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Culture techniques for isolation of HIV-1 from small amounts of whole blood (WB) treated with anticoagulant have been reported and gave results identical to those of culture of separated peripheral blood mononuclear cells. Some authors obtained much higher isolation rates when EDTA was used instead of heparin. We compared two previously described techniques for cultivation of HIV-1 from WB of adult HIV+ patients staged according to the CDC classification. In addition, we assessed the influence of the type of anticoagulant used for the collection of blood in viral replication in cell cultures from whole blood. Small volumes of WB treated with either heparin or EDTA were cocultivated with phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PHA-PBMC) from healthy donors. We used two procedures for WB culture: procedure I, based on the culture of 250 microliters of WB with 1 x 10(6) PHA-PBMC from donors; and procedure II based on the culture of 500 microliters of WB with 4 x 10(6) PHA-PBMC from donors. The cocultures were placed in 24-well plates and incubated for as long as 28 days in medium containing interleukin 2 (IL-2). Twice weekly half of the medium was replaced with fresh medium. In procedure II, one million fresh PHA-PBMC from donors was added on the 7th day of culture. The culture supernatant was assayed for the presence of HIV-1 p24 antigen in an enzyme immunoassay. The kinetics of HIV-1 replication in cultures of WB from 7 AIDS patients were similar using procedures I and II. In 8 HIV+ patients the isolation rate was higher with heparin- than with EDTA-treated samples. The isolation rate was higher in AIDS patients (n = 8) than in others with both methods. In stage IV patients without AIDS (n = 8) we failed to isolate HIV-1 in 1 patient with procedure I, whereas we succeeded with procedure II. In stage II, HIV-I was isolated in 1 of 4 patients with both methods. HIV was isolated in cultures of WB from patients receiving zidovudine or related nucleoside analogues and in cultures of WB from untreated patients. HIV-1 could not be isolated from WB of patients with more than 400 CD4+ T lymphocytes in their peripheral blood (n = 4); however, it was isolated from 14 of 16 patients with less than 400 CD4+ T lymphocytes. Our results suggest that procedure II is more sensitive than procedure I and that heparin is better than EDTA for collecting WB. We showed that the rate of HIV-1 isolation from WB increased in advanced-stage patients. Further studies are needed to define the clinical applications of WB culture.

Correlations between the in vitro biological properties of HIV strains isolated from patients and the prognosis of their disease have been reported. We developed a technique to study the phenotype of HIV strains isolated from patients. We used the P4 cell line, derived from HeLa cells, which has been transfected with receptor CD4 gene. HIV laboratory strain (HIVLAI) and peripheral blood leukocytes (PBLs) from donors infected with HIVLAI induce syncytium in P4 cell cultures in vitro. The presence of reporter gene (LacZ gene) under the control of the HIV-1 long terminal repeat (LTR) in these cells allows colorimetric visualization of syncytia in the cytoplasm using a beta-galactosidase (beta gal) assay in the presence of X-gal. We cocultivated 1 x 10(6) patient PBLs with 2 x 10(6) normal PHA-activated normal PBLs for 4 days in the presence of IL-2 in 24-well plates. Half of the medium was replaced twice a week and PHA-activated normal PBLs were added every 7 days. HIV-1 was isolated from cocultured PBLs of 18 patients with advanced-stage HIV infection as assessed by the production of HIV p24 detected with a commercially available HIV-1 p24 ELISA. Supernatant and 10(5) cells were collected twice a week from cocultured PBLs and were added to P4 cells in 96-well microtiter plates. The cultures were observed every day for 3 days and then the beta gal assay was performed. We did not observe any effect with cells and supernatant from 8 patients, harvested from cultures incubated for as long as 28 days. The phenotype of these isolates was called NC (noncytopathic). In cells from 2 patients, we obtained blue multinucleated giant cells; the phenotype of these strains was called SI (syncytium inducing). In cultures from 8 other patients, we obtained the death of P4 cells without syncytium formation, and the phenotype of these strains was called CI (cell-killing inducing). In every case, the cytopathic effect of HIV-1 isolates could be detected with cocultured PBLs collected as early as day 4 of culture. Cocultured PBLs from 13 healthy controls did not alter the P4 cells. We displayed the replication of CI strains of HIV-1, but not the one of NC strains in P4 cell line. Our micromethod allowed the detection of cytopathic effects of HIV isolates. Further investigations should define the clinical applications of this method.

The authors report 5 cases of pseudotumoral tuberculosis of the liver. They essentially involved young men from disadvantaged socio-economic backgrounds. Clinically, poor general condition with massive weight loss was constant, while pyrexia and hepatomegaly were sometimes absent. Hepatobiliary ultrasonography revealed hypoechogenic or even anechogenic lesions between 1.5 and 3.5 cm in diameter. Diagnostic confirmation was histological after needle biopsy. The outcome after antibiotic treatment with or without drainage, was recovery, with ultrasonographic normalization of the liver, in all patients, with a mean follow-up of 22 months.

