RESUMO
Within the Innovative Health Initiative (IHI) Inno4Vac CHIMICHURRI project, a regulatory workshop was organised on the development and manufacture of challenge agent strains for Controlled Human Infection Model (CHIM) studies. Developers are often uncertain about which GMP requirements or regulatory guidelines apply but should be guided by the 2022 technical white paper "Considerations on the Principles of Development and Manufacturing Qualities of Challenge Agents for Use in Human Infection Models" (published by hVIVO, Wellcome Trust, HIC-Vac consortium members). Where those recommendations cannot be met, regulators advise following the "Principles of GMP" until definitive guidelines are available. Sourcing wild-type virus isolates is a significant challenge for developers. Still, it is preferred over reverse genetics challenge strains for several reasons, including implications and regulations around genetically modified organisms (GMOs). Official informed consent guidelines for collecting isolates are needed, and the characterisation of these isolates still presents risks and uncertainty. Workshop topics included ethics, liability, standardised clinical endpoints, selection criteria, sharing of challenge agents, and addressing population heterogeneity concerning vaccine response and clinical course. The organisers are confident that the workshop discussions will contribute to advancing ethical, safe, and high-quality CHIM studies of influenza, RSV and C. difficile, including adequate regulatory frameworks.
Assuntos
Clostridioides difficile , Vacinas contra Influenza , Influenza Humana , Vírus , Humanos , Influenza Humana/prevenção & controle , Vírus/genéticaRESUMO
BACKGROUND: Human immunodeficiency virus (HIV) uses 2 distinct chemokine receptors, CCR5 (R5) or CXCR4 (X4), during entry. Viruses may be R5 tropic, X4 tropic, or dual/mixed (D/M) tropic. R5-tropic virus predominates at high CD4 cell counts, with the number of X4-tropic strains increasing as CD4 cell count decreases. METHODS: We investigated the relationship between tropism and decreases in CD4 cell count before antiretroviral therapy initiation, the frequency of clinical events, and responses to antiretroviral therapy in a cohort of treatment-naive patients. RESULTS: Four hundred two treatment-naive patients underwent tropism determination; 326 harbored R5-tropic virus, and 76 harbored X4- or D/M-tropic virus. After adjustment for baseline characteristics, the rate of decrease in CD4 cell count was significantly greater in patients infected with X4- or D/M-tropic virus at 12 months (P=.026). Two hundred twenty-nine individuals infected with R5-tropic virus and 60 individuals infected with X4- or D/M-tropic virus commenced antiretroviral therapy between tropism testing and the time of data analysis. Time to viral suppression and the proportion of patients achieving viral suppression were similar at 6, 12, and 24 months. CD4 cell count increases were similar. Clinical events were significantly more common in the group infected with X4- or D/M-tropic virus. Multivariate analysis demonstrated a relative risk of experiencing a clinical event of 2.56 (95% confidence interval, 1.37-4.76; P=.003) among patients infected with X4- or D/M-tropic virus. CONCLUSIONS: The presence of D/M- or X4-tropic virus has a deleterious effect on CD4 cell count decrease and risk of clinical disease. Response to standard antiretroviral therapy is not affected by viral tropism.
Assuntos
Antirretrovirais/uso terapêutico , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/fisiologia , Carga Viral , Contagem de Linfócito CD4 , Progressão da Doença , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/fisiopatologia , HIV-1/isolamento & purificação , Humanos , Masculino , Receptores CCR5/metabolismo , Receptores CXCR4/metabolismo , Resultado do TratamentoRESUMO
Generally physicians have a legal and ethical obligation of keeping confidentiality regarding their communication with patients and it is clear that we all have rights. The application of rights theorem, which usually refers to the recognition of individual human rights, to the deceased offers possible answers to the problematic question of patient confidentiality after death. Philosophical considerations broadly support utilitarian ideals concerning the 'common good'. However, it may be possible to rank rights according to a hierarchy of need and thus preserve individual rights where they do not impinge upon the public's right to protection from harm and the physician's right to tell the truth. This has broad implications for confidentiality, anonymity and health care information in general for patients, their families and healthcare workers. We discuss these issues, with specific reference to an individual case.
