density (LVD) and microvessel density (MVD), Vimentin and VEGF, but was positive correlated with E-cadherin. Maspin expression decreased, but ITGB1 and p-FAK expressions increased gradually in MKN-28 (well differentiated), SGC-7901 (moderate differentiated), and MKN-45 (poorly differentiated). Maspin CRISPR and ITGB1 siRNA increased E-cadherin with the decreased Vimentin, VEGF and bFGF, and the reductions of tube length. In comparison with the ITGB1 siRNA group, cells in the Maspin siRNA + ITGB1 siRNA group presented the more evident EMT and angiogenesis. Furthermore, ITGB1 siRNA reduced the malignancies of GC cells, which could be restored by Maspin siRNA. Maspin was downregulated in GC tissues, which could inhibit the EMT and angiogenesis by blocking the ITGB1/FAK pathway, thereby decreasing cell invasion and migration of GC.

CD31 and D2-40 Contribute to Peritoneal Metastasis of Colorectal Cancer by Promoting Epithelial-Mesenchymal Transition.

Background/AimsColorectal cancer (CRC) patients often exhibit peritoneal metastasis, which negatively impacts their prognosis. CD31 and D2-40 have recently been suggested to be predictors of breast cancer prognosis, but their role in colorectal peritoneal metastasis (CRPM) remains unknown.MethodsThe expression profiles of CD31 and D2-40 were analyzed in CRC patients with or without CRPM and in CRC cell lines with increasing metastatic potential. Overexpression and short hairpin RNA knockdown assays were performed in CRC cells, and the effects of these alterations on epithelial-mesenchymal transition (EMT) in vitro, growth of xenograft tumors in vivo, and peritoneal metastasis potential in a mouse model of CRPM were examined.ResultsThe expressions of CD31 and D2-40 were upregulated in CRC tumor tissues and was elevated further in tumor tissues from patients with CRPM. CD31 and D2-40 expression levels exhibited increasing trends parallel to the EMT potential of CRC cells. CD31 and D2-40 are essential for CRC cell EMT in vitro as well as for xenograft tumor growth and peritoneal metastasis in vivo.ConclusionsCD31 and D2-40 contribute to CRPM by promoting EMT and may serve as prognostic markers and therapeutic targets for CRC, particularly in patients with peritoneal metastasis.

Histopathological and Immunohistochemical Study of D2-40 Expression in Malignant Pleural Mesothelioma and Metastatic Adenocarcinoma

Abstract Background: Malignant pleural mesothelioma (MPM) is known to be an aggressive malignant tumor due to the difficulty of making early diagnosis and its rapid progression. Its incidence is increasing worldwide but no treatment plans are accepted. D2-40 is an immunohistochemical marker (mono-clonal antibody) that has been used as a lymphatic endothelial marker and used in the differential diagnosis of MPM (epi-thelioid type) versus metastatic adenocarcinoma. Fifty spec-imens of the pleural biopsy were viewed. They were diagnosed as 40 cases of MPM (epithelioid type) and 10 cases of meta-static adenocarcinoma (unknown primary origin). These diagnoses based on Hematoxylin and Eosin (Hx & E) stained sections and other immunohistochemical markers such as calretinin, carcinoembryonic antigen (CEA), thyroid transcrip-tion factor-1 (TTF-1). Aim of Study: To compare D2-40 immunostaining in MPM (epithelioid type) and metastatic adenocarcinoma with deter-mining its sensitivity and specificity in both types. Results: All cases of MPM epithelioid type were positive for D2-40 while all cases of metastatic adenocarcinoma were negative (both sensitivity and specificity= 100%). D2-40 staining result was considered positive or negative according to the presence or absence of membranous staining. Statistical analysis was done for assessment of D2-40 expression in both types of tumors by using SPSS version 21. Conclusion: Routine immunohistochemical work using D2-40 with calretinin is recommended. D2-40 would be superior to calretinin because of its membranous pattern of staining which does not obscure the cytological features of the tumor cells as compared to calretinin.

Clinical, histopathologic, subtype, and immunohistochemical analysis of jaw phosphaturic mesenchymal tumors.

