Expression of HBME-1 and CD56 in follicular variant of papillary carcinoma in children: An immunohistochemical study and their diagnostic utility

Papillary thyroid carcinoma (PTC) is the most common differentiated thyroid cancer in children; and the follicular variant is the second most common variant after the classic subtype. The histological appearance of follicular variant of papillary thyroid cancer (FVPTC), can be mimicked by benign follicular nodules. Pediatric pathologists encountering such lesions with FVPTC-like appearance may err on diagnosing the benign lesions as malignant. In adult patients, several immunohistochemical markers have emerged recently as a useful adjunct to distinguish differentiated thyroid carcinomas from benign follicular lesions. We undertook an inter-institutional retrospective study to establish the diagnostic utility of immunohistochemical staining for HBME-1, Galectin-3 and CD56 in differentiating FVPTC from its benign mimics, follicular adenoma and adenomatoid nodules, in children. Our specific aim of the project was to define the sensitivity and specificity of the three antibodies in FVPTC. Based on institutional diagnoses, a total of 66 cases were obtained: 32 FVPTC and 34 benign follicular nodules that comprised of 23 follicular adenoma and 11 adenomatoid nodules. Five investigators, who were blinded to the original diagnoses, independently reviewed the slides following pre-determined criteria and semi-quantitatively scoring the immunohistochemical staining. The immunohistochemical staining revealed that a combination of positive HBME-1 and negative CD56 result gave 100% specificity and positive predictive value in distinguishing FVPTC from benign follicular nodules. However, the antibody combination suffered from a lower sensitivity (50%). We used a cutoff of 25% positivity of tumor cells in determining positivity of tumor cells to an antibody. In conclusion, our study found a very high specificity and strong positive predictive value for the combination of HBME-1 and CD56 immunohistochemical stains in distinguishing FVPTC from benign follicular lesions.     

甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1和CD56的表达及意义

目的探讨结节性甲状腺肿合并甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1及CD56的表达及意义。方法采用免疫组化SP法检测10例结节性甲状腺肿合并甲状腺微小乳头状癌中Galectin-3、CK19、HBME-1和CD56的表达水平。结果 Galectin-3、CK19和HBME-1在甲状腺微小乳头状癌中均呈中至强阳性表达,而在结节性甲状腺肿中主要呈阴性或弱阳性表达。然而在10例甲状腺微小乳头状癌中有8例CD56的表达均为阴性,2例呈轻度阳性着色;周围结节性甲状腺肿组织9例均呈中至强阳性表达,仅1例呈轻度阳性着色。结论 Galectin-3、CK19、HBME-1及CD56联合检测将进一步提高甲状腺微小乳头状癌的准确性。     

Immunohistochemical expression of HBME-1 and galectin-3 in the differential diagnosis of follicular-derived thyroid nodules

BackgroundThyroid nodules are common among adults with only a small percentage being malignant and histologically mimic benign nodules. Accurate diagnosis of these thyroid nodules is critical for the proper clinical management. The determination of malignancy in follicular patterned thyroid lesions is based on postoperative histological findings. Therefore, affected patients are referred for surgery, although only 10% will have a final diagnosis of malignancy. The aim of this study was to investigate the ability of two immunohistochemical (IHC) markers; galectin-3 and Hector Battifora mesothelial-1 (HBME-1) individually or in combination, to distinguish between benign (non-neoplastic and neoplastic) and malignant (follicular and papillary carcinomas) thyroid lesions removed by surgical resection.MethodsWe investigated the immunoexpression of galectin-3 and HBME-1 in 50 cases of benign and malignant thyroid nodules. The benign group included 13 cases of thyroid nodular goiter (NG) and 9 cases of follicular adenoma (FA). The malignant group included 5 cases of follicular thyroid carcinomas (FC), 18 cases of classic papillary thyroid carcinoma and 5 cases of follicular variant papillary carcinoma (FVPC).ResultsThe staining results showed that malignant tumors expressed galectin-3 and HBME-1 significantly more than benign nodules. The sensitivity of these markers for the distinction between benign and malignant lesions ranged from 89.3% to 92.9%. Co-expression of galectin-3 and HBME-1 was seen in 82.1% of carcinomas, but in none of the benign nodules. Immunoexpression was usually diffuse in malignant tumors, and focal in the benign lesions.ConclusionOur findings indicate that these immunohistochemical markers are significantly more expressed in malignant tumors compared to benign lesions and may be of additional diagnostic value when combined with routine histology. Galectin-3 has higher sensitivity and specificity of immunoexpression in thyroid malignancy than HBME-1, and the combined use of galectin-3 and HBME-1 can increase the specificity of immunoexpression in malignant tumors.     