Assuntos
Atitude Frente a Morte , Direitos Humanos , Médicos/ética , Autopsia/ética , Confidencialidade , Ética Médica , Humanos , Relações Médico-Paciente , Relações Profissional-Família , Opinião PúblicaRESUMO
There is increasing recognition of the potential morbidity and mortality associated with HIV-1 and hepatitis C (HCV) co-infection. HIV appears to adversely affect HCV disease while the reciprocal effect of HCV on HIV remains controversial. We therefore studied the effect of co-infection on dendritic cell function versus HIV infection alone, as previous work has shown that HCV impairs dendritic cell (DC) function. HIV-1 positive individuals with HCV were matched for CD4 count, HIV-1 RNA viral load and therapy, to HIV-1 positive patients without HCV. Monocyte-derived DC were generated and mixed leukocyte reactions were performed. We assessed allostimulatory capacity with and without administration of exogenous Th1 cytokines, using thymidine uptake and cell division analyses with the vital dye CFSE. We found that monocyte-derived DC from co-infected individuals showed no significant differences in allostimulatory capacity to ex vivo generated DC from HIV-1 infected individuals without HCV. Unlike the situation with HCV infection alone, this impairment was not reversed by increasing concentrations of either interleukin-2 or -12. Monocyte-derived DC from HIV-1 and HCV co-infected individuals have a similar allostimulatory capacity to DC from matched patients with HIV-1 alone. These findings are compatible with results of prior clinical studies that found no evidence that HCV co-infection altered HIV disease progression and has implications for immunotherapeutic approaches in co-infected individuals.
Assuntos
Células Dendríticas/imunologia , Células Dendríticas/virologia , Infecções por HIV/virologia , Hepatite C/virologia , Adulto , Células Cultivadas , Estudos de Coortes , Células Dendríticas/citologia , Progressão da Doença , Infecções por HIV/complicações , Infecções por HIV/imunologia , HIV-1/genética , HIV-1/imunologia , Hepacivirus/genética , Hepacivirus/imunologia , Hepatite C/complicações , Hepatite C/imunologia , Humanos , Testes Imunológicos , Linfócitos T/imunologiaRESUMO
A 32-year-old HIV-1 positive man was diagnosed with Castleman's disease following a long history of constitutional symptoms. Primary therapy with single agent rituximab was associated with a near complete response. During this time, his KSHV (Kaposi's sarcoma-associated herpesvirus) viral load decreased and we also observed immediate, large and sustained decreases in interleukin-6 (IL-6) and tumor necrosis factor-alpha levels (TNF-alpha). This highlights the close association between circulating cytokines such as IL-6 and virally-induced malignancy.
Assuntos
Síndrome da Imunodeficiência Adquirida/complicações , Anticorpos Monoclonais/uso terapêutico , Antígenos CD20/imunologia , Hiperplasia do Linfonodo Gigante/complicações , Hiperplasia do Linfonodo Gigante/tratamento farmacológico , Interleucina-6/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/virologia , Adulto , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais Murinos , Hiperplasia do Linfonodo Gigante/imunologia , Hiperplasia do Linfonodo Gigante/virologia , HIV-1/fisiologia , Humanos , Imunoterapia , Interleucina-6/sangue , Masculino , RituximabRESUMO
The evolutionary dynamics of RAL resistance in the HIV-2 virus were examined through population and clonal sequence analysis of the IN from baseline, during treatment, and after stopping RAL therapy. The treatment failure of an RAL regimen in the HIV-2 patient studied was associated with the emergence of mutations via the N155H resistance pathway and subsequent switching to the Y143C mutational route. This study has also identified four novel secondary mutations, Q91R, S147G, A153G, and M183I, not previously reported in HIV-1 patients failing RAL therapy. Resistant variants involving the Y143C pathway were noted to have persisted beyond 4 weeks following the cessation of RAL therapy. All resistance-associated mutations were lost at 20 weeks after stopping RAL therapy. Our findings provide evidence supporting the supposition that substantial cross-resistance between strand transfer IN-Is is likely in HIV-2 as shown in HIV-1.
