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Primary gastrointestinal lymphoma (PGIL), an uncommon subtype of lymphoma, accounts for 1%-4% of gastrointestinal cancers. This study, therefore, aimed to investigate the current 10-year epidemiology and outcomes of PGIL.This retrospective study involved a hospital-based chart review to analyze the epidemiology, clinical features, predisposing factors, and clinical outcomes of patients diagnosed with, and treated for, PGIL. Data covering 10 years was collected of Thai patients aged ≥ 15 years who had been diagnosed as PGIL with pathological confirmation and treated at Siriraj Hospital, Thailand.A total of 175 PGIL patients were enrolled. Their median age was 60 years (range, 20-98), with a male predominance. The stomach was the most common site of gastrointestinal (GI) organ involvement by lymphoma (38.9%), followed by the small intestine (23.4%) and multiple sites of GI involvement (23.4%). Diffuse large B-cell lymphoma (DLBCL) had the highest proportion of PGIL, accounting for 61.1%. The median patient follow-up time was 13.9 months (range: 0-104.9 months). The median overall survival (OS) of PGIL patients was not reached during the 10 years, with a 5-year OS of 64.4%. The probability of having a better OS was demonstrated in patients with a good performance status who received a rituximab-containing regimen.The stomach was the most common site of lymphoma involvement in the GI tract, with DLBCL accounting for the highest proportion of those patients. The long-term survival outcome was significantly improved in patients with good performance status and rituximab exposure.

Primary gastrointestinal lymphoma (PGIL), an uncommon subtype of lymphoma, accounts for 1%–4% of gastrointestinal cancers. This study, therefore, aimed to investigate the current 10-year epidemiology and outcomes of PGIL. This retrospective study involved a hospital-based chart review to analyze the epidemiology, clinical features, predisposing factors, and clinical outcomes of patients diagnosed with, and treated for, PGIL. Data covering 10 years was collected of Thai patients aged ≥ 15 years who had been diagnosed as PGIL with pathological confirmation and treated at Siriraj Hospital, Thailand. A total of 175 PGIL patients were enrolled. Their median age was 60 years (range, 20–98), with a male predominance. The stomach was the most common site of gastrointestinal (GI) organ involvement by lymphoma (38.9%), followed by the small intestine (23.4%) and multiple sites of GI involvement (23.4%). Diffuse large B-cell lymphoma (DLBCL) had the highest proportion of PGIL, accounting for 61.1%. The median patient follow-up time was 13.9 months (range: 0–104.9 months). The median overall survival (OS) of PGIL patients was not reached during the 10 years, with a 5-year OS of 64.4%. The probability of having a better OS was demonstrated in patients with a good performance status who received a rituximab-containing regimen. The stomach was the most common site of lymphoma involvement in the GI tract, with DLBCL accounting for the highest proportion of those patients. The long-term survival outcome was significantly improved in patients with good performance status and rituximab exposure. Trial registrationNot applicable.

The distinction between subcutaneous panniculitis-like T-cell lymphoma (SPTCL) and lupus erythematosus (LE) panniculitis is remarkably challenging. Rimming by lymphocytes with an elevated Ki-67 cell proliferation index has been forwarded as a potential diagnostic finding in biopsies of SPTCL but has not been rigorously compared with biopsies from patients with LE panniculitis. Nineteen and 17 examples of SPTCL and LE panniculitis, respectively, were evaluated for periadipocytic rimming by lymphocytes expressing Ki-67, CD8, and βF1 and for attributes associated with LE, including clusters of CD123-positive cells. The identification of periadiopocytic rimming using Ki-67, CD8, and βF1 held sensitivity of 79%, 100%, and 89.5% and specificity of 100%, 52.9%, and 88.2%, respectively (P < 0.01). CD123-positive cells were in both disorders. LE-like histopathology was commonly encountered in SPTCL. In conclusion, an elevated Ki-67 cell proliferation index with rimming is useful for distinguishing SPTCL from LE panniculitis. Notably, many features of LE panniculitis can also be encountered in SPTCL.