Differential reactivity of HBME-1 and CD15 antibodies in benign and malignant thyroid tumours

We studied a wide range thyroid tumours and non-neoplastic conditions (total 463 cases) immunohistochemically to evaluate the possible diagnostic potential of HBME-1 and CD15 antibodies. HBME-1 monoclonal antibody recognizes a biochemically unknown epitope present in mesothelioma and variably present in some adenocarcinomas. CD15 antibodies recognize a sugar epitope also included in Lewis X blood group antigen. All papillary (145/145) and follicular carcinomas (27/27) were HBME-1 positive, usually in the the majority of tumour cells. In contrast, cases of nodular goitre and papillary hyperplasia either showed no reactivity or were focally positive (in a third of cases). The patterns of CD15 reactivity were generally similar, although smaller numbers of tumour cells were positive in papillary carcinomas, and only 50% of follicular carcinomas were positive. Because fetal thyroid also showed CD15 reactivity, this antigen appears to behave as an oncofetal antigen in relation to thyroid tissue. Anaplastic carcinomas were negative with both antibodies, indicating the loss of these epitopes upon high grade malignant transformation. We conclude that HBME-1 and CD15 immunohistochemistry may be helpful in the histological differential diagnosis between benign lesions and differentiated thyroid carcinomas, especially papillary tumours. Although the biochemical basis of HBME-1 reactivity is unknown, increased CD15 reactivity in malignant thyroid tumours probably reflects changes in thyroid follicular epithelial glycoconjugates related to malignant transformation.     

Diagnostic utility of CD56 immunohistochemistry in papillary carcinoma of the thyroid

Diagnosis of papillary thyroid carcinoma (PTC), in many but not all cases, is an easily achievable diagnosis with almost minimal interobservable variability between pathologists. However, some cases of PTC, particularly the follicular variant, are quite challenging and show wide interobservable variability even among expert thyroid pathologists. Since proper diagnosis of PTC is crucial as it affects patients’ clinical management and prognosis, indications of PTC must be clearly apparent to be an objective rather than a subjective diagnosis. Unfortunately, to date, immunohistochemistry and molecular studies have failed to fully solve this problem. In this study, we assessed the protein expression and loss using antibodies against CD56 in normal follicular thyroid epithelium, follicular thyroid lesions, and follicular thyroid neoplasms in an attempt to evaluate its diagnostic value. A total of 185 cases were studied with tissues from 75 carcinomas (72 papillary, 2 follicular, 1 Hürthle cell) and 35 adenomas (32 follicular and 3 Hürthle cell) evaluated by immunohistochemistry for the expression of this marker. Non-neoplastic thyroids included 65 cases: nodular hyperplasia (n=25), thyrotoxic hyperplasia (Grave's disease) (n=5), lymphocytic thyroiditis (n=19), and Hashimoto's thyroiditis (n=6). Ten cases of normal thyroids from radical laryngectomies for laryngeal squamous cell carcinomas were also studied.     

Prognostic significance of Leu-M1 immunostaining in papillary carcinomas of the thyroid gland

Summary Leu-M1 antigen is a monocyte/granulocyte-related marker known to be consistently expressed in the Reed-Sternberg cells of patients with Hodgkin's disease. Recently, however, the presence of Leu-M1 has also been noted in tumour cells of a variety of non-haematopoietic neoplasms, most of them adenocarcinomas. The biological significance of this aberrant reaction has not been clarified. We have been able to demonstrate marked epithelial Leu-M1 immunoreactivity (>15% tumour cells positively stained) in 24 out of 76 (32%) papillary carcinomas of the thyroid gland (PC). This phenomenon was more frequently observed among PCs at an advanced stage of disease (pT₄ vs. pT_1–3 and M₁ vs. M₀ p<0.05). The degree of epithelial Leu-M1 positivity was also shown to be significantly correlated to the clinical course of PC. Irrespective of other morphological and clinical features, death resulting from cancer occurred 17 times more frequently among PCs with marked Leu-M1 positivity (8/24) when compared with tumours with only slight or absent immunoreactivity (1/52) (p<0.00005). These findings suggest that Leu-M1 immunostaining provides significant prognostic information for patients with papillary carcinoma of the thyroid gland.     

