Xiaoge Zhou

Morphological, immunophenotypic and molecular characterization of mature aggressive B-cell lymphomas in Chinese pediatric patients

Abstract Differential diagnosis of Burkitt lymphoma (BL), diffuse large B-cell lymphoma (DLBCL) and B cell lymphoma, unclassifiable, with features intermediate between BL and DLBCL (intermediate BL/DLBCL) can be difficult. We studied 97 Chinese pediatric mature aggressive B-cell lymphomas including 81 BL cases, eight DLBCL cases and eight intermediate BL/DLBCL cases using immunohistochemistry, interphase fluorescence in situ hybridization (FISH) and Epstein–Barr virus (EBV) in situ hybridization. Our results showed that there were no significant differences in the expression of CD10 and BCL6 among cases of BL (91% and 86%, respectively), DLBCL (75% and 63%, respectively) and intermediate BL/DLBCL (75% and 63%, respectively) (p > 0.05). The expression of BCL2 (3% in BL, 50% in DLBCL, 50% in intermediate BL/DLBCL), expression of MUM1 (17% in BL, 63% in DLBCL, 63% in intermediate BL/DLBCL) and mean Ki-67 proliferation index (PI) (93% in BL, 83% in DLBCL, 80% in intermediate BL/DLBCL) were significantly different between BL and DLBCL and between BL and intermediate BL/DLBCL. The frequency of an extra copy of BCL6 (0% in BL, 37.5% in DLBCL, 25% in intermediate BL/DLBCL) and EBV-encoded RNA (EBER) positivity (48% in BL, 0% in DLBCL and intermediate BL/DLBCL) were also significantly different between BL and DLBCL and between BL and intermediate BL/DLBCL. The frequency of C-MYC rearrangement in BL (98%) was much higher than in DLBCL (37.5%) and intermediate BL/DLBCL (50%). Our findings suggest that diagnosis of the mature aggressive B-cell lymphomas in pediatrics should be based on the comprehensive review and integration of morphological, immunohistochemical and molecular genetic features. The expression of CD10 and BCL6 but not BCL2, a high Ki-67 PI (>90%) and a C-MYC rearrangement but not BCL2 or BCL6 rearrangement are the features of BL. MUM1 is not an exclusionary diagnostic marker for BL. As the immunophenotype and molecular genetic features of DLBCL and intermediate BL/DLBCL are similar, intermediate BL/DLBCL is more likely a subgroup of DLBCL in the pediatric population. Regardless of the expression of CD10 and BCL6, strong staining for BCL2, Ki-67 PI below 90% and the presence of extra copies of BCL6 favor a diagnosis of DLBCL or intermediate BL/DLBCL.

Ocular adnexal mucosa-associated lymphoid tissue lymphoma in Northern China: high frequency of numerical chromosomal changes and no evidence of an association with Chlamydia psittaci

Studies from different countries showed variations of genetic changes and association with Chlamydia psittaci in ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma. A total of 38 ocular adnexal MALT lymphoma cases from Northern China were studied. Genetic abnormalities were investigated in 28 cases by interphase FISH. C. psittaci and other infectious agents that are commonly-associated with chronic eye disease were screened in 38 cases by PCR. Genetic abnormalities were detected in 60.7% of cases. Among them, only one showed a break-apart of the IgH gene and all others showed numerical abnormalities, including trisomy 18 in 7 cases (25%), 3 copies of BCL6 gene in 12 cases (43%), and 3 copies of C-MYC gene in 2 cases (7%). C. pneumoniae was positive in two cases (5.3%), and C. psittaci, C. trachomatis, HSV1, HSV2, ADV8, and ADV19 were not detected in any cases. In conclusion, numerical abnormalities are frequent and the chromosomal translocations commonly associated with MALT lymphomas are rare in ocular adnexal MALT lymphoma of Northern China. C. psittaci and other infectious agents are not associated with ocular adnexal MALT lymphoma in these patients.

Extra copies of ALK gene locus is a recurrent genetic aberration and favorable prognostic factor in both ALK-positive and ALK-negative anaplastic large cell lymphomas

Systemic anaplastic large cell lymphoma (ALCL) is subtyped into ALK-positive ALCL and ALK-negative ALCL based on the presence or absence of ALK protein expression. ALK-positive ALCL is characterized by t(2;5)(p23;q35)/NPM-ALK or variant ALK-involved translocations, while little is known about the genetic changes in ALK-negative ALCL. We investigated the structural and numerical aberrations of the ALK gene using interphase fluorescence in situ hybridization (FISH) in 81 cases with ALCL and analyzed their association with clinical outcome of the patients. ALK gene rearrangement was found in 47 of 50 (94%) ALK-positive ALCLs but in none of 31 ALK-negative ALCLs. Extra copies of the ALK gene locus, representing mainly extra copies of chromosome 2, were seen in 19 ALK-positive (38%) and 15 ALK-negative (48%) cases (P=0.357). In 55 cases with follow-up information, the mean survival time of the 38 ALK positive cases (58 months) was significantly longer than that of 17 ALK-negative cases (22.5 months) (P=0.038). Interestingly, the cases with extra copies of ALK had a significantly longer mean survival time than those without (64.4 months vs 35.3 months) (P=0.023) and this difference was observed in both ALK-positive (72.3 vs 45.9 months) and ALK-negative (34.7 vs 9.9 months) cases. Multivariate analysis showed that both ALK protein expression and extra copies of ALK gene were independent predictors for better survival (P=0.008). Our results suggest that the extra copies of ALK gene locus are a frequent genetic aberration in both ALK-positive and ALK-negative ALCL and is a favorable prognostic marker for the patients.