Castleman’s disease (CD) is a rare lymphoproliferative disorder, and its prevalence in Thailand is not known. This 10-year period study investigated the prevalence of CD in Thailand, and the clinical characteristics and outcomes of Thai CD patients, with special focus on the existence and prevalence of TAFRO syndrome. TAFRO syndrome is defined as CD with thrombocytopenia, anasarca, fever, reticulin fibrosis, and organomegaly. Thirty-three CD patients diagnosed and treated at Siriraj Hospital during January 2007 to December 2016 were included. The prevalence of CD was 1.4 per 1,000,000 patients/10 years. Median age was 46 years, with slight female predominance. Six patients were assigned to the TAFRO group. A high proportion of TAFRO syndrome (18.2%) was found among Thai adult CD patients. In addition to routine TAFRO diagnostic criteria, significantly lower hemoglobin and albumin levels were observed in the TAFRO group than in the non-TAFRO group. Treatment outcomes of CD patients were complete remission (52%), stable disease (30%), and death (13%). Three-year overall survival in the non-TAFRO group and TAFRO group was 88 and 50%, respectively. While most CD patients had a good prognosis, severe cases with TAFRO syndrome had poor outcome.

Epitheliotropic intestinal T-cell lymphoma (EITL, also known as type II enteropathy-associated T-cell lymphoma) is an aggressive intestinal disease with poor prognosis and its molecular alterations have not been comprehensively characterized. We aimed to identify actionable easy-to-screen alterations that would allow better diagnostics and/or treatment of this deadly disease. By performing whole-exome sequencing of four EITL tumor-normal pairs, followed by amplicon deep sequencing of 42 tumor samples, frequent alterations of the JAK-STAT and G-protein-coupled receptor (GPCR) signaling pathways were discovered in a large portion of samples. Specifically, STAT5B was mutated in a remarkable 63% of cases, JAK3 in 35% and GNAI2 in 24%, with the majority occurring at known activating hotspots in key functional domains. Moreover, STAT5B locus carried copy-neutral loss of heterozygosity resulting in the duplication of the mutant copy, suggesting the importance of mutant STAT5B dosage for the development of EITL. Dysregulation of the JAK-STAT and GPCR pathways was also supported by gene expression profiling and further verified in patient tumor samples. In vitro overexpression of GNAI2 mutants led to the upregulation of pERK1/2, a member of MEK-ERK pathway. Notably, inhibitors of both JAK-STAT and MEK-ERK pathways effectively reduced viability of patient-derived primary EITL cells, indicating potential therapeutic strategies for this neoplasm with no effective treatment currently available.

Vascular endothelial growth factor (VEGF) is one of the key modulators of angiogenesis. The highly polymorphic promoter and 5' untranslated region of VEGF have been associated with susceptibility to and aggressiveness of several types of cancer. To examine the functional role of VEGF polymorphisms at -634 and -1498 positions, VEGF mRNA and protein in breast cancer tissues were analyzed by quantitative polymerase chain reaction and immunohistochemistry. A dual-luciferase assay was performed to determine promoter activity. The VEGF-634CC genotype demonstrated the highest VEGF mRNA expression. High VEGF mRNA expression was correlated with a tumor size of >2 cm, the presence of lymphovascular invasion and the presence of axillary nodal metastasis. The promoter containing the -1,498T/-634C haplotype exhibited the highest basal promoter activity. These findings suggest that the interaction between -1,498T and -634C polymorphisms increases VEGF expression and is involved in breast cancer aggressiveness.

Polymorphisms in the promoter and 5' untranslated region of vascular endothelial growth factor (VEGF) have been associated with VEGF levels. To investigate the role of VEGF polymorphisms in breast cancer, the VEGF ‑2578C/A, ‑1498C/T, ‑1154G/A and ‑634G/C polymorphisms were genotyped in 483 breast cancer patients and 524 healthy controls. VEGF mRNA levels in breast cancer tissue were determined using semi‑quantitative RT‑PCR. The genotypes, ‑634G/C and ‑634C/C, were associated with an increased risk for breast cancer when compared with the ‑634G/G genotype. The VEGF ‑634G/C genotype was associated with tumor size >20 mm, perineural invasion and stage II‑IV. Individuals with ‑634C/C had lower disease‑free survival. Patients with the VEGF ‑634C/C genotype exhibited the highest VEGF mRNA levels. High VEGF mRNA expression correlated with tumor size >20 mm, presence of lymphovascular invasion and axillary nodal metastasis. These observations suggested that VEGF ‑634G/C polymorphisms have a significant role in breast cancer susceptibility and aggressiveness.

