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We report a case of POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes) syndrome in a 55-year-old African American woman in which human herpesvirus-8 (HHV-8) was demonstrated within rare lymphocytes in a Castleman disease lymph node biopsy and numerous endothelial cells and lymphocytes in a microvenular hemangioma skin biopsy. Initial symptoms and findings of night sweats, weight loss, anorexia, generalized lymphadenopathy, and hemangiomas improved after chemotherapy with cyclophosphamide and prednisone. However, in the year following the initial diagnosis, the patient suffered from recurrent bouts of night sweats, gastroparesis, and lymphadenopathy, which required further treatment with plasmapheresis, cyclophos-phamide, prednisone, and rituximab. One year later, the patient is asymptomatic but has persistent gammopathy. Although HHV-8 has previously been detected in POEMS-associated Castleman disease tissue, to our knowledge, this is the first case report in which HHV-8 has been directly demonstrated within the endothelial cells of a POEMS-associated hemangioma. [ABSTRACT FROM AUTHOR]

In order to determine the role of human herpesvirus 6 (HHV-6) in human disease, several confounding factors, including methods of detection, types of controls, and the ubiquitous nature of the virus, must be considered. This is particularly problematic in the case of cancer, in which rates of detection vary greatly among studies. To determine what part, if any, HHV-6 plays in oncogenesis, a review of the literature was performed. There is evidence that HHV-6 is present in certain types of cancer; however, detection of the virus within tumor cells is insufficient for assigning a direct role of HHV-6 in tumorigenesis. Findings supportive of a causal role for a virus in cancer include presence of the virus in a large proportion of cases, presence of the virus in most tumor cells, and virus-induced in-vitro cell transformation. HHV-6, if not directly oncogenic, may act as a contributory factor that indirectly enhances tumor cell growth, in some cases by cooperation with other viruses. Another possibility is that HHV-6 may merely be an opportunistic virus that thrives in the immunodeficient tumor microenvironment. Although many studies have been carried out, it is still premature to definitively implicate HHV-6 in several human cancers. In some instances, evidence suggests that HHV-6 may cooperate with other viruses, including EBV, HPV, and HHV-8, in the development of cancer, and HHV-6 may have a role in such conditions as nodular sclerosis Hodgkin lymphoma, gastrointestinal cancer, glial tumors, and oral cancers. However, further studies will be required to determine the exact contributions of HHV-6 to tumorigenesis.

Aims: Mediastinal gray zone lymphoma (MGZL) is a rare entity with morphologic, immunophenotypic, and genetic features intermediate between classic Hodgkin lymphoma (CHL) and primary mediastinal large B-cell lymphoma (PMBL). It is challenging to differentiate from CHL and PMBL. A specific dendritic cell gene expression profile can distinguish CHL and MGZL from PMBL. We hypothesized that the dendritic markers fascin and CD123 may be helpful in distinguishing MGZL from CHL and PMBL. We also investigated programmed death-ligand 1 (PD-L1) expression in MGZL, which may have therapeutic significance in this difficulty to treat tumor., Methods: Representative sections from 89 CHL, 20 PMBL, and 7 MGZL cases were stained for fascin, CD123, and PD-L1, and scored on a scale from 0 to 3+. Most (71%) MGZLs stained for CD123, as well as some (23%) CHLs, and few (11%) PMBLs. All MGZLs stained for fascin, as well as most (90%) CHLs, and approximately half (53%) of the PMBLs. PD-L1 was positive in all MGZLs, most (77%) CHLs and most (66%) PMBLs., Conclusions: Our study is the first to show CD123 is positive in a subset of formalin-fixed, paraffin-embedded MGZLs and CHLs, in contrast to PMBL which is largely negative. Staining for fascin was not significantly different between the lymphomas, but was less likely to be positive in PMBL. These findings suggest a role for CD123 and fascin in supporting diagnoses of MGZL and CHL, and in ruling out PMBL. By immunohistochemistry, PD-L1 is positive in MGZL, pointing to its therapeutic potential.

Aims: Lymphadenopathy, haematological abnormalities and constitutional symptoms are among the non-specific manifestations seen in drug rash with eosinophilia and systemic symptoms (DRESS), an uncommon but potentially fatal cutaneous adverse drug reaction. The ubiquitous human herpesvirus 6 (HHV-6) plays a unique role in the pathogenesis of DRESS, with emerging data suggesting that reactivation occurs in most cases and contributes to the clinical manifestations, including lymphadenopathy. Further, in the appropriate clinical context, demonstration of HHV-6 reactivation may lend support to a diagnosis of DRESS. The histopathology of DRESS-associated HHV-6 lymphadenitis is reported rarely, with morphologic and immunophenotypic characteristics concerning for T cell lymphoma. The aim is to characterize the histopathology of HHV-6 lymphadenitis in the context of DRESS and to highlight this as an important cause of lymphadenopathy that may be a clinical, morphologic and immunophenotypic mimic of lymphoma., Methods and Results: We describe a case of lymphoma-mimicking lymphadenitis in which the histopathological demonstration of reactivation of HHV-6 infection lent support to the clinical diagnosis of DRESS., Conclusion: Lymph node biopsies concerning for T cell lymphoma should be evaluated for HHV-6 involvement in a clinical context suggestive of DRESS., (© 2016 John Wiley & Sons Ltd.)

Genetic analysis of classical Hodgkin lymphoma (cHL) has been hampered by the paucity of Hodgkin cells in biopsies and their poor growth in vitro. However, a wealth of information has been obtained from cHL cell lines. Here we report results of whole-exome sequencing and karyotypic analysis of five cHL cell lines. Four genes with potentially pathogenic single nucleotide variants (SNV) were detected in three cell lines. SNV were also detected in seventeen HL-related genes and three mitosis-related genes. Copy number variants were detected in four HL-related genes in all five cell lines. Given the high degree of aneuploidy in HL, mitosis-related genes were screened for defects. One mitotic gene (NCAPD2) was amplified in all five HL cell lines, and two genes (FAM190A, PLK4) were amplified in four cell lines. These results suggest that genomic instability of HL may be due to defects in genes involved in chromosome duplication and segregation.

