Increased incidence of IgA antibodies to the Epstein-Barr virus-associated viral capsid antigen and early antigens in patients with chronic lymphocytic leukemia

Antibody titers to Epstein-Barr virus (EBV)-associated early antigens (EA) and the viral capsid antigen (VCA) were determined by ELISA on 263 sera obtained from healthy donors, patients with Hodgkin's disease (HD), non-Hodgkin lymphomas (NHL), infectious mononucleosis (IM), Burkitt's lymphoma (BL), and nasopharyngeal carcinoma (NPC). As expected, most lymphoma patients showed markedly elevated anti-VCA IgG and anti-EA IgG antibody titers. Only one patient in the NHL group (n = 56) consisting of patients with lymphomas other than chronic lymphocytic leukemia (CLL) and hairy-cell leukemia (HCL), and 3 patients with HCL (n = 19) had high antibody titers of the IgA class to VCA and EA. Seventeen out of 48 patients (36%) with CLL had high IgA anti-VCA titers and 10 of these sera (21%) also contained IgA anti-EA. The geometric mean titer (GMT) of IgA anti-VCA was 2,510, the GMT of IgA anti-EA was 780. These antibody titers were about 10 times lower than the corresponding GMT of the NPC patients investigated in this study. The elevated IgG and IgA antibody titers to VCA and EA in CLL and HCL patients seem to reflect an immunodeficiency secondary to the malignant disease leading to reactivation of latent EBV infection. The possibility that at least some of these B-cell lymphomas are associated with EBV cannot be excluded.     

Serologic studies in hairy cell leukemia: high prevalence of Epstein-Barr and cytomegalovirus antibodies and absence of human T-cell lymphotrophic viruses antibodies

Serum from 60 patients with hairy cell leukemia (HCL) were studied for the presence and the titers of antibodies to Epstein-Barr virus (EBV), cytomegalovirus (CMV) and human T-cell lymphotrophic viruses (HTLV). Eighty-three percent of the patients were seropositive for EBV, with a (reciprocal) geometric mean titer (GMT) of 960. Seventy-eight percent of the patients had antibodies to CMV with a GMT of 435. All 21 patients tested for HTLV I and HTLV III were seronegative; only one patient showed detectable antibodies to HTLV II. The potential role of these infections in the physiopathology of HCL is discussed.     

Specific neutralizing antibodies in Epstein-Barr virus associated diseases

The results of Epstein-Barr virus (EBV) neutralization tests on 650 sera from patients with EBV-associated diseases and controls were compared with the titers of antibodies to EB viral capsid antigens (anti-VCA) and to EBV-induced early antigens (anti-EA). All anti-VCA negative sera failed to neutralize EBV. Among 546 anti-VCA positive sera, only eight yielded negative results in neutralization tests. Four of these came from patients in the early acute phase of infectious mononucleosis who subsequently developed neutralizing antibodies. The ratios between neutralizing and anti-VCA titers varied over a wide range indicating that different antibodies are involved in the two tests. In line with the regular presence of mostly elevated anti-VCA titers, the geometric mean neutralizing titers in Burkitt's lymphoma and nasopharyngeal carcinoma were also higher (5- to 7-fold) than in appropriate controls. In Hodgkin's disease, an over-representation of high neutralizing as well as anti-VCA titers was noted, although some patients had neither of these antibodies. All patients with infectious mononucleosis developed neutralizing antibodies with a geometric mean ultimately matching that of controls whereas in the early acute phase it was lower than in the controls. In contrast, the geometric mean anti-VCA titer in the early acute phase was higher than in controls and then declined in convalescence toward the control value. There was no evident relationship between neutralizing and anti-EA activities whether directed against the D or the R component of the EA complex.     

Epstein-Barr virus infections in Brazil. II. Hodgkin's disease.