Jaw phosphaturic mesenchymal tumors (PMTs) are a rare neoplasm with uncertain histogenesis. This study aimed to clarify the clinical and pathological features of jaw PMTs.We reviewed the clinical records of 39 patients diagnosed with PMTs in the jaws, and investigated clinical and morphologic characteristics, histologic subtypes, and immunophenotypes of all cases.Microscopic analyses revealed 2 major histologic tumor subtypes: “phosphaturic mesenchymal tumors of mixed epithelial and connective tissue” (PMTMECT), and “phosphaturic mesenchymal tumors of mixed connective tissue” (PMTMCT). PMTMECTs and PMTMCTs accounted for 29 and 10 cases of PMTs, respectively. Most PMTMECT diagnoses were made predominantly in males aged <45 years, and the incidence was similar in both the mandible and maxilla. In contrast, patients with PMTMCTs are predominantly females aged ≥45 years, and all tumors were in the mandible. Histologically, PMTMECT had lower cellularity and a more elongated and spindled mesenchymal component with less elaborate intrinsic microvasculature than PMTMCT. Immunohistochemically, the epithelia of all PMTMECTs was immunoreactive for AE1/AE3. Other immunohistochemical staining of PMTMECTs revealed positive expression of vimentin, SATB2, ERG, CD99, Bcl-2, CD56, S-100, D2-40, CD68, SMA, and CD34 in either one or both components. Immunohistochemical staining of PMTMCTs was diffusely positive for vimentin and a varied ratio of positivity for SATB2, ERG, CD99, Bcl-2, CD56, S-100, D2-40, CD68, SMA, and CD34, but negative for AE1/AE3. Most patients were cured by complete resection, except 2 patients who had repeated recurrences, one of which also had multiple metastasis.Jaw PMT can be divided into 2 major histological subtypes. PMTMECTs are more common than are PMTMCTs, and can transform into malignant PMTMCTs during the progression. PMTMECTs were more commonly observed in males and the incidence was similar in both the maxilla and mandible. PMTMCTs were almost always observed in the mandible of females. Compared with PMTMCTs, PMTMECTs have an admixture of epithelial components with less prominent vasculature and lower cellularity. There were no statistically significant differences in the expression of immunohistochemical markers except AE1/AE3 between PMTMECTs and PMTMCTs. However, immunohistochemical markers have great significance for differentiating other mesenchymal tumors.

The oral mucosa: Epithelial professional phagocytes, lymphatics, telocytes, and false telocytes

BackgroundThe oral mucosa protects the underlying tissue from mechanical damage as well as from the entry of exogenous particles and microorganisms. Telocytes (TCs) are disputed stromal cells featuring peculiarly long and thin processes with uneven calibre known as telopodes, which play a number of roles within the interstitia. The present study aimed to test the key markers recommended for discriminating between TCs and false TCs in samples of normal oral mucosa. MethodsArchived paraffin-embedded oral mucosa samples were tested by means of immunohistochemistry with the following markers: CD34, D2-40, CD31 and CD68. ResultsThe epithelial expression of CD68, D2-40 and CD34 was detected. Two subsets of CD34-expressing stromal cells were identified, large cells with telopodial processes, presumably of the hematopoietic lineage, and spindle-shaped TC-like cells. Macrophages and TC-like cells within the lamina propria expressed CD68. The lymphatic endothelia were found to express CD31 and D2-40, but not CD34. Sprouting lymphangiogenesis was demonstrated by the lymphatic endothelial tip cells, which were projecting thin processes within the connective stroma. ConclusionsThe epithelial expression of CD68 suggests the professional phagocytic potential of the oral epithelium. Regarding the TCs and TC-like cells in the oral mucosa they could not be accurately distinguished from other possible cell types, neither on morphological basis (evidence of telopodes) nor by use of panels of markers which include CD34.