This review examines handling and processing of spleen biopsies and splenectomy specimens with the aim of providing the pathologist with guidance in optimizing examination and diagnosis of splenic disorders. It also offers recommendations as to relevant reporting factors in gross examination, which may guide diagnostic workup. The role of splenic needle biopsies is discussed. The International Spleen Consortium is a group dedicated to promoting education and research on the anatomy, physiology, and pathology of the spleen. In keeping with these goals, we have undertaken to provide guidelines for gross examination, sectioning, and sampling of spleen tissue to optimize diagnosis (Burke). The pathology of the spleen may be complicated in routine practice due to a number of factors. Among these are lack of familiarity with lesions, complex histopathology, mimicry within several types of lesions, and overall rarity. To optimize diagnosis, appropriate handling and processing of splenic tissue are crucial. The importance of complete and accurate clinical history cannot be overstated. In many cases, significant clinical history such as previous lymphoproliferative disorders, hematologic disorders, trauma, etc, can provide important information to guide the evaluation of spleen specimens. Clinical information helps plan for appropriate processing of the spleen specimen. The pathologist should encourage surgical colleagues, who typically provide the specimens, to include as much clinical information as possible., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Classical Hodgkin lymphoma (HL) exhibits a bi-modal age distribution that suggests an infectious aetiology. However, most cases of nodular sclerosis HL (NSHL) are Epstein-Barr virus (EBV) negative (60-90%). Previous studies regarding human herpesvirus 6 (HHV-6) positivity of HL have led to conflicting results. In order to clarify this situation, we examined NSHL biopsies for the presence and distribution of HHV-6 by immunohistochemistry (IHC), polymerase chain reaction (PCR), and fluorescence in situ hybridization (FISH). PCR identified HHV-6 DNA in 86% of NSHL cases. As HHV-6 DNA was also identified in most cases of reactive lymphoid hyperplasia, we sought to localize the virus to specific cells by IHC, which detected HHV-6 in Reed-Sternberg (RS) cells of nearly half (48%) of NSHL cases. Dual CD30/HHV-6 immunostaining confirmed HHV-6 immunoreactivity in CD30+ RS cells, and HHV-6 PCR positivity was confirmed in laser capture microdissection-isolated CD30+ RS cells. FISH demonstrated multiple copies of HHV-6 genome in scattered cells. In contrast, EBV+ RS cells were identified in only 24% of the cases. HHV-6+ cases trended toward a younger age than EBV+ cases. These results conclusively demonstrate that RS cells in many cases of NSHL are HHV-6 positive, and suggest that HHV-6 may play a role in NSHL pathogenesis, particularly in younger patients with EBV-negative disease., (© 2012 Blackwell Publishing Ltd.)

Background: After primary infection, human herpesviruses (HHVs) maintain long-term latent persistence, often punctuated years later by sporadic episodes of symptomatic lytic activation. Also, blood-borne herpesvirus from healthy persistently infected blood donors can lead to active primary infection of immunocompromised transfusion recipients., Study Design and Methods: Utilizing a set of newly developed real-time polymerase chain reaction assays for detection and quantification of all eight human herpesviruses, the prevalence and viral DNA load of white cell-enriched blood from 100 randomly selected blood donors from the southeast Texas region are reported., Results: Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), and HHV-8 DNA were not detected in any donor sample. In contrast, Epstein-Barr virus (EBV) (72%) and HHV-7 (65%) were commonly detected, HHV-6 (30%) was often detected (Type B only), and cytomegalovirus (CMV; 1%) was rarely detected. Median viral loads of positive samples (per milliliter of blood) ranged from 4278 for HHV-6 to less than 46 for EBV., Conclusions: These results suggest that the potential for transfusion-mediated transmission of herpesviruses from healthy adult blood donors is high for EBV and HHV-7; moderately high for HHV-6; uncommon for CMV; and rare for HSV-1, HSV-2, VZV, and HHV-8. Perhaps the most remarkable finding in this study was the detection of a single donor sample with greater than 6.1 x 10(7) HHV-6 Type B genome equivalents per mL blood. Given that this extraordinarily high level of HHV-6 DNA was obtained from a healthy adult blood donor, this phenomenon is likely unrelated to active infection or immunodeficiency.

Chromosome 1 pericentromeric heterochromatin (1q) has been shown to play an important role in the pathogenesis of non-Hodgkin lymphoma and multiple myeloma. Myelodysplastic syndrome (MDS) results from marrow failure in two or more cell lineages. Although trisomy 1q has been reported in MDS, it is usually present with additional common abnormalities such as trisomy 8, monosomy 5 or monosomy 7, leading to speculation that 1q abnormalities are mostly secondary events representing clonal evolution. We report two cases of MDS in which consistent involvement of 1q heterochromatin is seen as the primary clonal abnormality. Both patients presented with fatigue and pancytopenia. Based on the published reports and our cases, we propose that the 1q heterochromatin plays a vital role in the pathophysiology of MDS. Abnormalities involving 1q result in aberrant heterochromatin/euchromatin junctions, leading to gene dosage abnormalities. Further studies of 1q abnormalities in MDS might provide specific insights as to the exact role of the excess 1q heterochromatin in the etiology of MDS.

Kikuchi-Fujimoto disease (KFD), or histiocytic necrotizing lymphadenitis, is a subacute inflammatory disorder most often seen in young women with clinicopathologic features suggestive of an infectious etiology. The most commonly suspected infectious agents in KFD are the human herpesviruses EBV, HHV6, HHV7 and HHV8. In order to identify herpesviruses in KFD, we have compared the frequency of detection of herpesvirus DNA with a recently developed real time PCR method, EBER in situ hybridization, and EBV latent membrane protein (LMP) immunostaining in 30 cases of KFD and 12 cases of reactive lymphoid hyperplasia (RLH). EBV DNA was commonly detected, while HSV2, CMV, HHV6, and HHV7 DNA were seldomly detected, and HSV1, VZV, and HHV8 DNA were not detected in KFD. EBV was also commonly detected in RLH. EBER-positive cells with apoptotic features were identified in necrotizing regions of many KFD cases, and LMP-positive cell debris was detected in one case. Viable EBER-positive cells were identified in four of twelve RLH cases, and rare LMP positivity detected in three cases. These data lend support to the notion that the necrotizing lesions in KFD may in some cases be due to a vigorous immune response to EBV-infected lymphoid cells.