The expression of podoplanin protein is a diagnostic marker to distinguish the early infiltration of esophageal squamous cell carcinoma

The esophageal squamous cell carcinoma (ESCC) is usually develped from low-grade intraepithelial neoplasia (LGIEN) and high-grade intraepithelial neoplasia (HGIEN) to infiltrative squamous cell carcinoma. Till now, it remains hard to screen for infiltration at earlier stages, especially the differentiation between HGEIN and early infiltrative carcinoma. The purpose of this study is to determine a role of podoplanin in differentiating between HGEIN and early infiltrative squamous cell carcinoma. Totally 133 patients pathologically diagnosed with early ESCC and/or precancerous lesions were enrolled.The EnVision two-step IHC staining technique was applied using the monoclonal mouse anti-human Podoplanin antibody (clone number: D2-40). The expressions of PDPN protein on the basal layer of squamous epithelium lesions could be divided into three different patterns: complete type, incomplete (non-continuous) type, or missing type. A diagnosis of HGEIN can be made if the basal layer showed non-continuous or complete expression of PDPN and a diagnosis of early infiltration can be made if the expression of PDPN is completely missing. Our study confirmed that PDPN was a potential biomarker to identify the presence of early infiltrative squamous cell carcinoma.

Hydroa vacciniforme present for 48 years with cytotoxic CD4+ T-cell infiltration and Epstein-Barr virus infection.

patients were included (male to female ratio 66 : 42; mean age 51Æ22 years, range 14–81) and 152 courses of biologic therapies were administered (adalimumab 15, efalizumab 55, etanercept 47, infliximab 35) from May 2003 to May 2011. Seventy-five patients received one biologic therapy and 33 received more than one biologic therapy, although not concomitantly (22 patients received two and 11 patients received three biologics, respectively). Patient characteristics, severity measures, mean duration of therapies, adherence to therapy at 6 and 24 months and reasons for withdrawal are presented in Table 1. Kaplan–Meier estimates of the cumulative probability of withdrawal from treatment were obtained. A multivariate Cox proportional hazard model was used to assess the independent risk estimates of withdrawal in each treatment group, adjusted for sex, presence of psoriatic arthritis (PsA), concomitant methotrexate treatment and previous antitumour necrosis factor (TNF)-a treatment. No statistically significant differences in age, sex and associated comorbidities were present between treatment groups. The independent predictors of drug survival were the biologic agent and the presence of PsA (Fig. 1a, b). Treatment with infliximab was significantly associated with a shorter adherence to therapy when compared with adalimumab [hazard ratio (HR) 6Æ58, 95% CI 1Æ78–24Æ4], with etanercept (HR 2Æ07, 95% CI 1Æ03–4Æ17) and with efalizumab (HR 2Æ97, 95% CI 1Æ40–6Æ29). Patients concomitantly affected by PsA treated with anti-TNF-a agents (Fig. 1b) had a longer adherence to therapy when compared with patients affected only by psoriasis vulgaris (P = 0Æ047). Regarding patients previously treated with anti-TNF-a agents, we found that etanercept-treated patients had a lower adherence to therapy mainly related to lack of efficacy (46Æ2%, P < 0Æ001). Comparison of drug survival rates between anti-TNF-a-naive patients and those previously treated with anti-TNF-a agents are shown in Figure 1c and d. Other tested parameters, including sex and concomitant methotrexate, did not have any statistically significant association with drug survival. In our experience, adalimumab-, etanerceptand efalizumab-treated patients appear to have a better long-term adherence to therapy than patients treated with infliximab (Fig. 1a and Table 1). Short-term adherence to therapy is directly related to efficacy and safety, but other factors determine the long-term drug survival. In contrast to Gniadecki et al. we found that infliximab had the worst patient retention, with < 10% of patients still on the drug after 4 years. The reason for the discrepancy with our results may be due to several factors regarding our cohort of patients, such as poor compliance (related to the route of administration), a less favourable safety profile, the loss of response (17%), the lower frequency of PsA (17Æ1% vs. 48Æ8% reported by Gniadecki et al.) and the infliximab schedules followed [between 2003 and 2004, 12 patients followed an interrupted schedule of treatment, were treated with five infusions without concomitant methotrexate and were retreated after relapse (mean 5 months) with a lower efficacy and a higher rate of SAEs]. It is worth noting that no loss of response during infliximab treatment and a lower frequency of immunogenicity were seen in patients treated concomitantly with methotrexate, but the clinical response was shortened in three cases. After analysing the data presented by Gniadecki et al., we believe that the continuous schedule of infliximab therapy in association with low doses of methotrexate can improve the drug survival of the most efficacious available anti-TNF-a agent for psoriasis vulgaris.

