Infection of T but not B lymphocytes by cytomegalovirus in lymph node. An immunophenotypic study.

A 45-year-old black man presented with fever, night sweats, weight loss, and cervical lymphadenopathy. Biopsy of an enlarged node showed partial effacement of the nodal architecture by numerous cells with typical cytomegalovirus (CMV) inclusions. These were also seen by electron microscopy. CMV titers were not elevated, and he tested negative for HIV. An immunophenotypic study done on both paraffin-embedded and frozen tissue showed that the infected cells were of T-lymphocyte phenotype. These cells were remarkable for the lack of HLA-DR and IL-2R antigens, which were expressed by numerous neighboring apparently uninfected cells. The lack of HLA-DR and IL-2R expression by CMV-infected T lymphocytes may be one of the mechanism by which CMV causes immunosuppression.     

Hepatitis B virus (HBV) genotype and YMDD motif mutation profile among patients infected with HBV and untreated with lamivudine.

OBJECTIVES: A few reports exist on hepatitis B virus (HBV) genotype distribution in Iran; however the sample sizes of these studies are insufficient. The first objective of this study was to determine the HBV genotype distribution with a large sample size (147 specimens). The second objective was to determine the incidence of the lamivudine-resistant YMDD mutant profile among HBV-infected patients not treated with lamivudine; some studies have reported that YMDD mutants are detectable even before antiviral treatment. METHODS: We used two cost-effective PCR-based methods that have been developed in-house: gap-PCR and artificially created restriction site-PCR (ACRS-PCR). Also, 11 samples were randomly selected and bi-directionally sequenced and subjected to phylogenetic analysis. RESULTS: Gap-PCR results revealed genotype D of HBV in all patients, and ACRS-PCR results disclosed the absence of mutation within the YMDD motif before antiviral therapy in the study population. Phylogenetic analysis supported the former genotyping results with the segregation of all Iranian HBV isolates in the genotype D branch with a high bootstrap value (99%, 1000 replicates). CONCLUSIONS: The present study using two cost-effective methods showed that genotype D of HBV is dominant among Iranian HBV-infected subjects, and HBV lamivudine-resistant strains do not exist naturally among Iranian patients not treated with lamivudine.     

Growth inhibition of follicular small-cleaved-cell lymphoma cells in short-term culture by interleukin-3

BACKGROUND: The role of interleukin-3 (IL-3) in stimulating the growth ofearly myeloid progenitor cells is very well established. Therefore,IL-3 has been incorporated into many post-bone-marrow transplantationand intensive chemotherapy programs for the treatment of solidtumors and hematologic malignancies, including lymphomas. However,the effect of IL-3 on normal and malignant lymphocytes has notbeen well studied. PURPOSE: The purpose of this study was to evaluate the in vitro effectof IL-3 on the growth of follicular small-cleavedcell lymphoma(FSCCL). MATERIALS AND METHODS: IL-3 receptor expression on the surface of CD19⁺ cells was determinedby two-color flow cytometry measuring the receptor-binding ofbiotinylated IL-3 to CD19⁺ B-cells. Seven cases of FSCCL werecompared to six normal controls. Cell proliferation was evaluatedby [³H]thymidine incorporation into cells grown in suspensioncultures. RESULTS: All seven cases of FSCCL expressed the IL-3 receptor on thesurface of CD19⁺ cells, whereas all six cases of CD19⁺ cellsisolated from the peripheral blood of normal donors did notexpress IL-3 receptors. IL-3 had antiproliferative activityagainst FSCCL as manifested by a decrease in [³H]thymidine incorporationand a decrease in the total number of cells after 72 hours ofculture. CONCLUSION: IL-3 inhibits the growth of FSCCL cells in vitro. Clinical trialsto evaluate the in vivo effect of IL-3 in patients with FSCCLare warranted. interleukin-3, lymphoma, flow cytometry     

YMDD Motif Mutation Profile Among Patients Receiving Liver Transplant Due to Hepatitis B Virus Infection With Long Term Lamivudine/Immunoglobulin Therapy

Background:

Recurrence of Hepatitis B Virus infection in patients undergoing liver transplanted (LT) is a serious and often fatal problem. Lamivudine (LAM) and Hepatitis B Immunoglobulin (HBIG) are widely used to manage hepatitis B recurrence after liver transplantation. However, the outcomes in patients are less elucidated.

Objectives:

The current study aimed to evaluate the YMDD motif mutations profile among the patients undergoing LT infected with HBV and treated with LAM/HBIG at least for one year.

Patients and Methods:

Thirty patients with liver transplantation due to HBV were enrolled, while DNA level remained under detection limit of 50 IU/mL before transplantation and abnormal higher levels of liver enzymes after LT. The HBV genome detection was performed by two different Polymerase Chain Reaction methods following viral quantification by commercial Real-Time PCR. HbsAg detection, besides liver function tests were conducted as complementary assays. To assess nucleotide analogue mutations, the major part of polymerase gene (aa 80 - 240) was amplified by Nested-PCR, introduced to sequencing and subjected to phylogenetic analysis.

Results:

Totally, according to the laboratory criteria there were 13 cases with detectable HBV genome, while the mean liver enzyme levels were higher in recurrent patients and HBsAg was detected only in four out of the 13 cases. Phylogenetic analysis demonstrated that all isolated genomes belonged to genotype D. Critical M204I mutation, as a proof for resistance to LAM, was detected among 46% of the subjects and natural entecavir resistance (S202I) was also distinguished in one subject. Viral quantification showed higher titer in LAM resistant group in comparison to the group with undetectable drug resistance mutant (P > 0.05).

Conclusions:

Although the patients carrying M204I mutation were more likely to show lack of responses to LAM therapy, LAM replacing by other nucleoside/tide analogs plus HBIG maybe still effective in decreasing hepatitis B recurrence after liver transplantation. However, it is suggested that drug resistance test should be considered by clinicians during therapeutic management to avoid the following viral breakthrough.     

Pentostatin,cyclophosphamide and rituximab for previously untreated advanced stage,low‐grade B‐cell lymphomas

We conducted a prospective phase II trial of pentostatin, cyclophosphamide and rituximab as initial therapy for patients with previously untreated advanced stage low‐grade or indolent B‐cell lymphomas (iNHLs). Of 83 evaluable patients, 91·6% attained an overall response and 86·8% a complete or unconfirmed complete response. The 3‐year progression‐free survival (PFS) and overall survival rates were 73% and 93%, respectively. The 3‐year PFS rate was significantly different for different diagnoses (P = 0·01): 83% [95% confidence interval (CI): 0·72, 0·96] for follicular lymphomas, 73% (95% CI: 0·54, 1·0) for marginal zone lymphomas and 61% (95% CI: 0·46, 0·81) for small lymphocytic lymphomas. The most common adverse events were haematological. Of 509 cycles of chemotherapy administered, grade 3 or 4 neutropenia was reported in 68 cycles (13% of cycles administered) and most frequently occurred during cycles 4–6. This is the first report demonstrating the effectiveness of pentostatin, cyclophosphamide and rituximab in patients with previously untreated iNHLs, including those over 60 years of age.