A longitudinal study of phagocyte function in HIV-infected patients.

OBJECTIVE: To study the influence of HIV infection on phagocyte function. To date, the results of phagocyte function studies in HIV-infected patients have been contradictory. This is the first longitudinal study of these functions in HIV infection. DESIGN: We followed 50 individuals with HIV infection for 2-51 months (mean, 28 months) and examined polymorphonuclear leukocyte (PMNL) and monocyte functions at intervals of 0.5-1 years. METHODS: PMNL random migration and chemotaxis were assessed using an under-agarose method, and PMNL and monocyte oxidative metabolism by chemiluminescence production during phagocytosis of opsonized zymosan. RESULTS: PMNL random migration and chemotaxis were impaired at entry into the study by 15 and 19%, respectively. After 3 years the reduction was 35 and 32%, respectively. The mean chemiluminescence production by PMNL was reduced by 6% at entry into the study. After 4 years a decrease of 18% was observed. The decrease in PMNL function was most marked in patients with lymphadenopathy syndrome or AIDS. No significant change in monocyte chemiluminescence production was detected at any time during the study. CONCLUSIONS: A distinct and progressive decrease of PMNL function occurs during HIV infection. This may contribute to increased susceptibility to opportunistic infections in HIV-infected patients. For monocytes, chemiluminescence production is not influenced by HIV infection.

Negative selection of human monocytes using magnetic particles covered by anti-lymphocyte antibodies.

We have developed a standardized procedure for the isolation of monocytes from peripheral blood by negative selection using magnetic polymer particles coated with monoclonal antibodies against T and B lymphocytes. The average purity of the monocyte suspension was 85%, and monocyte recovery was 72% from Ficoll-Hypaque gradient separated mononuclear cells and 32% from whole blood. In a lucigenin enhanced chemiluminescence assay there was no significant difference between cells separated immunomagnetically and those separated on a gradient. Nor did electron microscopy show any significant difference in morphology between such monocytes. Negative selection using magnetic polymer particles is an efficient method for the separation of monocytes with intact morphology and function as measured by chemiluminescence.

Isoprinosine stimulates granulocyte chemiluminescence and inhibits monocyte chemiluminescence in vitro.

Isoprinosine may delay disease progression in human immunodeficiency virus infection, presumably through modulation of lymphocyte function. However, the influence of isoprinosine on phagocyte function is largely unknown. This study describes the effects of isoprinosine and azidothymidine on phagocyte chemiluminescence and migration. Incubation with isoprinosine concentrations of 250 micrograms/ml and above increased the chemiluminescence of granulocytes. Random migration of granulocytes was decreased at isoprinosine concentrations of 50 micrograms/ml and higher, but chemotaxis was not affected. Azidothymidine exerted no effect on the chemiluminescence or migration of granulocytes. For monocytes, luminol-enhanced chemiluminescence was reduced at isoprinosine concentrations of 250 micrograms/ml and above, whereas migration was not affected. These findings suggest that the immunomodulatory properties of isoprinosine may extend to phagocytic cells. This may be of significance in the treatment of immunodeficiency states.

Serum antibodies to viral pathogens and Toxoplasma gondii in HIV-infected individuals.

Sera from 38 HIV-infected individuals were examined longitudinally for antibodies to viruses that may increase morbidity in HIV infection, as well as commensal viruses and Toxoplasma gondii. HTLV infection was seen in Norway for the first time as four patients had antibodies to HTLV-II and one had antibodies to HTLV-I. Antibodies to hepatitis B virus (HBV) were found in 47.2%, while 21.6% of the patients had antibodies to hepatitis C virus (HCV). There was no evidence of acquisition of HBV or HVC during the mean observation period of 2 years. A titre increase in CMV antibody with time was observed for 7 out of 21 patients and a decrease for 2 patients. For Epstein-Barr virus, herpes simplex, varicella-zoster, rubella and measles viruses, human polyomavirus BK as well as for Toxoplasma gondii, antibody prevalences and titres were within the range seen in normal populations. Also, no longitudinal changes were observed in titres of these antibodies, indicating that humoral immunity remained intact during the study period. The high prevalences of HTLV-I/II, HBV and HCV antibodies in HIV-infected patients reflect common modes of virus transmission, and the fluctuations in CMV antibody titre are indicative of reactivations. Such coinfections may influence disease progression.

Monocyte phagocytosis of opsonized Neisseria meningitidis serogroup B.

The chemiluminescence (CL) was examined when peripheral blood monocytes were incubated with opsonized Neisseria meningitidis, serogroup B, serotype 15:P1.16 or serotype 2a:P1.2. The monocytes were separated from a mononuclear cell suspension by an immunomagnetic negative selection technique using magnetic polystyrene microspheres coated with monoclonal antibodies specific for T and B lymphocytes. More than 90% of the lymphocytes were removed, yielding a suspension containing 93% monocytes. Optimal sensitivity for phagocytosis was obtained using 1% serum (10 microliters), 72 bacteria per monocyte cell, and 7.5 min opsonization and incubation time during continuous agitation at 37 degrees C. The CL was amplified by lucigenin. Preliminary experiments suggest that convalescent sera from patients with group B meningococcal disease induced increased CL responses compared to acute sera. Sera from volunteers immunized with an outer membrane complex vaccine from serogroup B, serotype 15:P1.16 or 2a:P1.2 meningococci also induced increased CL activity compared to preimmune sera. No such response was shown when a group B capsular polysaccharide vaccine was given. This response pattern was also demonstrated by a flow cytometric phagocytosis technique (FCM). Internalization of meningococci by monocytes was demonstrated by a FCM quenching technique and by transmission electron microscopy. CL and FCM represent rapid and reproducible methods for the measurement of opsonophagocytosis of meningococci by monocytes and may be performed with minute amounts of sera.