Background: Hodgkin lymphoma (HL) is characterized by relatively few malignant Reed-Sternberg (RS) cells admixed within a reactive T cell rich inflammatory infiltrate. There is growing recognition that the HL-associated inflammatory milieu may enhance rather than inhibit growth of RS tumor cells. Since little is known of the immunophenotype of the HL inflammatory infiltrate we have performed a detailed retrospective comparison of the flow immunophenotype of HL and reactive lymphoid hyperplasia (RLH) to identify HL-specific immunophenotypic features., Methods: Single cell suspensions from 59 lymph nodes involved by HL (at initial diagnosis) and 38 lymph nodes involved by RLH were subjected to a battery of 3-color combinations of well-characterized fluorochrome-conjugated monoclonal antibodies (DAKO) to a number of lymphocyte subsets. Cells were analyzed on a FACSCalibur flow cytometer with CellQuest software (Becton Dickinson)., Results: Overall, CD4+ T cells are increased and CD19+ B cells decreased in HL vs. RLH, yielding median T:B cell (CD3:CD20) ratios of 2.3:1 and 1.6:1, resp. There is no difference in median CD8+ T cell % (16% in HL & RLH). The T:B cell ratio is highest in nodular sclerosis (NSHL) and lymphocyte depletion (LDHL) subtypes, and lowest in mixed cellularity HL (MCHL). There is no significant difference in CD4:CD8 ratio in any comparison. NKT cells were slightly increased in HL vs. RLH, especially in MCHL. CD4+CD25+ regulatory T cells are significantly increased in HL (9%) vs. RLH (2%), especially in MCHL (29%) and NSHL (12%). EBV positivity in NSHL is associated with older age, decreased CD4+CD25+ regulatory T cells, CD4:CD8 ratio, and CD19/CD20+ B cells, and increased NKT cells, and CD14+ low forward-side scatter-gated monocytes., Conclusion: The cellular composition of the reactive lymphocytic infiltrate in HL differs significantly from that seen in RLH, with significant differences also noted between HL subtypes. In general, the HL infiltrate contains increased T cells (CD4+ and NKT subsets), decreased B cells, and increased regulatory T cells in comparison with RLH. The major difference between HL subtypes is decreased CD4+ T cells in MCHL as compared with NSHL and NLPHL. The most notable EBV-related difference in NSHL is increased regulatory T cells in EBV negative cases. While many differences in the reactive lymphocytic infiltrate of Hodgkin lymphoma and reactive lymphoid hyperplasia were identified, the sole difference that may prove to be of differential diagnostic value in flow cytometric analysis of HL versus RLH is the increased percentage of CD4+ bright CD25+ regulatory T cells in HL., ((c) 2007 Clinical Cytometry Society)

We report the detailed features of a unique case of a 33-year-old male in whom a large percentage of marrow (68%) and blood (31%) cells carried the Philadelphia chromosome by cytogenetics and FISH, as well as the p210 BCR-ABL transcript by RT-PCR, for 15 months prior to development of chronic myelogenous leukemia. The patient was closely followed throughout the preleukemic period with repeat blood and bone marrow analysis with no hematologic or pathologic evidence of leukemia, despite harboring a large number (65%) of Ph+ marrow cells. we report the detailed history of a 33-year-old male who carried a large number of BCR-ABL-positive cells in the marrow and blood for more than 15 months before finally presenting with classic chronic myelogenous leukemia. This study is unique in that it provides the most detailed documentation of the hematologic, pathologic, and cytogenetic features of the preleukemic phase of chronic myelogenous leukemia ever reported.

Several lines of evidence indicate that cytokines can affect adrenal function. To date most of these cytokines have been shown to be pro-inflammatory, such as interleukin (IL)-1, tumor necrosis factor (TNFalpha), and IL-6. However, we have previously shown that IL-10-/- (IL-10 knockout) mice have higher serum corticosterone levels than IL-10+/+ (wild type) mice following acute immune and physiologic stress, implying that IL-10, an anti-inflammatory cytokine, regulates glucocorticoid synthesis in a negative manner. Here, we show that IL-10 knockout mice produce more corticosterone under basal conditions as well (shown by ELISA). We further support this contention by showing that in Y-1 adrenocortical cells IL-10 inhibits steroid production (StAR) (measured by the production of the corticosterone precursor, progesterone), the expression of steroidogenic acute regulatory protein (semi-quantitative RT-PCR), as well as the activity of the proximal steroidogenic enzymes P450scc and/or 3beta-hydroxysteroid dehydrogenase (3beta-HSD) (measured by progesterone production in 22(R)-hydroxycholesterol-treated cells). Interestingly, all of the above-mentioned effects of IL-10 occur through its inhibition of ACTH effects, but not by IL-10 alone. Furthermore, immunocytochemistry data shows that the region of the adrenal gland responsible for the vast majority of corticosterone synthesis, the zona fasciculata, predominantly expresses the IL-10 receptor 1 (IL-10R1), with little expression in the zona glomerulosa and reticularis. These data demonstrate that IL-10 could play an important role in the regulation of glucocorticoid biosynthesis and in maintenance of homeostasis and immunity during periods of stress.

Following primary infection, all eight human herpesviruses persist lifelong in the human host. However, a mapping of all anatomic sites of human herpesvirus persistence is lacking. Fresh tissue specimens representing approximately 40 major anatomic sites from eight autopsies were screened using a recently developed real-time PCR method for detection of all eight human herpesviruses. Patients with evidence of active herpesvirus infection (herpes simplex 1 (HSV-1), herpes simplex 2 (HSV-2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpesvirus 6 (HHV-6), herpesvirus 7 (HHV-7), and herpesvirus 8 (HHV-8)) at the time of death were excluded to avoid detection of widely disseminated infection. Despite this precaution, widespread HSV-1 positivity (with blood positivity) was detected in one case-an elderly male who died of cardiac arrest. In a middle-aged male with HIV-AIDS, HSV-1 was found in neural and pharyngeal tissues, skin, cartilage, bone, and urinary bladder, whereas in two other cases, HSV-1 was restricted to neural tissues. HSV-2 was detected in a single site, the anus, in the male with HIV-AIDS. VZV was detected only twice, once in the adrenal gland and once in the small intestine. CMV was detected in three cases, most commonly in nasal mucosa, trachea, thyroid, intestine, and liver. EBV was detected in all eight cases, especially in nasal mucosa, tonsil, spleen, lymph node, tongue, and intestine, but in only two of six whole-blood specimens. HHV-6, like EBV, was detected in all eight cases, most commonly in salivary glands, thyroid, stomach, intestines, liver, and pancreas. HHV-7, like EBV and HHV-6, was detected in all eight cases, most commonly in salivary glands, tonsil, lymph nodes, and bone marrow. HHV-8 was detected in only two sites (both lymph nodes) from two cases. Herpesviruses were detected in three of six whole-blood specimens, including HSV-1, EBV, HHV-6, and HHV-7. These results represent the most comprehensive mapping of herpesvirus tissue distribution in humans reported to date.