Variations in cytokine production in patients with human immunodeficiency virus (HIV) infection could be involved in the physiopathology and in the progression of the disease. Therefore we studied the level of granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF alpha) produced in patients with HIV infection at stage II (asymptomatic seropositives) and stage IV (AIDS) of the CDC classification, by using an enzyme amplified sensitivity immunoassay. We measured the level of GM-CSF and TNF alpha in supernatant of phytohemagglutinin-activated peripheral blood mononuclear cells from patients and healthy individuals. In one out of 10 stage II patients and 4 out of 14 stage IV patients, we obtained higher levels of GM-CSF than the mean + 2 S.D. of controls, but in 3 stage IV patients with very low CD4+ T lymphocyte counts (< 50/mm-3) compared to other patients, the GM-CSF values were very low. High levels of TNF alpha were detected in 3 out of 10 stage II and 6 out of 11 stage IV patients. The high values of TNF alpha were associated with high values of GM-CSF in stage II and in most of AIDS patients except those with very low CD4+ T cell counts, who produced low levels of GM-CSF. Plasma levels of cytokines were evaluated in 10 stage II, 22 stage IV patients and 20 controls. Increased levels of GM-CSF (more than 9 pg/ml) were observed in the plasma from 8 out of 10 stage II patients and 17 out of 22 stage IV patients.(ABSTRACT TRUNCATED AT 250 WORDS)

The level of sCD23 produced in the course of human immunodeficiency virus (HIV) infection was measured in patients grouped according to the Centers for Disease Control by using an immunoradiometric assay. Soluble CD23 was evaluated in supernatants of peripheral blood mononuclear cell (PBMC) (10(6) cells/ml) stimulated by phytohemagglutinin (PHA). Compared with healthy controls (m +/- S.D. = 1.0 +/- 0.34 U/ml, n = 7), higher values were observed in some of the patients of group II (asymptomatic) (m +/- S.D. = 2 +/- 1.33, n = 9) and some of the patients of group IV (AIDS) (m +/- S.D. = 1.3 +/- 1.40, n = 8). Those results prompted us to compare the plasma levels of sCD23 in group II and group IV HIV-infected patients and in healthy individuals. Soluble CD23 plasma levels in healthy patients (n = 42) ranged from 0 to 1.5 U/ml (m +/- S.D. = 0.9 +/- 0.33), in group II patients (n = 17) from 0 to 3 U/ml (m +/- S.D. = 0.92 +/- 0.83) and in group IV patients (n = 73) from 0 to 2.9 U/ml (m +/- S.D. = 1.15 +/- 0.71). The differences between the patients and the healthy individuals were not statistically significant but individual sCD23 values higher than 2 U/ml were obtained in 6% of the group II patients and 16.7% of the group IV patients. Increased values of sCD23 were obtained in plasma from patients with secondary infectious diseases (groups IV-Cl and IV-C2) and from patients without secondary infectious diseases (group II, group IV-A and group IV-B).(ABSTRACT TRUNCATED AT 250 WORDS)

We report five cases of varicella pneumonia among ten otherwise healthy pregnant women who were admitted in our hospital between 1986 and 1991 with chickenpox. The precise frequency of this rare complication is not well known actually but analysis of the literature shows that the mortality rate is about 20%. Beside the problem of the fetal varicella syndrome, the other complication is the severe varicella of the neonate which can appear when varicella occurs in the mother within 5 days before, and 2 days after delivery. When primary varicella infection occurs during pregnancy clinical examination must be repeated for a week after occurring of the exanthema to find elements of severity significance. Acyclovir is the drug of choice (10 to 15 mg/kg every 8 hours) for 7 days when pneumonia is present. Varicella-zoster immunoglobulin is useful for prophylaxis and for neonates with high risk of severe varicella.

Malaria is an old but still current parasitosis. Transmitted by the bite of the female anopheles, it can be revealed by more or less serious symptoms according to the species of Plasmodium. Plasmodium falciparum is responsible for the serious forms. It is the most frequent species, resistant to 4 amino-quinolein and sulfamides in some areas. P. vivax and P. ovale, usually harmless, are not so widely geographically spread as P. falciparum; and apparently are not drug resistant but they expose patients to the risk of relapses. During pregnancy, the choice of a curative and prophylactic therapy must take into account the supposed or confirmed species, the place of the stay and the duration of the exposure. On this case the authors call to mind the epidemiological criteria necessary for early diagnosis, the antimalarial drugs that can be used in pregnant women and draw attention to immuno-allergic and the classical secondary effects of quinine, which can also occur with chloroquine.

