D2-40 Antibody Research Articles

Lymphatic fluctuation in the parenchymal remodeling stage of acute interstitial pneumonia, organizing pneumonia, nonspecific interstitial pneumonia and idiopathic pulmonary fibrosis

Because the superficial lymphatics in the lungs are distributed in the subpleural, interlobular and peribroncovascular interstitium, lymphatic impairment may occur in the lungs of patients with idiopathic interstitial pneumonias (IIPs) and increase their severity. We investigated the distribution of lymphatics in different remodeling stages of IIPs by immunohistochemistry using the D2-40 antibody. Pulmonary tissue was obtained from 69 patients with acute interstitial pneumonia/diffuse alveolar damage (AIP/DAD, N = 24), cryptogenic organizing pneumonia/organizing pneumonia (COP/OP, N = 6), nonspecific interstitial pneumonia (NSIP/NSIP, N = 20), and idiopathic pulmonary fibrosis/usual interstitial pneumonia (IPF/UIP, N = 19). D2-40+ lymphatic in the lesions was quantitatively determined and associated with remodeling stage score. We observed an increase in the D2-40+ percent from DAD (6.66 ± 1.11) to UIP (23.45 ± 5.24, P = 0.008) with the advanced process of remodeling stage of the lesions. Kaplan-Meier survival curves showed a better survival for patients with higher lymphatic D2-40+ expression than 9.3%. Lymphatic impairment occurs in the lungs of IIPs and its severity increases according to remodeling stage. The results suggest that disruption of the superficial lymphatics may impair alveolar clearance, delay organ repair and cause severe disease progress mainly in patients with AIP/DAD. Therefore, lymphatic distribution may serve as a surrogate marker for the identification of patients at greatest risk for death due to IIPs.

D2-40 immunohistochemical overexpression in cutaneous squamous cell carcinomas: A marker of metastatic risk

Approximately 4% of cutaneous squamous cell carcinomas (cSCCs) develop lymphatic metastases. The value of lymphatic endothelial markers to enhance the detection of lymphatic tumor invasion in cSCC has not been assessed previously. We sought to evaluate the use of the antibody D2-40, a podoplanin immunohistochemical marker, to identify tumor lymph vessel invasion in cSCC and to assess its expression in tumor cells. This was a retrospective case-control study. A series of 101 cSCC, including 51 cases that developed lymphatic metastatic spread (metastasizing cSCC [MSCC]) and 50 cases that resolved definitely after surgical excision (non-MSCC) were included in the study. Lymph vessel invasion using D2-40 was evaluated on all primary biopsy specimens. The percentage of tumor cells showing D2-40 positivity and intensity scoring were recorded. All the immunohistochemical findings were correlated with the clinicopathological features. Lymph vessel invasion was observed in 8% of non-MSCCs and in 25.5% of MSCCs (P = .031). D2-40 expression was significantly increased, both in intensity (odds ratio 4.42 for intensity ++/+++) and in area (odds ratio 2.29 for area >10%), in MSCC when compared with non-MSCC. Interestingly, almost half (49%) of the MSCC had moderate to intense D2-40 positivity compared with 16% of non-MSCC. D2-40 immunohistochemical expression was increased in tumors with an infiltrative pattern of extension. In the multivariate analysis, histologically poorly differentiated tumors, recurrent lesions, and cSCC showing D2-40 overexpression (in intensity) were significantly associated with lymphatic metastases development (odds ratios 15.67, 14.72, and 6.07, respectively). This was a retrospective study. The expression of podoplanin associates with high metastatic risk in cSCC.

