High levels of discordance between sequencing and serological subtyping in a predominantly non-B subtype HIV-1 infected cohort.

Samples from 457 randomly selected HIV-1 infected patients attending King's College Hospital were analysed using a subtype specific enzyme immunoassay. All serotyped non-Bs that provided unambiguous sequence and for which sufficient sample was available (n=100), which included three serotyped subtype B samples were further analysed by env sequencing and subtyping using neighbour joining phylogenetic analysis, the NCBI Retrovirus Genotyping tool and the Los Alamos BLAST search tool. Of the serotyped viruses, 45% (n=204) samples were subtype B. Specifically serotyped non-B strains (n=130) accounted for 28% of the total, of which the largest proportion were subtype C (n=66). Twenty-seven samples (6%) were classified as non-B, 9% (n=40) were multiply-reactive and 12% were non-reactive (n=56). Of the 100 samples subtyped by sequencing the majority were subtype C (n=32), followed by subtype A (n=20). There was little concordance between the two methods. Although a 100% match was found among the serotyped and sequenced non-B viruses (n=13), only 16 of the sequenced subtype C specimens matched the 29 obtained by serotyping. Of the 20 multiply-reactive samples analysed by serotyping, only 1 sample consisted of a subtype mixture by sequencing. Of the 14 serologically non-reactive samples analysed, all were successfully sequenced, with subtype B strains (57%) the most common. Sequencing 15 samples in both env and pol regions revealed differences in subtype assignment for the same sample in some cases. Only 1/6 env subtype A and 4/5 env subtype C samples were concordant in pol sequence subtype. Differences were also found in subtyping by the different methods used. The overall agreement between the three methods was 89%. Four out of 11 samples agreed between the phylogenetic and Los Alamos methods, 1/11 between phylogenetic and BLAST and 2/11 between Los Alamos and BLAST.     

Intracellular cytokines may model immunoregulation of abacavir hypersensitivity in HIV-infected subjects

BACKGROUND: The clinical treatment of patients with HIV and adverse drug events may be enhanced by an understanding of the underlying mechanisms. About 4% of patients with HIV receiving the potent antiretroviral drug abacavir develop a hypersensitivity reaction. This idiosyncratic reaction appears to have an immunologic component that has yet to be defined. Given that the T-cell type 2 cytokine IL-4 may be overproduced by patients with allergy or other immunologic dysregulation, an index cytokine profile could help elucidate the character of a drug-specific hypersensitivity reaction. OBJECTIVE: Quantitation of the production of the type 2 IL-4 and the counterregulatory type 1 cytokine IFN-gamma in patients with abacavir-related hypersensitivity. METHODS: Intracellular cytokines were enumerated in blood T cells by flow cytometry. Subjects were grouped for evaluation as patients with a hypersensitive response after abacavir treatment, patients initiating abacavir who also were evaluated again after 1 month on abacavir, patients on abacavir for 6 months without hypersensitivity, and HIV-naive control individuals. RESULTS: There was a significant association between increased IL-4 production by CD4 and CD8 T lymphocytes and hypersensitivity reactions to abacavir. Lymphocytes from hypersensitive subjects expressed CD28 and the anti-HIV chemokine macrophage inflammatory protein 1beta with a frequency comparable with HIV-naive control cells, suggesting the possibility that the activated T cells from patients with hypersensitivity are functional. CONCLUSION: The expansion of type 0 and type 2 T cells phenotyped by IL-4 production may correlate with abacavir-associated hypersensitivity. The data suggest a cytokine bias that may facilitate B-cell differentiation and downregulate T-cell cytotoxic responses.     

Does an HIV clinical trial information booklet improve patient knowledge and understanding of HIV clinical trials?

Objectives To evaluate the impact of an information booklet on HIV clinical trials, Clinical Trials in HIV and AIDS: Information For People Who Are Thinking About Joining a Trial, in addition to the standard trial information (SI) on patients' knowledge; understanding and attitudes about clinical trials; and to investigate patients' motivations and reasons for enrolling or not enrolling in a clinical trial. Methods Fifty HIV‐1 positive patients who attended the HIV clinic at a west London hospital were randomized to receive either SI alone (n = 27) or SI and a 16 page information booklet explaining the principles and procedures of HIV clinical trials (n = 23). A self‐administered questionnaire was used at baseline to assess past experience and attitudes to clinical trials (10 questions), knowledge and understanding of HIV treatments (8 questions) and clinical trials (11 questions). At 2–6 months after randomization, a second interviewer‐administered questionnaire addressed the patient's assessment of the usefulness and comprehensiveness of the information provided by the SI and information booklet, whether or not the patient had enrolled in a clinical trial and reasons for enrolling/not enrolling, knowledge of specific aspects of the trial protocol the patient was eligible to join (13 questions) and general knowledge of clinical trial procedures (repeat of 11 baseline questions). Changes in the attitudes and scores on knowledge and understanding of clinical trials were compared for the two groups. Results In both groups, patient knowledge of clinical trial procedures improved significantly over the study period. The median score increased from 30 at baseline to 35/44 at follow‐up (SI only) vs. 24–31/44 (SI plus booklet), but this did not differ significantly between the two groups. However, knowledge of the specific trial protocol was poor [median score 13/25, interquartile range (IQR) 8–14], and there was no difference in the scores for the two groups. The prime motivations for joining a clinical trial were to benefit personal health and to gain access to new treatments. Potential side‐effects were the main concern of prospective trial participants. Conclusions This small trial shows that, while the patients' general knowledge and understanding of clinical trials improved over time, this was not improved by the information booklet and recollection of the details of the relevant trial protocol remained poor.     