Assuntos
Farmacorresistência Viral/genética , Infecções por HIV/virologia , Inibidores de Integrase de HIV/uso terapêutico , HIV-2 , Pirrolidinonas/uso terapêutico , Substituição de Aminoácidos , Evolução Molecular , Infecções por HIV/tratamento farmacológico , Integrase de HIV/análise , Integrase de HIV/genética , HIV-2/efeitos dos fármacos , HIV-2/genética , RNA Viral/análise , RNA Viral/genética , Raltegravir Potássico , Análise de Sequência de RNA , Falha de TratamentoRESUMO
BACKGROUND: CXCR4-using virus is associated with higher viral load and accelerated human immunodeficiency virus (HIV) disease progression. Additionally, CCR5 antagonists may not reduce the HIV-1 RNA load when mixed/dual-tropic or CXCR4-using virus is present. The determination of coreceptor tropism may be required before CCR5 or CXCR4 antagonists are initiated, unless reliable predictive markers of coreceptor use are established. METHODS: Samples from treatment-naive and -experienced HIV-1-positive individuals with date-matched CD4 and CD8 cell counts and HIV-1 RNA, clade, and pol sequences were assessed for coreceptor usage, by use of the ViroLogic PhenoSense assay. RESULTS: Coreceptor use determination was successful in 563 of 861 samples. Non-clade B virus was present in 18.6% of samples. CXCR4 or mixed/dual-tropic CCR5/CXCR4 virus was present in 112 of samples (19.9%). Only 4 samples (0.7%) showed exclusive CXCR4 use. In a multivariate model, higher CD4 cell count, lower viral load, and higher natural killer cell counts were significantly associated with CCR5 usage. No associations with treatment experience, clade, or pol gene mutations were observed. CONCLUSION: The prevalence of CCR5-tropic HIV-1 phenotype declines with decreasing CD4 cell count and increasing viral load. Mixed/dual tropic virus is found in all CD4 cell count and viral load strata. Coreceptor usage is not influenced by viral clade.
Assuntos
Infecções por HIV/epidemiologia , HIV-1/patogenicidade , Kit de Reagentes para Diagnóstico , Receptores de Quimiocinas/metabolismo , Adulto , Contagem de Linfócito CD4 , Feminino , Infecções por HIV/virologia , HIV-1/metabolismo , HIV-1/fisiologia , Humanos , Células Matadoras Naturais/imunologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prevalência , RNA Viral/sangue , Receptores CCR5/metabolismo , Receptores CXCR4/metabolismoRESUMO
Cluster of differentiation (CD) antigens are expressed on cells of myeloid and lymphoid lineages. As most disease processes involve immune system activation or suppression, these antigens offer unique opportunities for monitoring host responses. Immunophenotyping using limited numbers of CD antigens enables differentiation states of immune system cells to be determined. Extended phenotyping involving parallel measurement of multiple CD antigens may help identify expression pattern signatures associated with specific disease states. To explore this possibility we have made a CD monoclonal antibody array and scanner, enabling the parallel immunophenotyping of leukocyte cell suspensions in a single and rapid analysis. To demonstrate this approach, we used the specific example of patients infected with human immunodeficiency virus type-1 (HIV-1). An invariant HIV-induced CD antigen signature has been defined that is both robust and independent of clinical outcome, composed of a unique profile of CD antigen expression levels that are both increased and decreased relative to internal controls. The results indicate that HIV-induced changes in CD antigen expression are disease specific and independent of outcome. Their invariant nature indicates an irreversible component to retroviral infection and suggests the utility of CD antigen expression patterns in other disease settings.