Sixty-seven cases of Hodgkin's disease (HD) occurring in S?o Paulo, Brazil, were studied. Males with HD predominated over females 2.3 to 1. Sixty-six percent of the cases occurred in patients under 30 years of age, 31.7% under 20 years of age, and only 7.5% after 50 years of age. Lymphocyte predominance and mixed cellularity histologic types were most common in patients less than 15 years old, and nodular sclerosis was most common in the 15- to 19-year-old group. Sera from all patients had antibody to the viral capsid antigen (VCA) of Epstein-Barr virus (EBV). The geometric mean titer (GMT) of VCA antibody with the use of Jijoye cells as antigen was 1:162, and 31.3% of patients had titers of 1:320 or more; in controls, the GMT was 1:67 and 3.8% had titers of 1:320 or more. Similar results were obtained when EB-3 cells were used as antigen. The highest titers occurred in males, in mixed cellularity and lymphocyte depletion forms, and in stage 2 of illness. EBV-specific IgM antibody and heterophile antibody levels were not elevated, but 20.5% of the HD patients had antibody to the early antigen of EBV present in their sera. Antibody levels for herpes simplex virus, cytomegalovirus, rubella, measles, parainfluenza viruses, and papovavirus were not significantly elevated over those in matched controls.     

Antibodies to Epstein-Barr virus-associated nuclear antigen and to other viral and non-viral antigens in Hodgkin's disease.

Antibody reactivity to Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) was investigated by means of the anticomplement immunofluorescence technique on sera from patients with Hodgkin's disease (HD) and from appropriate controls. Antibody levels to other EBV-determined antigens, i.e. viral capsid (VCA) and early antigens (EA), and to measles and rubella viruses, to cytomegalovirus (CMV), and to toxoplasma gondii were also measured. The results of anti-EBV antibody titrations demonstrated that anti-VCA, anti-EA and anti-EBNA reactivity was significantly higher in HD patients than in healthy subjects. There was no significant difference between the distribution of high rubella and measles antibody titers in HD and control sera. The GMT and the incidence of high titer anti-CMV and toxoplasma antibodies were greater in HD patients than in controls. The analysis of the data, according to histological subtypes, showed that the condition of lymphocyte depletion was associated in HD patients with the highest anti-EBNA antibody levels and the lymphocyte predominance with the lowest. This pattern seemed to be peculiar for anti-EBV reactivity, since anti-CMV and anti-toxoplasma antibody levels in the lymphocyte-depleted group of patients did not significantly differ from those of controls. No correlation was found between anti-VCA and anti-EBNA in individual sera of HD patients. This observation suggests that different mechanisms are probably responsible in HD for the release of EBV-related antigen from infected cells.     

Antibody to Epstein-Barr virus and cellular immunity in Hodgkin's disease and chronic lymphatic leukemia

Antibodies to Epstein-Barr virus capsid antigen (VCA), early antigen (EA) and cellular immunity as measured by skin reactivity to keyhole limpet hemocyanin (KLH) and Brucella antigen (BA) were measured in 15 cases of Hodgkin's disease (HD) and 14 cases of chronic lymphatic leukemia (CLL) before treatment and one year later. Both groups of diseases were associated with elevated VCA and EA antibody levels when compared with 18 controls, and in both groups the mean titers were unchanged after therapy. A negative skin test to KLH was associated with a short survival in the HD group and was found more frequently in CLL patients with active disease. In the young HD patients, a higher EBV titer was found in pretreatment sera of non-survivors compared with those who are still alive. There was no correlation, however, between tests for cell-mediated immunity and humoral antibodies against EBV. The finding of similar titers in patients with depressed and normal skin reactivity indicates that the elevated titers are probably not the result of a non-specifically depressed immune defense.     

Longitudinal study of Epstein-Barr virus antibody titers and excretion in pediatric patients with Hodgkin's disease.

Nineteen pediatric patients with Hodgkin's disease (HD) who had experienced primary Epstein-Barr virus (EBV) before, or in one case after, diagnosis, were studied longitudinally for changes in the titers and spectra of EBV-related antibodies, excretion of EBV into the oropharynx, the number of EBV-carrying lymphoid cells in the peripheral blood, and clinical signs and symptoms suggestive of reactivation of the latent virus. The incidence and geometric mean titers of IgG antibodies to viral capsid antigen (VCA) in the HD patients at the time of diagnosis and in the controls were similar. The anti-VCA titers of the patients rose above control levels during and after therapy and remained elevated for up to 7 years of observation. At no time were heterophil or VCA-specific IgM antibodies detected. Antibodies to EBV-induced early antigens were more common in patients (ultimately 80%) than in controls (9%). In contrast, antibody levels to EBV-associated nuclear antigen were disproportionally low in the patients. Excretion of EBV was noted at increased frequency in the patients but the number of circulating, EBV-carrying lymphoid cells was the same as in controls. No discrete clinical syndrome was associated with rising antibody titers or viral excretion. While these results are best explained by a presumed reactivation of the persistent EBV infection by immunosuppressive effects of HD or its therapy, they have not provided direct evidence for this suggestion.     