[Pneumococcal vaccine]. / Pneumokokkvaksinen.

Streptococcus pneumoniae is a common cause of morbidity and mortality in children, immunodeficient individuals and elderly people. As antibiotic resistant strains become more prevalent, pneumococcal infections will become more difficult to manage. Pneumococcal vaccination is inexpensive, and serious side effects are rare. However, the efficacy and safety of the vaccine have been questioned, and the use of vaccination is limited. This article discusses factors to be considered when assessing the indications for pneumococcal vaccination. Immunisation together with a strict policy on the use of antibiotics are our most efficient means for preventing pneumococcal disease and spread of antibiotic resistant strains. Norwegian physicians are encouraged to use pneumococcal vaccine according to the present indications.

Effect of fusidic acid on migration and chemiluminescence of polymorphonuclear leukocytes.

Incubation of polymorphonuclear leukocytes (PMNs) in fusidic acid resulted in a decrease of random migration and chemiluminescence. The effects were dose-dependent but moderate, with statistical significance only at concentrations of 50 mg/l or more. At concentrations used for therapy of bacterial infections and AIDS, fusidic acid was not shown to be deleterious to these aspects of PMN function.

Longitudinal study of cytomegalovirus antibodies in individuals infected with the human immunodeficiency virus.

Cytomegalovirus (CMV) antibody profiles were studied in 25 HIV-infected patients over periods of up to 56 months. Specific antibodies against CMV antigen components were monitored by complement-fixation (CF) test, EIA, Western blot and a neutralization assay. Three subjects remained CMV seronegative throughout the study. Marked fluctuations were observed in anti-CMV antibodies assayed by the CF test as compared to a control group. Fluctuations on immunoblots of purified virion antigens were also observed in the HIV-infected patients; neutralizing antibodies and anti-CMV nucleocapsid antibodies showed less variability. Seven of 22 individuals exhibited an increase in CF-test titre of up to 64-fold without clinically apparent CMV disease. On Western-blot testing of IgG reactivity with disrupted virions, ten individuals exhibited increasing reactivity to pp65, and only three of these also showed a titre rise in the CF test. In contrast, 7 of 22 showed low reactivity to the pp28 antigen. The homosexual patient group exhibited the highest levels of anti-CMV antibody. In conclusion, many asymptomatic HIV-infected subjects showed fluctuations at different levels of their antibody response to CMV, thought to be indicative of CMV reactivation/reinfection. Western-blot findings indicated that some CMV antibodies increased in level while others were lost.

Diagnosis of infection with human T-lymphotropic virus type II (HTLV-II) in Norwegian HIV-infected individuals.

Sera from 298 HIV-infected individuals from Southern Norway were examined for antibodies against HTLV. 30 sera (10.1%) were HTLV-II positive and 1(0.3%) HTLV-I positive. 25 of the HTLV-II infected subjects were intravenous drug abusers (IVDAs), giving a prevalence of HTLV-II infection of 24.5% in this group. Examination of blood samples by polymerase chain reaction followed by restriction enzyme analysis or sequencing confirmed the serological diagnosis. To evaluate current screening and verification HTLV tests, 44 sera were examined using a gelatin particle agglutination test, 5 different enzyme-linked immunoassays (ELISA) and 4 Western blots (WB). While earlier ELISAs and WBs were inadequate, a recent ELISA and WB including recombinant envelope glycoproteins from both viruses permitted serological diagnosis and distinction between HTLV-I and HTLV-II. Thus, HTLV-II now spreads among IVDAs in a North-European country. Health authorities in other countries should estimate the magnitude of the problem to decide upon measures to avoid transmission through blood transfusion.

[Research training--the work situation of research trainees at the School of Medicine, University of Bergen]. / Forskerutdanning--forskerrekruttenes arbeidssituasjon ved Det medisinske fakultet, Universitetet i Bergen.

In an investigation in spring 1989 on research training at the Medical Faculty, University of Bergen, 111 questionnaires were returned of a total of 180 distributed. 35% of the trainees were satisfied with the training, 31% not, while 30% did not know as yet. Trainees who had been full-time research fellows for two years or more (n = 33), were split in two groups with different opinions. Half of them were satisfied with the training programme, and the other half were not. The most important factor linked with the trainees' opinion of the training was personal supervision. The satisfied ones were content with supervision they had received, whereas the unsatisfied ones were not. By and large, office and laboratory conditions were adequate for the trainees, but data equipment had been supplied by many of the trainees themselves. The overall impression of this investigation is that the research training programme is working fairly well, but there is remove for improvement in the supervision of the individual trainee.

[Research training--what trainees think about future research education]. / Forskerutdanning--hva forskerrekruttene mener om fremtidens forskerutdanning.

A research training programme should have a clearly defined aim and a framework for accomplishment and contents. In Norway no such programme exists for the medical PhD. Present research training depends to a large extent on the individual student. The aim of this paper is to contribute to a discussion on the goals for a research programme and to report what research trainees and research fellows at the Medical Faculty, University of Bergen think is necessary as regards framework and content. 111 of 180 questionnaires were returned. On the average, the students preferred a research programme to last 4.0 years. 91 persons wanted the programme to contain a compulsory part lasting, on average, 5.5 months. The majority preferred no examination during this part of the programme. Most students wanted to maintain the present Norwegian standard for the medical PhD.