Role of cd56 and e-cadherin expression in the differential diagnosis of papillary thyroid carcinoma and suspected follicular-patterned lesions of the thyroid: the prognostic importance of e-cadherin

Introduction: The pathological diagnosis of papillary thyroid carcinoma (PTC) is generally easy on routine sections stained with hematoxylin and eosin (H&E). However, the differentiation of the follicular variant of PTC (FVPTC) from other suspected follicular-patterned lesions of the thyroid is highly difficult. Among these, the lesions for which FVPTC cannot be excluded are classified as well-differentiated tumors of uncertain malignant potential (WDT-UMP). The most common immunohistochemical (IHC) markers used in the differential diagnosis include HBME-1, galectin-3, and CK19. However, none of these markers provide a 100% differential diagnosis. Objective: The present study compared the diagnostic value of CD56 and E-cadherin for the differentiation of FVPTC from the other benign follicular-patterned lesions, with HBME-1, galectin-3, and CK19. Using these markers, the controversial cases within the WDT-UMP group were reclassified. Additionally, the relationship between the reductions in E-cadherin expression with poor prognostic factors was investigated. Materials and methods: The IHC expressions of CD56, E-cadherin, HBME-1, galectin-3, and CK19 were evaluated in 181 thyroid lesions, including 101 PTCs (45 classical variant PTCs and 56 FVPTCs), 20 WDT-UMPs, 20 follicular adenomas (FAs), 20 hyperplastic nodules (HN), and 20 hyperplastic foci of lymphocytic thyroiditis. The results were statistically compared via SPSS. Results: The expressions of all of the markers were statistically significantly different in PTC and follicular-patterned lesions (P<0.05). It was found that the only marker with both sensitivity and specificity above 90% was CD56 negativity (sensitivity 91.1%, specificity 91.7%). The most sensitive and also the most specific double panel was CD56 negativity and galectin-3 positivity (sensitivity 96%, specificity 85%), and the most sensitive and specific triple panel was CD56 negativity, HBME-1 positivity, and galectin-3 positivity (97% and 70%, respectively).     

Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1

The histopathology of papillary thyroid hyperplasia and papillary thyroid carcinoma is similar enough to cause a diagnostic dilemma in a few cases. Both lesions may have papillary fronds with fibrovascular cores, nuclear crowding, and nuclear anisocytosis. Formalin-fixed paraffin-embedded tissues from 30 randomly selected patients with papillary thyroid hyperplasia and an equal number from patients with papillary thyroid carcinoma were analyzed for expression of cytokeratin 19 (CK19), galectin-3, and HBME-1. Cases of papillary thyroid carcinoma had moderate to strong CK19, galectin-3, and HBME-1 reactivity although both CK19 and galectin-3 showed positive staining in a significant number of nonneoplastic thyroid cases. HBME-1 was uncommon in the nonneoplastic cases. These results indicate that HBME-1 may be useful in helping to distinguish papillary thyroid carcinoma from hyperplasia in diagnostically difficult cases.     

Galectin-3 and HBME-1 expression in oncocytic cell tumors of the thyroid

Oncocytic cell tumors (OCTs) of the thyroid include oncocytic cell adenomas (OCAs) and oncocytic cell carcinomas (OCCs). Oncocytic variant of papillary carcinoma (OVPC) has also been described. These tumors may present similar diagnostic problems as their non-oncocytic counterparts, in both conventional histology and fine-needle aspiration biopsies. Several markers were shown able to distinguish benign from malignant thyroid follicular tumors, galectin-3 and HBME-1 being the most promising ones. Controversial data have been reported on their discriminatory potential in the small series of OCTs so far analyzed. We aimed to assess the role of galectin-3 and HBME-1 in a large series of 152 OCTs (including 50 OCAs, 70 OCCs and 32 OVPCs). The expression of PPAR protein was also evaluated. Using a biotin-free detection system, the sensitivity of galectin-3 was 95.1%, while that for HBME-1 was nearly 53%. The combination of galectin-3 and HBME-1 increased the sensitivity up to 99%. However, for both markers, the specificity was 88%, lower than that reported for non-oncocytic follicular tumors. PPAR protein overexpression was absent in all OCAs tested and present in only 10% of OCCs, confirming previous reports on the low prevalence of PAX8-PPAR translocations in OCT and ruling out its role as a potential diagnostic marker of malignancy.     