Assuntos
Antígenos CD/análise , Infecções por HIV/patologia , Imunofenotipagem/métodos , Anticorpos Monoclonais , Regulação da Expressão Gênica , Infecções por HIV/imunologia , Humanos , Leucócitos/química , Leucócitos/patologia , Leucócitos/virologia , Análise Serial de Proteínas/métodosRESUMO
Ritonavir-boosted saquinavir (SQV/r) is currently licensed as a twice-daily regimen. Reducing the pill burden with once-daily dosing may improve adherence. Intracellular concentrations of drugs must be related to the clinical efficacy of protease inhibitors. The aims of the study were to determine the cellular and plasma saquinavir and ritonavir concentrations, to determine the half-lives (t(1/2)s) of the drugs in each compartment, and to examine relationships between drug accumulation and lymphocyte subset P glycoprotein (P-gp) expression. Venous blood samples from 12 human immunodeficiency virus-infected patients receiving a hard-gel formulation of SQV/r (1,600/100 mg once daily) were collected at 2, 6, 12, and 24 h after dosing. Peripheral blood mononuclear cells were separated by density gradient centrifugation, and P-gp expression was measured by dual-color flow cytometry. Plasma and intracellular (cell-associated) drug concentrations were measured by high-performance liquid chromatography-tandem mass spectrometry. The ratio of the intracellular drug area under the concentration-time curve from 0 to 24 h (AUC(0-24 h)) to plasma drug AUC(0-24 h) was calculated to determine cellular drug accumulation. The median (range) AUC(0-24 h) of saquinavir in plasma was 16.2 (5.7 to 39.3) mg. h. liter(-1), and that in cells was 46.3 (24.7 to 114.6) mg. h. liter(-1). Corresponding ritonavir values were 7.5 (1.5 to 14.6) mg. h. liter(-1) and 10.4 (3.2 to 13.7) mg. h. liter(-1), respectively. The median accumulation ratios of cellular AUC to plasma AUC for saquinavir and ritonavir were 3.31 (range, 1.49 to 6.69) and 1.46 (range, 0.83 to 4.15), respectively. Significant differences between the plasma and intracellular saquinavir t(1/2)s (4.5 h [range, 2.5 to 9.3 h] and 5.9 h [range, 4.0 to 17.7 h]; P = 0.034) and between the plasma and intracellular ritonavir t(1/2)s (4.1 h [range, 2.6 to 8.3 h] and 6.2 h [range, 3.9 to 18.6 h]; P = 0.032) were observed. No relationship was observed between the accumulation of saquinavir or ritonavir and lymphocyte subset P-gp expression. The intracellular t(1/2)s of saquinavir and ritonavir were longer than the plasma t(1/2)s, indicating that intracellular drug may be available at a time when concentrations in plasma are below the minimum effective concentration.
Assuntos
Infecções por HIV/metabolismo , Inibidores da Protease de HIV/farmacocinética , Ritonavir/farmacocinética , Saquinavir/farmacocinética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Adulto , Área Sob a Curva , Cromatografia Líquida de Alta Pressão , Combinação de Medicamentos , Feminino , Citometria de Fluxo , Inibidores da Protease de HIV/administração & dosagem , Meia-Vida , Humanos , Subpopulações de Linfócitos/efeitos dos fármacos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Ritonavir/administração & dosagem , Saquinavir/administração & dosagemRESUMO
A small proportion of patients with human immunodeficiency virus type 1 (HIV-1) remains asymptomatic for a long period after infection. It is thought that a vigorous immune response may contribute to long-term nonprogression, though studies are confounded by heterogeneity among patients. We studied the levels of HIV-1 receptors, costimulatory T-cell molecules, and dendritic cell (DC) numbers in 18 patients with long-term infection, CD4 counts greater than 400 cells/mm(3), and HIV-1 viral loads lower than 50 copies/mL. These patients were further differentiated through the presence or absence of 2-LTR DNA circles, a possible marker for residual ongoing HIV-1 replication. A statistically significant increase in levels of CD91, the heat-shock protein (HSP) receptor, was observed in therapy-naive patients who had no evidence of ongoing viral replication (P =.01). This difference was most notable on their monocytes. High levels of CD91 may be a host factor that contributes to the maintenance of long-term nonprogression. The ability of CD91 to internalize alpha-defensins and to cross-present exogenous antigen to cytotoxic T lymphocytes through major histocompatibility complex (MHC) class 1 may maintain CD8(+) responses in these patients.