Human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV) antigens and genome in lymph nodes of HIV-positive patients affected by persistent generalized lymphadenopathy (PGL)

The presence of human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV) antigens and genome has been investigated in 50 lymph nodes involved by persistent generalized lymphadenopathy (PGL). All the patients were HIV infected and most of them (42 of 50) also had anti-EBV serum antibodies. At lymph node level, HIV and EBV antigens were studied by immunohistochemistry using monoclonal antibodies directed against viral core proteins. The HIV p24 protein was detected in 43 of 50 lymph nodes within the B-cell germinal centers with a reticular pattern. Few cells with positive results for EBV antigens were found in only 2 of 50 lymph nodes. These rare EBV-positive centrocyte-like cells were mainly located in the germinal centers. The presence of HIV and EBV genome was also studied in lymph nodes involved by PGL, with the use of in situ and Southern blot hybridization. A positive reaction for HIV genome was detected in only 1 of 14 lymph nodes with the Southern blot hybridization, and the presence of EBV genome was never demonstrated in these lymph nodes with the use of both in situ and Southern blot hybridization. The expression of EBV antigens and genome was also investigated in the peripheral blood of 15 patients with PGL in which cells with positive results for EBV antigens were detected in a single case with a frequency of 1 X 10(-4). No evidence of EBV genome was found with the use of the in situ hybridization. These results suggest that EBV is not present in lymph nodes during the PGL phase and that its possible implication in the pathogenesis of acquired immune deficiency syndrome (AIDS)-associated lymphoma might be a late event.     

Tissue localization of complement component 3 receptor-bearing cells in lymphoid tissue after injection with complete Freund adjuvant.

Localizations of complement component 3 (C3) receptor (C3R)-bearing cells in lymph nodes obtained from normal guinea pigs or from guinea pigs inoculated with complete Freund adjuvant were examined by staining with fluorescein-labeled anti-guinea pig C3 antibody after treatment with aggregated rabbit immunoglobulin M bound with guinea pig complement. In normal lymph nodes, a small number of C3R-positive cells were observed in the cortical and medullary areas. Non-granulomatous lymph nodes from complete Freund adjuvant-inoculated animals showed a number of C3R-positive lymphocytes in a mantle zone of the secondary follicles between the follicles and medullary cords, whereas in the paracortical areas and germinal centers, only a few positive cells were scattered. Long-lasting existence of positive cells was seen in the epithelioid cell granuloma, although the staining patterns were different from those of the lymphocytes. The appearance of a number of C3R-bearing lymphocytes in lymph nodes from complete Freund adjuvant-inoculated animals might be an expression of adjuvant activity.     

CD95 (Fas) ligand expression of Epstein-Barr virus (EBV)-infected lymphocytes: a possible mechanism of immune evasion in chronic active EBV infection

The Epstein-Barr virus (EBV) induces infectious mononucleosis (IM) and can be associated with chronic active EBV infection (CAEBV). Cytotoxic T lymphocytes (CTL) play an important role in excluding EBV-infected cells. Two cytotoxic mechanisms of CTL have been demonstrated: one perforin/granzyme-based and the other Fas (CD95)/Fas ligand (FasL)-based. To clarify these two pathways in CAEBV, we analyzed six patients with CAEBV and four patients with IM using immunohistochemical staining of the lymph nodes. In both CAEBV and IM, CD8⁺ T-cells increased in number, but CD56⁺ natural killer cells were rare. In four of six cases with CAEBV, approximately half the lymphocytes were positive for T cell-restricted intracellular antigens (TIA-1), which were recognized by the cytolytic granules of CTL. In IM, the number of TIA-1 positive cells was smaller than that in CAEBV. Fas-positive lymphocytes were frequently encountered in both CAEBV and IM. However, FasL-positive lymphocytes increased in three of six patients with CAEBV, but not in patients with IM. Except for one case with CAEBV, the number of perforin- and/or granzyme-positive cells was small in number in both CAEBV and IM cases. In double-staining FasL and EBV in situ hybridization, FasL-positive EBV-infected lymphocytes were detected in CAEBV but not in IM. In CAEBV, the Fas/FasL pathway and not perforin pathways appears to play an important role in the pathogenesis. The data suggest that EBV-infected lymphocytes may evade immune attack through the expression of FasL.     

Lymphocyte function in experimental endemic syphilis of Syrian hamsters.