Clinicopathological significance of lymphatic vessel density marked by D2-40 and E-cadherin expression in non-small-cell lung cancer

The aim of our study is to investigate the relationship between lymphatic vessel density (LVD) marked by D2-40 and E-cadherin expression and the clinicopathological characteristics of patients with non-small-cell lung cancer (NSCLC). Immunohistochemical analysis was used to detect the expression of D2-40 and E-cadherin in 46 archival surgical specimens of human NSCLC and 10 cases of benign pulmonary disease. The LVD positively stained with D2-40 was observed and counted. The LVD was mainly distributed in the tumor borderline. The LVD in NSCLC tissues was significantly higher than in benign lesions (p < 0.001), and the LVD marked by D2-40 was significantly associated with lymph node metastasis (p < 0.001). The reduced expression of E-cadherin was present in NSCLC tissues. The positive rate of E-cadherin expression was negatively correlated with TNM stage (p = 0.027), pathological grade (p = 0.032), and lymph node metastasis (p = 0.014), and not significantly correlated with histologic classification (p = 0.714) in NSCLC tissues. A correlation analysis showed that high LVD marked by D2-40 was correlated with reduced E-cadherin expression in NSCLC tissues (t = 36.476, p < 0.001). The new monoclonal antibody D2-40 can specifically recognize lymphatic endothelial cells in NSCLC tissue. The relationship between LVD marked by D2-40 and the reduced expression of E-cadherin and lymph node metastasis in NSCLC was demonstrated. The detection of D2-40 and E-cadherin may be used as an indicator of lymph node metastasis in NSCLC.

The lymphatic system in clinically localized urothelial carcinoma of the bladder: Morphologic characteristics and predictive value

ObjectiveTo assess the lymphatic vessel density (LVD) and lymphangiogenesis in urothelial carcinoma of the bladder (UCB) and to identify predictors of progression in patients treated by transurethral resection (TUR). Materials and methodsOne hundred eleven patients who underwent TUR for UCB were retrospectively included. Lymphatic endothelial cells were stained immunohistochemically [D2-40 (podoplanin) antibody in all samples; Prox-1, LYVE-1, and VEGFR-3 (Flt-4) in subgroups]. LVD was measured in representative intratumoral (ITLVD), peritumoral (PTLVD), and nontumoral (NTLVD) areas using standardized criteria. Double-immunostainings with D2-40/CD-34 were performed to distinguish between blood and lymphatic vessels, and D2-40/Ki-67 stainings were done to detect lymphangiogenesis. Lymph-specific parameters were correlated with pathologic and clinical characteristics. In patients with non-muscle-invasive UCB (n = 76) univariable and multivariable analyses were performed to identify predictors of progression. ResultsThe PTLVD was significantly higher than ITLVD and NTLVD (P < 0.001). Proliferating lymphatic vessels were observed in all specimens assessed with D2-40/Ki-67. Characteristic suburothelial D2-40 positivity was observed in noninvasive pTa tumors. LYVE-1-stainings revealed the existence of tumor-associated macrophages. The presence of intratumoral lymphatic vessels was significantly associated with higher tumor stage, high grade, and sessile growth (all P < 0.001). Muscle-invasive tumors (P = 0.020), higher grade (P = 0.026), the presence of lymphovascular invasion (P < 0.001), and concomitant carcinoma in situ (CIS) (P = 0.020), sessile growth (P = 0.004), and loss of suburothelial D2-40 positivity (P = 0.031) were associated with disease progression in univariable analysis. LVD values in any area were not significantly associated with progression despite detection of proliferating lymphatic vessels. The presence of concomitant CIS was identified as an independent predictor of progression on multivariable analysis (P = 0.041; hazard ratio 4.620). ConclusionsA high peritumoral LVD is present in clinically localized UCB. The presence of intratumoral lymphatic vessels correlates with characteristics of aggressive disease. Lymphangiogenesis occurs; however, the lymph-specific parameters tested in this study cannot be used to predict progression following TUR. The presence of concomitant CIS is an important risk factor for later disease progression in patients with non-muscle-invasive UCB. Our results contribute to the understanding of metastatic tumor spread in UCB.