We report a 51-year-old woman with cutaneous involvement by extranodal NK/T-cell lymphoma (TCL) of the colon that microscopically mimicked mycosis fungoides (MF). She had a history of fever of unknown origin for 2 months and then developed multiple erythematous papules on her trunk and extremities. A skin biopsy revealed superficial infiltration by atypical small to medium-sized lymphocytes with epidermotropism and Pautrier collections. Immunohistochemical studies showed expression of CD3 and TIA-1 with lack of expression (double negative) of CD4 and CD8. Initially, we reported the diagnosis as MF, cytotoxic variant. Thereafter, computerized tomography scan incidentally identified a colonic mass. A colonic biopsy revealed infiltration of atypical lymphoid cells with the same morphology and immunophenotype as those found in the skin. Additionally, CD56 and Epstein-Barr virus-encoded RNA in situ hybridization in both skin and colonic biopsies were diffusely positive. Thus, extranodal NK/TCL was diagnosed. Delta T-cell receptor (TCR) gene rearrangement was documented in the skin biopsy by polyacrylamide gel electrophoresis and fluorescence capillary gel electrophoresis methods. There was no TCR gene rearrangement detected in the colonic biopsy. Unfortunately, the patient died within 2 months of diagnosis. Sitthinamsuwan P, Pongpruttipan T, Bunyaratavej S, Karoopongse E, Kummalue T, Sukpanichnant S. Cutaneous involvement by colonic extranodal NK/T-cell lymphoma mimicking mycosis fungoides: a case report.

B-cell lymphomas generally do not express T-cell intracellular antigen-1 (TIA-1), a cytotoxic granule-associated protein. We report a case of EBV-positive diffuse large B-cell lymphoma (DLBCL) of the elderly with aberrant expression of TIA-1. The patient was an 84-year-old man with a history of progressively enlarged left parotid mass and mild facial paresis for 2 months. The incisional biopsy revealed sheets of medium to large lymphoma cells with focal angiocentricity and extensive tumor necrosis; only a few scattered small lymphocytes were noted among the lymphoma cells. Immunohistochemistry in formalin-fixed paraffin-embedded tissue demonstrated that the lymphoma cells marked with CD20, CD30, CD45 (focal), CD79a, BCL2 (focal and weak), PAX5, Oct2, BOB.1, IRF4/MUM1, EBV-LMP1, and TIA-1, but they did not mark with CD3, CD4, CD5, CD7, CD8, CD10, CD15, CD23, CD56, CD57, CD138, kappa and lambda immunoglobulin light chains, BCL6, βF1, granzyme B, or perforin. The aberrant TIA-1 expression was then confirmed by two different autostainers. EBV-encoded RNA (EBER) in situ hybridization was positive in the lymphoma cells. Gel electrophoresis for immunoglobulin heavy chain (IgH) and T-cell receptor (TCR) gene rearrangements revealed no definite clonal band, but fluorescence capillary electrophoresis demonstrated positive results for both IgH and TCR genes. Finally, we concluded this case as an EBV-positive DLBCL of the elderly with dual genotype (rearrangements of both IgH and TCR genes) and aberrant TIA-1 protein expression.

Primary mediastinal neuroendocrine tumors are a rare malignancy that accounts for < 10% of all mediastinal tumors. The case presented here involves a 52-year-old man who had been suffering for 3 mo from chronic cough, anorexia and substantial weight loss, as well as 2 wk of jaundice prior to his admission. A computed tomography scan showed a 4.3 cm × 6.6 cm mediastinal mass with multiple liver nodules scattered along both hepatic lobes. Endoscopic ultrasound showed a large heterogeneous hypoechoic mass at the mediastinum with multiple target-like nodules in the liver. Fine-needle aspiration specimens revealed numerous, small, round cells with hyperchromatic nuclei, scarce cytoplasm, and frequent mitotic features. Immunohistochemical study revealed positive results for AE1/AE3, CD56 and chromogranin A, with negative findings for synaptophysin, CK20, vimentin, CK8/18 and CD45. The patient was subsequently diagnosed with a poorly differentiated neuroendocrine carcinoma, small cell type. A bone marrow biopsy also revealed extensive involvement by the carcinoma.