We have studied the changes in the lymph nodes, spleen and thymus that occur in inbred LSH Syrian hamsters infected with Treponema pallidum Bosnia A, the causative agent of endemic syphilis, as well as the B-cell responses of these infected animals to helper T-cell independent and dependent antigens. The lymph nodes increased significantly in weight up to 6 weeks after infection, and contained viable treponemes. No significant changes in the spleen weight were observed, and no viable treponemes could be recovered from the spleen. However, the size of the thymus decreased steadily during the course of the disease. The relative number of Ig+ cells (B cells) increased in the spleen and regional lymph nodes, whereas the relative number of T cells decreased during the course of infection. In both the spleen and lymph nodes, the relative number of macrophages increased initially and decreased thereafter in the form of a bell-shaped curve showing a peak at 4-6 weeks of infection. The ability of splenic lymphocytes from infected hamsters to mount a primary PFC response to pneumococcal polysaccharide type III (SIII), a helper T-cell independent antigen, was elevated throughout the course of infection. However, the splenic PFC response to sheep erythrocytes (SRBC), a helper T-cell dependent antigen, was increased only during the first 4 weeks of infection and progressively decreased thereafter. The PFC responses of infected lymph node lymphocytes to both SIII and SRBC were increased during the first 4 weeks and decreased thereafter. These data suggested that atrophy of the thymus seen in syphilitic infection is accompanied by the complex losses of subsets of T cells and altered B-cell functions. An early loss of suppressor T cells in both the lymph nodes and spleen occurs concomitantly with a loss of T helper cells and heterologous (treponema-unrelated) B-cell functions in the lymph nodes. Helper T cells are lost from the spleen only in the later stages of infection, whereas splenic B-cell functions remain intact throughout the course of the disease. These findings were further tested by in vitro methods where splenic and lymph node lymphocytes from infected hamsters were examined for their ability to respond to Con A in terms of the induction of antigen non-specific suppressor T cells. The mixing of Con A stimulated splenic or lymph node lymphocytes from infected hamsters was unable to inhibit the primary antibody responses of SRBC as compared to the normal control.(ABSTRACT TRUNCATED AT 400 WORDS)     

Differential cellular susceptibility to Epstein-Barr virus infection in a patient with X-linked lymphoproliferative disease

Epstein-Barr virus (EBV) is often associated with lethal lymphoproliferative diseases in immunologically compromised individuals. Recently, we have studied a 20-month-old boy with X-linked lymphoproliferative disease (XLP) who had succumbed to infectious mononucleosis (IM) complicated by fulminant hepatitis and virus-associated hemophagocytic syndrome following EBV infection. EBV genomes were detected in peripheral blood lymphocytes (PBL), cervical and mesenteric lymph nodes, liver, spleen, thymus, and bone marrow. According to restriction endonuclease analyses, the EBV-DNA pattern was similar in all samples except for the EBV-DNA from the bone marrow. Additionally, circular EBV-DNA (suggesting a latent infection) predominated in spontaneously established lymphoblastoid cell lines (LCLs) derived from both the lymph node and cord lymphocytes co-cultured with PBL. In contrast, both circular and linear EBV-DNA (suggesting a lytic infection) were noted in spontaneously established LCLs derived from his PBL. Furthermore, LCLs derived from both the lymph node and cord lymphocytes co-cultured with PBL expressed fewer reactive cells for early antigen (EA) and viral capsid antigen (VCA) than spontaneous LCLs from his PBL, thus providing evidence for different B cellular susceptibility to EBV infection in this patient with XLP. Finally, defective EBV-specific cytotoxic T cell activity was observed in this patient. Latent EBV infected cells may easily escape immunosurveillance by the host. These findings may explain the fatal course of EBV infection in this patient.     

Simultaneous occurrence of Hodgkin's lymphoma and Kaposi's sarcoma within the same lymph nodes of a non-AIDS patient

We report a case of simultaneous occurrence of Hodgkin's lymphoma and Kaposi's sarcoma within the same lymph nodes of a 61-year-old woman without human immunodeficiency virus (HIV) infection. Epstein-Barr virus (EBV) was detected in the Hodgkin cells and Reed-Sternberg cells by EBV LMP-1 immunostaining and Epstein-Barr virus encoded early RNA (EBER) in situ hybridization. In contrast, Kaposi's sarcoma cells were positive for human herpes virus 8. This case is documented because the occurrence of 2 independent tumors infected by 2 unrelated viruses within the same lymph nodes of a patient without HIV infection has rarely been observed.     