Objective assessment of blood and lymphatic vessel invasion and association with macrophage infiltration in cutaneous melanoma

The aims of this study were to investigate the role of vascular invasion (blood and lymphatic), vessel density and the presence of tumour-associated macrophages as prognostic markers in 202 cutaneous melanoma patients. Sections of primary melanoma were stained with lymphatic-specific antibody D2-40 to assess lymphatic vessel invasion and density in intratumoural and peritumoural areas; an antibody against endothelial marker CD34 was used to determine blood vessel invasion and density, and an antibody against CD68 was used to determine macrophage counts. Immunohistochemically determined vascular invasion (combined blood and lymphatic) was compared with that determined using haematoxylin and eosin (H&E) staining. The use of immunohistochemistry increased detection of vascular invasion from 8–30% of patients, and histological exam of H&E-stained tissue was associated with a false positive rate of 64%. Lymphatic vessel invasion occurred at a much higher frequency than blood vessel invasion (27 and 4% of patients, respectively). Although immunohistochemically detected vessel invasion was significantly associated with histological markers of adverse prognosis, such as increased Breslow thickness, ulceration and mitotic rate (all P<0.001), no associations with relapse-free or overall survival were observed. High macrophage counts were significantly associated with markers of aggressive disease, such as Breslow thickness, ulceration and mitotic rate (P<0.001, P<0.001, P=0.005, respectively), and lymphatic vessel invasion and high microvessel density (P=0.002 and P=0.003, respectively). These results suggest that vascular invasion is more accurately detected using immunohistochemistry and occurs predominantly via lymphatic vessels. The association of vessel characteristics with histological characteristics of the primary melanoma provides evidence for their biological importance in melanoma, but that they were not associated with clinical outcome attests to the value of existing histological prognostic biomarkers. We note that a high macrophage count may be associated with neovascularisation and primary tumour growth, and may also promote invasion through lymphatic vessels.

Intratumoral Lymphatic Vessel Density in Vulvar Squamous Cell Carcinomas

Lymphatic vessels serve as major routes for regional dissemination, and therefore, lymph node status is a key indicator of prognosis. To predict lymph node metastasis, tumor lymphatic density and lymphangiogenesis-related molecules have been studied in various tumor types. To our knowledge, no previous studies have evaluated the role of intratumoral lymphatic vessel density (LVD) in the behavior of vulvar carcinomas. The aim of this study was to analyze intratumoral LVD in relation to patient survival and well-characterized prognostic factors for cancer. Thirty-five patients with vulvar squamous cell carcinoma underwent vulvectomy and dissection of regional lymph nodes. Clinical records were reviewed, in addition to histological grade, peritumoral lymphatic invasion, and depth of infiltration for each case. Tissue microarray paraffin blocks were created, and lymphatic vessels were detected using immunohistochemical staining of podoplanin (D2-40 antibody). Intratumoral LVD was quantified by counting the number of stained vessels. Higher values for intratumoral LVD were associated with low-grade and low-stage tumors, and with tumors without lymphatic invasion and reduced stromal infiltration. In a univariate analysis, high intratumoral LVD was associated with a higher rate of overall survival and a lower rate of lymph node metastasis. Our results suggest that increased intratumoral LVD is associated with favorable prognosis in vulvar squamous carcinomas.

Characterization of lymphatic vessels in human peripheral neuropathies

Immunohistological studies on lymphatics’ topography in human peripheral nerve are few and recent. By D2-40 immunohistochemistry, we previously described lymphatics in epineurium of human sural nerve. Lymphangiogenesis is described in inflammation. In angiopathic and vasculitic neuropathies proliferation of epineurial blood capillaries is reported. The aim of our study is therefore to investigate the topography and the density of lymphatics in human peripheral nerve and to search possible correlation with blood capillary neovascularization in different neuropathies. We examined biopsied sural nerves of patients suffering from CIDP (chronic inflammatory demyelinating polyneuropathy), vasculitic neuropathy and non inflammatory axonal neuropathy. Immunohistochemistry for detection of lymphatic marker podoplanin (D2-40 antibody) and for general endothelial marker CD31, as well as for Schwann cells protein S-100, was carried out on serial cryostat sections. Morphometric analysis was performed. Lymphatic capillaries were detected in epineurium, most consistently in adjacency of main blood vessels. Occasionally lymphatics were dilated and repleted with mononuclear cells. Podoplanin was also expressed by perineurium and by Schwann cells. No lymphatics were observed endoneurially. Lymphatics showed a far lesser density than blood capillaries and increase of epineurial vascularization resulted significantly associated with higher density of lymphatics. Density variations of epineurial lymphatics, accompanying blood capillaries proliferation, suggest that lymphangiogenesis may occur in neuropathies, in response to inflammation/ischaemia. Lymphatics’ responsiveness to molecular microenvironment is indicated by their expression of growth factor receptors, such as VEGFR3. Lack of lymphatics in closed endoneurial environment is in agreement with analogous findings in brain. Non lymphatic expression of podoplanin is reported in several cell types of nervous system: normal and tumoral ependymal cells, perineurium and Schwann cells. As biological functions of podoplanin, involved in cell migration and cytoskeletal reorganization, are uncompletely understood, its localization on non lymphatic structures of peripheral nerve needs to be defined.

