HIV infection among Brazilian transvestites in a prison population.

Eighty-two male transvestites imprisoned in Casa de Detenção (São Paulo, Brazil) were tested for HIV antibodies, and completed a questionnaire investigating their demographics, arrest and imprisonment records, sexual practices, and drug use. Data were then analyzed to evaluate the incidence of HIV infection and its association with various behavioral and other factors. Sixty-four of 82 (78%, 95% confidence interval [CI], 67-87%) transvestites were positive for HIV infection. The factors associated with significant differences in positivity among these individuals were the time spent in prison and the number of sexual partners during the previous year. It appears that the high rate of infection in this group obscured the importance of other risk factors and behavioral patterns potentially associated with infection. Given the social environment and the high rate of HIV infection among imprisoned transvestites, their role as "vectors" for dissemination of HIV in urban areas of Brazil may be significant.

Rapid detection of HSV with an enzyme-linked virus inducible system (ELVIS) employing a genetically modified cell line.

BACKGROUND: Infections with herpes simplex viruses (HSV) are common and may cause severe disease in immunocompromised hosts and in neonates. Isolation of infectious HSV in tissue culture is the most sensitive method of detection, but is not the most rapid. Recently, however, an Enzyme-Linked Virus Inducible System (ELVIS) for rapid detection of HSV in culture has been developed. The system employs genetically engineered baby hamster kidney (BHK) cells (ELVIS cells) whose DNA bears and HSV inducible promoter gene chimerically linked to an E. coli LacZ "reporter" gene. Induction of the promoter by HSV leads to the production of LacZ product, beta-galactosidase, which is readily detected histochemically. OBJECTIVE: To evaluate these ELVIS cells, as a test for HSV, in comparison with HSV detection in MRC-5 cells in shell vial cultures confirmed by staining with fluorescent antibodies. STUDY DESIGN: Over a period of one month, 167 specimens submitted to the laboratory for detection of HSV were evaluated. Specimens were inoculated onto MRC-5 cells growing on glass coverslips in each of two shell vials and into two wells of a 24-well cluster plate containing ELVIS cells. MRC-5 shell vial cultures were observed daily for cpe for up to 7 days. With the appearance of cpe, the coverslips were fixed and the cells were typed for HSV-1 and HSV-2 with monoclonal antibodies. Specimens inoculated onto ELVIS cells were incubated for 16-24 h, then substrate was added to stain for beta-galactosidase. ELVIS cells, induced by HSV infection to express beta-galactosidase, stained blue upon reaction with substrate. RESULTS: Of 167 specimens inoculated onto MRC-5 cells, 13 were excluded because of contamination or toxicity. Among the remaining 154 specimens, 24 were positive for HSV in the MRC-5 shell vials. Of 166 specimens inoculated into the ELVIS cell, all were completed within 24 h. Twenty-three (23) of the 24 shell-vial-positive cultures also were positive on the ELVIS cells. All 23 specimens detected in the ELVIS cells were positive within 24 h, whereas only nine were positive within 24 hours in MRC-5 shell vial cultures. The remaining 15 became positive after 24 h. Specimens positive for viruses other than HSV-1 or HSV-2 were not positive on the ELVIS cells. CONCLUSIONS: The ELVIS assay for HSV is simple to perform, is rapid, sensitive, and specific. The assay detects both HSV-1 and HSV-2. No antibodies are required unless typing, which can be done on the ELVIS cells, is necessary.

Laboratory diagnosis of human immunodeficiency virus infection.

HIV infection and AIDS will continue to grow as a major medical and social problem. The incidence of heterosexual transmission is rising, and it will become increasingly difficult for physicians and counselors to assess an individual's risk of infection. In coming years, physicians can expect to see patients who are infected, but whose risk may not be apparent, and who may not present with conditions immediately suggestive of their infection. The majority of these patients may not even suspect they are infected. Many of the tests currently available for diagnosing HIV infection are very good. They are both highly sensitive and highly specific and their predictive values are good when their limits are understood. But, as HIV infection expands to an ever larger number of women and children and as the ability to sharply define risk groups fades, demand for a broader range of tests that can reliably confirm infection and are easy to perform can only increase. We have attempted to describe some of the common serologic tests currently used to diagnose HIV infection and some of the limitations of these tests. We also have pointed out that criteria used by laboratories for interpreting tests such as the Western blot may not always be uniform, and physicians should know the criteria used by their reference laboratory and the quality control measures taken. We also have attempted to describe some of the tests available to detect the virus, its genes, or its gene products. PCR and other rapidly evolving gene amplification techniques hold great promise as highly sensitive and specific tests for the diagnosis/confirmation of HIV infection. As with the serologic tests, it is important for those who may use these tests to understand their value as well as their limitations.