S100 T-cell lymphomas are infrequent, and except 1 all have been CD4 negative. On the basis of an index case of CD4 S100 T-cell prolymphocytic leukemia (T-PLL), we studied S100 protein expression in 19 additional T-PLLs and 56 other T-cell lymphomas that are usually CD4, including 15 angioimmunoblastic T-cell lymphomas, 24 anaplastic large cell lymphomas (16 ALK and 8 ALK), 7 mycosis fungoides/Sezary syndrome, and 10 peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS). Two additional S100 CD4 PTCL, NOS cases were also reviewed. Thirty percent (6/20) of T-PLLs were S100 compared with 0/56 other T-cell lymphomas with previously unstudied S100 reactivity (40 CD4, 2 CD8, 11 CD4/CD8, 3 unknown) (P=0.0007). There were no significant differences between the S100 and S100 T-PLLs with regard to the male:female ratio (2:1 vs. 1:1), age (71.6±7.7 vs. 65.4±9.3), peripheral blood lymphocyte count (67.2±116.6 vs. 101.1±159.7×10/L), or median survival (463 vs. 578 d, where known). The 2 S100 PTCL, NOS cases occurred in a 7-year-old boy and a 45-year-old woman. Both had involvement of the bone marrow and peripheral blood but were morphologically unlike T-PLL and lacked TCL1 gene rearrangement. These results demonstrate that S100 T-cell lymphomas include a subset that are CD4 and most often, but not exclusively, are T-PLL. Although having diagnostic implications, there were no documented clinical differences between the S100 and S100 T-PLLs.

Breast cancer is the most frequent type of cancer among women worldwide. Vascular endothelial growth factor (VEGF), the key modulator of angiogenesis, has been implicated in breast cancer susceptibility and aggressiveness. VEGF expression was determined in 99 breast cancer tissue samples using reverse transcription-polymerase chain reaction and the human epidermal growth factor receptor 2 (HER2) status was determined by immunohistochemistry. Subsequently, the associations of VEGF, HER2 and hormone receptor status with clinicopathological data were evaluated. High VEGF expression was found to be significantly correlated with the presence of lymphovascular invasion. In hormone receptor-positive/HER2-positive, HER2-positive and triple-negative breast cancer, high VEGF expression was correlated with the presence of axillary nodal metastasis and lower overall survival rates. Therefore, the assessment of the VEGF status along with the hormone receptor and HER2 status may help identify high-risk patients who may benefit from anti-VEGF treatment.

Background Lymphomas are common malignancies that have various subtypes with many overlapping histologic, immunophenotypic and genetic features. Therefore, discordance in classifying lymphoma among pathologists may be encountered. But this issue is not well characterized. We conducted the present study to demonstrate discordances among Thai hematopathologists as well as to highlight common arguing points for classifying lymphomas. Methods The 117 lymphoma cases were randomly retrieved and individually reviewed by 7 hematopathologists, members of the “Thai Hematopathologist Group,” without knowing the original diagnoses. The consensus diagnoses were given from a discussion by all members. In each case, the diagnosis from each participant was compared with the consensus diagnosis and classified into 4 categories as follow: 1) concordance, 2) minor discordance, 3) major discordance and 4) serious discordance. Results There were approximately 11% discordances between original and consensus diagnoses. The average discordances among all pathologists according to minor, major and serious discordances were 10%, 3.5% and 0.3%, respectively. Diffuse large B-cell lymphoma had the least discordance (7%). Small biopsies had been found to increase discordances in some lymphoma subtypes. Conclusions The present study reveals some degrees of interobserver variation in classifying of lymphoma by using the 2008 WHO classification among hematopathologists. Some types of lymphomas on small biopsies were found to have a significant higher discordance rate. This study also described some common diagnostic discordances regarded as potential pitfalls in classifying lymphomas. Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_162