Although Epstein-Barr virus (EBV) is often found in human tonsils, it remains to be precisely determined in what cells and microenvironment the virus is present. Although generally regarded as a B lymphotropic virus, EBV is associated with non-B-cell tumors, for example, NK/T-cell lymphoma, carcinoma, and leiomyosarcoma. To provide a basis for understanding the origin and biology of EBV-infected non-B cells, the immunophenotype of all EBV-infected cells in reactive human tonsils was determined by subjecting tonsil sections to dual/triple EBER in situ hybridization and immunohistochemistry with monoclonal antibodies to T cells (CD3, CD4, CD8, CCR3), B cells (CD20), plasma cells (CD138), natural killer (NK) cells (PEN5), and epithelial cells (cytokeratin), as well as frozen section immunostaining with antibodies to EBV latent proteins EBNA1, EBNA2, LMP1, and EBV early protein BZLF1. Most tonsils contained nearly equal numbers of EBNA1- and LMP1-positive cells (latency program) while only a few contained EBNA2-positive cells (growth program). More than 1000 EBER-positive cells from six tonsils were detected in the interfollicular zone (59%), tonsillar crypts (26%), and follicles (15%). Most (82%) EBER-positive cells are CD20-positive B cells, 7% are CD3-positive T cells, and 11% are cells of indeterminate lineage, often with plasmacytoid morphology. However, no EBER-positive plasma cells were identified. Rare EBER-positive NK cells and EBER/BZLF1-positive epithelial cells were identified. The direct demonstration of EBV within rare T cells, NK cells, and epithelial cells in reactive human tonsils provide a basis for further understanding of the origin of EBV-associated tumors of non-B-cell type.

We report a case of nodal nevus present in enlarged lymph nodes with changes of dermatopathic lymphadenopathy sampled by fine-needle aspiration (FNA) cytology prior to clinical evaluation of the patient. This lymph node pathology was established later by lymph node excisional biopsy, by which along with a skin biopsy the dermatopathic lymphadenopathy was tentatively attributed to early mycosis fungoides. The FNA revealed fairly atypical melanotic tissue from the dermatopathic lymphadenopathy along with nodules of uniform melanocytic nevoid cells, the presence of which in combination with the dermatopathic atypical tissue provided a tentative diagnosis of metastatic melanoma of unknown primary, with the diagnosis of nodal nevus presented as a less likely possibility. This is to our knowledge the first cytologic report on FNA of nodal nevus, which besides presenting cytologic findings of this entity highlights some of the problems related to providing an accurate diagnosis, if this exceptionally unusual pathologic entity is encountered in lymph nodes sampled for enlargement from pathologies unrelated to this entity. The subject of nevus changes in lymph nodes is briefly discussed.

Infectious mononucleosis (IM) due to all causes is characterized by atypical lymphocytosis. We sought to compare hematologic parameters of infectious mononucleosis due to Epstein-Barr virus (EBV) infection (heterophile antibody (HA) positive) with mononucleosis due to other causes. Mono-Latex Slide Agglutination Test results and complete blood counts (CBC) of 147 patients with mononucleosis were retrospectively analyzed. Leukocyte count, absolute lymphocyte count, and presence of atypical lymphocytes in EBV-positive and EBV-negative groups were statistically compared. We analyzed 68 EBV-positive and 79 EBV-negative cases. EBV-positive patients were significantly younger than EBV-negative patients were. Mean total WBC count and mean absolute lymphocyte count were significantly higher in EBV-positive patients. Absolute lymphocytosis, absolute leukocytosis, and atypical lymphocytosis were also significantly more frequent in EBV-positive patients. Leukopenia was more frequently seen in EBV-negative patients., (Copyright 2004 Wiley-Liss, Inc.)

The association of interleukin-10 (IL-10) promoter single-nucleotide polymorphisms (SNPs) as risk factors for certain inflammatory diseases, viral infections, cancers, and transplant rejection have been the subject of recent studies. The SNPs -1082 G --> A, -819 C --> T, and -592 C --> A, which have been associated with differential IL-10 production, are strongly linked with ethnicity. In this study, we determined the ethnic distribution of IL-10 promoter SNPs and their haplotype rates among Hispanics, African Americans, and Caucasians from Texas and Ashkenazi Jews from New York. Significant differences in prevalence rates of IL-10 SNPs (and their haplotype distribution) were found. African Americans and Hispanics have a lower rate of putative high-producer SNPs and a higher rate of low IL-10 producers when compared to Caucasians or Ashkenazi Jews. No statistically significant differences in allelic frequencies and haplotype rates were observed between Caucasians and Ashkenazi Jews. This study provides critical new information on the ethnic distribution of IL-10 promoter SNPs in a regional U. S. population and is the first to analyze the rate of SNPs in an unstudied ethnic population, Ashkenazi Jews. Knowledge of IL-10 promoter polymorphisms may prove useful in prediction of immunization responses, disease severity, and in the intelligent design of customized immunotherapy.

There are eight currently known human herpesviruses, all of which are capable of latent persistence and reactivation following primary infection. Herpesvirus induced disease is common, widespread, and associated with significant morbidity, particularly in the immunocompromised human host. Current methods of herpesvirus detection include viral culture and polymerase chain reaction (PCR). A robust PCR method based upon amplification of the highly conserved herpesvirus DNA polymerase gene that is capable of detection of all eight human herpesviruses, including EBV and HHV-6 subtypes, from clinical material is described. Species identification of PCR products is accomplished by either of two methods, chemiluminescent dot blot hybridization and heteroduplex mobility shift assay, both of which allow for simultaneous detection of multiple herpesviruses. This method should prove useful for rapid and accurate species identification of all eight human herpesviruses from clinical material.

Background: Atypical lymphocytosis is a common peripheral blood abnormality seen not only in Epstein-Barr virus (EBV)-associated acute infectious mononucleosis but also in other conditions, including other viral infections, cancer, immune reactions, etc. Despite numerous reports of individual immunophenotypic alterations in EBV-positive infectious mononucleosis, a detailed comparative analysis of the immunophenotypic changes of peripheral blood lymphocyte subsets in infectious mononucleosis and other forms of atypical lymphocytosis is lacking., Methods: Multiparametric flow immunocytometry with 26 monoclonal antibodies was performed on peripheral blood lymphocytes from 97 cases of atypical lymphocytosis and 37 normal controls. Atypical lymphocytosis was defined as absolute lymphocytosis with >10% atypical lymphocytes. Absolute or relative mean values of various lymphocyte subsets from EBV-positive cases, EBV-negative cases, and normal controls were compared with the Student's t-test., Results: Highly significant abnormalities detected in atypical lymphocytosis include increases in CD3+/CD8+, CD3-/CD16/56+, CD3+/gammadelta+, CD8+/CD48-, CD8+/CD57-, CD8+/CD95+, CD4+/CCR5+ CD4+/CD7-, CD4+/CD43-, CD4+/CD48-, and CD4+/CD62L- subsets. In contrast, no change in absolute CD4+ T cell and CD19+ B cell counts is seen. When compared with EBV-negative cases, EBV-positive cases are characterized by younger age, and increased numbers of absolute lymphocytes, atypical lymphocytes, CD8+ cells, NK cells, gammadelta T cells, CD8+/CD45RO+ cells, CD8+/CD57- cells, and CD8+/CD28+ cells., Conclusions: All forms of atypical lymphocytosis are characterized by a marked increase in activated CD8-positive T cells, a moderate increase in NK cells, and no increase in CD4-positive T cells and B cells. Although morphologically indistinguishable, EBV-associated mononucleosis is characterized by several significant differences in peripheral blood lymphocyte subsets when compared with EBV-negative atypical lymphocytosis, most notably increased numbers of CD57-negative CD8 T cells and gammadelta T cells., (Copyright 2003 Wiley-Liss, Inc.)