Evaluation of the Genetic Systems Corp. Integra HIV-1 Pageblot system.

The Genetic Systems Corp. Integra HIV-1 Pageblot system was evaluated as a supplementary assay to confirm the presence of antibodies to human immunodeficiency virus type 1 in 57 specimens from individuals at high risk of infection with the virus. Forty-one specimens identified as reactive in the Genetic Systems Integra HIV-1 Pageblot system were likewise identified as reactive in a U.S. Food and Drug Administration-licensed (Biotech/Dupont) Western blot (immunoblot). Six specimens identified as indeterminate in either or both immunoblot assays were all identified as nonreactive in a U.S. Food and Drug Administration-licensed enzyme immunoassay with recombinant antigens.

Lymphocytic alveolitis in primary HIV infection.

Primary infection with the human immunodeficiency virus (HIV-1) has been associated with a self-limited illness resembling acute infectious mononucleosis. Pulmonary manifestations have been notably absent in published reports. The authors describe a 28-year-old homosexual male who presented with primary HIV-1 infection associated with CD8+ lymphocytic alveolitis. Diagnosis was delayed because HIV antibody was not detected by the Abbott ELISA, although the same and subsequent specimens were later found to be positive by Genetic Systems' ELISA and Western blot analysis. Lymphocytic alveolitis must be added to the expanding clinical spectrum of acute HIV-1 infection. The time to detection of seroconversion may vary with different immunoassays.

Evaluation of the Recombigen HIV-1 Latex Agglutination Test.

The Recombigen HIV-1 Latex Agglutination (LA) Test was recently licensed by the U.S. Food and Drug Administration for use as a rapid screening assay for human immunodeficiency virus type 1 (HIV-1) antibodies. However, its performance in various settings and in different populations has not been firmly established. Consequently, we evaluated the test in the Cleveland Clinic Retrovirus Laboratory, a regional reference laboratory for HIV diagnostic testing and a testing laboratory for the Ohio Department of Health Anonymous HIV Testing and Counseling Program. Serum samples from 93 individuals presumed to be at high risk for HIV infection were evaluated. The sera were initially tested for HIV antibodies by enzyme-linked immunosorbent assay (ELISA). All repeatedly reactive sera were subjected to confirmatory Western blot (WB; immunoblot) testing. Of 97 serum specimens tested (5 were from one seroconverter), 44 were repeatedly reactive by ELISA and 53 were nonreactive. Of the reactive serum specimens, 31 were confirmed positive and 12 were indeterminate by WB. All of the sera were coded and then retested by the LA test. Of 53 serum specimens nonreactive by ELISA, 51 were also nonreactive in the LA test. Of the 44 serum specimens reactive by ELISA, 16 were nonreactive by LA; however, 3 of the latter were WB positive. No serum specimen with an ELISA ratio (specimen optical density/cutoff optical density) of less than 2.1 scored reactive in the LA test. The LA test was positive for only two of five consecutive serum specimens from a seroconverter despite the fact that all but the earliest of these were ELISA reactive and WB positive. Although the LA test appears to be an adequate first-line screening test when appropriately used according to the directions of the manufacturer, our data suggest that occasional sera with low levels of reactivity by ELISA may not be readily detected as reactive by the LA test.

Systemic vasculitis in association with human immunodeficiency virus infection.

Fourteen cases of vasculitis associated with human immunodeficiency virus infection have thus far been described. Five of these cases may be classified as angiocentric immunoproliferative disorders, including benign lymphocytic angiitis, lymphomatoid granulomatosis, and angiocentric lymphoma. We report a case of benign lymphocytic angiitis of T cell lineage. Extensive studies found no evidence of viral antigens in the inflammatory infiltrates, and immunologic evaluation of the pathologic lesions revealed the infiltrating cells to be predominantly CD3+, CD8+, CD4-. A significant number of these lymphocytes demonstrated a deletion of T cell antigen receptor determinants. We believe that in certain cases of human immunodeficiency virus disease, there occurs a spectrum of lymphoproliferative disorders with angiocentric features that lead to the clinical picture of systemic necrotizing vasculitis. Clinicians should be aware of this association.

Interferon gamma production by human intestinal mucosal mononuclear cells. Decreased levels in inflammatory bowel disease.