Elevated antibody levels to Epstein-Bbarr virus antigens in patients with hairy cell leukemia compared to controls in relation to exposure to pesticides, organic solvents, animals, and exhausts

Epstein-Barr virus (EBV) is a B-lymphotropic human herpes virus infecting B-cells, which has been associated with lymphoid malignancies, above all non-Hodgkin lymphomas (NHL). Severe immunosuppression is the best recognized risk factor for NHL. Many factors in the environment that have been described as risk factors for NHL cause measurable changes in immune functions. Hairy cell leukemia (HCL) is a rare, indolent non-Hodgkin lymphoma, originating from B-lymphocytes. This was a case-control study including 111 male cases with HCL and 400 controls. In a subgroup of 57 cases and 65 controls analysis of antibodies to EBV early antigen, viral capsid antigen, and EBNA-1, measured as P107, was performed. In this study, we confirm other studies describing elevated levels of antibodies to the EBV early antigen (EA) in patients with HCL compared to controls. We found only minor differences in the levels of antibodies to the viral capsid antigen (VCA) and EBNA-1. measured as P107. We found a positive association of a titer to EA IgG > or =40 (OR 4.1; CI 1.9-9.5). The ORs were further elevated when subjects with high levels of EA IgG and exposed to environmental agents such as organic solvents, certain pesticides, impregnating agents, animals, and exhausts were compared to those subjects with low levels that were not exposed. Antibody reactivity against the EBV EBNA 1-alanine-glycine repeat (P107 IgG) above the median gave an increased OR for HCL, which further increased in subjects exposed to organic solvents, certain pesticides. impregnating agents, animals, and exhausts. However, the numbers of exposed cases and controls were small in some of the calculations.     

EBV infections in Brazil. I. Occurrence in normal persons, in lymphomas and in leukemias

Comparisons of IF antibody tests with EB3 and Jijoye cells indicated an overall agreement of 80%. However, the Jijoye line results in a higher frequency of positives at 1:10, a higher frequency of elevated titers of 1:160 or more, and a higher geometric mean antibody titer than with EB3 cells. At titers of 1:320 or higher the results with the two lines were quite comparable and are taken as elevated. Antibody to EBV is widely present in sera from Brazilian residents. Infection occurs early in life in the lower socioeconomic groups. In the middle and upper economic groups over 50% have antibody by age 12. In 69 lymphoma cases tested, all except one (Hodgkin's disease) had EBV antibody demonstrable at 1:10 titer or higher. Elevated EBVantibody titers at levels of 1:320 or higher were found in 41.9% of sera from the 43 cases of Hodgkin's disease having antibody and 4 of 8 cases of Burkitt's lymphoma; the geometric mean titers (GMT) were 1:173.4 and 1:246.8 respectively as compared to 1:51.4 in controls. In 13 cases of lymphosarcoma 23.1% had titers of 1:320 or higher and the GMT was 1:104.4. ERV antibody titer was present but not elevated in four cases of reticulosarcoma. Of 43 leukemia patients, one lacked EBV antibody at 1:10. Of the 42 possessing antibody, 31.0% had elevated titers with a GMT of 1:142.5. The highest titers occurred in mute myeloid leukemia.     

