A study to improve the identification of pancreatobiliary adenocarcinoma utilizing fine-needle aspiration cytology and immunohistochemical application for KOC and S100P.

INTRODUCTION: The identification of pancreatic adenocarcinoma by fine-needle aspiration (FNA) cytology is a difficult, yet critical, task. This study uses a panel of two immunohistochemical (IHC) markers, KOC and S100P, to augment the interpretation of pancreatic adenocarcinoma using cytopathology specimens and to compare these to corresponding surgical specimens. MATERIALS AND METHODS: We retrospectively reviewed 33 surgical specimens with pancreatic adenocarcinoma and 33 corresponding, preceding FNA cytology specimens. IHC studies for KOC and S100P were performed on both the surgical specimens and cytology cell blocks. Three pathologists reviewed the staining intensity and amount of tumor cell staining within these blocks. The findings were then analyzed for sensitivity, specificity, and combined sensitivity and specificity for the 2 markers. RESULTS: KOC performed similarly to S100P in sensitivity for surgical specimens (90.9% for both) and better for FNA specimens (92.3% versus 82.7%, respectively). The specificity of KOC was significantly better than S100P for surgical and FNA specimens (100% for KOC in both specimens versus 72.7% and 89.7% for S100P in both specimens, respectively). The combined sensitivity of the panel of KOC and S100P was 99.2% for surgical specimens and 98.7% for FNA specimens. The combined specificity was 72.7% for surgical specimens and 89.7% for FNA specimens. CONCLUSIONS: We found using KOC and S100P on FNA cell block cytology specimens to be a useful adjunct for interpretation when an interpretation of atypical or suspicious for pancreatic ductal adenocarcinoma is being considered and there are atypical epithelial cell groups in the cell block.

Parvovirus b19 persistence in abnormal thyroid tissue of a mature cystic ovarian teratoma: a case report.

Ovarian teratomas represent the most common neoplasm derived from germ cells and can contain mature ectodermal, mesodermal, and endodermal tissues. In rare cases, these teratomas can be composed predominantly or solely of thyroid tissue. These thyroid cells often function similarly to normal thyroid tissues. This laboratory and others have previously shown that parvovirus B19 (B19V) persists in primary and metastatic thyroid tissues. No reports exist on possible B19V persistence in thyroid tissues that may arise de novo outside the thyroid gland proper. In this case report, the detection of B19V (genotype 1) in the thyroid epithelial cells of a mature teratoma is reported. Nested PCR and immunohistochemistry were used to detect viral nucleic acids and proteins, respectively. Viral genomes were amplified in lesion DNA, confirming persistence of B19V. Positive immunohistochemical staining was seen for B19V capsid proteins in the thyroid epithelial cells within the mature teratoma, but not in surrounding ovarian tissue or in the non-thyroidal elements of the mature teratoma. These results demonstrate for the first time that thyroid epithelial cells, derived from non-thyroid tissue, are capable of supporting B19V infection and persistence.

Infection and persistence of erythrovirus B19 in benign and cancerous thyroid tissues.

Human erythrovirus B19 (EVB19) is a small, pathogenic DNA virus that has been associated with a wide range of illnesses. The primary site of replication is in bone marrow-derived erythroid progenitor cells, but EVB19 DNA has been detected in a wide range of organs. Recently, studies have linked EVB19 to thyroid cancers and other thyroid diseases. Previous studies from multiple laboratories have detected EVB19 capsid proteins in Graves' disease, Hashimoto's thyroiditis, and thyroid cancer tissues. Data on viral gene expression and mechanism of infection in the thyroid are lacking. To investigate EVB19 infection and persistence in the thyroid, previously archived adult and pediatric tissue sections were examined for EVB19 DNA, RNA, and capsid proteins, as well as EVB19 receptor P-antigen and co-receptor α5ß1 integrin. EVB19 DNA and protein were detected in a majority of tissues examined (87% and 68%, respectively). Detection was similar in adult and pediatric samples. Quantification of viral genomes revealed no significant difference in the amount of viral DNA in benign, cancerous, or metastatic thyroid tissues. EVB19 capsid RNA was detected in 67% of the tissues examined, confirming at least low-level viral gene expression. Immunohistochemical staining for P-antigen and α5ß1 detected the receptor and co-receptor most frequently on normal thyroid epithelial cells. EVB19 capsid staining could be detected in tumors lacking viral receptors. These results suggest that normal thyroid epithelial cells are the initial target for EVB19 infection in the thyroid and allow for continued persistence in both normal and cancerous thyroid tissues.