Background: To correlate breast cancer subtypes with prognostic factors, microvessel density (MVD), vascular endothelial growth factor (VEGF) expression and clinical features. Materials and Methods: One hundred cases of primary breast carcinoma were classified using biomarkers on tissue microarray as: luminal A [estrogen receptor (ER)+, HER2-, Ki-67≤14%], luminal B [ER+, HER2+ or ER+, HER2-, Ki-67>14%], HER2, triple negative basal-like (TNB) [any basal cytokeratins (CKs, 5, 14, 17) and/or endothelial growth factor receptor (EGFR) expression], and TN without such markers [TNN, null], and assessed for p53, vimentin, VEGF and CD31 immunoperoxidase. Results: Of the 100 cases (mean age, 51 years; mean tumor size, 3.2cm; 56% with nodal metastasis; 89 invasive ductal carcinomas, not otherwise specified, 4 invasive lobular carcinomas, 3 metaplastic carcinomas, and 4 other types) there were 39 luminal A, 18 luminal B, 18 HER2, 15 TNB and 10 TNN. The positivities of basal-like markers in the basal-like subtype were 78.3% for CK5, 40% for CK14, 20% for CK17, 46.7% for EGFR. There was no significant difference in age distribution, tumor size, degree of tubular formation, pleomorphism, lymphovascular invasion, nodal metastasis, MVD, VEGF expression and survival among subgroups. TNs demonstrated significantly higher tumor grade, mitotic count, Ki-67 index, p53 and vimentin and decreased overall survival compared with nonTN. Conclusions: The distribution of breast cancer subtypes in this study was similar to other Asian countries with a high prevalence of TN. The high grade character of TN was confirmed and CK5 expression was found to be common in our basal-like subtype. No significant elevation of MVD or VEGF expression was apparent.

Extranodal NK/T-cell lymphoma (ENKTL), nasal type, may be of NK or T-cell origin; however, the proportion of T-ENKTLs and whether they are of αβ or γδ type remains uncertain. To elucidate the cell of origin and detailed phenotype of ENKTL and assess any clinicopathologic associations, 67 cases of ENKTL from Thailand were investigated, together with 5 γδ enteropathy-associated T-cell lymphomas (EATLs) for comparison. In all, 70% of the ENKTL were T-cell receptor (TCR) β,γ and, in cases tested, δ negative (presumptive NK origin); 5% were TCR γδ, 3% were TCR αβ, 1% were TCR αβ/γδ, and 21% were indeterminate. Out of 17 presumptive NK-ENKTLs tested, 3 had clonal TCR rearrangements. All cases were EBV and TIA-1; >85% were positive for CD3, CD2, granzyme B, pSTAT3, and Lsk/MATK; and all were CD16. Presumptive NK-ENKTLs had significantly more frequent CD56 (83% vs. 33%) and CXCL13 (59% vs. 0%) but less frequent PD-1 (0% vs. 40%) compared with T-ENKTLs. Of the NK-ENKTLs, 38% were Oct-2 compared with 0% of T-ENKTLs, and 54% were IRF4/MUM1 compared with 20% of T-ENKTLs. Only αβ T-ENKTLs were CD5. Intestinal ENKTLs were EBV and had significantly more frequent CD30, pSTAT3, and IRF4/MUM1 expression but less frequent CD16 compared with γδ EATL. Significant adverse prognostic indicators included a primary non-upper aerodigestive tract site, high stage, bone marrow involvement, International Prognostic Index ≥2, lack of radiotherapy, Ki67 >40%, and CD25 expression. The upper aerodigestive tract ENKTLs of T-cell origin compared with those of presumptive NK origin showed a trend for better survival. Thus, at least 11% of evaluable ENKTLs are of T-cell origin. Although T-ENKTLs have phenotypic and some possible clinical differences, they share many similarities with ENKTLs that lack TCR expression and are distinct from intestinal γδ EATL.

Background: Pathological evaluation of sentinel lymph nodes of breast cancer patients is a critical factor in determining the treatment strategy including patients’ outcome. Multiple factors have been known to correlate with disease free survival and clinical outcomes such as sentinel lymph node metastases. Recently, real-time RT-PCR which is a very useful, rapid, high sensitivity and specificity tool has been reported to detect micrometastases in sentinel lymph nodes which could be helpful to identify high risk patients. Objectives: The purpose of this study are: To evaluate the sensitivity, specificity, and accuracy of CK19 detection by multiplex real-time RT-PCR and to elucidate the correlation of multiplex real-time RT-PCR results and clinicopathological parameters in sentinel lymph nodes of Thai breast cancer patients. Material and methods: 119 patients diagnosed as breast cancer stage I or II at Siriraj Hospital during October 2008 – June 2010 were enrolled in this study. RNAs were extracted from their sentinel lymph nodes and cDNAs were synthesized. Multiplex real-time RT-PCR for detection of CK19 was performed and analysed. Patients’ characteristics and pathological parameters were compared with multiplex real-time RT-PCR results. Results: Significant correlation between angiolymphatic and perineural invasion status and CK19 expression was demonstrated. The overall concordance of CK19 gene with pathological results was 72.3%. Sensitivity and specificity for CK19 detected by multiplex real-time RT-PCR were 78.3%, and 66.1%, respectively. Conclusion: Significant correlation between angiolymphatic and perineural invasion status and CK19 expression in sentinel lymph nodes of breast cancer patients was found in this study.