Local immunity in lung-associated lymph nodes in a murine model of pulmonary histoplasmosis.

Local immunity against acute pulmonary histoplasmosis was studied in the lung-associated lymph nodes of normal nonimmune mice infected intratracheally with live Histoplasma capsulatum yeasts. The phenotypes and distribution of cells in lung-associated lymph nodes and spleens were determined by flow cytometry. In addition, the immune responsiveness of these cells was evaluated by in vitro blastogenesis. Anti-H. capsulatum antibodies in serum and H. capsulatum antigen in tissue were measured by immunoassays. Cellular immune responses were greater in the lymph nodes than in the spleens. In lymph nodes 7 days after infection, a marked increase in the number of B lymphocytes caused the percentage to rise to 43%, compared with 26% in controls, and it remained elevated throughout the course of infection. A CD3+ cell that did not express CD4 or CD8 increased in number until it constituted 21% of lymph node cells, compared with 5% in controls, by day 14. The numbers of CD4+ and CD8+ T lymphocytes were modestly increased from days 7 to 35, but their percentages dropped because of the greater numbers of B lymphocytes and CD3+4-8- cells. Macrophages consistently constituted 2 to 3% of lymph node cells during the study. In spleens 7 days after infection, the percentage of macrophages in infected mice rose to 21%, compared with 9% in controls, but the total spleen cell number did not increase until day 14, when all cell subsets were nearly double in number. The in vitro blastogenic response of lymph node cells to H. capsulatum peaked at day 7, but spleen cell response was minimal during the course of infection. Histoplasma-specific serum immunoglobulin G antibodies reached peak levels by day 21 and remained high to the end of the study. In contrast, levels of antigen-specific immunoglobulin M antibodies were very low. These data suggest that antigen-specific immune responses occur in lung-associated lymph nodes and that this draining lymph node response may be an important component in host defense against Histoplasma lung infection.     

Migration of newly formed small lymphocytes from bone marrow to lymph nodes during primary immune response

Selective DNA labelling of bone marrow cells in vivo was used to determine the effect of antigenic stimulation on the migration of small lymphocytes from bone marrow to popliteal lymph nodes. Following footpad injection of keyhole limpet haemocyanin (KLH) in guinea-pigs the regional nodes showed an early increase in weight and cellularity together with a progressive increase in cell proliferation. When [³H]thymidine was injected into tibial and femoral marrow 2 days before KLH administration the DNA radioactivity of the KLH-stimulated nodes increased rapidly and always exceeded that of contralateral nodes. Simultaneously, in radioautographic sections of lymph nodes labelled small lymphocytes, indicative of an origin from marrow precursors, appeared throughout the cortex, post-capillary venules, subcapsular sinus, medullary cords and sinuses. In KLH-stimulated nodes the number of labelled small lymphocytes per section was higher than in contralateral nodes, especially in the cortex, and some of these cells appeared in germinal centres. Labelled large blast cells and macrophages were also increased in numbers. Similar changes were observed in lymph nodes of parental strain rats following intramyeloid [³H]thymidine administration and footpad injection of lymphoid cells from F1 hybrid rats. The results demonstrate that, during the early response of lymph nodes to various antigens, local changes in cell traffic include an enhanced accumulation of newly formed small lymphocytes, putative virgin B lymphocytes, generated in the bone marrow prior to the antigenic stimulation.     

HHV-6 and EBV DNA quantitation in lymph nodes of 86 patients with Hodgkin's lymphoma