The promising role of mucin 4 in association with D2-40 immunohistochemistry in differentiating pleural sarcomatoid mesothelioma from lung sarcomatoid carcinoma

Background Malignant mesothelioma is considered as a rare, aggressive malignant tumor that arises from mesothelial cells of the pleura. Pulmonary spindle cell carcinoma and pleomorphic carcinomas are considered as subtypes of sarcomatoid carcinoma, with an incidence of 2 and 3% of all lung malignancies. The differential diagnosis between sarcomatoid mesothelioma and lung sarcomatoid carcinoma by immunohistochemical markers has not been reached and requires further study. The aim of this study was to investigate the role of mucin 4 (MUC4) and D2-40 in differentiating sarcomatoid mesothelioma from lung sarcomatoid carcinoma. Materials and methods The cases were divided into 20 cases of sarcomatoid mesothelioma and 14 cases of lung sarcomatoid carcinoma and were immunostained for MUC4 and D2-40 using streptavidin-biotin methods. Results MUC4 expression was seen in tumor cells of 11 (79%) lung sarcomatoid carcinoma cases. All cases of sarcomatoid mesothelioma showed negative immunostaining for MUC4 (0%). D2-40 immunostaining was positive in 16 (80%) cases of sarcomatoid mesothelioma. D2-40 expression was observed in two (14%) lung sarcomatoid carcinoma cases. The negative expression of MUC4 showed 78.57% sensitivity; however, specificity was 100%. The positive expression of D2-40 showed 80% sensitivity and 85.71% specificity. MUC4-negative expression and D2-40-positive expression were statistically significant. Conclusion We conclude that MUC4 and D2-40 immunohistochemical markers can differentiate sarcomatoid mesothelioma from lung sarcomatoid carcinoma. The combination of the two markers increases sensitivity and specificity for both.

Immunohistochemical Analysis of Endothelial Cells in Vascular Transformation of Lymph Node Sinuses: Vascular or Lymphatic Differentiation?

Vascular transformation of sinuses (VTS) is an uncommon and benign lesion, defined by conversion of lymph node sinuses into complex, anatomizing and endothelial-lined channels. Despite the name of VTS, which implies a change in differentiation from lymphatic to vascular endothelium, very few studies have systematically examined VTS with modern immunohistochemical markers commonly used in clinical laboratories. It is unclear whether endothelial cells in VTS display pure vascular or lymphatic differentiation, or both. A total of 11 cases with a diagnosis of VTS (identified in the tissue archives of the Cleveland Clinic between 1992 and 2015) were reviewed and confirmed. Twenty cases of benign lymph nodes without specific diagnoses were used as control tissues. Immunohistochemical stains were performed on formalin-fixed, paraffin-embedded lymph node tissue using an automated immunohistochemistry platform with antibodies against CD31, CD34, D2-40, and ERG. Positivity in the VTS lesions was defined as distinct expression in the appropriate cell compartment in ≥20% of cells. In control cases, staining was evaluated in both vascular and lymphatic channels-vascular structures were identified by presence of red cells or well-formed vascular walls and lymphatics by anatomic location and absence of vascular features. In the VTS lesions, D2-40 expression was absent in the lesional endothelial cells of 5/11 (45%) cases. In the cases lacking D240 expression, uninvolved lymphatic endothelium maintained expression. CD34 expression was also seen in 6/11 (54%), CD31 was seen in 10/11 (90%), and ERG expression was seen in all cases. In all the control cases, D240 expression was exclusively seen in lymphatic endothelial cells and not seen in vascular endothelial cells (eg, vascular channels in the hilum). CD34 was weakly positive in the lymphatic endothelium of only 7/20 (35%) control cases, but expressed in 20/20 (100%) control cases in the vascular endothelium. 20/20 (100%) of control cases showed expression of CD31 and ERG in both vascular and lymphatic endothelium. VTS lesional endothelial cells demonstrate patterns of vascular markers that show mixed blood vascular and lymphatic features. There appears to be a degree of alignment toward endothelial differentiation with decreased expression of D2-40 in some cases.

An Immunohistochemical Study of Gastric Mucosa and Critical Review Indicate that the Subepithelial Telocytes are Prelymphatic Endothelial Cells.