Diagnostic value of galectin-3, HBME-1, cytokeratin 19, high molecular weight cytokeratin, cyclin D1 and p27(kip1) in the differential diagnosis of thyroid nodules

The distinction between benign and malignant thyroid tumors is critical for the management of patients with thyroid nodules. We applied immunohistochemical staining for galectin-3, HBME-1, cytokeratin 19 (CK19), high molecular weight cytokeratin (HMWCK), cyclin D1 and p27(kip1) in 295 thyroid lesions to determine their diagnostic accuracy. The expression of all markers was significantly associated with differentiated thyroid carcinoma (DTC).The sensitivity for the diagnosis of DTC was 94.7% with galectin-3, 91.3% with HBME-1, and 90.3% with CK19. The specificities of these markers were 95.5%, 69.7%, and 83.1%, respectively. Combining these markers, co-expression of galectin-3 and CK19 or galectin-3 and HBME-1 was seen in 93.2% of carcinomas but in none of the benign nodules. Comparing follicular variant of papillary carcinoma (FVPC) with follicular carcinoma (FC), the expression of galectin-3, CK19, and HMWCK was significantly higher in FVPC. When comparing FC with FA, the expression of galectin-3 and HBME-1 was significantly higher in FC. These results suggest that 1) galectin-3 is a useful marker in the distinction between benign and malignant thyroid tumors, 2) the combined use of HBME-1 and CK19 can increase the diagnostic accuracy, and 3) the use of CK19 and HMWCK can aid in the differential diagnosis between PC and FC.     

The meanings of markers: ancillary techniques in diagnosis of thyroid neoplasia

This review article primarily discusses immunohistochemical markers used to aid the diagnosis of thyroid neoplasia. The review concentrates on high molecular weight cytokeratins, cytokertain 19, HBME-1, and galectin-3. Diagnostic uses of proliferation markers such as p27^kip1 and Ki-67 are discussed along with the oncogene ret in relation to papillary carcinomas and PAX8-peroxisome proliferator-activated receptor γ1 in relation to follicular carcinoma. The diagnostic use of retinoblastoma tumor suppressor gene product is also discussed. The sensitivities and specificities as well as the pitfalls of these techniques are reviewed. On the basis of the literature, the most highly recommended of these markers to aid in diagnosis are HBME-1 and galectin-3.     

Immunohistochemical analysis of nm23 protein expression in thyroid papillary carcinoma and follicular neoplasm

Introduction

We aimed at assessing the significance of nm23 gene expression in papillary and follicular carcinomas, the two most common differentiated thyroid carcinomas.

Materials and methods

During a cross-sectional study, 173 paraffin blocks, including 131 papillary thyroid carcinomas, 12 follicular carcinomas and 30 follicular adenomas were stained with nm23 marker by immunohistochemistry method. Cytoplasmic staining in more than 10% of the tumor cells was considered as positive, and α<0.05 was established as the level of statistical significance for the evaluation of the correlation between nm23 expression and age, sex, tumor size, vascular /capsular invasion and lymph node involvement.

Results

nm23 was positive in 40% of the follicular adenoma, 67.2% of the papillary carcinoma and 66.7% of the follicular carcinoma. p value was more than 0.05 in the assessment of the relationship between nm23 and all of the above-mentioned parameters in differentiated thyroid carcinomas. nm23 expression did not significantly differentiate between follicular adenoma and carcinoma.

Conclusion

According to our results, there is no relationship between nm23 immunoreactivity and age or sex of the patients. Also, nm23 cannot be considered as a useful marker for the evaluation of invasion in differentiated thyroid carcinomas or in distinctions between follicular adenoma and carcinoma.     

Immunohistochemical markers in diagnosis of papillary thyroid carcinoma: Utility of HBME1 combined with CK19 immunostaining.

Papillary thyroid carcinoma and its variants can be difficult to distinguish from cellular adenomatous nodules. Prior studies have advocated various antibodies to aid in the differential diagnosis, but there is little agreement on their utility. We undertook this study to evaluate immunohistochemical markers in the diagnosis and differential diagnosis of papillary thyroid carcinoma. Ten cases of papillary thyroid carcinoma were initially stained for HBME1, CK19, fibronectin1, Ki-67, Calretinin, p16, SFTPB and CITED1. Additionally, two previously untested antibodies to molecules that have been found to be upregulated in papillary thyroid carcinoma (CST6 and EPS8) were also evaluated. Of these, only HBME1, CK19 and fibronectin1 showed diagnostic utility. These three markers were then further evaluated in 51 papillary thyroid carcinomas and 57 benign thyroids. HBME1 was the most sensitive and specific marker, staining 49/51 papillary thyroid carcinomas and only 4/57 benign thyroids. CK19 was equally sensitive staining all 51 papillary thyroid carcinomas, but it was nonspecific staining 39 of 57 benign thyroids. A negative result, however, was helpful in excluding papillary thyroid carcinoma. Fibronectin1 was positive in 35/51 papillary thyroid carcinomas (69%) and 4/57 (7%) benign thyroids, but its utility was hampered by high background staining. These findings suggest that the combination of HBME1 and CK19 has the greatest diagnostic utility in the differentiation of papillary thyroid carcinoma from its benign mimics.     

