Immune changes in male homosexuals--predisposing factors for HIV seroconversion.

Israel is still a low incidence country for AIDS and thus the temporal relationships between various cofactors and HIV infection can be more easily determined. We have studied a cohort of 243 asymptomatic male homosexuals (MHS) during the last 4 years. At the beginning of the study, 40% had significant decreases in the peripheral blood lymphoid populations, signs for reactivation of CMV and EBV infections, and elevated serum levels of interferon. There was no difference in the prevalence of these immune derangements between seropositive and seronegative individuals. In the course of 4 years' follow-up, the prevalence of these immune derangements has remained the same though being significantly more severe in the HIV seropositive subjects. HIV seroconversion did not exceed 8% (about 2% annually) and was related to the immune impairments found when first seen at the beginning of the study. The results suggest that immune impairments are common among asymptomatic MHS, precede HIV infection, and probably contribute to the increased risk for AIDS among male homosexuals.

Lymphomatous T-cell leukemia in two Arab children. Is there a role for an environmental effect.

Two Arab children from the Gaza strip presented with fever, weakness, hepatosplenomegaly, lymphadenopathy and leukocytosis. The peripheral and bone marrow blasts had an immunophenotype compatable with T-cell acute lymphoblastic leukemia, and exhibited unusual markers (CD2+, CD3+, CD4-, CD8-). Cytogenetic studies revealed t(8;14)(q24;q11), possibly involving the alpha/delta locus of the T-cell receptor gene on chromosome 14 rather than the immunoglobulin heavy-chain locus usually involved in the t(8;14)(q24;q32), which is typical for Burkitt's leukemia/lymphoma. One of the children had a brother who died of T-cell acute lymphoblastic leukemia a few years later, however, his blasts showed deletion of chromosome 12. The possible role for environmental factors associated with low socioeconomic status, as well as of genetic factors in leukemogenesis are discussed.

Inhibition of the reverse transcriptase activity and replication of human immunodeficiency virus type 1 by AS 101 in vitro.

In a search for compounds active against human immunodeficiency virus type 1 (HIV-1), it was found that the novel low-molecular weight immunoenhancer ammonium trichloro(dioxyethylene-O,O'-) tellurate (AS101) suppresses production of HIV-1 in vitro. Treatment of HIV-1-infected peripheral blood mononuclear cells (PBMC) with increasing concentrations of AS101 resulted in substantial inhibition of virus production as measured by both reverse transcriptase (RT) activity and antigen presence in supernatants of treated cells. AS101 had no effect on PBMC viability, growth, or morphology up to a concentration of 15 microM for 14 days. To elucidate a possible mechanism for the inhibition of AS101, we have analyzed the effect of the drug on the catalytic functions associated with HIV RT, namely the RDDP, DDDP, and RNase H activities. RDDP and DDDP activities were impaired by the drug with calculated IC50 value of about 4 microM. On the other hand, the RNase H activity was less sensitive to AS101, with an apparent IC50 value of about 30 microM. The anti-HIV-1 activity of AS101 as reflected by inhibition of the different catalytic functions associated with viral RT, in the absence of drug-related toxicity to lymphocytes, together with its immunomodulating activity strongly argues in favor of its evaluation, as a therapeutic agent for patients with HIV infection.

Detection of CMV in urine: comparison between DNA-DNA hybridization, virus isolation, and immunoelectron microscopy.

Rapid detection of CMV-DNA in urine specimens by dot-blot hybridization was compared to conventional virus isolation and to virus identification using solid-phase immunoelectron microscopy (SPIEM). To detect viral DNA, 32P-labeled EcoR1 J fragment of CMV-DNA was used as a probe in the hybridization assay. In addition, DNA extracted from infected human embryo fibroblasts (amplified DNA) was also hybridized to the same probe. Urine specimens were obtained from 10 renal transplanted patients, seven premature infants, three family members, and five children suspected of CMV infection. CMV was isolated from 10 urine specimens and SPIEM detected viral particles in nine specimens. Ten positive samples were identified as such by hybridization with DNA extracted directly from urine specimens, while hybridization with amplified DNA yielded 17 positives. Only in one urine specimen, positive by virus isolation and SPIEM, DNA was not detected by the hybridization assays. Elevated IgG or IgM-specific antibodies were found in 10 patients. Hybridization with amplified DNA proved the most sensitive and relatively rapid assay, as compared with direct DNA detection in urine, tissue culture isolation, SPIEM, or serologic tests.

'Silent' HIV infection among wives of seropositive HIV carriers in the Ethiopian community in Israel.