Clinical significance of lymphatic and blood vessel invasion in oral tongue squamous cell carcinomas

Although vascular invasion (VI) is recognized as an important predictor of lymph node metastasis and a significant prognostic factor in head and neck squamous cell carcinoma (HNSCC), there is currently no common definition for the pathological evaluation of VI status. We reviewed the medical records of 63 consecutive resected primary oral tongue SCCs (OTSCCs) without preoperative treatment between June 1999 and April 2008, and evaluated VI status by investigating lymphatic vessel invasion (LVI) and blood vessel invasion (BVI) by using immunohistochemistry (IHC) with monoclonal antibody D2-40 (D2-40) and Elastica van Gieson (EVG) staining, respectively. Subsequently, we analyzed their correlations with cervical lymph node metastasis and prognosis. LVI was found in 16 of the 63 tumors (25.4%) and BVI was in 32 tumors (50.8%). Univariate analysis revealed that the presence of LVI is statistically correlated with lymph node metastasis. Moreover, multivariate logistic regression analysis revealed that LVI is an independent risk factor of nodal metastasis (odds ratio=4.262, 95% confidence interval=1.262-14.397, p=0.020). In contrast, Kaplan-Meier survival analysis revealed that patients with BVI had a significantly shorter disease-free survival (DFS) and overall survival (OS) rates than those without BVI (68.6% versus 90.3%, p=0.028 and 68.6% versus 93.5%, p=0.013, respectively). The present study clearly demonstrated that LVI at primary OTSCC had significant correlation with lymph node metastasis, and that BVI was significantly associated with recurrence and poor prognosis. Evaluation of VI status, as LVI and BVI status separately, using IHC with D2-40 and EVG staining may be useful in predicting lymph node metastasis and poor prognosis in OTSCCs.

Pleurabefunde beim Yellow-Nail-Syndrom

A 76-year-old man presented clinically with coughing and shortness of breath and was diagnosed radiologically to have massive pleural effusion as a combined feature of yellow nail syndrome. A lung biopsy was taken and revealed histologically: chronic non-specific inflammation in the pleuropulmonary border, intrapleural edema with eightfold pleural thickening in comparison to normal, angiogenesis in both the nutritive and functional intrapleural blood vessels, no abnormalities of lymphatic vessels with normal topographical distribution as detected by immunohistochemistry for antibody D2-40, granulomatous chronic foreign body reaction as a consequence of pleural effusion therapy by talcum pleurodesis.The histopathological findings of chronic non-specific pleuritis with angiogenesis and increased permeability of blood vessels led to massive intrapleural edema with pleural effusion. Abnormalities of lymphatic vessels could not be confirmed. Considering the features of this disease, they are probably secondary to chronic r infectious or immunological inflammation or paraneoplastic complications with angiogenesis (in about 19%).