Background Extranodal NK/T-cell lymphoma, nasal type (ENKTL) is not common worldwide, but it is the most common T- and NK-cell lymphomas in many Asian countries. Immunophenotypic profiles were studied based on limited series. The authors, therefore, studied on ENKTL according to characterize immunophenotypic profiles as well as the distribution of EBV subtype and LMP-1 gene deletion. Methods By using tissue microarray (TMA), immunohistochemical study and EBV encoded RNA (EBER) in situ hybridization were performed. T-cell receptor (TCR) gene rearrangement, EBV subtyping, and LMP-1 gene deletion were studied on the available cases. Results There were 22 cases eligible for TMA. ENKTL were positive for CD3 (91%), CD5 (9%), CD7 (32%), CD4 (14%), CD56 (82%), TIA-1 (100%), granzyme B (95%), perforin (86%), CD45 (83%), CD30 (75%), Oct2 (25%), and IRF4/MUM1 (33%). None of them was positive for βF1, CD8, or CD57. TCR gene rearrangement was negative in all 18 tested cases. EBV was subtype A in all 15 tested cases, with 87% deleted LMP-1 gene. Cases lacking perforin expression demonstrated a significantly poorer survival outcome (p = 0.008). Conclusions The present study demonstrated TIA-1 and EBER as the two most sensitive markers. There were a few CD3 and/or CD56 negative cases noted. Interestingly, losses of CD45 and/or CD7 were not uncommon while Oct2 and IRF4/MUM1 could be positive in a subset of cases. Based on the present study in conjunction with the literature review, determination of PCR-based TCR gene rearrangement analysis might not be a useful technique for making diagnosis of ENKTL.

To identify the distribution of mature T-cell and NK-cell lymphomas in Siriraj Hospital and compare to the other world regions, using the latest WHO classification (2008).Newly diagnosed cases of such lymphomas between January 1, 2004 and December 31, 2006 at Siriraj Hospital were recruited and reviewed. Additional immunohistochemical studies and/or EBV-encoded RNA (EBER) in situ hybridization were performed from the formalin-fixed paraffin-embedded tissue. Then, lymphomas were reclassified according to the WHO classification (2008).Seventy-one cases including extranodal NK/T-cell lymphoma, nasal type (ENKTL, 31.0%), anaplastic large cell lymphoma (18.3%), angioimmunoblastic T-cell lymphoma (14.1%), peripheral T-cell lymphoma, not otherwise specified (12.7%), mycosis fungoides (MF, 8.5%), subcutaneous panniculitis-like T-cell lymphoma (SPTCL, 7.0%), primary cutaneous anaplastic large cell lymphoma (PCAL, 5.6%), primary cutaneous gamma-delta T-cell lymphoma (PCGDTL, 1.4%), and enteropathy-associated T-cell lymphoma (1.4%) were included in this study. In terms of changing version of the WHO classification from 2001 to 2008, only one case had the diagnosis changed from MF to PCGDTL, a newly proposed entity in the 2008 version.ENKTL was the most common in the present series and it had a significantly higher frequency than those reported in other previous studies. The frequency was relatively higher in SPTCL, PCAL, and MF when compared to the other series. Furthermore, changing the WHO classification from the 2001 version to the recently published2008 version may not affect the proportion of NK/T-cell lymphoma.

Cutaneous extranodal NK/T-cell lymphoma, nasal type (NK/T) is relatively rare, associated with aggressive behavior and poor prognosis. Histopathological findings, immunohistochemical study and EBV-encoded RNA (EBER) in situ hybridization are essential for the diagnosis.A 54-year-old Thai man with NK/T of the nasal cavity initially presented with cutaneous NK/T mimicking granulomatous panniculitis. The skin biopsies were performed twice due to the marked necrosis in the first one. The second biopsy revealed small, medium, and large atypical lymphoid cells infiltrating fat lobules with necrotic foci and granulomatous reaction. Within the granulomatous inflammation, the atypical lymphoid cells showing involvement of the blood vessel (angiocentricity) were noted. Immunostaining demonstrated that the atypical lymphoid cells marked with CD3, CD56, and TIA-1, but they did not mark with CD5, CD20, CD15, or CD30. EBER in situ hybridization was positive.Cutaneous NK/T can produce granulomatous panniculitis. The recognition of atypical lymphoid cells showing angiocentricity together with immunohistochemistry and EBER in situ hybridization are crucial for the correct diagnosis.