Human herpesvirus (HHV-6) and Epstein-Barr virus (EBV), are two ubiquitous human herpesviruses which share many common features although they belong to different sub-families. In particular, both viruses are found in lymph nodes of patients suffering from Hodgkin's lymphoma. The aim of this study was to detect and to quantify independently HHV-6 and EBV by a real-time PCR in lymph nodes from 86 patients with Hodgkin's lymphoma. EBV quantitative method was compared with LMP-1 protein detection among the same samples. EBV genome was detected for 61.6% of the patients (53/86) and the highest prevalence of this virus was observed in Hodgkin's lymphoma with mixed-cellularity histopathological type (80%). In contrast to that, HHV-6 genome was detected for 79.1% of the patients (68/86) and was most observed in the nodular-sclerosis group (83.6%). Among the 68 HHV-6 positive samples, 63 belonged to the B subtype. A large number of biopsies (47.7%) were positive for both viruses whereas a little number (7%) was negative for both. EBV quantitation and LMP-1 immunohistochemistry were correlated statistically but this latter technique was less sensitive. Among the nodular-sclerosis patients, HHV-6-/EBV+ patients were significatively older than HHV-6+/EBV- patients. Patients infected dually had higher values of quantitation for each virus than those positive for one virus. Data of the clinical follow-up obtained by diagnosis and during the treatment of 83 patients, were correlated with the virological findings.     

In Hodgkin's disease Reed-Sternberg cells and normal B-lymphocytes are infected by related Epstein-Barr virus strains

In Hodgkin's disease (HD), both neoplastic Reed-Sternberg (RS) cells and bystander B-lymphocytes may be infected by Epstein-Barr virus (EBV). We postulated that if tumorigenic EBV strains did exist, they would be preferentially found in consistently EBV-associated tumors, such as RS cells, and differ significantly from the strains present in other, non-pathological sites of the same patients. In the present study we have compared LMP1-BNLF1 polymorphism of EBV strains infecting RS cells and B-lymphocytes in lymph nodes effected by HD on the one hand, and bystander B-lymphocytes in reactive lymph nodes on the other. It appeared that viral strains detected in HD tissues including RS cells and bystander B-lymphocytes were infected by different, but related EBV strains and were four times more polymorphic than EBV strains infecting bystander B-lymphocytes of reactive lymph nodes. The question arises as to the biological significance of these observations and the origin and chronology of multiple infections in the same patient. Since RS cells are derived from B-lymphocytes it is conceivable that the latter events could have occurred during the proliferation of bystander B-lymphocytes and their EBV episome following an antigenic stimulation.     

Selective labeling of mesenteric lymph nodes: cell production and emigration of newly formed lymphocytes to other organs

Lymphocyte production by mesenteric lymph nodes of normal young pigs was studied by intranodal injections of either tritiated thymidine or tritiated deoxycytidine as DNA precursors. One or two days after selective labeling of the mesenteric lymph nodes the relative and absolute number of lymphocytes derived from mesenteric lymph nodes were determined autoradiographically in the following organs: mesenteric, cervical and inguinal lymph nodes, spleen, thymus, bone marrow, Peyer's patches, tonsil, different regions of the gut, lung and liver. The overall cell production of mesenteric lymph nodes, as derived from the sum of all labeled cells one day after labeling, was estimated to be about 7 X 10(9) lymphocytes. Up to 40% of all newly formed lymphocytes had already left the lymph nodes within one day and were found in all organs studied. There was a preferential homing to the mucosa of the small intestine, but a considerable number migrated to the spleen and even to the thymus and bone marrow. In lymphoid organs all labeled cells were small and medium-sized lymphocytes one and two days after labeling. In cervical lymph nodes, spleen, tonsil and Peyer's patches the relative distribution to T and B cell areas was determined. There was an obvious preference of newly formed lymph node cells to home to T cell areas. The differences of labeling between thymidine or deoxycytidine were surprisingly low.     

Epstein-Barr virus interactions with human lymphocyte subpopulations: virus adsorption, kinetics of expression of Epstein-Barr virus-associated nuclear antigen, and lymphocyte transformation.