Infectious mononucleosis-like lesions, a rare manifestation of angioimmunoblastic T-cell lymphoma

Angioimmunoblastic T-cell lymphoma (AILT) is a peripheral T-cell lymphoma characterized by systemic disease, a polymorphous infiltrate involving lymph nodes, and extensive proliferation of high endothelial venules and follicular dendritic cells, and is commonly associated with Epstein–Barr virus (EBV) [1]. Early lesions of AILT usually contain hyperplastic follicles with ill-defined borders and characteristic clear cells in the perifollicular areas [1]. We report a successive biopsy case in which an infectious mononucleosis-like lesion with EBV infection was followed by a typical AILT lesion. A 69-year-old man had had lower-extremity edema for 5 years without a significant clinical cause. Twenty days before admission for lymph node biopsy (in late January 2009), the patient felt that the edema had suddenly become more severe. The lymph nodes in the inguinal area were enlarged and were approximately 4 cm in diameter. One week later, he had a fever with a temperature of 388C. Routine blood tests showed a white blood cell count of 7.616 10/L, red blood cell count of 3.16 10/L, and platelet count of 3406 10/L. A differential count revealed 11.6% lymphocytes, 12.0% monocytes, and 71.2% neutrophils. Levels of b2-microglobulin, lactate dehydrogenase (LDH), C-reactive protein, and erythrocyte sedimentation rate (ESR) were normal. A computed tomography (CT) scan revealed generalized lymphadenopathy. Therefore, one lymph node in the left neck was removed for biopsy. Histologically, the lymph node showed complete absence of a normal architecture. The lesion was diffuse with extensive polymorphous infiltrate, including mature small lymphocytes, mediumto large-sized lymphoid cells, plasma cells, histiocytes, and follicular dendritic cells, in addition to scattered tingible body macrophages. The mediumto large-sized lymphoid cells were sparse but clustered. The cells had abundant cytoplasm with an indistinct cell margin, round to ovoid-shaped nuclei with clumped and dispersed chromatin, prominent single centralized nucleoli, or multiple small nucleoli [Figure 1(A)]. The cells in the lesion showed a B-cell differentiation with activated cells, immunoblasts, plasma blasts, plasma-like cells, and plasma cells. The mitotic figures were easy to visualize. Immunohistochemical staining showed that the small to medium lymphocytes scattered in the background were positive for CD3. A subpopulation of mediumto large-sized lymphoid cells was positive for CD20, but with varying intensity. Several cells were positive for CD30. There was a tendency for immunoglobulin light chain restriction with lambda and kappa staining. Moreover, the cells in the lesion had a high proliferation index (Ki-67 labeling index480%). EBV-encoded small RNA (EBER) was detected in the nuclei of many lymphoid cells of varying sizes using in situ hybridization [Figure 1(B)]. Based on these findings, the pathological diagnosis was infectious mononucleosis-like prominent B-cell atypical lymphoproliferation with EBV infection, and with recommendations for regular follow-up. After 2 months of follow-up, pleural effusion, ascites, and splenomegaly were detected with spiral CT scanning. Increased glucose metabolism was revealed in the bilateral neck, mediastinum, and

The degree of overlap between the follicular dendritic cell meshwork and tumor cells in mantle cell lymphoma is associated with prognosis

This study concerning mantle cell lymphoma (MCL) investigated retrospectively an association between patient prognosis and the percentage of the total number of lymphoma cells found in the follicular dendritic cell (FDC) meshwork, that is, the degree of overlap of lymphoma cells. Two hundred and nine MCL patients were apportioned to grades I-III, in which the CD21-positive FDC meshwork covered ≤50%, 51%-89%, and ≥90% of the tumor area, respectively. Significant differences among the grades (all, P < 0.01) were found in the following: duration of disease (from onset of clinical manifestation to diagnosis); clinical staging; extranodal involvement (non-lymphoid organs); histological subtype; and Ki-67 proliferation index (PI). After removing the aggressive variants, the overall survival rates of grade I (n = 92) and II (n = 57) patients were similar. The overall survival rates of grade III (n = 46) patients differed from that of grade I + II patients (P < 0.01). The grades negatively correlated with the Ki-67 PI value (r = -0.234, P = 0.001). At each grade the OSR of patients with Ki-67 PI ≤30% was similar to that of patients with Ki-67 >30%. In the Ki-67 PI ≤30% group, the OSRs of the patients differed significantly among the grades. In the Ki-67 >30% group the OSRs of the grades were similar. The results of multivariate Cox regression analysis showed that the degree of overlap, age and Ki-67 PI was the independent prognostic factors of the OSRs of MCL patients. Our data suggests that MCL patients in whom there was a high degree of overlap between the FDC meshwork and tumor area have a better clinical prognosis. The degree of overlap correlates well with the Ki-67 PI, which can be used to predict the prognosis of patients.