Niels Obel

Randomized Trial to Evaluate Indinavir/Ritonavir versus Saquinavir/Ritonavir in Human Immunodeficiency Virus Type 1–Infected Patients: The MaxCmin1 Trial

This trial assessed the rate of virological failure at 48 weeks in adult human immunodeficiency virus (HIV) type 1-infected patients assigned indinavir/ritonavir (Idv/Rtv; 800/100 mg 2 times daily) or saquinavir/ritonavir (Sqv/Rtv; 1000/100 mg 2 times daily) in an open-label, randomized (1:1), multicenter, phase 4 design. Three hundred six patients began the assigned treatment. At 48 weeks, virological failure was seen in 43 (27%) of 158 and 37 (25%) of 148 patients in the Idv/Rtv and Sqv/Rtv arms, respectively. The time to virological failure did not differ between study arms (P=.76). When switching from randomized treatment was counted as failure, this was seen in 78 of 158 patients in the Idv/Rtv arm, versus 51 of 148 patients in the Sqv/Rtv arm (P=.009). A switch from the randomized treatment occurred in 64 (41%) of 158 patients in the Idv/Rtv arm, versus 40 (27%) of 148 patients in the Sqv/Rtv arm (P=.013). Sixty-four percent of the switches occurred because of adverse events. A greater number of treatment-limiting adverse events were observed in the Idv/Rtv arm, relative to the Sqv/Rtv arm. In conclusion, Rtv-boosed Sqv and Idv were found to have comparable antiretroviral effects in the doses studied.

Serological and clinical findings in patients with serological evidence of reactivated Epstein‐Barr virus infection

IgM directed against Epstein‐Barr virus (EBV) early antigen (IgM‐EA) has been established as an early marker of EBV infection and IgG directed against Epstein‐Barr virus nuclear antigen 1 (IgG‐EBNA‐1) as a late marker. Simultaneous seropositivity to IgM‐EA and IgG‐EBNA has therefore been proposed as indicating reactivation of latent EBV infection. We have studied 191 patients with serological evidence of reactivated EBV infection with regard to clinical presentation, antibodies directed against EBV viral capsid antigen (IgM‐VCA, IgG‐VCA), cytomegalovirus (IgM‐CMV), human herpesvirus 6 (IgM‐HHV‐6), IgM rheumatoid factor (IgM‐RF), anti‐nuclear or anti‐cytoplasmic antibodies (ANA/ACA), and total IgM. The clinical manifestations varied considerably, but a diagnosis was established in 121 of the patients. The diversity of the clinical diagnosis probably reflects common reasons for requesting an EBV serological test rather than clinical manifestations of reactivated EBV infection. Only 5.8% of the patients with “serological EBV reactivation” gave a positive result for IgM‐VCA. We conclude that “serological EBV reactivation” does not represent an entity relating to clinical manifestations, but probably reflects a non‐specific activation of the immune system.

Changing demographics in an HIV-infected population: results from an observational cohort study in Western Denmark.

We present demographic data from an observational database of HIV and AIDS in the Western part of Denmark, a region with a population of 2,935,156 individuals (55.1% of the population of Denmark). Five centers in the region treat HIV-positive adults; all patients attached to these centers since 1995 are included in this study. In total, 749 adult HIV-infected individuals were enrolled as of 31 December, 1999. Estimates of prevalence and incidence of HIV infection in the area were 25.9/100,000 and 2.6/100,000, respectively, which are lower than average for the country. The number of newly diagnosed HIV-infected patients remained constant during the period 1995?99, with an average of 62 diagnoses per year. The number of HIV-related deaths declined from 43 in 1995 to 15 in 1999. Of the enrolled patients, 70.9% were of Danish origin, 75% were Caucasians, 69.7% were male and 47.2% had heterosexual contact as their primary risk behavior. There seems to have been a shift in the HIV epidemic in recent years, with a higher proportion of newly diagnosed HIV patients having contracted the infection through heterosexual contact, a higher proportion being immigrants from less developed countries and newly diagnosed individuals getting older.

Impaired Neutrophil Chemotaxis after Cardiac Surgery

Chemotaxis of circulating peripheral neutrophils was studied in 12 patients undergoing elective cardiac surgery. Tests of neutrophil chemotaxis were made preoperatively and 16-20 hours postoperatively, using a multipore filter assay. Statistically significant postoperative decrease was demonstrated in both stimulated and random neutrophil migration (p = 0.012 and p = 0.009, respectively). Although the peripheral neutrophil count showed a statistically significant postoperative increase (p = 0.001), the increase did not correlate to the decrease in neutrophil chemotaxis. Nor could correlation be demonstrated between the altered neutrophil chemotaxis and any per- or postoperative variables.