Malignant lymphoma, especially non-Hodgkin lymphoma, is one of the most common hematologic malignancies in Thailand. The diagnosis of malignant lymphoma is often problematic, especially in early stages of the disease. Detection of antigen receptor gene rearrangement including T cell receptor (TCR) and immunoglobulin heavy chain (IgH) by polymerase chain reaction followed by heteroduplex has currently become standard whereas fluorescent fragment analysis (GeneScan) has been used for confirmation test. In this study, three techniques had been compared: thermocycler polymerase chain reaction (PCR) followed by heteroduplex and polyacrylamide gel electrophoresis, GeneScan analysis, and real time PCR with High Resolution Melting curve analysis (HRM). The comparison was carried out with DNA extracted from paraffin embedded tissues diagnosed as B- cell non-Hodgkin lymphoma. Specific PCR primers sequences for IgH gene variable region 3, including fluorescence labeled IgH primers were used and results were compared with HRM. In conclusion, the detection IgH gene rearrangement by HRM in the LightCycler System showed potential for distinguishing monoclonality from polyclonality in B-cell non-Hodgkin lymphoma. Introduction Malignant lymphoma, especially non-Hodgkin lymphoma, is one of the most common hematologic malignancies in Thailand. The incidence rate as reported by Ministry of Public Health is 3.1 per 100,000 population in female whereas the rate in male is 4.5 per 100,000 population 1. At Siriraj Hospital, the new cases diagnosed as malignant lymphoma were 214.6 cases/year 2. The diagnosis of malignant lymphoma is often problematic, especially in early stages of the disease. Therefore, detection of antigen receptor gene rearrangement including T cell receptor (TCR) and immunoglobulin heavy chain (IgH) by polymerase chain reaction (PCR) assay has recently become a standard laboratory test for discrimination of reactive from malignant clonal lymphoproliferation 34. Analyzing DNA extracted from formalin-fixed, paraffin-embedded tissues by multiplex PCR techniques is more rapid, accurate and highly sensitive. Measuring the size of the amplicon from PCR analysis could be used to diagnose malignant lymphoma with monoclonal pattern showing specific and distinct bands detected on acrylamide gel electrophoresis. However, this technique has some limitations and some patients might require a further confirmation test such as GeneScan or fragment analysis 56. GeneScan technique or fragment analysis reflects size and peak of DNA by using capillary gel electrophoresis. This technique is highly sensitive and can detect 0.5-1% of clonal lymphoid cells. It measures the amplicons by using various fluorescently labeled primers at forward or reverse sides and a specific size standard. Using a Genetic Analyzer machine and GeneMapper software (Applied Bioscience, USA), the monoclonal pattern revealed one single, sharp and high peak at the specific size corresponding to acrylamide gel pattern, whereas the polyclonal pattern showed multiple and small peak condensed at the same size standard. This technique is the most sensitive and accurate technique; however, it usually requires high technical experience and is also of high cost 7. Therefore, rapid and more cost effective technique are being sought. LightCycler PCR performs the diagnostic detection of amplicon via melting curve analysis within 2 hours with the use of a specific dye 89. This dye consists of two types: one known as SYBR-Green I which is non specific and the other named as High Resolution Melting analysis (HRM) which is highly sensitive, more accurate and stable. Several reports demonstrated that this new instrument combined with DNA intercalating dyes can be used to discriminate sequence changes in PCR amplicon without manual handling of PCR product 1011. Therefore, current investigations using melting curve analysis are being developed 1213. In this study, three different techniques were compared to evaluate the suitability of LightCycler PCR with HRM as the clonal diagnostic tool for IgH gene rearrangement in B-cell non-Hogdkin lymphoma, i.e. thermocycler PCR followed by heteroduplex analysis and PAGE, GeneScan analysis and LightCycler PCR with HRM.