We have previously described the phenomenon of 'silent HIV carriers', i.e. individuals with HIV specific immunity and a positive PCR for HIV-1, yet HIV seronegative. In the present study, we have looked for such 'silent' carriers among wives of individuals infected with HIV in Africa (Ethiopia). In addition to determining HIV serology, peripheral blood mononuclear cells (PBMC) were tested by PCR for HIV-1 and for their ability to generate specific antibodies to HIV upon polyclonal B-cell activation (P-BAT). Out of 16 wives so tested, three were HIV seropositive and among the 13 seronegatives, eight were P-BAT positive and five were both P-BAT and PCR positive. These findings suggest that (1) 'silent' HIV carriers may indeed be present in African populations; (2) interpretation of the 'silent' carrier phenomenon is not clear and will depend on clinical follow-up and the ability to culture virus from such carriers; and (3) results of HIV serology in this population and probably in other African populations should be viewed with caution.

Detection of CMV-DNA in cells from peritoneal fluid of IPD/CAPD patients by polymerase chain reaction.

Documented viral peritonitis in peritoneal dialysis patients is rare, although up to 20% of all cases are culture negative (non-fungal, non-bacterial). CMV-infected peritoneal cells may serve as a reservoir for reinfection and/or reactivation of CMV after renal transplantation. CMV-Polymerase Chain Reaction (CMV-PCR) amplification analysis identified CMV-DNA in cells from the peritoneal dialysate of 8 patients (3 culture negative peritonitis from a total of 5 examined) and 5 asymptomatics) out of 17 potential kidney transplant recipients (6 on IPD 16.6 +/- 6 months, range 10-29 months; 11 on CAPD 28.1 +/- 25 months, range 2-81 months). Serum titers (10/17 patients analyzed) of anti-CMV IgG antibodies ranged from < 1:20 to 1:320 (no correlation with CMV-DNA) while anti-CMV IgM antibodies were undetectable. Detection of CMV specific sequences in peritoneal cells in peritoneal dialysis patients by the PCR assay is sensitive (amplification of a 133 bp immediate early CMV gene sequence allows detection of 10 CMV infected cells in a background of 10(5) uninfected peritoneal cells), rapid (1 day visual, 3 days with confirmation by Southern hybridization), specific (no amplification of human embryo and kidney cell DNA, or HSV, EBV, or VZV infected cells) and is non-invasive in IPD/CAPD patients since no additional invasive technique is required.

Human immunodeficiency virus (HIV) inactivation of banked bone by gamma irradiation.

OBJECTIVE: The increasing number of patients with acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus(HIV)-positive carriers, poses difficulties when musculo-skeletal tissues are considered for banking in readiness for future clinical application. This study was conducted to test the actual yield of gamma irradiation on HIV infectivity, within HIV-infected bones. METHODS: The effect of gamma irradiation on bones containing T-cells chronically infected with HIV type I (HIV-I) was studied, in respect to inactivation of the virus. RESULTS: After exposure of the cell-free virus or infected T-cells to 2.5 megarads of gamma irradiation, the authors were able to demonstrate complete inactivation of the virus. CONCLUSIONS: It would appear from this study that gamma irradiation at this dose is sufficient to achieve clinical sterilisation of bones and facilitate their use for reconstructive procedures by eliminating the risk of HIV transmission to the recipient. Furthermore, when preparing bones for banking, this would also seem to be the method of choice in preventing the transmission of various strains of bacteria, fungi and other viruses.

Antibodies to Epstein-Barr virus in subacute sclerosing panencephalitis patients.

Blood samples from 45 cases of subacute sclerosing panencephalitis in Israel were tested for the presence of antibodies to Epstein-Barr virus capsid antigen and early antigen by the indirect immunofluorescence method. Significantly higher titers were found in patients as compared to healthy controls with higher titers especially in Jews of Asian-African origin. Mean geometric titer was 1:42.8 in this group as compared to 1:10.8 in the control group. Antibodies to Epstein-Barr Virus early antigen were found in 50% of the patients and in none of the controls tested. Elevated titers of antibodies to Epstein-Barr Virus antigens seem to be related to impaired immunological mechanisms in subacute sclerosing panencephalitis patients, especially of a certain ethnic group.

Evidence for type-C retrovirus production by Burkitt's lymphoma-derived cell line.