Background: Human herpesvirus 8 (HHV-8) is widely suspected to be a human tumor virus because it is associated with Kaposi's sarcoma and primary effusion B cell lymphoma. Report of a case of HHV-8-positive donor blood in the US has led to concern for the safety of donor blood from HHV-8-seropositive donors., Study Design and Methods: The findings of HHV-8 seroprevalence and virus load from 100 randomly selected blood donors from the Houston, Texas, area are reported. Serology with serial titration was performed using a highly sensitive indirect immunofluorescence assay to lytic and latent HHV-8 antigens. For detection of blood-borne virus, buffy-coat DNA was subjected to two ultrasensitive nested PCR-dot blot assays to HHV-8 orf26 and orf72 regions., Results: At a screening titer of 1 in 10, nearly one-quarter (23%; 95% CI, 15-33) of the blood donors are HHV-8 seropositive with a geometric mean titer of 1 in 53. Seroreactivity to lytic antigens (23%) greatly exceeded that to latent antigens (5%). There was a significant association between seropositivity and older age (p < 0.02), white ethnicity (OR, 3.33; 95% CI, 1.40-7.95) and ABO blood group B (OR, 6.44; 95% CI, 2.46-16.80). No association with sex or CMV seropositivity was demonstrated. No HHV-8 viremia was detected, even though 64 percent of tested donor blood samples were EBV DNA positive., Conclusions: Despite a relatively high HHV-8 seroprevalence in this cohort of Houston area blood donors, HHV-8 DNA was not detected in any sample of donor whole blood using a highly sensitive PCR assay. Thus, at least in the southeast Texas region, large-scale screening of blood donor units for HHV-8 antibody or DNA seems unwarranted.

Plasma cell granuloma of the thyroid is an uncommon lesion; only 6 cases have been reported in the English literature to date. All reported cases occurred in women, mostly after the age of 50 years. We report a case of plasma cell granuloma of the thyroid in a 46-year-old woman with a 20-year history of euthyroid goiter and a positive family history of goiter in 3 close relatives. The lesion was composed of sheets of plasma cells involving the entire parenchyma that histologically resembled plasmacytoma. Plasmacytoma was excluded by demonstration of polyclonal kappa/lambda light chain immunostaining and by lack of evidence of clonal bands by polymerase chain reaction for immunoglobulin heavy-chain gene rearrangement. Similarly, the predominant histologic pattern in all previously reported cases is that of marked plasma cell infiltration. A family history of thyroid disease (goiter, thyroiditis) was associated with diffuse involvement of the thyroid. Prognosis after surgery is excellent, and to our knowledge no cases of malignant transformation or recurrence have been described.

The importance of hyperhomocysteinemia, birth defects, and vascular diseases has been the subject of intense investigations. The polymorphic MTHFR mutations (C677T and A1298C) cause mild hyperhomocysteinemia, especially in homozygotes for C677T, but also in compound heterozygotes for C677T/A1298C. The subject of this report is the frequency of the polymorphic mutations in the MTHFR gene C677T, C1298A, and newly discovered mutation G1793A, as well as the association with MTRR polymorphic site A66G in different ethnic groups. Four ethnic groups were studied: African-Americans, Caucasians, Hispanics, and Ashkenazi Jews. There are statistically significant differences in the frequency of these alleles in the different populations studied, which impacts compound heterozygosity for such alleles in these populations. DNA samples obtained from the blood of healthy individuals of African-Americans, Hispanics, and Caucasians from south Texas were analyzed and compared to those obtained from Ashkenazi Jewish individuals. The polymorphic site, the G1793A allele, is least frequent among Ashkenazi individuals, 1.3%, compared to 6.9% among Caucasians (P = 0.001), 5.8% among Hispanics (P = 0.012), and 3.1% among African-Americans. The MTRR polymorphic site shows the lowest allele frequency among Hispanics, 28.6%, compared to 34% among African-Americans, 43.1% among Ashkenazi Jews (P = 0.002), and 54.4% among Caucasians (P < 0.0001). Statistically significant differences in allele frequencies of C677T and C1298A polymorphisms were also observed in these populations. Compound heterozygosity for multiple polymorphic alleles may play a role in birth defects and vascular diseases., (Copyright 2001 Wiley-Liss, Inc.)

Productive Epstein-Barr virus (EBV) replication characterizes hairy leukoplakia, an oral epithelial lesion typically occurring in individuals infected with human immunodeficiency virus (HIV). Serial tongue biopsy specimens were obtained from HIV-infected subjects before, during, and after valacyclovir treatment. EBV replication was detected by Southern hybridization to linear terminal EBV genome fragments, reverse-transcriptase polymerase chain reaction amplification of EBV replicative gene transcripts, immunohistochemical detection of EBV replicative protein, and in situ hybridization to EBV DNA. EBV replication was detected in both hairy leukoplakia and normal tongue tissues. Valacyclovir treatment completely abrogated EBV replication in vivo, resulting in resolution of hairy leukoplakia when it was present. EBV replication returned in normal tongue epithelial cells after valacyclovir treatment. These data suggest that normal oral epithelium supports persistent EBV infection in individuals infected with HIV and that productive EBV replication is necessary but not sufficient for the pathogenesis of oral hairy leukoplakia.

The association of Hodgkin disease with monoclonal gammopathy has rarely been reported. We present a case of a 48-year-old woman with a history of autoimmune hemolytic anemia and Graves disease who presented with hepatosplenomegaly and a gamma-heavy-chain paraprotein. Histopathology of lymph node and bone marrow revealed nodular lymphocyte-predominance Hodgkin disease, while examination of the spleen revealed plasmacytosis consistent with gamma-heavy-chain disease. Following splenectomy, the patient has remained in complete remission for both conditions with no further treatment. To our knowledge, this is the first report of a patient with both gamma-heavy-chain disease and nodular lymphocyte-predominance Hodgkin disease. Given recent data indicating the B-cell nature of this form of Hodgkin disease, the authors propose that in this unique case there may be a clonal relationship between these 2 concurrent B-cell lymphoproliferative processes.