Activation of the respiratory burst by Pneumocystis carinii. Efficiency of different antibody isotypes, complement, lung surfactant protein D, and mannan-binding lectin

The effect of opsonization of Pneumocystis carinii with different antibody classes, complement, mannan‐binding lectin (MBL), and lung surfactant protein D (SP‐D) on respiratory burst activation was studied. Antibodies were obtained by affinity chromatography, complement from a hypogammaglobulinaemic patient, and phagocytic cells from blood donors. Respiratory burst activation was measured by chemiluminescence (CL). With freshly isolated neutrophils the combination of antibodies and complement but not antibody alone, had opsonizing properties. With neutrophils cultured for 20 h, however, IgG increased the CL response. In macrophages P. carinii opsonized with IgG alone induced a CL response proportional to the antibody titre used. With IgA an effect, albeit lower, was also seen, whereas IgM alone was inefficient. The combined effect of antibodies and complement increased the response significantly for all three antibody classes, IgG and complement giving the largest response. Binding of MBL to P. carinii and Candida albicans was demonstrated; however, only the former stimulated activation of the respiratory burst. SP‐D did not bind to either microorganism and had no effect on the respiratory burst. It is concluded that IgG, IgA and complement are important opsonizing factors in infections involving P. carinii. The relative importance varies with the type of phagocytic cell studied.

The use of calcium carbonate in nelfinavir-associated diarrhoea in HIV-1-infected patients.

To evaluate the efficacy of oral calcium supplements in HIV‐infected patients with nelfinavir (NFV)‐associated diarrhoea, and to investigate the influence on the pharmacokinetics of nelfinavir and the active metabolite M8.

Evaluation of a method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes.

A modified method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes is described. After phagocytosis of S. aureus the extracellular bacteria were sufficiently removed by repeated centrifugations and washings of the granulocytes. The optimal conditions for incubation of granulocytes for measurement of intracellular killing were found to be 37°C in the presence of 5% CO2. Under these conditions, stable pH, the viability and the capacity of the granulocytes for superoxide anion generation were preserved. The number of intracellular viable bacteria was determined after lysis of the granulocytes, which should be done in H2O at pH 11 to ensure sufficient cell lysis. The kinetics and individual variation of the intracellular killing are described. The intra‐ or extracellular location of surviving bacteria was studied. After approximately 8 h incubation we observed intracellular growth of S. aureus followed by lysis of granulocytes and extracellular growth of bacteria. Consequently, the incubation period should not be extended beyond 5 to 8 h when the assay is used to study the effects of antibiotics on intracellular killing.

Normal immunological status in four patients with ectrodactyly-ectodermal dysplasia-clefting syndrome (EEC-syndrome)

Lobster‐claw deformity of the extremities, clefting of the primary and secondary palate, ectodermal dysplasia, and atresia of the lacrimal system are common features of the ectrodactyly‐ectodermal dysplasia‐clefting syndrome (EEC‐syndrome). The patients often suffer from repeated infections of eyes, upper respiratory tract and urogenital system. To exclude an immunodeficiency as cause of the infectious predisposition in patients with EEC‐syndrome, we screened the immunosystem in four related patients with EEC‐syndrome. All patients were found to present normal immunoglobulin production, complement activity, lymphocyte‐, and granulocyte function. We conclude that recurrent infections observed in the EEC‐syndrome are not caused by an immunological defect, but seem to result solely from anatomical anomalies.

Danish patients with untreated multiple myeloma do not harbour human herpesvirus 8

The role of human herpesvirus 8 (HHV‐8) in multiple myeloma (MM) remains controversial. We examined 15 Danish MM patients before cytoreductive therapy. Mononuclear cells isolated from peripheral blood and bone marrow aspirates, as well as long‐term cultured bone marrow stromal cells, were assayed for the presence of HHV‐8 DNA. All material was tested by three simple unnested polymerase chain reaction (PCR) assays (amplifying regions of ORF26, ORFK1 and ORF75) and two nested PCR assays (amplifying regions of ORF26). HHV‐8 was not demonstrated in any of the samples. Our findings do not suggest an association between HHV‐8 and MM in the Danish population.

Quantification of neutrophil chemotaxis: A comparison of stereological and 51Cr‐labelling methods

A stereological method has been used to estimate the number of migrated polymorphonuclear leukocytes in a multipore filter assay. The estimates have been compared to those obtained with conventional 51Cr‐labelling methods. The precision of the estimates in individual subjects with the two methods was similar, and in both cases the contribution to the interindividual variance made by the methods was small compared to the biological variance. However, the 51Cr‐labelling method resulted in a significant reduction in the fraction of neutrophils that migrated as well as a delay in the chemo‐tactic response. Optimal assay conditions with regard to sample size, cell concentration, incubation time and chemoattractant fMLP concentration have been established for the stereological method and the virtues of the two techniques discussed.