Background and Objectives: There are only a few studies regarding gut subepithelial telocytes (TCs). The telopodes, namely peculiar TCs’ prolongations described on two-dimensional cuts, are not enough to differentiate this specific cell type. Subepithelial TCs were associated with the intestinal stem niche but a proper differential diagnosis with lymphatic endothelial cells (LECs) was not performed. In this study, we will also critically review studies suggesting that distinctive TCs could be positioned within the lamina propria. Materials and Methods: We performed an immunohistochemical study of human gastric mucosa to test the expression of D2-40, the lymphatic marker, as well as that of CD31, CD34, CD44, CD117/c-kit, α-smooth muscle actin (α-SMA) and vimentin in the gastric subepithelial niche. Results: The results support the poorly investigated anatomy of intramural gastric lymphatics, with circumferential collectors located on both sides of the muscularis mucosae (mucosal and then submucosal) and myenteric collectors in the muscularis propria. We also found superficial epithelial prelymphatic channels bordered by D2-40+ but CD31–TC-like cells. Deep epithelial lymphatic collectors drain in collectors within the lamina propria. Blood endothelial cells expressed CD31, CD34, CD44, and vimentin. Conclusions: Therefore, the positive diagnosis of TC for subepithelial CD34+ cells should be regarded with caution, as they could also be artefacts, resulting from the two-dimensional examination of three dimensional structures, or as LECs. Lymphatic markers should be routinely used to discriminate TCs from LECs.

Elevated blood/lymphatic vessel ratio in pterygium and its relationship with vascular endothelial growth factor (VEGF) distribution.

Pterygium is a conjunctival fibrovascular tissue growth on the cornea. The pathogenesis of pterygium involves several factors such as the presence of active angiogenic factors. Expansion of the lymphatic microvasculature has also been hypothesized. This study examines the activity of the angiogenic/lymphangiogenic factor VEGF and the expression of vascular and lymphatic endothelial proteins in pterygia and normal conjunctival tissues. Primary grade 2 pterygium (n=20) and normal conjunctiva (n=20) biopsies were obtained during surgery after written informed consent. mRNA expression for CD31, podoplanin, and VEGF (isoforms VEGF-A and VEGF-165) were determined by qRT-PCR. Tissue samples were also processed for immunohistochemical techniques to examine the lymphatic and vascular endothelium (anti-D2-40, anti-CD31 respectively) and VEGF-A and VEGF-C levels and distribution. VEGF-A gene expression levels failed to differ between the healthy and pterygium tissues. However, expression of its more angiogenic isoform, VEGF-165, was significantly higher in the pterygia. Immunohistochemistry revealed the greater presence of VEGF-A, compared to VEGF-C, in pterygium than conjunctiva, both in blood vessels and extracellular matrix. In addition, pterygia showed higher expression levels of the endothelial junction protein CD31. Lymphatic marker D2-40 expression was slightly augmented in this pathological tissue. The ratio between blood and lymphatic vessel counts was 1.05 in the normal conjunctiva and 3-fold this value in pterygium. In pterygium, while both lymphangiogenesis and angiogenesis take place, the formation of new blood vessels is the most relevant event, correlating with the increased expression of vascular endothelial CD31 and an elevated blood/lymphatic vessel ratio. The presence of high levels of VEGF-A in both vessel networks and extracellular matrix in human pterygium tissue may have a major impact on angiogenesis in this pathological tissue.

Expression of Angiopoietin and VEGF in Cervical Cancer and its Clinical Significance.