A clonal T-gamma rearrangement was found in peripheral blood and bone marrow in a 57-year-old female who presented with 6-week history of fevers, night sweats, and weight loss. Splenomegaly, hemolytic anemia, atypical lymphocytosis, a marrow lymphoid aggregate, and elevated LDH had suggested lymphoproliferative disease. However, IgM serology for cytomegalovirus (CMV) was positive. With observation alone, her clinical features improved over 4 weeks with normalization of the blood count and disappearance of CMV viremia and the aberrant T-gamma clone. Acute CMV infection may mimic lymphoproliferative disease. T-gamma gene rearrangement may be part of the immune response to CMV infection and is not specific to lymphoid neoplasia.

The expression of myelomonocytic-associated antigens in anaplastic large cell lymphomas (ALCLs), particularly those presenting in extranodal sites, can make their distinction from extramedullary myeloid cell tumors (EMCTs) or histiocytic tumors problematic. Yet, this distinction is clinically significant because of its therapeutic and prognostic implications. Herein, we describe a case of extranodal anaplastic lymphoma kinase-positive CD30-positive ALCL of T-cell origin in a 12-year-old boy, which was initially called an EMCT because of the expression of CD13 and HLA-DR detected by flow cytometry and the absence of other T-cell-related surface markers. However, the detection of cytoplasmic CD3 by flow cytometry prompted further studies. The tumor was composed of large cells with abundant slightly eosinophilic vacuolated cytoplasm and ovoid or reniform nuclei with a few small nucleoli. Using immunohistochemistry, the tumor was positive for CD45, CD30, CD45RO, and CD43 with a strong cytoplasmic and nuclear anaplastic lymphoma kinase stain. The tumor cells showed a T-cell clonal genotype. Electron microscopy revealed no ultrastructural features of myelomonocytic or histiocytic origin. The patient responded well to the chemotherapy and was in complete remission for 10 months at the time of submission of this manuscript. Review of the literature showed inconsistencies regarding the diagnosis, nomenclature, and, therefore, treatment and prognosis of these tumors. In addition, the CD13 expression in ALCL raises some histogenetic questions and may indicate origin from a pluripotent stem cell, misprogramming during malignant transformation, or a microenvironmental effect on lymphoid cell expression of surface antigens. Therefore, ALCL should be considered in the differential diagnosis of EMCTs or histiocytic tumors, particularly when surface marker lineage assignment is ambiguous.

Background: Estrogen receptor (ER) levels in tumor cells are important for determining the outcome of treatment and the prognosis of breast cancer patients. Flow cytometry is a convenient tool for quantifying the ER in cells, but a more sensitive, reproducible method for immunostaining the ER with anti-ER antibody is needed. Materials and Methods ER-positive human breast cancer cells MCF-7 and T47D, and ER-negative MDA-MBA-321 cells, were fixed and permeabilized by three different protocols. The cells were then stained by indirect immunofluorescence, using two commercial antibodies to ER (MA1-310 and DAKO 1D5), or by direct immunofluorescence using FITC-labeled anti-idiotypic antibody clone 1D(5). The stained cells were analyzed by flow cytometry., Results: The fixation of cells with a mixture of 0.25% paraformaldehyde and 70% methanol, permeabilization with 0.05% Triton X-100, and increasing antibody and antigen reaction time led to 80-99% of cells being stained with anti-ER antibodies. The relative brightness of ER immunostaining was as follows: anti-idiotypic antibody ID5 > MA1-310 > DAKO 1D5., Conclusions: Direct immunofluorescence with the FITC-labeled anti-idiotypic antibody of permeabilized cells resulted in improved specific staining of the ER, as compared to indirect immunofluorescence with anti-ER antibodies of fixed and permeabilized cells. Increasing the length of staining, and treatment of cells with Triton X-100, are both necessary to improve the staining of intracellular antigen for flow cytometric analysis., (Copyright 2000 Wiley-Liss, Inc.)

Hemophagocytic syndrome (HS) is a histiocytic reactive process often associated with infections and/or malignancies. Clonal karyotypic abnormalities have been the hallmark of several hematological malignancies and have been shown to be of clinical significance in terms of both diagnosis and prognosis. While there are limited reports of both clonal and nonclonal abnormalities in HS, their clinical significance has not been established. Detection of such clonal abnormalities, as seen in some cases of HS, may indicate the presence of an occult malignant process, even when there is no microscopic evidence of a hematological malignancy. We report a case of HS in a child with clonal t(4;7)(q21;q36) which later progressed to acute myeloid leukemia (AML) with further clonal evolution. Our case strengthens the argument that cytogenetic studies in HS may be important in identifying the underlying occult malignant process.

Muscle biopsy homogenates contain GLUT-3 mRNA and protein. Before these studies, it was unclear where GLUT-3 was located in muscle tissue. In situ hybridization using a midmolecule probe demonstrated GLUT-3 within all muscle fibers. Fluorescent-tagged antibody reacting with affinity-purified antibody directed at the carboxy-terminus demonstrated GLUT-3 protein in all fibers. Slow-twitch muscle fibers, identified by NADH-tetrazolium reductase staining, possessed more GLUT-3 protein than fast-twitch fibers. Electron microscopy using affinity-purified primary antibody and gold particle-tagged second antibody showed that the majority of GLUT-3 was in association with triads and transverse tubules inside the fiber. Strong GLUT-3 signals were seen in association with the few nerves that traversed muscle sections. Electron microscopic evaluation of human peripheral nerve demonstrated GLUT-3 within the axon, with many of the particles related to mitochondria. GLUT-3 protein was found in myelin but not in Schwann cells. GLUT-1 protein was not present in nerve cells, axons, myelin, or Schwann cells but was seen at the surface of the peripheral nerve in the perineurium. These studies demonstrated that GLUT-3 mRNA and protein are expressed throughout normal human skeletal muscle, but the protein is predominantly found in the triads of slow-twitch muscle fibers.

The detailed immunophenotype of peripheral blood lymphocytes from a neonate with pertussis was determined by flow cytometry and compared with results from cord blood from healthy newborns. Most (72%) of the lymphocytes were CD3+ T cells with a normal CD4/CD8 ratio (2.5). The T cells were largely HLA-DR negative and CD45RA+, consistent with unstimulated naïve T cells. Almost all of the CD4+ T cells were Leu8 (L-selectin, CD62L) negative, while almost all of the CD8+ T cells were CD28+. There was no increase in CD7- CD4+ T cells (Th2-like). No relative increase in CD16/56+ NK cells (5%) or CD19/20+ B cells was seen. The most dramatic finding in this case was the remarkable lack of expression of L-selectin by the T cells. L-selectin expression is associated with homing of peripheral blood lymphocytes to lymph nodes. The dramatic reduction in L-selectin expression of the T lymphocytes in pertussis, perhaps induced by pertussis toxin, likely prevents homing of the T cells to peripheral lymphoid tissues and provides a likely explanation for the marked lymphocytosis noted in this disease.