The Predictive Effect of Cadherin-17 on Lymph Node Micrometastasis in pN0 Gastric Cancer

Previous studies identified cadherin-17 (CDH17) as one of the most upregulated genes in node-positive gastric cancer. However, the prognostic significance of CDH17 in pN0 gastric cancer and its association with lymph node micrometastasis (LNMM) have not been investigated. Clinicopathologic features of 191 patients with node-negative gastric cancer were studied retrospectively. All dissected lymph nodes were immunostained by cytokeratin to detect micrometastasis. CDH17 and lymphatic invasion (LVI) in primary carcinoma were evaluated by immunostaining of monoclonal CDH17 and D2-40 antibody. Correlation of CDH17 with clinicopathologic characteristics was subsequently assessed. Risk factors of LNMM were analyzed by univariate and multivariate logistic regression. Cox's proportional hazard model was applied to investigate independent prognostic factors of pN0 gastric cancer. Overall survival rates of patients with positive and negative CDH17 were compared, stratifying by pT stage, Lauren grade, and LNMM status. CDH17 was observed in 126 patients (66.0%). Positive expression of CDH17 was significantly associated with the age, tumor size, pT, Lauren grade, LVI, and LNMM, and identified as one of the independent risk factors of LNMM. Negative predictors of pN0 gastric cancer included pT, Lauren grade, LNMM, and CDH17. Furthermore, in tumors of pT2-3, intestinal histotype, and negative-LNMM, the survival rate of patients with CDH17 was significantly lower than that of patients without CDH17. CDH17 was positively associated with larger tumor size, deeper invasion, diffuse/mixed histotype, LVI, and LNMM, predicting a poor prognosis in pN0 gastric cancer. Additionally, CDH17 may also serve as a potential indicator of LNMM.

Lymph vessel density in seminomatous testicular cancer assessed with the specific lymphatic endothelium cell markers D2-40 and LYVE-1: Correlation with pathologic parameters and clinical outcome

ObjectivesTo evaluate the role of lymph vessel density (LVD) and lymphangiogenesis in seminomatous testicular cancer (STC) by using the lymphatic endothelial cell (LEC) markers LYVE-1 and D2-40. Methods and materialsParaffin embedded tumor specimens from 40 patients with STC were stained by specific D2-40 and Lyve-1 antibodies. LVD was measured in different representative and standardized areas. Fluorescence double immunostaining for Lyve-1 and Ki-67 was performed and results were correlated with clinicopathologic data. The median follow-up period was 55 (range 10–135) months. ResultsMean intratumoral LVD (D2-40: 1.30 ± 1.99; Lyve-1: 1.82 ± 2.34) was significantly lower than peritumoral LVD (D2-40: 4.94 ± 2.58; Lyve-1: 4.62 ± 2.73) and LVD in nontumoral areas (D2-40: 4.81 ± 3.79; Lyve-1: 4.22 ± 3.19). There was no significant difference between LVD measures when using D2-40 or LYVE-1. Detection rates of lymphatic vascular invasion (LVI) were significantly higher than in conventional HE-stained sections (77.5% vs. 52.5%). No proliferating lymphatic vessels were found. ConclusionsWe found that LVD is decreased within tumor areas of STC. Despite a higher peritumoral LVD, no signs of proliferating endothelial cells were observed, suggesting a lack of lymphangiogenesis in STC. Detection of LVI can be optimized by specific D2-40 or LYVE-1 staining.

Microvascular Lymphatic Density Analysis in Cutaneous Regressive and Nonregressive Superficial Spreading Melanomas Using the Lymphatic Marker D2-40

The prognostic significance of spontaneous regression in melanoma, especially thin lesions, has been a controversial issue for the past 20 years, although recent studies suggest that extensive and late regression may be related to worse prognosis. Many data suggest that lymphangiogenesis predicts metastatic spread in melanoma. We have quantified lymphatic microvascular density (LMVD) in thin (≤ 1.0 mm) superficial spreading melanomas comparing regressive and nonregressive melanomas, regressive and nonregressive areas from the same tumor, and early and late histological stages of regression in the same tumor. In addition, we tried to correlate lymphangiogenesis and tumor growth phase. We conducted histological examinations and immunohistochemical analyses using monoclonal antibody D2-40 with subsequent quantification by image analysis of 37 melanomas, 16 regressive and 21 nonregressive (controls). We found higher LMVD in the late stage of regression compared with nonregressive area (internal control) of regressive melanomas. Our study suggest that the late stage of spontaneous regression in thin melanomas may be related to worse prognosis as it showed higher LMVD, and evidence shows that this is related with increased risk of metastatic spread. But this supposition must be confirmed by a longer follow-up for detection of lymph node metastases.