In order to further understand Epstein-Barr virus (EBV)-lymphocyte interactions, we investigated a chain of events including: (i) EBV binding to human lymphocyte subpopulations; (ii) the earliest appearance of EBV-determined nuclear antigen (EBNA) in the lymphocytes after EBV infection; and (iii) establishment of continuous lymphoblastoid cell lines (LCL) by infecting with EBV different types of lymphocyte preparations from the same as well as from different donors. By using direct membrane immunofluorescence assay, we found that only a small fraction of human peripheral blood and cord blood lymphocytes (CBL), and possibly less than 31% of the T cell-depleted lymphocyte population, carry receptors for P3HR-1 strain of EBV. The number of cells carrying receptors for EBV did not vary considerably among different blood lymphocyte populations from several normal donors. EBV adsorption on lymphocyte subpopulations showed that purified thymus-dependent (T) cells and thymocytes did not adsorb EBV, in contrast to T cell-depleted lymphocyte populations and lymphoid cells from fetal liver and spleen. In CBL infected with EBV strain B95-8, EBNA was detected by anti-complement immunofluorescence as early as 18 h after infection. This indicates that EBNA is the earliest detectable EBV-determined intracellular antigen to appear after infection and before or during lymphocyte transformation by EBV. Transformation was observed only in lymphocyte cultures containing detectable thymus-independent B cells but not in cultures of purified T cells. With one exception (es-b-1), all the EBV-transformed LCL from different origins carried surface-bound immunoglobulins (a B cell marker). These included also the 10 LCL obtained by infecting cultures of adherent cells from different donors. With regard to its surface markers, ES-B-1 appeared to be an exceptional EBV genome-carrying line, and it also lacked the ability to form spontaneous rosettes with sheep erythrocytes (a T cell marker). Therefore, it is possible that ES-B-1 was derived from an atypical B cell or B cell precursor or from a so-called "null cell" transformed by EBV.     

T cell-rich B cell lymphoma bearing Epstein-Barr virus in tumor cells: A case of IBL-T-like lesion following Lennert's lesion

A case of T cell-rich B cell lymphoma (TCRBCL) with Epstein-Barr virus (EBV) infection in tumor cells is reported. A 50 year old male developed right cervical lymph node swelling in July 1988. Initial biopsy in April 1989 demonstrated many scattered Hodgkinoid atypical cells with Lennert's lesion. After partial remission following chemotherapy, the lymph nodes enlarged again, and a second biopsy in February 1991 showed an IBL-T-like lesion. Only a small number of Hodgkinoid atypical cells were still observed. After apparently, complete remission, the lesion soon recurred and the patient died in November 1992. Immunohistochemically the Hodgkinoid cells were positive for L26, but negative for LN2, LN3, UCHL-1, MT1, lysozyme, Ber-H2 and Leu-M1. Reactivity for immunoglobulins showed falsepositive because of poly-clonal staining. IgH monoclonality was detected by the poly-merase chain reaction method in the first biopsied specimen, and by Southern blotting in the second biopsied snap-frozen specimen. Monoclonal TCRβ rearrangement was not detected. The Hodgkinoid atypical cells were positive for EBVencoding RNA by in situ hybridization, and LMP-1 by immunostaining. Occasionally, EBV-bearing immunoblastic, medium sized, or small lymphocytic cells were also observed. This case indicates the possibility that EBV is related to the pathogenesis of TCRBCL.     

Epstein-Barr virus (EBV) -induced CD30 + natural killer cell-type malignancy resembling malignant histiocytosis: Malignant transformation in chronic active EBV infection associating hypogammaglobulinemia

A 27-year-old male suffered from Epstein-Barr virus (EBV)-related liver dysfunction with persistent hypogammaglobulinemia. IgG titers to EBV antigens were significantly high, while other hepatitis markers were negative. Liver biopsy disclosed active intralobular inflammation. Two years later, he manifested persistent fever, leukopenia, effusions and hypopmteinemia. and his general condition worsened progressively. the peripheral blood small lymphocytes predominantly expressed natural killer (NK)-like phenotypes (CDP, CD7+, CD16+, CD56+). Hepatosplenomegaly and marked elevation of serum lactic dehydrogenase were observed. He died of respiratory failure at the age of 29. At autopsy, the liver (2190 g), spleen (860 g), small bowel and mesenteric lymph nodes showed massive Infiltration of large atypical lymphoid cells in close association with hemophagocytic histiocytes. Involvement was mildly noted also in the bone marrow, lungs, gall-bladder and kidneys. The atypical cells belonged to CD30+ activated NK-type cells expressing CD2, cytoplasmic CD3, CD7, CD45RO, CD56, HLA-DR and HLA-DQ.T cell receptors (TCR), surface CD3, CD4, CD5 and CD8 were not expressed. Epstein-Barr virus-related small nuclear RNA (EBER1) and Epstein-Barr virus-associated nuclear antigen 1 were detected In the nuciel of a significant number of atypical cells, while EBV-related latent membrane protein-1 was negative. EBER1 was also Identified In the nuclei of non-neoplastic small lymphocytes at both biopsy and autopsy. Monoclonal integration of the EBV genome into the lymphoma cells was shown by Southern blot analysis. Clonal rearrangement of TCR was undetectable. Roles of chronic active EBV infection in the development of NK cell-type malignancy resembling malignant histiocytosis are discussed.     