Increased IL-6 detection in adult and pediatric lymphoid tissue harboring parvovirus B19.

BACKGROUND: Parvovirus B19 (B19V) is a common pathogenic virus infecting humans. Previous studies have shown evidence of B19V infection in patients with non-Hodgkin's lymphoma (NHL) and Hodgkin's lymphoma using ELISA and PCR on serum specimens. B19V nonstructural protein is known to alter the expression of cellular factors including interleukin-6 (IL-6), which can increase the risk for and worsen the prognosis of lymphomas. OBJECTIVE: The objective of this study was to detect B19V capsid protein and IL-6 expression in normal and malignant lymphoid tissue, as well as any correlation between the two. STUDY DESIGN: IHCs for B19V capsid protein, IL-6, and B19V co-receptors P-antigen and α5ß1 integrin were performed on a tissue array containing 70 duplicated pediatric and adult lymphoma tissues and 5 duplicated benign lymph node sections. Cases were identified as normal, B-cell NHL, diffuse large B-cell NHL, Hodgkin's lymphoma, extranodal NK/T cell lymphoma, anaplastic large cell lymphoma, or mantle cell lymphoma. IL-6 and B19V capsid staining were quantified using a positive pixel count algorithm, and P-antigen and α5ß1 staining using a membrane quantification algorithm. RESULTS: B19V capsid protein was detected in both benign and malignant lymphoid tissue. The Spearman rank correlation coefficient analysis was performed to determine the relationship between the level of positivity for B19V and IL-6 staining, yielding an overall correlation coefficient of 0.679 (p-value<0.0001). CONCLUSIONS: Our results show a moderate correlation between the levels of positive B19V and IL-6 staining by IHC, indicating a possible role for B19V in the pathogenesis of lymphomas.

Chlamydia induces anchorage independence in 3T3 cells and detrimental cytological defects in an infection model.

Chlamydia are gram negative, obligate intracellular bacterial organisms with different species causing a multitude of infections in both humans and animals. Chlamydia trachomatis is the causative agent of the sexually transmitted infection (STI) Chlamydia, the most commonly acquired bacterial STI in the United States. Chlamydial infections have also been epidemiologically linked to cervical cancer in women co-infected with the human papillomavirus (HPV). We have previously shown chlamydial infection results in centrosome amplification and multipolar spindle formation leading to chromosomal instability. Many studies indicate that centrosome abnormalities, spindle defects, and chromosome segregation errors can lead to cell transformation. We hypothesize that the presence of these defects within infected dividing cells identifies a possible mechanism for Chlamydia as a cofactor in cervical cancer formation. Here we demonstrate that infection with Chlamydia trachomatis is able to transform 3T3 cells in soft agar resulting in anchorage independence and increased colony formation. Additionally, we show for the first time Chlamydia infects actively replicating cells in vivo. Infection of mice with Chlamydia results in significantly increased cell proliferation within the cervix, and in evidence of cervical dysplasia. Confocal examination of these infected tissues also revealed elements of chlamydial induced chromosome instability. These results contribute to a growing body of data implicating a role for Chlamydia in cervical cancer development and suggest a possible molecular mechanism for this effect.

Detection of parvovirus B19 capsid proteins in testicular tissues.