Periacinar Retraction Clefting and D2-40 Expression in Prostatic Adenocarcinoma

Retraction clefting is known to appear in various types of tumors, but it has only recently been recognized as a specific histological phenomenon. Previously, it was considered merely a laboratory procedure artifact, but lately, there have been some assumptions that peritumoral retractions actually represent lymphatic spaces. In our study, we analyzed neoplastic glands in 52 specimens of prostatic adenocarcinoma. Immunohistochemical analysis was performed using D2-40 antibody, to highlight lymphatic endothelium and thereby differentiate actual lymph vessels or lymphovascular invasion from periacinar retractions. Our results showed that the number of lymph vessels was significantly lower in tumorous tissue compared to adjacent normal prostatic tissue. On the other hand, the number of lymph vessels in tumorous tissue was significantly higher than the number of lymph vessels mimicking periacinar retractions. Overall, the number of lymph vessels mimicking periacinar clefts was particularly low. These results are in accordance with our previous studies, which had shown that periacinar clefting appears due to lack of basal cells and stromal changes around tumorous acini. Also, these results support our hypothesis that retractions do not represent lymph vessels but should be considered a distinct entity, which is proven to be helpful both as diagnostic and predictive factor.

Targetoid hemosiderotic hemangiomas (hobnail hemangiomas) are vascular lymphatic malformations: A study of 12 pediatric cases

Targetoid hemosiderotic hemangioma (THH), also called hobnail hemangioma, is a benign vascular lesion and thought to be of lymphatic origin. We sought to perform a clinicopathologic analysis of cases diagnosed as THH in a tertiary care children's hospital. Clinical and histopathologic data were obtained from a chart review of 12 confirmed pediatric cases of THH. To determine the presence or absence of lymphatic vessels in lesional biopsy specimens, we evaluated the expression of the lymphatic endothelial cell marker podoplanin using the D2-40 antibody. Wilms tumor-1 gene immunostaining and Ki-67 proliferation index were also performed to evaluate the proliferative nature of these lesions. Three children had a lesion since birth and 4 had a history of trauma before appearance of the THH. D2-40 immunostaining was positive in every case. Wilms tumor-1 gene immunostaining was negative in 9 cases, focally positive in two cases, and not performed in one case. The Ki-67 proliferation index was very low in all cases studied. The small number of cases and restriction to a pediatric population were limitations. Our findings suggest that THH should be classified as a lymphatic vascular malformation.

Increased lymphatic vessel density and lymphangiogenesis in inflammatory bowel disease

Involvement of the lymphatic system in inflammatory bowel disease (IBD) has been suggested. To examine the density and distribution of lymphatic vessels (LV) within inflamed and non-inflamed wall sections of IBD patients compared with controls, and to evaluate expression of major lymphangiogenic factors. Ileal and colon specimens of 22 patients with Crohn's disease (CD), 16 patients with ulcerative colitis (UC) and 11 controls were studied. Quantification of LV was performed using immunohistochemistry with podoplanin and D2-40 antibodies on seven randomly selected fields. Mucosal expression of podoplanin and lymphangiogenic factor mRNA was measured using PCR. In CD patients, lymphatic density was significantly increased in non-inflamed and inflamed ileal (P < 0.01 and P < 0.001) and colonic (P < 0.01 and P < 0.001) mucosa compared to controls. Podoplanin mRNA levels were similar in non-inflamed mucosal areas and controls, whereas a four- and sixfold increase was seen in inflamed ileal and colonic areas (P < 0.05). In UC, lymphatic density increased fourfold in non-inflamed (P < 0.001) and fivefold in inflamed colonic mucosa (P < 0.001) compared with controls. An increase in podoplanin mRNA levels was seen in both non-inflamed and inflamed areas (P < 0.01) compared with controls. In CD and UC, lymphatics were found throughout the inflamed mucosa, including the upper half of the lamina propria. Expression of lymphangiogenic factors was similar in patients and controls. Increased density of lymphatic vessels is a constant feature of IBD and is present in non-inflamed areas. It is transmural in CD and confined to the mucosa in UC. Its origin remains unclear.