Burkitt's lymphoma cell line, P3HR-I, was found to secrete virions with properties of known type-C RNA tumor viruses. The viral particles had a buoyant density of 1.16 g/ml in sucrose gradients and contained a high-molecular-weight RNA and an RNA-instructed DNA polymerase. The viral polymerase was active in an endogenous reaction requiring the presence of the four deoxyriboside triphosphates and manganese ions, and was sensitive to RNase. The DNA product of the endogenous reaction specifically hybridized to P3HR-I viral 60 to 70S RNA. Electron microscopic examination of ultrathin sections of P3HR-I cells revealed immature, mature and budding virions typical of type-C retroviridae. Nucleic acid hybridization assays showed no sequence homoblastosis virus, murine oncornaviruses, simian sarcoma virus or RD114 virus.

Immediate immunosuppression caused by acute HIV-1 infection: a fulminant multisystemic disease 2 days post infection.

A healthy 19-year-old woman had vaginal intercourse on a single occasion with an HIV-1 positive male from Gambia. Two days later she developed an acute HIV infection presenting as a fulminant multisystem disease that lasted for 35 hospital days and included: immediate immunosuppression with extreme CD4+ lymphocytopenia and combined with CD8+ lymphocytosis, neutropenia and hypogammaglobulinemia; intermittent spiking fever; pneumonitis; hepatitis; changing skin rashes; peripheral neuropathy with myopathy, and panencephalitis. P24 antigen was detected by Western blot on day 23 and seroconversion was detected by ELISA on day 25. Cultured lymphocytes from peripheral blood and cerebrospinal fluid grew HIV-1.

Stimulation of antibodies to Epstein-Barr virus (EBV) in acute viral infections.

Acute and convalescent sera from 77 patients with serologically confirmed influenza, measles and adenovirus infections and from 36 healthy controls were tested for the level of antibodies to Epstein-Barr (EB) virus. In the three groups of patients significantly higher titers of antibodies to EB viral capsid antigen (VCA) were found as compared to the controls. In 19 patients twofold or higher rise in antibody titers between the first and second blood sample was demonstrated. It is suggested that in patients with influenza, measles or adenovirus infections, involvement of lymphocytes leads to reactivation of EB virus and antibody formation is stimulated.

Rearrangement of the immunoglobulin heavy chain gene in juvenile chronic myeloid leukaemia.

Five patients with clinical and laboratory features typical for juvenile chronic myeloid leukaemia (JCML) are presented. Rearrangement of the j joining region of the immunoglobulin heavy chain (Jh) was demonstrated in three children out of five analysed. As no Vh to DhJh nor kappa light chain rearrangements were demonstrated, it is reasonable to speculate that the transforming event of the stem cell happened at the stage when Dh to Jh rearrangement took place. As the monocytic lineage is prominent in JCML, it is suggested that the transforming event happens in a unique stem cell with intermediate differentiation towards the myelomonocytic as well as the B-lymphatic lineage. This stem cell, which is present at a certain stage of embryogenesis, disappears later. Such an early 'hybrid' cell is sometimes involved in leukaemias of early infancy, and may be the transformed cell in some cases of infantile leukaemia.

Infection and persistence of varicella-zoster virus in lymphoblastoid Raji cell line.

Infection of Raji cells by varicella-zoster virus (VZV) resulted in permissive infection with establishment of a persistently infected lymphoblastoid cell line. VZV antigens of the membrane and nuclear type, as detected by the indirect immunofluorescence membrane antigen (IFAMA) and anticomplement immunofluorescence (ACIF) tests, were observed. Minute amounts of infectious virus were detected by co-cultivation of VZV-infected Raji cells (Raji-VZV), with permissive human embryo fibroblasts (HEF). The virus isolated was found to be similar to the parent strain. Transient induction of Epstein-Barr viral capsid antigen (EB-VCA) was also observed. The persistently infected Raji-VZV cell line, when free of EB-VCA, was found suitable for measuring antibodies to varicella-zoster virus. The possible interaction in the infected Raji cells between EBV, which is implicated in human malignancy, and VZV which belongs also to the herpes group of viruses, is discussed.

Blastogenic response of lymphocytes derived from patients with hematopoietic malignancies to antigens of a type C retrovirus isolated from a Burkitt's lymphoma cell line.