Epstein-Barr virus-associated antibody pattern in untreated non-Hodgkin lymphoma patients. Relationship to clinical variables and lymphocyte functions

Sera from 296 unselected and untreated patients with non-Hodgkin lymphoma (NHL) classified according to the Rappaport and the Kiel systems were analzyed for antibodies to Epstein-Barr virus (EBV). The aim of the study was to determine whether antibody spectra and liters to EBV-coded antigens correlated to clinical and immunological variables and whether the liters were of any prognostic significance. Increased antibody titers to EB viral capsid antigen (VCA) and slightly raised titers to early antigens (EA) of the diffuse (D) and restricted (R) types were noted frequently. Anti-VCA antibody titers correlated to clinical stage and age of the patients but not to histological. subgroups according to the Rappaport or the Kiel classification systems. However, and-VCA titers ? 1:2560 were seen only in diffuse lymphomas according to the Rappaport and in non-follicle cell-derived lymphomas according to the Kiel classifications. Patients with complement-receptor-positive diffuse lymphomas had higher anti-VGA titers than complement-receptor-positive nodular cases. Anti-VCA titers also correlated positively to serum IgG levels (p <0.01). Total number of lymphocytes separated from peripheral blood and mitogen induced (ConA, PWM) DNA synthesis were recorded before treatment in 54 of the patients. The patients exhibited a significant lymphocytopenia as well as a significantly reduced lymphocyte response to mitogens (p <0.001) compared to healthy controls. Elevated anti-VCA titers and anti-EA titers correlated to a good mitogen-induced lymphocyte response (p <0.05). Only anti-D 1:40 at diagnosis predicted a poor prognosis.     

The reliability of IgA antibody to Epstein-Barr virus (EBV) capsid antigen as a test for the diagnosis of nasopharyngeal carcinoma (NPC)

Since patients with nasopharyngeal carcinoma were first reported to have elevated levels of IgA antibody to Epstein-Barr virus (EBV) in their sera, workers in a number of countries have studied the possibility that this assay could be used in the diagnosis and monitoring of patients with this disease. In the United States, a collaborative project involving seven centers has been established to investigate the potential value of IgA antibody to EBV viral capsid antigen (VCA) as a clinical tool. In this report, we will summarize the results obtained from three studies: a comparison of EBV serology in three laboratories; a retrospective study of 37 nasopharyngeal carcinoma (NPC) patients and controls, and a prospective study of 126 NPC patients and 683 controls, including 149 patients with other malignancies involving the head and neck. The study of testing comparability in three laboratories demonstrated the feasibility of using this assay in a number of laboratories. The retrospective study confirmed the difference in IgA antibody titers between NPC patients and matched controls. The prospective study showed a relationship between IgA antibody titers and histopathology but not disease stage. IgA antibody titers were elevated more frequently in patients with nonkeratinizing or poorly differentiated types of NPC than for the well-differentiated squamous cell carcinomas. While IgA antibodies to EBV VCA appear to be of value in the early detection and diagnosis of NPC, it is possible that additional serologic tests for immunity to EBV, such as IgG antibody to VCA or early antigen (EA), will improve even further the clinical value of EBV serology in the management of NPC.     

Clinical and laboratory observations in a lymphoma-prone family

Three sisters developed non-Hodgkin's lymphoma (NHL). Thirteen maternal relatives had adenocarcinomas of various types, predominantly breast and large bowel. Detailed laboratory assessment of this family revealed a variety of immunologic abnormalities. Two lymphoma patients had elevated anti-early antigens (EA) Epstein-Barr virus (EBV) titers. One of the two also had a markedly elevated anti-viral capsid antigen (VCA) EBV titer. The two lymphoma patients were human leukocyte antigen (HLA)-identical, but two unaffected siblings shared the same HLA haplotypes. One of the lymphoma patients and four unaffected family members were anergic; and three unaffected family members had a decreased percentage of T-cells. These data suggest that a subtle disturbance in cellular immune functions may play a causal role in some familial aggregations of malignant lymphoma.     

The relation between Epstein-Barr virus antibodies and clinical symptomatology and immunodeficiency in patients with Hodgkin's disease.

Anti-Epstein-Barr virus (EBV) titers were measured in the sera of 37 patients with Hodgkin's disease and in 40 normal controls. The patients were grouped according to histologic type, clinical symptomatology (relapse or remission), and their immune state (immunodeficient or non-immunodeficient). Anti-Epstein-Barr nuclear antigens (EBNA) and antiviral capside antigens (VCA) titers were higher in patients with Hodgkin's disease than in the controls. Anti-EBNA titers were significantly higher in patients with lymphocyte predominance, and anti-VCA titers were significantly higher in patients with mixed cellularity. Patients in clinical relapse had higher anti-EBV antibody titers than patients in remission or those in the control group. Immunodeficent pateints had significantly higher anti-VCA titers than either the non-immuno-deficient or the control cases. We believe high anti-EBV titers are related to immunodeficiencies. The relationship between Hodgkin's disease and EBV is discussed.     