OBJECTIVE: To detect B19 capsid proteins, VP1 and VP2, in testicular tissues, both normal and tumor, using immunohistochemistry. METHODS: Samples of normal, fetal, and tumor testicular tissue (n = 31) and normal testicular DNA (n = 1) were tested for the presence of B19. Immunohistochemistry staining was used for the detection of viral capsid proteins VP1 and VP2. Polymerase chain reaction with 4 primer sets was used to test for the presence of B19 DNA in a normal testicular sample. RESULTS: B19 capsid protein VP1 and VP2 was detected by immunohistochemistry in 6 (85.7%) of 7 normal testicular samples and 17 (73.9%) of 23 tumor samples. The findings from a normal fetal testicular sample were equivocal. B19 DNA was detected in normal testicular DNA with 4 of the 4 primer sets used. CONCLUSION: In contrast to previous reports, B19 capsid proteins VP1 and VP2 have now been detected in both normal and tumor testicular tissue. The persistence of B19 in a diverse range of tissues, including the testes, requires more research into the molecular mechanisms by which B19 can enter these cells, as well as the possible etiologic roles in chronic diseases, including cancer.

Parvovirus B19 infection in Hashimoto's thyroiditis, papillary thyroid carcinoma, and anaplastic thyroid carcinoma.

BACKGROUND: The human pathogenic parvovirus B19 (B19) has recently been detected in papillary thyroid carcinoma (PTC) and Hashimoto's thyroiditis (HT) tissues at a high frequency in two studies of a Chinese cohort. We wanted to extend these data to include another cohort and expand the thyroid tumor tissue types assessed. In particular, we were interested to find whether B19 also infects anaplastic thyroid carcinoma (ATC), one of the most aggressive human cancers. METHODS: Commercially available thyroid tumor tissue arrays were used to detect B19 capsid protein by immunohistochemistry in various types of thyroid tumors and disorders. The arrays were representative of the four main types of thyroid tumors, as well as other thyroid autoimmune disorders such as HT and Graves' disease, and adenomas, goiters, lymphomas, and normal thyroid tissue. In total, at least 12 different types of thyroid conditions as well as normal tissue were represented, many with multiple subjects. RESULTS: Twenty-one of the 24 (88%) PTC tumors, 3 of the 3 ATC/undifferentiated tumors, and 3 of the 3 HT tissue samples were positive for B19 capsid protein by immunohistochemistry. The localization of the protein differed based on pathological disease type, with a nuclear to cytoplasmic shift seen from unaffected to tumor tissue. CONCLUSIONS: We extend the data available on B19 detection in the thyroid to show a high correlation of virus in another cohort of PTC and HT at the protein level. We also show, for the first time, B19 infection of much more highly aggressive ATC/undifferentiated tumors. Nuclear to cytoplasmic shift in B19 protein in cancer tissue suggests a possible link between B19 and thyroid cancer pathogenesis/progression.

Enlarged benign-appearing cervical lymph nodes by ultrasonography are associated with increased likelihood of cancer somewhere within the thyroid in patients undergoing thyroid nodule evaluation.