We report the case of a 41-year-old black man with acquired immunodeficiency syndrome who developed a severe chronic anemia due to parvovirus infection. Bone marrow biopsy revealed erythroid aplasia. The infectious nature of the anemia was not recognized, and the patient was treated with erythropoietin. The patient's reticulocyte response was inadequate, however, and he remained anemic. A second bone marrow biopsy showed erythroid hyperplasia and prominent intranuclear parvovirus inclusions within erythroid progenitors. Erythropoietin was discontinued and was followed by a course of intravenous immunoglobulin, which resulted in rapid correction of anemia. To our knowledge, this is the first reported case of fulminant human parvovirus infection exacerbated by erythropoietin administration and documented by sequential bone marrow histologic examination. This case illustrates the critical importance of considering parvovirus in the etiology of chronic anemia with erythroid aplasia in immunocompromised patients.

Human herpesvirus 8 (HHV-8) has been causally linked to Kaposi's sarcoma (KS). There is significant homology between some HHV-8 genes and cellular genes including D-type cyclin (vCYC), G protein coupled receptor (vGCR), macrophage inflammatory proteins (vMIP-I, vMIP-II), bcl-2 (vBCL2), interferon regulatory factor-1 (vIRF1), interleukin-6 (vIL6), and complement-binding protein (vCBP). In this study, we analyzed expression of these viral homologs and HIV-1 Tat by reverse-transcriptase polymerase chain reaction (RT-PCR) coupled with Southern blot hybridization in AIDS-KS (AKS) tissue, classic KS tissue(CKS), and peripheral blood mononuclear cells, and phorbol ester (TPA)-treated and untreated HHV-8 positive lymphoma cells (BCBL1). While vCYC (AKS 6 of 6; CKS 3 of 3), vMIP-I (AKS 5 of 6, CKS 3 of 3), vBCL2 (AKS 6 of 6; CKS 3 of 3), and vIRF1 (AKS 5 of 6, CKS 3 of 3) transcripts were detected in both AKS and CKS, vGCR and HIV-1 Tat were expressed only in AKS samples (vGCR: AKS 3 of 6, CKS 0 of 3; Tat: AKS 4 of 6, CKS 0 of 3). vMIPII, vCBP, and vIL6 expression were not detected in any KS samples. Since vGCR expression is limited to AKS, it is possible that vGCR is activated by HIV-1 Tat. These results suggest that HIV-1 Tat may contribute to AKS pathogenesis through the tumorigenic and angiogenic effects of vGCR., (Copyright 2000 Academic Press.)

The polymorphic mutation C677T in the gene of MTHFR is considered a risk mutation for spina bifida and vascular disease. Another common mutation on the MTHFR gene, A1298C, has also been described as another risk mutation. We studied the frequencies of these two mutations on DNA samples from healthy Jewish individuals and compared them to the frequency of these mutations in DNA samples obtained from healthy individuals in South Texas. The presence of the C677T allele was determined by PCR and Hinf I digestion, and mutation A1298C by PCR and Mbo II digestion. A total of 310 alleles was examined for C677T in the Ashkenazi samples and 400 alleles in the non-Jewish samples. The rate of C677T among the Ashkenazi Jewish alleles was 47.7% as compared to 28.7% among the alleles from the non-Jewish population. The difference is statistically significant, P < 0.0005. Mutation A1298C was examined in 298 alleles of Jewish individuals and 374 alleles of non-Jewish counterparts from Texas. The rate of the A1298C mutation in the Jewish samples was 27.2% whereas in the non-Jewish was 35%. This was also statistically significant, P < 0.031. No individuals were homozygous for both mutations or were found to be homozygous for one mutation with heterozygosity of the other mutation, and that the C677T and the A1298C alleles did not occur in cis position. This study shows a unique distribution of C677T and the A1298C alleles among the Ashkenazi Jews. In spite of high frequency of C677T mutation, spina bifida is less common among Ashkenazi Jews. Further studies are needed to establish whether the C677T and the A1298C mutations have an impact on vascular disease in the Ashkenazi Jewish population., (Copyright 1999 Wiley-Liss, Inc.)

Cutaneous biopsies of five eschars and two rash lesions from five patients from New York City with documented rickettsialpox were examined by immunohistochemical methods with a monoclonal antibody directed against spotted fever group rickettsial lipopolysaccharide for the presence and cellular location of Rickettsia akari Rickettsiae were identified in all of the five patients, with good concordance of results for the same biopsy tissues with previously reported results by the direct immunofluorescence method. In contrast with immunofluorescence, which did not reveal the location of the organisms, immunohistochemical examination demonstrated R. akari to be in perivascular cells, morphologically resembling macrophages. Evaluation with double staining for rickettsiae and either CD68 or Factor VIII-related antigen revealed that the predominant infected cell type was CD68-positive macrophages, and only a rare rickettsia was detected in vascular endothelium, the major target cell for other rickettsioses. These results provide a diagnostic method for rickettsialpox and other spotted fever group rickettsioses and indicate that the elucidation of the pathogenesis of rickettsialpox must take into account that its target cell differs from that of Rocky Mountain spotted fever, boutonneuse fever, louse-borne typhus fever, and murine typhus.

Background: Patients undergoing chronic steroid therapy for organ transplantation are at increased risk for development of human herpes virus 8(HHV-8)-associated Kaposi's sarcoma (KS). It has also been reported that following steroid withdrawal, KS lesions often undergo partial or complete regression., Methods: We have examined the effect of corticosteroid treatment on HHV-8 replication, gene expression, and lytic protein expression in BCBL-1 cells in vitro. BCBL-1 cells were collected after culture for 24-72 hr with hydrocortisone (HC) 1-5 microM, phorbol ester 20 ng/ml (positive control), and culture medium only (negative control). HHV-8 genomic conformation was examined by Gardella gel analysis. mRNA expression of viral cyclin (v-Cyc), viral Bcl-2 (v-Bcl-2), viral macrophage inflammatory protein-I (v-MIP-I), viral interferon regulatory factor-1(v-IRF-1), and viral tegument protein (TP) was examined by RT-PCR Southern blot. Viral protein expression within the cells was examined by indirect immunofluorescence using 5 different HHV-8 positive antisera from 4 renal transplant recipients and 1 patient with classic KS., Results: Gardella gel analysis revealed that HC induced an accumulation of the linear replicative genomic form of the virus in a time-dependent fashion. Southern blot analysis of the RT-PCR products revealed that HC induced increased expression of v-IRF-1, v-Bcl-2, and TP mRNA, with little discernible effect on v-Cyc, and v-MIP-I. Immunofluorescence revealed that HC induced increased numbers of cells expressing lytic antigens., Conclusions: These data indicate that hydrocortisone acts directly on BCBL-1 cells to activate the lytic cycle of HHV-8 and provide further support for the hypothesis that HHV-8 is activated in corticosteroid-treated immunocompromised patients.