Preventing congenital toxoplasmosis is fundamental, and in France screening tests for primary infection are obligatory for pregnant women. All nonimmunized women should be encouraged to follow good dietary and general health rules until delivery. Nevertheless, the risk of seroconversion does exist but can be detected early through monthly serum tests. Spiramycin as initial treatment of maternal primary infection is an essential step in preventing parasite transmission to the fetus. Fetal antitoxoplasmosis antibodies and indirect signs of congenital abnormalities should be detected early by amniotic fluid and fetal blood tests before echographic evidence confirms the diagnosis. Pyrimethamine-sulfonamide therapy should be undertaken and modulated according to laboratory results in order to prevent or treat any possible fetopathy. Congenital toxoplasmosis can be prevented in utero through biological diagnosis and specific therapy.

Experimental infection of mice by Toxoplasma gondii is the best way for antigen production or direct diagnosis. Early serological response from sera and peritoneal exudate of mice infected by T. gondii has been studied. In this report the authors show that specific antibodies (IgM) are found in the serum at 24 hours after intraperitoneal inoculation. Serological methods were unable to detect free antibodies in the peritoneal exudate. Specific antibodies coating the parasite surface were detected by direct immunofluorescence as soon as 48 hours post infection. Biochemical treatment usually used for antigen production (trypsin, pepsin) were unable to destroy this antibodies which where no longer detected after 2 mercapto-ethanol treatment. Control experiments showed that, in these conditions, 2 mercapto-ethanol treatment had no effect on the reactivity of surface antigens with antibodies. Therefore, this results show that checking for specific antibodies absorbed at the surface of T. gondii is a prerequisite of the use of antigen produced in vivo.

For the empiric management of community-acquired pneumonia, ciprofloxacin (750 mg b.i.d.) was compared with two antibiotics frequently used in this indication, i.e., amoxicillin + clavulanic acid and erythromycin. One hundred and forty-two patients were randomized in this prospective study. Among them, the 63 patients with bacteriologically documented disease were evaluated. Clinical recovery was achieved in 73.3% of patients in the ciprofloxacin group (22/30) versus 81.8% of patients in the amoxicillin + clavulanic acid or erythromycin group (27/33) (non-significant difference). Clinical failures seen with ciprofloxacin were found to be correlated with recovery of Streptococcus pneumoniae. Ciprofloxacin is effective in pneumonia but should be used only in cases where Streptococcus pneumoniae can be excluded as the causative agent.

An acute polyradiculoneuropathy occurred in a 30 years homosexual male. E.L.I.S.A. test and Western Blot showed recent infection by H.I.V. Besides, endogenous reinfestation by cytomegalovirus was found: high concentrations of specific IgG antibodies and presence of the virus in the blood. T4 helper cells were severely reduced, without any other sign of cellular immunity failure. None of the two viruses was found in the nervous biopsy. This Guillain-Barre syndrome with a subsequent cellular reaction in the CSF, is probably to be related to an immunoallergic mechanism. Brief increase of antibodies specific for HBsAg and Borrelia Burgdorferri and the beneficial effect of plasmapheresis, supported this view. Two months later, the patient showed superficial lymph nodes hyperplasia, without any other symptom of pre-Acquired Immuno-Depression Syndrome.

Amphotericin B (Am. B), a polyene heptaene, is an antifungal antibiotic substance produced by Streptomyces nodosus, a telluric actinomycetal from Venezuela. Although it is a very toxic substance and its pharmacokinetic is not completely known, Am. B is yet the former antifungal substance utilised against number of pathogenic agents of systemic mycoses. Am. B binds irreversibly to sterols of fungal cytoplasmic membranes causing a leak of potassium and other impairments leading the fungal cell to death. Further, Am. B might to induce an enhancement of humoral and cellular immunity. After intravenous perfusion, 95 per cent of Am. B binds to plasma lipoproteins. Only a low proportion of the Am. B serum level is detected in the CSF. Distribution of Am. B to extravascular inflammatory fluids and secretions might be letter. Am. B might be eliminated essentially by biliary way. Am. B toxic effects are very frequent. Generalized reactions are observed to the earlier doses. Toxic visceral, above all, nephrotoxic manifestations, appearing later. Recent results, from experimental and human infections suggest that Am. B encapsulated in liposomal vesicles is more active, less toxic and more easily administered.

Two cases of hepatosplenic candidiasis (HSC) are reported occurring after protracted episodes of neutropenia, induced by chemotherapy for acute leukemia in one case and drug hypersensitivity in the other. The disease presented with persistent or recurrent fever after correction of the neutropenia and with splenomegaly. The alkaline phosphatases were elevated. The diagnosis was strongly suggested by abdominal ultrasonography, CT scan and MRI, which showed multiple hepatosplenic defects. It was confirmed by serologic tests for candidiasis, the presence, in 1 case, of circulating candida antigens, and the rapid response to amphotericin B. The diagnosis of HSC should be considered in patients with persistent fever after an episode of neutropenia. Ideally, histologic confirmation is desirable, but this is often obtainable only by open liver biopsy, an aggressive procedure in such patients. Failing this, our 2 cases stress the diagnostic value of noninvasive imaging techniques, serological testing (in particular the discovery of circulating candida antigens) and the response to amphotericin B.