甲状腺乳头状癌中CK19、Galectin-3、HBME-1和TPO的表达及意义

目的 探讨CK19、Galectin-3、HBME-1和TPO在甲状腺良恶性病变中的表达及联合应用在甲状腺乳头状癌鉴别诊断中的价值.方法 采用免疫组化EnVision法检测68例甲状腺乳头状癌、31例甲状腺腺瘤、19例结节性甲状腺肿和15例桥本甲状腺炎中CK19、Galectin-3、HBME-1和TPO的表达.结果 在甲状腺乳头状癌中CK19、Galectin-3和HBME-1的阳性表达率分别为98.5%、98.5%、80.9%,TPO的阴性表达率为89.7%;在甲状腺良性病变中CK19、Galectin-3和HBME-1阳性表达率分别是24.6%、21.5%、1.5%,TPO的阴性表达率是1.5%.CK19、Galectin-3、HBME-1和TPO在甲状腺乳头状癌与良性病变中的表达差异有显著性(P<0.001).联合应用四种抗体在鉴别甲状腺乳头状癌与良性病变时的敏感性、特异性、准确度分别为95.6%、98.5%、97%.结论 CK19、Galectin-3、HBME-1和TPO是诊断甲状腺乳头状癌的重要标志物.联合应用四种抗体在甲状腺乳头状癌与良性病变的鉴别诊断中具有重要价值.     

Intranodal benign thyroid tissue: significance of HBME-1 in differentiation from metastatic papillary thyroid carcinoma

The aim of this study was to determine the significance of HBME-1 immunostaining in the differentiation between intranodal benign thyroid tissue and metastatic papillary thyroid carcinoma in the lymph node. Immunohistochemically we examined normal-appearing intranodal thyroid tissue in four patients who did not show evidence of papillary carcinoma histologically or clinically. We also examined follicular-pattern-predominant papillary carcinoma with metastatic foci in the lymph nodes. Normal-appearing intranodal thyroid tissue and normal thyroid showed no immunopositivity for HBME-1. In contrast, all papillary carcinomas in both the lymph nodes and thyroid demonstrated strong positivity for HBME-1. HBME-1 was predominantly positive for the luminal surface of the tumor cells. The immunopositivity of the cuboidal and low columnar carcinoma cells was more intensive than that of the flat-shaped cells in the lymph nodes and thyroid. The results probably indicate that HBME-1 immunostaining is helpful in distinguishing between intranodal benign thyroid tissue and metastatic papillary carcinoma in lymph nodes. We emphasize that the HBME-1 reactivity should be evaluated in connection with the histological findings, and that positive and negative controls stained in parallel are necessary.     

甲状腺乳头状癌与良性乳头状增生中p16、CD56蛋白表达特点及诊断价值

目的探讨p16与CD56标记在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)和良性乳头状增生(benign papillaryhyperplasia,BPH)中的表达特点与诊断价值。方法采用免疫组化EnVision两步法检测46例PTC及22例BPH中p16与CD56蛋白的表达。结果乳头状癌中p16阳性率以(+～)为主,分布呈癌巢中央区阳性率低、浸润性边缘阳性率高的特点,癌周组织(-)。CD56在80%的癌组织中表达缺失,其余病例呈(+～),癌周组织(+)。p16、CD56对PTC和BPH的诊断敏感度为98%、80%;特异度为82%、64%;准确度为93%、75%。两者平行联合检测,敏感度达100%,特异性降为55%,如串联联合,特异性升高为91%,敏感度率为78%。结论 p16蛋白是鉴别PTC与BPH的良好标记物,CD56对这两种病变的诊断价值有限,p16、CD56蛋白联合检测并不优越于p16单独标记。     

Immunohistochemical demonstration of lactoferrin in follicular adenomas and thyroid carcinomas

Summary By immunohistochemistry, the presence of lactoferrin was investigated in follicular adenomas (10 cases) and carcinomas of the thyroid gland (23 cases). Normal thyroid tissue was also tested as control.Follicular adenomas showed a consistant negativity, whereas follicular and papillary carcinomas exhibited various degrees of positivity for lactoferrin. Incorporated organoid structures observed in anaplastic carcinomas were strongly stained; the spindle cell parts of these cancers were always negative for this iron-binding protein. Medullary carcinomas were also unstained.These findings are discussed in relation to the distribution pattern of thyroglobulin.The authors emphasize the possibility that lactoferrin may be useful in clarifying some diagnostic problems in neoplastic thyroid pathology.