Immune (gamma) interferon is a substance produced by immunologically activated mononuclear cells. Besides its antiviral activity, interferon gamma has a crucial role in immunoregulation, by acting directly upon lymphocytes and monocytes, and interacting with other soluble mediators of the immune response. Studies of the interferons system in inflammatory bowel disease have been limited, and little information is available on the generation of interferon during immunological events occurring in the human gut. To investigate the capacity of intestinal mucosal mononuclear cells to produce interferon gamma, lamina proprial mononuclear cells, isolated from Crohn's disease, ulcerative colitis, and control patients, were incubated with interleukin 2 or phytohemagglutinin, and the amounts of interferon gamma present in the culture supernatants were measured by a virus cytopathic effect inhibition assay. Under identical stimulatory conditions, culture supernatants of cells derived from actively involved mucosa of inflammatory bowel disease specimens contained two- to fivefold less interferon gamma than those of cells from control tissue. However, the amount of interferon gamma present in supernatants of cells from uninvolved inflammatory bowel disease mucosa was similar to that found in control supernatants. These results indicate that, in patients with active Crohn's disease and ulcerative colitis, mononuclear cells produce decreased amounts of interferon gamma in the intestinal mucosa. The exact significance of these findings is unclear, but because of the importance of interferon gamma in a variety of cell-mediated immune phenomena, its impaired availability might be relevant to the pathogenesis of inflammatory bowel disease.

Clonal diversity of the Kirsten-ras oncogene during tumor progression in athymic nude mice: mechanisms of amplification and rearrangement.

Single-cell clones from primary and lung metastatic tumors have been evaluated for the state of the viral-Kirsten-ras oncogene (v-Ki-ras) by Southern blot analysis after injection of Kirsten sarcoma virus-transformed BALB/c 3T3 cells (KiMSV, with a replication-defective provirus) into athymic nude mice by four different injection routes. While all clones of early-passage KiMSV cells contained an EcoRI-generated 5.3-kilobase DNA fragment at high dosage level, most clones of late-passage cells had lost this v-Ki-ras fragment or had greatly diminished levels. However, all clones of all tumors (greater than 90 tested) obtained after injection of these late-passage cells contained a dosage of the 5.3-kilobase v-Ki-ras band similar to that of the early-passage KiMSV cells, suggesting either a very strong selection for v-Ki-ras-bearing cells of the early-passage type in tumor formation and/or the ability of a subset of late-passage cells to amplify this gene to some minimal dosage level. Both flow cytometric analyses for DNA content and quantitation of chromosomes showed that all primary and lung metastatic tumors had more than twice the number of chromosomes as the late-passage KiMSV cells; however, four of 80 late-passage cells had a chromosome count in the range of tumors, consistent with their importance in tumor generation and possibly amplification of the v-Ki-ras-bearing chromosome. Clonal analyses of lung micrometastatic tumors revealed a v-Ki-ras blot pattern identical to that of the s.c. primary tumors. However, two of five lung metastases from the footpad (as large rapidly growing nodules) and i.v. routes had multiple copies of v-Ki-ras in new sites; a second injection round led to even greater complexity in v-Ki-ras patterns in clones of lung tumors. Two assays were used to demonstrate that these new v-Ki-ras integrations were generated by superinfection with a "helper" retrovirus, not sarcomagenic by itself in the nude mice, that led to rescue/reinfection of tumor cells with the defective Kirsten sarcoma proviral genome--cellular transformation of 3T3 or C3H10T1/2 cells and RNA dot blot analyses for medium-secreted retrovirus specific for LTR or v-Ki-ras sequences. This "helper" retrovirus could not be detected in early- or late-passage KiMSV cells used for inoculation but could be detected in certain tissues of normal nude mice, demonstrating its in vivo origin.(ABSTRACT TRUNCATED AT 400 WORDS)

Acute infection with the human immunodeficiency virus (HIV) associated with acute brachial neuritis and exanthematous rash.

Clinical descriptions of acute or primary infection with the human immunodeficiency virus (HIV) are rare. Among cases previously reported, most describe an acute illness resembling infectious mononucleosis. We describe the case of a 32-year-old homosexual man with an acute illness associated with strong serologic evidence of a primary infection with HIV. This case illustrates two new clinical features: an acute, bilateral brachial neuritis, and a vesicular, pustular exanthematous and enanthematous rash. Studies of HIV-related serologic results show differential sensitivities for enzyme-linked immunosorbent assay, Western blot, immunofluorescence, and viral antigen techniques in the acute phase of HIV infection. There appears to be significant clinical heterogeneity of the acute phase of HIV infection.

Comparison of Plasmagel with LeucoPREP-Macrodex methods for separation of leukocytes for virus isolation.