Detection of the Epstein-Barr virus in primary gastric lymphoma by in situ hybridization

The Epstein-Barr virus (EBV) has been shown to be associated with numerous human malignancies including Burktt's lymphoma and nasopharyngeal lymphoepithelioma. In addition, some typical gastric adenocarcinomas were also recently reported to demonstrate EBV relevance. The present study was designed to detect EBV in primary gastric lymphoma, using the in situ hybridization (ISH) method, in which oligo-nucleotide probes for the EBERl RNA and the EBV DNA W region have been used. Of the 49 cases of primary gastric lymphoma studied, which all showed B cell immunopheno-type, EBER1 sequences could only be found in four cases, including two low-grade cases and two high-grade cases of histological subtypes while the number of positive cells was less than 50% of the tumor cells. In one case of low-grade mucosa associated lymphoid tissue (MALT) lymphoma, the EBER1 -positive neoplastic cells were found in the regional lymph node, but the primary site of the stomach showed no positive signals. The EBV presence was further confirmed by the EBV DNA ISH. Using the ISH method, rare or occasional positive lymphoid cells (probably non-tumorous bystander cells) could be detected in 10 other cases including all histological subtypes. The present study shows that only a small proportion of primary gastric lymphoma is associated with EBV, and such positive cases could be found in both high- and low-grade histological subtypes. It is also suggested that the EBV presence in the neoplastic cells of some cases of primary gastric lymphoma is most likely a secondary phenomenon.     

A pathological and immunohistological case report of fatal infectious mononucleosis,Epstein-Barr virus infection,demonstrated by in situ and Southern blot hybridization

We present an autopsy case of 20-month-old boy who had a fulminant course of infectious mononucleosis, with severe hepatic failure. Autopsy revealed marked infiltration of immunoblasts in the lymph nodes, liver, spleen, thymus and kidneys. We identified a large number of Epstein-Barr virus (EBV) genomes in the immunoblasts of the lymph nodes, liver and spleen by in situ hybridization. EBV genomes were also detected in the liver and spleen by Southern blot hybridization. Histology of the liver revealed diffuse feathery degeneration of the hepatocytes. However, EBV genomes were not detected in the hepatocytes by in situ hybridization and monoclonal antibody studies. Immunostaining of the autopsy liver specimen revealed a large number of suppressor/cytotoxic T cells (Leu2a positive) in the portal areas and of natural killer (NK) cells (Leu7 positive) in the portal areas and sinusoids of the liver. We therefore suggest that the hepatocellular damage was not caused by the viral replication in the hepatocytes but was mainly caused by the abnormal killer cell activity of the suppressor/cytotoxic T cells and NK cells.     

Carcinoma of stomach and breast with lymphoid stroma: localisation of Epstein-Barr virus.

AIMS--To determine the presence of Epstein-Barr virus (EBV) genome in six patients (three with gastric and three with breast carcinoma) with severe small lymphoid cell infiltration. METHODS--The polymerase chain reaction and in situ hybridisation were used to detect EBV genome. The number and distribution of T and B lymphocytes were evaluated by immunohistochemistry. RESULTS--Histologically all of the patients had poorly differentiated tumours. Immunohistochemistry showed that T cells predominated in three cases, B cell in two, and almost equal numbers in one case. PCR showed that the EBV genome was present in two cases each of gastric and breast carcinoma. In situ hybridisation for EBV genome provided positive signals only in the small lymphoid cells in one gastric and two breast carcinomas giving a positive reaction for EBV genome by PCR. The gastric and breast cancer cells did not give positive signals. CONCLUSIONS--Severe lymphoid cell infiltration in gastric and breast carcinoma does not necessarily indicate that these tumours are associated with EBV. Larger numbers of cases will need to be studied to confirm this.     

Encoded latent membrane protein 1 of Epstein-Barr virus on follicular dendritic cells in residual germinal centres in Hodgkin's disease.