Detection of early lymphangiogenesis by lymphatic microvascular density and endothelial proliferation status in preneoplastic and neoplastic lesions of the uterine cervix

The characteristics of lymphangiogenesis in preneoplastic and neoplastic lesions of the uterine cervix are not well known and the role of this process in tumor progression and metastasis is not well understood. The aim of the present study was to characterize the morphology and distribution of lymphatic vessels and lymphatic proliferative status and to evaluate the value of lymphatic microvascular density (LMVD) in premalignant and malignant lesions of the uterine cervix. One hundred and twenty-eight paraffin-embedded cervical specimens were immunostained with D2-40 antibody specific for lymphatic endothelial cells. Colocalization of D2-40 and Ki67 for the proliferative characterization of lymphatic vessels was obtained by performing double immunostaining. A low density of lymphatic vessels was detected in normal cervix and squamous metaplasia. Intense and particular lymphangiogenic response was found in low and high grade squamous intraepithelial lesions and microinvasive carcinoma. Lymphatic proliferation occurred early in cervical lesions, being more active in premalignant lesions and microinvasive carcinomas than in invasive lesions. Our results suggest an early initiation of an active lymphangiogenesis in cervical lesions. These findings support the hypothesis that cervical preneoplastic lesions represent a critical point in the development of the lymphatic network vasculature. Early lymphangiogenesis could explain lymph node metastasis associated with cervical invasive carcinomas at preliminary diagnosis.

Hepatocyte Growth Factor is a Potential Lymphangiogenic Factor; Clinical Implications

Hepatocyte growth factor (HGF) is a known angiogenic factor that has profound effects on vascular endothelial cells. In recent years, it has been reported that the factor also acts as a lymphagiogenic factor that induces lymphangenesis from lymphatic endothelial cells. The current article debates some of the recent evidence showing such an effect and also discusses the clinical studies that link to lymphatic related conditions, together with clinical implications. Keywords: Hepatocyte growth factor, lymphangiogenesis, endothelial cells, VEGF, melanoma, cMET, lymph node metastases, prostate cancer, lymphatic invasion, metastasis, squamous cell carcinoma, lymphatic microvessel density, LYVE-1, Prox-1, VEGF-R3, breast cancer, HGF antagonist, in vivo tumour models, D2-40 antibody, MW 40K, ERK, akt, PECAM

Adrenal lymphangioma: clinicopathologic and immunohistochemical characteristics of a rare lesion

Adrenal lymphangiomas, also known as cystic adrenal lymphangiomas, are rare, benign vascular lesions that usually remain asymptomatic throughout life. Although previously adrenal lymphangioma lesions were primarily found at autopsy, they are currently detected during imaging work-up for unrelated causes and are likely to imitate other adrenocortical or adrenal medullary neoplasms. We aimed to retrospectively review all adrenal lymphangioma cases at our hospital and further document their lymphatic origin by immunohistochemical staining. A search of surgical pathology records (1984-2008) was conducted. All hematoxylin and eosin sections were retrieved from archives and reviewed by 2 pathologists in the study. Clinical information was gathered from electronic medical records. Representative paraffin-embedded sections from each case were selected for immunohistochemical analysis using monoclonal antibodies D2-40 and AE1/AE3. A total of 9 adrenal lymphangioma cases were identified (6 women and 3 men). All 9 patients were adults at time of diagnosis with a mean age of 42 years (range, 28-56 years). There were 7 white patients, 1 African American patient, and 1 Asian patient. The average size of an adrenal lymphangioma lesion was 4.9 cm (range, 2.0-13.5 cm). Adrenal lymphangioma was twice more frequently located on the right side (6 right-sided and 3 left-sided). Clinically, 4 (44%) of the 9 lesions presented with abdominal, flank, or back pain. One lymphangioma was found during work-up for labile hypertension. The remaining 4 lesions (44%) were asymptomatic and incidentally found during imaging studies for unrelated causes. Surgical removal was achieved by total adrenalectomy in 8 of the 9 lesions and by partial adrenalectomy in the remaining case. No evidence of recurrence or development of a contralateral lesion was encountered in any of the patients. Histologically, our adrenal lymphangiomas showed a typical multicystic architecture with dilated spaces lined by flattened, bland, simple lining. The cystic channels/spaces occasionally contained proteinaceous material and lacked red blood cell content. On immunohistochemical stains, D2-40 cytoplasmic staining was positive in all 9 examined lesions, whereas AE1/AE3 was negative, thus, confirming their lymphatic nature. D2-40 staining was diffuse in 2 and focal in the 7 remaining lesions. Adrenal lymphangiomas are very rare, benign lymphatic neoplasms with a female, right-sided predominance in our current series. They may clinically present with abdominal pain or can be incidentally found during adulthood as a mass, necessitating surgical removal to rule out other types of adrenal neoplasms.