Plasmagel (Cellular Products, Inc., Buffalo, NY), which can separate both polymorphonuclear leukocytes (PMN) and mononuclear cells from other blood components, and LeucoPREP (Becton Dickinson Immunocytometry Systems, Mountain View, CA), which can separate mononuclear cells from other blood components, were used to harvest leukocytes from whole blood for the purpose of virus isolation. Macrodex was combined with the later, in a second step, for recovery of PMN. Of 90 peripheral blood specimens examined, cytomegalovirus was recovered from 10: in six by both methods, in three from Plasmagel prepared cells only, and in one from cells from the LeucoPREP-Macrodex preparation only. Total leukocyte counts, differential counts, and leukocyte viability did not differ significantly for the two methods. Plasmagel provided an efficient, inexpensive means of harvesting leukocytes from whole blood for virus isolation.

Augmentation of skin test reactivity and lymphocyte blastogenesis in patients with AIDS treated with transfer factor.

Nine patients with the acquired immunodeficiency syndrome (AIDS) were administered four doses of pooled transfer factor obtained from the lymphocytes of three healthy controls and three homosexuals with stable lymphadenopathy and serum antibody to the human immunodeficiency virus. Before receiving transfer factor, all patients exhibited anergy to skin test antigens. After four weeks of transfer factor therapy, six of seven patients tested had at least one skin test response. Lymphocyte blastogenic responses to phytohemagglutinin rose from a stimulation index of 6.77 +/- 1.31 before treatment to 19.77 +/- 6.24 after four weeks of transfer factor therapy. Smaller but significant increases were also seen in blastogenic responses to antigens. Improvements in immune responses diminished after administration of transfer factor was halted. Thus, administration of transfer factor to patients with AIDS resulted in partial immune reconstitution. Further studies are indicated to examine the clinical efficacy of this immune response modifier in the treatment of AIDS.

Persistence of high risk sexual activity among homosexual men in an area of low incidence for acquired immunodeficiency syndrome.

A survey of 305 homosexual men was performed in an area of relatively low incidence for AIDS and low seroprevalence for antibody to the human immunodeficiency virus (HIV). The objective of the study was to investigate current knowledge and practice of sexual behavior designed to limit transmission of HIV. The results showed that while the majority of the study group believed that they had made changes in their life styles that would reduce the likelihood of transmitting or acquiring the virus, nearly half admitted to persistently engaging in active and passive anal intercourse without condoms. These data suggest the need for more widespread and effective forms of education to help prevent the continuing spread of the AIDS epidemic.

Immunologic mechanisms in the pathogenesis of virus-induced leukemia. IV. Mechanism of target cell recognition by autoreactive thymocytes.

Neonatal infection of mice with Moloney murine leukemia virus (MuLV-M) results in the establishment of a chronic virus-carrier state. Such MuLV-carrier mice exhibit several immunologic abnormalities including generalized immunosuppression and autoimmunity. Previously, we found thymocytes from MuLV-M-carrier mice to be cytotoxic for normal syngeneic and allogeneic fibroblasts but not for xenogeneic (hamster) target cells. However, when the same syngeneic or allogeneic target cells were infected with MuLV-M, they were "spared" from the autoreactivity, leading us to speculate that the MuLV receptor on the target cell membrane was involved in the autoreactivity. To address this question, we tested MuLV-carrier thymocytes for their ability to lyse hamster/mouse-hybrid target cells; some of which possessed chromosome 5 (which codes for the ecotropic MuLV receptor). Of the nine hybrid cell lines initially tested, only the five clones that carried chromosome 5 were killed by the autoreactive thymocytes. In additional experiments, we noted that the cytotoxic reaction was inhibited in the presence of a monoclonal antibody that reacts with an MuLV-M gp70 epitope. The results suggest that the autoreactive cytotoxicity is mediated, at least in part, through the formation of a "bridge" between MuLV budding from the membrane of the thymocytes and the ecotropic MuLV receptor on the target cells.

Small noncleaved B cell Burkitt-like lymphoma with chromosome t(8;14) translocation and Epstein-Barr virus nuclear-associated antigen in a homosexual man with acquired immune deficiency syndrome.

This case report describes new manifestations of the acquired immune deficiency syndrome (AIDS) in a promiscuous homosexual man. Investigation of upper gastrointestinal bleeding in the patient lead to discovery of a high-grade, small, noncleaved cell (Burkitt-like) gastroduodenal lymphoma with visceral and extralymphatic extension. Specific phenotyping of the lymphoma revealed that it was a monoclonal B cell lymphoma of mu kappa isotype. An in vitro cell line was established that was Epstein-Barr virus nuclear-associated antigen-positive. The lymphoma cells displayed a t(8;14) translocation similar to endemic African Burkitt lymphoma. Epstein-Barr virus genomes were identified in the lymphoma and an axillary lymph node biopsy specimen by molecular hybridization. These data strongly suggest that Epstein-Barr virus actively infected this patient. However, he showed normal Epstein-Barr virus-specific serologic responses, indicating an immune defect against the virus.