Although the WHO classification (2001) requires a great deal of morphologic, immunophenotypic, genetic, and clinical features for classifying lymphomas, it is still feasible to misdiagnose under limited resources, especially a limited panel of antibodies used for immunophenotyping. To identify pitfalls in classifying lymphomas among hematopathologist, general pathologists, and pathology residents under this situation.Newly diagnosed lymphoma cases from 1 July 2002 to 30 June 2003 at Siriraj Hospital were included for two rounds of individually blinded review by a hematopathologist, two general pathologists, and three pathology residents. Final diagnoses were given by consensus. Pitfalls were determined from misdiagnosis, in each case analyzed in terms of frequency.One hundred and four lymphoma cases included 61 diffuse large B-cell lymphoma (DLBCL, 58.6%), 12 MALT lymphoma (11.5%), eight follicular lymphoma (FL, 7.7%), seven classical Hodgkin lymphoma (HL, 6.7%), four unspecified peripheral T-cell lymphoma (PTCL, 3.8%), three Burkitt lymphoma (BL, 2.9%), two subcutaneous panniculitis-like T-cell lymphoma (SPTCL, 1.9%), and seven other uncommon types (1% each). Pitfalls were low infrequency on diagnosis of DLBCL, nodular sclerosis HL, and SPTCL (8% each), but not different among the participants only in DLBCL. Pitfalls in diagnosis of MALT lymphoma, mixed cellularity HL, BL, unspecified PTCL, and FL were 60%, 50%, 33%, 29%, and 24%, respectively. However, considering hematopathologist and non-hematopathologist groups, pitfalls in the former were lower, especially in the uncommon types of lymphoma.Pitfalls in classifying lymphomas are common. Interest in hematopathology reduces misdiagnosis in lymphomas other than DLBCL.

Epstein-Barr virus-induced 3 (EBI3) encoded by the EBI3 gene was identified by the induction of its expression in B lymphocytes by EBV infection. It encodes a secreted glycoprotein belonging to the hematopoietin receptor family, and heterodimerizes with a 28-kDa protein to form interleukin 27 (IL-27). Given this function, we hypothesized that EBI3 could serve as …

Patients with immunodeficiency disorders have an increased incidence of lymphoproliferative disorders; however, only 4 such patients with DiGeorge/chromosome 22q11.2 deletion syndrome have been reported. We report a case of a pulmonary Epstein-Barr virus-negative extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in a child with this syndrome.

Thirteen Thai patients, seven with pathologic diagnosis of T-cell lymphoma, four with clinical and/or pathological suspicion of T-cell lymphoma, one with lymphadenitis and one with chronic hypertrophic tonsillitis, were investigated for T-cell receptor gene rearrangement (TCR-GR) by polymerase chain reaction (PCR). Clonal TCR-GR was demonstrated in each T-cell lymphoma whereas polyclonal TCR-GR was found in the rest of the patients, including the four cases that were suspected of T-cell lymphoma. Aim: To identify T-cell receptor gene rearrangement in malignant lymphoma in Thai patients. Material and method: Thirteen specimens of formalin-fixed, paraffin-embedded tissue were collected from Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University. After conventional morphological evaluation and immunohistochemical studies, DNA in each specimen was extracted from 10μm thick paraffin sections using QIAamp DNA mini kit (QIAGEN) for TCR-GR analysis by multiplex PCR. Two pairs of primers were selected for Vγ11 (VγI), Vγ101(VγIII/IV), Jγ12(Jγ1/2) and Jp12(JPγ1/2). DNA of the four cases suspected of malignant lymphoma were repeated by using IdentiClone™ TCRG Gene Clonality Assay kit (InVivoScribe Technologies). Results: According to the WHO classification (2001), the seven cases of T-cell lymphoma were angioimmunoblastic type (3 cases) and unspecified type (4 cases). All cases showed monoclonal TCR-GR. Only one of the four cases suspected of T-cell lymphoma was doubtful for nasal NK/T-cell lymphoma while the others were suspected clinically. All of these four cases and the two reactive conditions revealed polyclonal TCR-GR. After using IdentiClone™ kit twice, the four cases suspected of T-cell lymphoma still showed polyclonal TCR-GR. Conclusion: The molecular analysis of TCR-GR performed in thirteen cases demonstrated the monoclonal TCR-GR in all seven cases of T-cell lymphoma and polyclonal TCR-GR in the four cases suspected of T-cell lymphoma and the two cases of reactive condition. Therefore, this technique is very useful to confirm the diagnosis of T-cell lymphoma and to provide information for further management in patients suspected of malignant lymphoma.