ObjectiveThe aim of this study was to evaluate the expression of Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2) and vascular endothelial growth factor (VEGF) in cervical cancer and its clinical significance.MethodsImmunohistochemical assay was used to examine the expression of Ang-1/2 and VEGF in tumor tissue from 56 cervical squamous cell carcinoma patients treated with operation only (SCC-O group), as well as 51 subjects with cervical squamous cell carcinoma treated with neoadjuvant radiotherapy (SCC-RCO group, n=28) or neoadjuvant chemotherapy (SCC-CO group, n=23). Both microvessel density (MVD) and lymphatic vessel density (LVD) were examined in the three groups through detection of CD34 and D2-40 expression in respective tissue samples.ResultsWith the progression of cervical cancer, the positive expression scores of Ang-2 and VEGF were significantly increased (p<0.05). Compared with surgical intervention, neoadjuvant chemoradiotherapy significantly reduced the positive expression scores of Ang-1, Ang-2, and VEGF in cervical cancer tissues (p<0.05). The MVD values of the SCC-CO and SCC-RO groups were significantly reduced as compared to the SCC-O group (p<0.05). Similarly, the LVD values of the SCC-CO and SCC-RO groups were also significantly reduced when compared to those of the SCC-O group (p<0.05). However, LVD values of the SCC-CO and SCC-RO groups were not statistical different (p>0.05).ConclusionAng-1, Ang-2 and VEGF may play an important role in the development of cervical cancer. Mutual synergism of Ang-2 and VEGF demonstrated a close relationship with the generation of cervical blood and lymphatic vessels. Cervical cancer radiotherapy and chemotherapy could significantly inhibit the formation of blood vessels and lymphatic vessels in tumor tissue.

Immunohistochemical characterization of lymphangiogenesis-related biomarkers in primary and recurrent gliomas: A STROBE compliant article.

Glial tumors constitute the majority of primary intracranial brain tumors. The expression of specific markers of lymphangiogenesis in gliomas still remains unclear.A total of 40 surgical specimens from 20 patients with recurrent gliomas were included in the study. The expression of D2-40, vascular endothelial growth factor (VEGF)-C, VEGF-D, and VEGF receptor-3 (VEGR-3) was detected by immunohistochemistry (IHC). The clinicopathologic data (p53 and Ki67) were also collected and analyzed.At relapse malignant transformation rate was 65% (13/20 cases). D2-40, VEGF-C, VEGF-D, and VEGFR-3 were expressed in 20%, 30%, 60%, and 20% of primary and 45%, 30%, 75%, and 35% of recurrent glioma tumors (P < .01, P = 1.00, P = .03, P = .03). In 13 cases with increased malignancy grade, the expression of Ki67 and p53 were higher at relapse compared with the primary tumors (P = .001, P = .045). Multivariate survival analysis showed VEGF-D was an independent prognostic factor for malignant transformation (HR = 0.376, P = .045).Glioma is easy to relapse with tumor progression. VEGF-D was an independent prognostic factor for malignant transformation.

Utility of Immunohistochemistry in Distinguishing Primary Adnexal Carcinoma From Metastatic Breast Carcinoma to Skin and Squamous Cell Carcinoma.

The distinction between primary adnexal carcinoma (PAC) from metastatic breast carcinoma (BrCa) to skin and squamous cell carcinoma (SCC), particularly those with ductal differentiation (SCCDD), can be quite challenging, despite adequate history. The aim of the study was to determine the utility of an immunohistochemistry (IHC) panel to differentiate these entities and apply them to ambiguous tumors. Twenty-seven PAC, 7 metastatic BrCa, 28 SCC, and 16 ambiguous cases (SCCDD vs. PAC, n = 13 and metastatic BrCa vs. PAC, n = 3) were analyzed using CD23, PAX5, D2-40, P63, and CD117 immunohistochemistry. A total of 9 (33%) PAC were CD117 positive, whereas all metastatic BrCa and SCC were negative (P = 0.0002). D2-40 was expressed in 16 (59%) PAC and 16 (57%) SCC cases, but none of the metastatic BrCa cases (P = 0.0041). Of the 13 ambiguous tumors with a differential diagnosis of SCCDD versus PAC, all were positive for P63, 10 were positive for D2-40, and 1 was positive for CD117. Of the 3 ambiguous tumors with a differential diagnosis of PAC versus metastatic BrCA, 2 were positive for CD117, whereas none showed reactivity for D2-40 or P63. All cases were negative for CD23 and PAX5. Our study indicates that CD117 reactivity favors a PAC with a sensitivity and specificity of 33% and 100%, respectively. D2-40 and P63 expression highlighted both PAC and SCC and seems to be useful in excluding metastatic BrCa with a sensitivity and specificity of 58% and 100%, and 98% and 100%, respectively. Despite previous reports, CD23 and PAX5 do not seem to be useful.