Ovarian cancer-associated fibroblasts contribute to epithelial ovarian carcinoma metastasis by promoting angiogenesis, lymphangiogenesis and tumor cell invasion

Cancer-associated fibroblasts (CAFs) are thought to play an essential role in cancer initiation and development. However, little research has been done to evaluate the role of CAFs in epithelial ovarian cancer (EOC) development. To address this issue, ninety-one specimens were immunostained with α-smooth muscle actin (α-SMA) and fibroblast activation protein (FAP) antibodies to quantify CAFs, and antibodies D2-40 and CD34 to evaluate the lymphatic vessel density (LVD) and microvessel density (MVD) of the lesions. We found there were no α-SMA or FAP positive fibroblasts in normal ovary tissues. More CAFs were found in EOC than in borderline tumors and benign tumors ( P < 0.01). Abundant CAFs in EOC were associated with advanced-stage disease ( P = 0.002), the occurrence of lymph node metastases ( P = 0.02) and omentum metastases ( P < 0.0001), and increased LVD ( P = 0.002) and MVD ( P = 0.0004). CAFs isolated from EOC tissues induced more cancer cells to invade ( P = 0.003) and migrate ( P = 0.005) compared with normal fibroblasts (NFs) isolated from normal ovary tissues in vitro. Our data indicate that CAFs play a vital role in ovarian cancer progression and metastasis. Targeting CAFs as a therapeutic strategy against ovarian cancer is an intriguing concept that needs further study.

D2-40 lymphatic marker for detecting lymphatic invasion in thin to intermediate thickness melanomas: Association with sentinel lymph node status and prognostic value—A retrospective case study

Sentinel lymph node (SLN) status is the greatest prognostic factor of morbidity in melanoma. D2-40 antibody specifically marks lymphatic endothelium and has been used for identifying lymphatic invasion (LI) in multiple cancers. We sought to determine the relationship between melanoma lymphatic invasion (as detected using D2-40 on primary melanoma biopsies/excisions) and the presence or absence of melanoma in subsequent SLN biopsy. We retrospectively evaluated LI using D2-40 on primary biopsies/excisions from patients with thin to intermediate thickness (Breslow thickness: ≤2.0 mm) melanomas, who underwent lymphatic mapping and SLN biopsy, and whose SLN status was known. Sixty-four cases met the criteria and were available for analysis. We analyzed patient age, patient sex, mitotic rate, ulceration, tumor depth, and D2-40 detected LI as predictors of SLN status. Lymphatic invasion detection increased from 3.1% using hematoxylin and eosin only to 21.9% using D2-40. Twelve of 14 patients with D2-40 LI were SLN positive (positive predictive value, 85.7%). D2-40 LI was detected in the primary biopsy specimen of 12 of 18 patients with a positive SLN (sensitivity 66.7%). Of 50 patients without D2-40 LI, 44 were SLN negative (negative predictive value, 88.0%). Of 46 SLN-negative patients, 44 did not have D2-40 LI (specificity, 95.7%). Results are retrospective and limited to SLN biopsy performed at one institution. On univariate and multivariate analysis, D2-40-detected LI was the most significant predictor of SLN status. D2-40 antibody staining to detect lymphatic invasion should be incorporated in routine melanoma biopsy evaluation.