AIMS--To determine if there is an association between Epstein-Barr virus (EBV) infection and Hodgkin''s disease. METHODS--Fifty cases of Hodgkin''s disease and 25 reactive lymph nodes were screened for the presence of EBV-RNA (EBER) using in situ hybridisation, and for the expression of EBV encoded latent membrane protein 1 (LMP-1) by immunohistochemistry. RESULTS--In 42% of the cases of Hodgkin''s disease, EBER was detected in the nuclei of the malignant cells, and in LMP-1 expression was found 36%. Both EBER and LMP-1 positivity were seen in 34% of the cases. An additional finding was the presence of LMP-1 on follicular dendritic cells in residual germinal centres in two cases of Hodgkin''s disease. EBER was not detected in these germinal centres. In reactive lymph nodes only occasional EBER positive, small, lymphoid cells were found, without LMP-1 expression. CONCLUSIONS--These results show a strong correlation between the presence of EBER and the LMP-1 expression in the Reed-Sternberg cells. They corroborate a role for EBV in at least some cases of Hodgkin''s disease. LMP-1 is probably presented as an immune complex in the germinal centres, as part of an immune response against EBV.     

Scanning electron microscopy of lymphoid cells from leukemia virus-infected mice

Spleen, lymph node, bone marrow, and thymus cells from Friend leukemia virus (FLV)-infected mice were examined by scanning electron microscopy. Whereas splenocytes from normal, noninfected animals showed the expected morphological classes of lymphocytes, including cells with numerous villous projections and smoother cell types, spleen cells from mice infected with FLV showed a rapid alteration of surface morphology. Shortly after infection, a decrease in the number and percentage of villous cells occurred, with a concomitant increase in the number of cells that were larger and smoother. Within 10 to 20 days after infection, the majority of splenocytes were smooth, large cells showing many distinct morphological charges, including surface "holes" and a "spongy" appearance. By days 25 to 35 after infection, most splenocytes were abnormal in appearance. Similar changes occurred in the lymph nodes after FLV infection, but the rate of change was much lower. Abnormal and larger smooth-surfaced cells did not become prominent until after week 2 or 3 infection. Thymus and bone marrow cells showed little if any change in surface morphology until late in the infectious process. However, even at that time only a few of the cells were abnormal in appearance. The changes in cell population in the spleen but not the lymph nodes paralleled the rapid decrease in the percentage of cells which stained positive for surface immunoglobulin and theta antigen. Futhermore, FLV antigen rapidly appeared on spleen cells after infection; fewer lymph node cells were positive, and only low numbers of marrow and thymus cells stained positive for FLV antigen. The marked immunosuppression induced by FLV infection paralleled and in some instances preceded the marked morphological changes.     

鼻咽原发性腺样囊性癌及其与EB病毒感染的关系

目的 探讨鼻咽癌高发的广州地区鼻咽原发性腺样囊性癌的临床病理特征及其与EB病毒感染的关系。方法 对17例鼻咽原发性腺样囊性癌进行临床资料分析。除HE、阿辛蓝及过碘酸Schiff(PAS)常规染色外,采用标记链酶素卵白素生物素(LSAB)免疫组织化学法检测一系列上皮性标志物、CD21和EB病毒编码的潜伏膜蛋白(LMP)1的表达。采用原位杂交法检测EB病毒编码的小RNAs(EBER)。用巢式聚合酶链反应(PCR)法检测EB病毒DNA W片段。结果 17例鼻咽原发性腺样囊性癌患者的男女之比为7：10,年龄30-63岁,中位年龄46岁。17例中14例的肿物呈明显局部侵袭性生长,病变属T3或T4期,仅1例有同侧颈淋巴结转移。大部分肿瘤细胞呈基底细胞样,核小而深染、仅有少许胞质;细胞之间常可见伊红染色同质性或黏液样物质;16例可见筛孔状的特征性结构。鼻咽原发性腺样囊性癌均表达广谱细胞角蛋白和上皮膜抗原。9例(9／17)可见少数癌细胞呈EBER阳性,采用巢式PCR法同时检测到这9例活检组织192bp的EB病毒W片段阳性带,EBER阳性癌细胞的病例中,5例的癌细胞表达LMPI;其中3例的肿瘤间质中又可见少数EBER阳性的浸润性淋巴细胞。所有17例的癌细胞,包括EBER阳性的癌细胞均不表达CD21。结论 鼻咽原发性腺样囊性癌患者以女性为多见。肿瘤生长呈明显的侵袭性,常侵犯鼻咽及其周围组织。鼻咽原发性腺样囊性癌与EB病毒感染的关系并不密切。