BACKGROUND: Benign-appearing cervical lymph nodes (CLN) are easy to assess during an ultrasonography (US) evaluation for a guided fine-needle aspiration biopsy of a suspicious thyroid nodule, but their clinical significance regarding thyroid cancer risk is not known. Non-malignant-appearing nodes may be an indicator of early malignancy in the thyroid. We hypothesize that there is an increased prediction of thyroid cancer when benign-appearing enlarged CLN (ECLN) > 1 cm in any dimension are present during an US evaluation of thyroid nodules. METHOD: A review of 269 consecutive patients' charts sent for thyroid nodule assessment that underwent thyroidectomy was conducted to compare ECLN, with the presence of thyroid cancer during an ultrasound-guided fine-needle aspiration biopsy of the thyroid nodule. Surgical excision pathology confirmed all abnormal cytology reports. RESULTS: From the final 265 charts reviewed, 213 had benign thyroid pathology and 52 had thyroid cancer. Sex, number, and size of the biggest thyroid nodule were not different between groups. Patients with cancer were on average 10 years younger and had higher thyroid-stimulating hormone (TSH) values (p < 0.003) as well as a 10-fold increase in enlarged non-malignant-appearing lymph nodes than their peers without cancer. The presence of ECLN had an 82% sensitivity, 90% specificity, and a 68% positive predictive value for thyroid cancer. There was also an 80% negative predictive value when enlarged lymph nodes were not present. In 8 of the 37 (21.6%) patients with malignancy and ECLN, the primary dominant thyroid nodule was negative on cytologic evaluation, but malignancies were confirmed on surgical specimen, in contralateral nodules on the same side as the ECLN. These nodules were mostly subcentimeric, ranging from 0.2 to 1.14 cm and were not biopsied due to their inconspicuous appearance. After multiple logistic regression analysis, enlarged lymph nodes had a 53.8 odds ratio for cancer (20.49-141.33, p < 0.01). CONCLUSION: Discovering the presence of ECLN in routine assessment of thyroid nodules is an easy and fast surveillance technique that increases the predictive value in diagnosing thyroid cancer, especially when the enlarged lymph nodes are on the same side as the thyroid nodule.

Application of immunohistochemistry to cytology.

CONTEXT: The uses of monoclonal antibodies via immunochemistry have been reported frequently within the literature using various methodologies with applications to cytology specimens. The direct application of immunochemistry to cytology may have a variety of pitfalls that the general pathologist familiar with its application to histology may be unaware of when applying it prospectively to patient specimens. OBJECTIVE: To review common pitfalls when applying immunochemistry to cytology specimens and to suggest approaches to the more common differential dilemmas that apply to a variety of cytology specimens that could be seen in a general pathology practice. DATA SOURCES: The authors' own experiences of applying immunochemistry to cytopathology specimens within an academic setting along with supportive data from the literature. CONCLUSIONS: Immunochemistry can be used to increase the predictability of a cytology diagnosis if care is taken with the cytology sample preparation methodology and there is judicious use of select monoclonal antibody panels to support a specific cytology diagnosis. Up-to-date evidence-based antibody databases should be used when selecting antibody panels.

Fine-needle aspiration in PreservCyt: a novel and reproducible method for possible ancillary proteomic pattern expression of breast neoplasms by SELDI-TOF.

Proteomic profiles of tumor protein expression by the surface enhanced laser desorption-ionization time of flight (SELDI-TOF) methodology have been shown to have a potential usefulness for protein discovery as well as screening, diagnosis, prognosis and therapeutic considerations of cancer from several organ systems. Fine-needle aspiration (FNA) specimens from tumor samples is an accepted method to diagnose the cells of interest but often can be a limited assessment due to quantity of the sample. The current use of fresh or rapidly frozen specimens for proteomic profiling can be burdensome for clinicians to collect and submit specimens. The current study tests the hypothesis that placement of FNA and other cytological material in PreservCyt may be an acceptable method of sample handling for protein profiling evaluation by this method though it may not be suitable for true protein discovery or characterization. Excised fresh breast tissue for evaluation and/or treatment of a variety of breast lesions were sampled by FNA technique and placed into PreservCyt. These samples were then homogenized under denaturing conditions and evaluated by the SELDI-TOF methodology. Most samples collected showed a satisfactory quantity of protein for analysis by the SELDI-TOF methodology. Protein patterns from a variety of benign and malignant lesions revealed reproducible patterns on triplicate testing. Benign lesions had similar protein patterns across age groups in this limited series that may have potential diagnostic significance. In conclusion, FNA of breast tissue placed in PreservCyt is a potentially acceptable method of sample handling for evaluation by the SELDI-TOF methodology for establishment of reproducible protein patterns. Preliminary results from a spectrum of breast lesions suggest these patterns may have potential for ancillary testing for diagnostic consideration of breast lesions. This collection methodology could simplify sample gathering for further testing of all types of cytological specimens by the SELDI-TOF methodology. Larger studies will be needed to assess this methodology as a diagnostic aid.