This study was designed to determine whether there is serologic or molecular evidence of human herpesvirus 8 (HHV-8) activation in renal transplant patients, an immunocompromised population at risk for development of Kaposi's sarcoma. Indirect immunofluorescence for detection of HHV-8 serum antibody and Southern blot polymerase chain reaction (PCR) for detection of viral DNA in whole blood were used. Seroprevalence and geometric mean titer (GMT) were significantly increased in the transplant group compared with healthy adults and were comparable to those in human immunodeficiency virus (HIV)-positive adults (transplant patients, 50% [GMT 1:210]; healthy adults, 7% [GMT 1:44]; HIV-positive patients, 73% [GMT 1:172]). Viral DNA was present in the blood of some renal transplant patients (3/33 PCR-positive) but in none of 20 healthy adults. Thus, there is both serologic and molecular evidence of HHV-8 activation in the renal transplant population compared with healthy adults (P<.01). The serologic results approximate those obtained for HIV-positive adults.

We report the clinicopathologic characteristics of pulmonary lymphomatoid granulomatosis (LYG) in 11 patients (identified from a series of 330 consecutive patients who underwent autopsy between 1984 and 1995 at the University of Texas Medical Branch at Galveston, Texas) with a diagnosis of acquired immunodeficiency syndrome (AIDS). We used immunohistochemical stains, RNA in situ hybridization (ISH), and gene rearrangement studies to identify the immunophenotype and the presence or absence of Epstein-Barr virus (EBV) infection. All of the patients were men ranging in age from 27 to 65 years (mean age, 38.6 yr). Autopsy lungs of 21 age-matched controls were examined for EBV using ISH; these included 9 patients with AIDS who did not have pulmonary lesions and 12 HIV-negative individuals who died accidentally (mean age, 38.6 yr). All of the 11 pulmonary lesions showed the gross and microscopic characteristics of LYG, with zonal necrosis and prominent angioinvasion. The tumor nodules consisted of a mixture of atypical large lymphocytes, with vesicular nuclei and prominent nucleoli and with a background of small and intermediate-size lymphocytes, histiocytes, and plasma cells. The large lymphocytes were CD20 positive, consistent with a B-cell phenotype. Ten of the 11 cases demonstrated EBV1-encoded RNA and CD20 positivity in the large, atypical lymphocytes by double labeling. One patient showed EBV positivity in CD20-negative, CD45RO-positive large cells, but these cells were CD3 negative and showed a monoclonal heavy chain gene rearrangement by polymerase chain reaction, indicating that these were of B-cell origin. Aberrant CD43 coexpression was identified in four cases. EBV latent membrane protein was demonstrated in 9 of 11 cases by immunohistochemical stains. The lungs of all of the 21 control patients were negative for EBV by ISH. We conclude that, in our series, AIDS-associated LYG is a B-cell neoplasm and that it has a strong association with EBV infection.

Background: Kaposi's sarcoma (KS) accounts for more than 5% of malignancies in immunosuppressed organ transplant patients (OKS). A new herpesvirus (HHV-8) was identified with high prevalence in biopsy specimens of AIDS-KS, endemic KS, and classic KS and in OKS. KS has also been associated with other underlying diseases in patients treated with corticosteroids, but this subset of KS has been reported to contain HHV-8 in only a few case reports., Objective: In this larger study, we determined the prevalence of HHV-8 in seven patients of Jewish origin in whom KS developed during immunosuppressive therapy for different primary diseases (ISKS)., Methods: The study included HHV-8 DNA detection by polymerase chain reaction (PCR) coupled with Southern blot and sequence analysis as well as by in situ hybridization., Results: HHV-8 sequences were detected by PCR with confirmation by Southern blot and sequence analysis in 100% of the ISKS samples. Direct sequencing revealed several previously unknown base changes within the 208 bp region from open reading frame 26 (ORF26[208]) of HHV-8 in ISKS., Conclusion: Ours is the largest known study describing the presence of HHV-8 in iatrogenic KS from immunosuppressed nontransplant patients and provides data of previously unknown sequence variations within the ORF26 of HHV-8 DNA.

The clinical and pathologic findings of Kikuchi's histiocytic necrotizing lymphadenitis may mimic those of malignant lymphoma. We describe a 6-year-old boy with generalized lymphadenopathy, spiking fever, chills, myalgias, malaise, and erythematous, crusted papules. Although cutaneous manifestations have been noted in 16% to 40% of patients with histiocytic necrotizing lymphadenitis, only three publications described skin lesions. The skin lesions and affected lymph nodes revealed histiocytic aggregates, atypical lymphoid cells, karyorrhectic debris, and patchy necrosis. Spontaneous resolution occurred in 2 months. Results of serologic studies, Epstein-Barr virus (EBV) latent membrane protein immunoperoxidase staining, EBER-1 RNA in-situ hybridization, and EBV EBNA-1 DNA polymerase chain reaction implicate EBV as the causative agent.

Background and Objective: Hypoplastic myelodysplastic syndromes (MDS) are being reported with increasing frequency. Aplastic anemia (AA) needs to be differentiated from hypoplastic MDS particularly primary hypoplastic refractory anemia (PHRA) because of the impact on management and prognosis. This distinction may be morphologically difficult even with careful marrow examination which may provide insufficient material due to extreme hypocellularity. The value of peripheral blood (PB) parameters in making the distinction between AA and PHRA is not well studied. In this work, we attempt to examine peripheral blood findings as an additional tool for differentiating PHRA from acquired idiopathic AA., Methods: PB findings in ten cases of PHRA, which are selected based on the following: less than 30% cellularity, multilineage dysplasia and/or clonal cytogenetic abnormality, are compared to ten cases of classic AA. The PB is examined for automated parameters, differential white cell count, morphologic changes in red cells, white cells, platelets, and the presence of circulating blasts, megakaryocytic fragments and micromegakaryocytes., Results: AA patients tend to have lower platelet and monocytic counts and higher lymphocytic percentages. The following morphologic findings are seen only in PHRA but not in AA: hypochromic red cells, left shift, circulating blasts, hypersegmentation with long filaments, hypogranular, ring, and pelgeroid neutrophils, Dohle bodies, circulating micromegakaryocytes and megakaryocytic fragments., Interpretation and Conclusions: We conclude that careful examination of peripheral blood may provide sufficient information to allow for the distinction between PHRA and AA early in the course of the disease. Similarly, patients with classic AA who subsequently develop unusual blood findings during routine follow up should be suspected of having a clonal evolution which needs to be confirmed by marrow examination and cytogenetic analysis.