Human nasopharyngeal carcinomas positive for Epstein-Barr virus DNA in North America

Nasopharyngeal carcinomas (NPC) from 2 black patients and 1 Caucasian patient were positive for Epstein-Barr virus (EBV) DNA. Of the tumors, 2 were lymphoepitheliomas (undifferentiated NPC) and 1 was a moderately differentiated NPC. All 3 patients had high IgG titers against EBV early antigen and high IgG and IgA titers against virus capsid antigen (VCA). In one patient, the levels of anti-VCA IgA were different than those of anti-VCA IgG over the course of the disease. Our data support the association of EBV and NPC in North America.     

Antibodies to Epstein-Barr viral capsid and early antigens associated with Burkitt's lymphoma and lymphoblastic lymphosarcoma in Israel.

Before and/or after chemotherapy was administered to patients with Burkitt's lymphoma (BL) or lymphoblastic lymphosarcoma (LLS), their sera and those of matched controls were tested for antibodies to Epstein-Barr virus (EBV) capsid antigen (VCA) and early antigen by the indirect immunofluorescence method. Ten of the 16 BL patients were Arab children and 8 of the 11 LLS patients were Jews of Asian-African origin. Although half the BL patients did not have elevated antibody titers when their disease was diagnosed, significantly higher ones were detected in the BL group as compared with the LLS patients and their matched controls; Arab patients had the highest titers. IgM antibodies specific for VCA were found in 2 patients concurrently with elevated titers. We found no correlation between the clinical course of BL and the patients' antibody titers to EBV.     

Lack of correlation between EBV serology and presence of EBV in the Hodgkin and Reed-Sternberg cells of patients with Hodgkin's disease

Epstein-Barr virus (EBV) is detected in Hodgkin and Reed-Sternberg (HRS) cells in up to 50% of patients with Hodgkin's disease (HD). HD patients have been reported to express high serum titers against EBV antigens, even prior to the diagnosis of HD. Patients with high serum titers have a poorer prognosis. The aim of this study was to examine the relationship between the presence of EBV in HRS cells and the antibody titers reactive with different EBV antigens. Frozen serum and histopathological tissues were available from 107 untreated HD patients diagnosed between 1979 and 1991. The presence of EBV in the HRS cells was evaluated with immunohistochemistry directed against the LMP-1 antigen and/or with in situ hybridization of EBER-1. Analyses were performed of serum titers against early antigen (EA), diffuse (IgA and IgG) and restricted (IgG), virus-capsid antigen (VCA) (IgA and IgG), and EBV-encoded nuclear antigens (EBNA, EBNA 1, EBNA 2A, EBNA 2B, EBNA 6). EBV was detected in 27/107 (25%) tumor specimens, with a higher proportion in the MC group 8/13 (62%) (p < 0.01). IgG VCA and EBNA were detected in 99/107 (93%), evidence of a previous EBV infection. There were no significant relationships between antibody titers reactive with different EBV antigens and detectable EBV in HRS cells. Furthermore, there did not appear to be any relationship between EBV serology or the presence of EBV in HRS cells and clinical outcome. The role of EBV in the development of HD, especially its relationship to the immunological response, remains unclear. Int. J. Cancer 72:394–397, 1997. © 1997 Wiley-Liss, Inc.     

Epstein-Barr virus-specific antibodies in patients with adult T-cell leukemia (ATL) and healthy ATL virus-carriers

Antibodies to Epstein-Barr virus (EBV)-capsid antigen (VCA), early antigen (EA) and EBV-associated nuclear antigen (EBNA) in the sera of 103 patients with adult T-cell leukemia (ATL) and the sera of 243 age- and sex-matched healthy adults, namely 99 anti-ATLA-positive (antibodies to ATL virus-associated antigen) and 144 anti-ATLA-negative individuals, were determined by indirect immunofluorescence. The anti-VCA titers in the sera from ATL patients were within the range observed in healthy controls. Anti-EA was found in 27% of the sera from ATL patients, but in only 8% of the sera from anti-ATLA-negative healthy controls. Antibodies to EBNA, which were present in almost all healthy adults, were not found in 30% of the sera from ATL patients. Of the sera of anti-ATLA-positive healthy adult donors 11% were negative for EBNA antibody. These results suggest that functional impairment of the T-cell system in most ATL patients, and also to a lesser extent in anti-ATLA-positive adults who might be healthy ATLV-carriers, may cause an unusual immune response of antibodies to EBV-associated antigens.     