Cellular immune response to antigens associated with a type C retrovirus derived from Burkitt's lymphoma (BL) lymphoblastoid cells was studied in patients with hematopoietic malignancies, noncancer patients and healthy controls. Response was determined by lymphocyte blastogenesis assay measuring [3H]-thymidine incorporation, thereby enabling the calculation of stimulation indices (SI). Positive response (SI greater than 2.0) was demonstrated in patients with multiple myeloma (MM), chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML) and BL. No response was demonstrated in the non-cancer and healthy controls. Specific blastogenic response was obtained towards antigen extracted from three different cell lines infected with the tested type C retrovirus. No response was evident towards purified whole or purified disrupted virions or antigen extracted from murine myeloma MOPC-315 cells secreting a murine type C retrovirus or an antigen extracted from Mason-Pfizer monkey virus-infected NC-37 cells. The correlation between human hematopoietic malignancies and the in vitro lymphocyte blastogenic response to the BL-derived type C retrovirus was statistically highly significant.

Identification of feline- and canine-like rotaviruses isolated from humans by restriction fragment length polymorphism assay.

Restriction fragment length polymorphism assay of reverse-transcribed and polymerase chain reaction-amplified rotavirus gene segment 9 was developed to differentiate human serotype 3 rotaviruses from animal serotype 3 rotaviruses. On the basis of similarities or differences in HinfI and DdeI restriction profiles, unusual group A serotype 3 human rotaviruses that belonged to subgroup I were shown to be of feline and canine origin. By this approach, the new human rotavirus isolates 5193, AU-387, AU-720, AU-785 and AU-1115 were shown to resemble certain feline-like human rotaviruses. Similar results were previously obtained by Nakagomi et al. (O. Nakagomi, A. Hoshima, Y. Aboudy, I. Shif, M. Mochizuki, T. Nakagomi, and T. Gotlieb-Stematsky. J. Clin. Microbiol. 28:1198-1203, 1990) by using RNA-RNA cross hybridization with established feline rotaviruses. The restriction fragment length polymorphism assay can provide fast and valuable information on the interspecies transmission of rotaviruses in nature.

EBV genome and immunoglobulin gene rearrangement in the differential diagnosis of nasopharyngeal carcinoma and lymphoma.

Head and neck tumors include nasopharyngeal carcinoma (NPC) and lymphoma. The differential diagnosis of these tumors is based on histology, immunocytochemical staining, and EBV serology. In rare cases it might be difficult to distinguish between NPC and lymphoma in HE section or biopsies. DNA hybridization with cloned EBV and human immunoglobulin gene fragments allows the detection of EBV-related sequences and immunoglobulin gene rearrangements. The presence of EBV genome supports the diagnosis of NPC or EBV related BL, while rearrangement of immunoglobulin genes points to B-cell lymphoma. The diagnosis in 11 patients suspected of head and neck tumors was carried out by hybridization of DNA extracted from the tumors and assayed with cloned EBV and IgHCJ DNA probes. One patient proved to have EBV-associated BL based on positive hybridization with EBV probes and immunoglobulin rearrangement, presenting a unique hybridization with cloned EBV DNA BamHI W fragment, with bands of 3.2 and 3.9 kb. BL was confirmed in this patient by demonstration of c-myc rearrangement. A second patient was negative in hybridization with EBV, and positive for immunoglobulin rearrangement, and therefore was diagnosed as having B-cell lymphoma. In seven patients NPC was confirmed by hybridization with EBV-DNA probes. In two patients, both NPC and B-cell lymphomas were excluded.

HIV-2 infection in Israel: a new epidemic?

AIDS may be caused by two different retroviruses, HIV-1 and HIV-2. Hitherto only HIV-1 has been reported in Israel. We recently discovered HIV-2 as a solitary pathogen in the blood of two foreign workers from West Africa. In view of the relative ease of travel to Israel, it is essential to perform screening for both HIV viruses in all subjects with an enhanced risk, including visitors from countries with a high incidence of HIV-1 or HIV-2 infection and their contacts.

Antibodies to Epstein-Barr virus in patients with Hodgkin's disease and leukemia.

Sera from 67 Hodgkin's disease patients, 71 leukemia patients, and 186 healthy subjects were tested for antibodies to Epstein-Barr (EB) viral antigens by immunofluorescence methods. In both disease categories, in particular Hodgkin's disease patients, levels of antibodies to the viral capsid antigen (EBV-VCA) and MGT were higher than in the healthy controls. Significantly higher titers were found in Jewish patients of Asian-African origin, as compared to Jews of European origin, with Arab patients as intermediates. The effect of ethnic origin was independant of age and histopathologic type. Sex had no effect on titer. Inconsistent differences in titer were found between age groups in the various ethnic-histopathologic type groups. Some of the leukemia patients had no detectable antibodies to EBV, while all Hodgkin's disease patients showed previous contact with EB virus. Antibodies to the early antigen (EBV-EA) were found in 27% of Hodgkin's and 37% of leukemia patients, and in none of the healthy controls tested.