Protection against a lethal H5N1 influenza challenge by intranasal immunization with virus-like particles containing 2009 pandemic H1N1 neuraminidase in mice

Highly pathogenic H5N1 influenza shares the same neuraminidase (NA) subtype with the 2009 pandemic (H1N1pdm09), and cross-reactive NA immunity might protect against or mitigate lethal H5N1 infection. In this study, mice were either infected with a sublethal dose of H1N1pdm09 or were vaccinated and boosted with virus-like particles (VLP) consisting of the NA and matrix proteins, standardized by NA activity and administered intranasally, and were then challenged with a lethal dose of HPAI H5N1 virus. Mice previously infected with H1N1pdm09 survived H5N1 challenge with no detectable virus or respiratory tract pathology on day 4. Mice immunized with H5N1 or H1N1pdm09 NA VLPs were also fully protected from death, with a 100-fold and 10-fold reduction in infectious virus, respectively, and reduced pathology in the lungs. Human influenza vaccines that elicit not only HA, but also NA immunity may provide enhanced protection against the emergence of seasonal and pandemic viruses.     

Detection and prevention of maculopathy associated with antimalarial agents.

Rheumatologists use both cholorquine and hydroxychloroquine in the treatment of systemic arthritic and immune disease. Hydroxychloroquine is much more expensive but is better tolerated by patients. My experience in watching patients being switched from one drug to another suggests that chloroquine is more effective in some patients than is hydroxychloroquine. Reynes thought that a review of the literature suggests that chloroquine is more toxic at 250 mg/day as compared to 400 mg of hydroxychloroquine if dose is based on these dosages. This theory may be related, in part, to the observation by Raines and associates that chloroquine crosses the blood-retinal barrier whereas hydroxychloroquine does not. Patients should be assessed 6 months after starting antimalarials. Routine automated perimetry is not indicated. An appropriate examination would include visual acuity testing, color-vision testing, Amsler grid testing, and corneal assessment. Patients should be dosed on the basis of ideal body weight (not actual body weight) to reduce the incidence of macular toxicity. Patients with no risk factors should be examined no more than once a year.     

Decreased morbidity and use of hospital services in English HIV-infected individuals with increased uptake of anti-retroviral therapy 1996-1997. National Prospective Monitoring System Steering Group.

OBJECTIVE: To investigate the relationship between changing morbidity patterns, the use of hospital services by HIV-infected patients and the uptake of antiretroviral therapy (ART) in England. DESIGN: Prospective serial cross-sectional analyses based on data collected through the National Prospective Monitoring System (NPMS), a multi-centre prospective monitoring system. SETTING: HIV-infected patients seen in 10 clinics, five London and five non-London, during the three semesters, 1 January 1996 to 30 June 1997. MAIN OUTCOME MEASURES: The mean use of hospital services per patient-year, mean new HIV-related opportunistic illnesses per 1000 patient-years and percentage uptake of ART. RESULTS: The use of inpatient services changed particularly among AIDS patients. The mean number of inpatient days for AIDS patients decreased from 19.7 [95% confidence interval (CI) 13.7-25.7] in 1996 to 11.2 (95% CI 6.1-15.6) per patient-year in 1997. Concurrently the number of new AIDS-defining events decreased significantly from 567 (95% CI 529-607) to 203 (95% CI 183-225) per 1000 patient-years. The overall uptake of ART increased significantly from 33% (95% CI 31-35%) to 50% (95% CI 48-52%), and a switch from mono or dual to triple therapy or quadruple or more therapy was observed. However, by mid-1997 only 29% (95% CI 26-32%) of asymptomatic patients and 51% (95% CI 49-54%) of patients with symptomatic non-AIDS were on ART, compared with 69% (95% CI 66-71%) of AIDS patients. CONCLUSION: The observed reduction in new AIDS-defining events has led to a reduction in the need for inpatient hospital care and has been associated with an increased uptake of ART, including a switch to triple therapy. All of these factors are likely to have contributed to the observed reduction in mortality among English AIDS patients. As the overall uptake of ART remained relatively low in English centres further improvements can be anticipated. However, the medium to long-term effects of these treatment regimens will need to be closely monitored.