Increased antibody responses to Herpes virus papio (HVP) ANTIGENS IN PRE-LYMPHOMATOUS BABOONS (Papio hamadryas) of the sukhumi high lymphoma stock

Antibody responses to Herpes virus papio (HVP) antigens were studied in 21 pre-lymphoma baboons (which subsequently died of malignant lymphoma), 21 paired controls, i.e. age-, sex- and population-matched healthy baboons, and 185 randomly selected healthy baboons of the same population. The sera were all collected at the same time and were tested blind in the fixed-cell indirect immunofluorescence test against HVP viral capsid antigen (VCA)-positive, early antigen (EA)-positive cell targets before and after absorption with HVP. Eleven of the pre-lymphoma sera were anti-EA-positive whereas none of the paired controls contained anti-EA. Anti-VCA titers of pre-lymphoma sera were higher than those of paired controls in thirteen cases. Only in four cases were anti-VCA titers of pre-lymphoma sera lower than those of paired controls. Qualitatively, the same results were obtained when anti-VCA and anti-EA titers of pre-lymphoma sera were compared with respective mean population values. The differences between pre-lymphoma group and control groups, especially in the case of anti-EA, were statistically highly significant. Thus, elevated anti-HVP titers in healthy baboons of the Sukhumi lymphoma-prone stock can be considered as a marker of high risk for development of malignant lymphoma.     

Anti-EBV antibody titers in non-Hodgkin lymphomas.

Antibody titers to Epstein-Barr virus (EBV)-related antigens, i.e. viral capsid antigen (VCA), the D and R components of the early antigen (EA) complex and the EBV-associated nuclear antigen (EBNA), were determined in a series of 86 patients with non-Hodgkin lymphomas and in 150 matched control subjects. The lymphoma patients belonged to four histological groups: diffuse, nodular, hyperbasophilic malignant lymphoma (HML) and unclassified. The EBV-related serological data were compared to the incidence of antibodies to other herpes viruses, i.e. cytomegalovirus (CMV), herpes simplex virus (HSV) and varicella zoster virus (VZV), and correlated with immune disorders, which are particularly frequent in the HML type of lymphoma. The results revealed a significantly higher incidence of anti-EA-D titers in lymphoma patients and slight but significant increases in the geometric mean anti-VCA titers in the HML and unclassified group of patients. These elevated anti-EBV titers in patients were not associated with an increase in titres of antibodies to other herpes viruses. They did not correlate with the signs of immune deficiency observed or with the incidence of auto-antibodies.     

A population-based case-control study of EBV and other viral antibodies among persons with Hodgkin's disease and their siblings

The Epstein-Barr virus (EBV) has been consistently found to be associated with Hodgkin's disease (HD) in two ways: cases generally have elevated titer distributions of antibodies against the viral capsid antigen, and the occurrence of HD among persons with a history of EBV infectious mononucleosis is two or three times higher than expected. We evaluated this association by measuring the prevalence and level of antibodies against EBV and related viruses among 304 cases of HD interviewed in a population-based study in comparison to 285 of their siblings. The most significant finding was that antibody titers to the viral capsid antigen of EBV were elevated (± 1:320) in 39% of the cases and in only 14% of the sibling-controls; the relative risk adjusted for age and sex was 4.1. The geometric mean titer was three-fold higher among cases (175.6vs. 58.1) Subjects who reported a history of IM had a higher distribution of titers than those who did not. Cases also had elevated titers against the early antigen of EBV - the D Component being most prominent. A significantly higher proportion of cases has elevated titers against CMV, relative risk = 3.4, but the prevalence of CMV antibody was relatively low and not consistently higher among cases. The findings support the hypothesis that EBV may play a role in the pathogenesis of HD among persons with elevated titers. The findings neither confirm nor deny a possible role of CMV.