Clinical Outcomes of Third-Generation Cephalosporin Definitive Therapy for Bloodstream Infections Due to Enterobacterales with Potential AmpC Induction: A Single-Center Retrospective Study.

The recommended therapy for severe infections caused by AmpC-inducible Enterobacterales (AmpC-E) typically involves cefepime or carbapenems. In an era of emerging resistance to these antimicrobials, we aim to assess the impact of third-generation cephalosporins (3GCs) vs. alternative antibiotics on clinical outcomes in bloodstream infections (BSIs) due to AmpC-E. We retrospectively included hospitalized adult patients with BSIs caused by 3GC-susceptible AmpC-E between 2012 and 2022, comparing the outcomes of 3GC and non-3GC definitive therapies. The primary outcome was overall treatment failure (OTF), encompassing 90-day all-cause mortality, 90-day reinfection, and 90-day readmission. Secondary outcomes comprised components of the OTF, in-hospital all-cause mortality, and length-of-stay. Within a total cohort of 353 patients, OTF occurred in 46.5% and 41.5% in the 3GC- and non-3GC-therapy groups, respectively (p = 0.36). The 3GC-therapy group exhibited a longer length-of-stay (38 vs. 21 days, p = 0.0003) and higher in-hospital mortality (23.3% vs. 13.4%, p = 0.019). However, the 90-day mortality, 90-day reinfection, and 90-day readmission were comparable between the therapy groups. Subgroup analyses involving high-risk AmpC-E and 3GC vs. standard-of-care yielded similar conclusions. Overall, our findings suggest that 3GC definitive therapy may not result in poorer clinical outcomes for the treatment of BSIs caused by AmpC-E.

Mycobacterium avium complex and Cryptococcus neoformans co-infection in a patient with acquired immunodeficiency syndrome: a case report.

INTRODUCTION: Opportunistic infections (OI) are common in patients with acquired immunodeficiency syndrome (AIDS). Cryptococcus neoformans and Mycobacterium avium complex (MAC) are frequently responsible of such infections. However, concurrent infection with these two pathogens is uncommon and underreported in the literature. CASE DESCRIPTION: We describe the case of a 28-year-old Caucasian Belgian patient with no travel history, who presented with low-grade fever, headache and wasting syndrome. He was diagnosed with human immunodeficiency virus (HIV) infection at AIDS stage, with a HIV viral load of 506,000 viral copies/mL and a CD4 + T-cells count of 10 cells/µL. Diagnosis of disseminated Cryptococcus neoformans infection was made by positive serum cryptococcal antigen and positive culture for Cryptococcus neoformans in blood and in cerebrospinal fluid. Diagnosis of disseminated Mycobacterium avium complex infection was made by positive culture on a biopsy of a mediastinal lymph node. With adequate anti-retroviral therapy (ART) and treatment of these OIs, the patient recovered well and had a good clinical evolution. DISCUSSION AND CONCLUSION: To our knowledge, this is the second case of coexistence of these two dangerous OIs reported in the post ART era. Clinicians should be aware that such co-infections still happen in high-income countries, in patients with severe immunodeficiency. Early detection and treatment of HIV is of paramount importance to prevent AIDS and its complications. We highlight the importance of thoroughly excluding all opportunistic infections in patients with newly diagnosed AIDS.Abbreviations: ABC: abacavir; AIDS: acquired immunodeficiency syndrome; AFB: acid-fast bacilli; ART: antiretroviral therapy; CM: cryptococcal meningitis; CrAg: cryptococcal antigen; CSF: cerebrospinal fluid; CT: computed tomography; EACS: European AIDS Clinical Society; FTC: emtricitabine; HIC: high-income countries; HIV: human immunodeficiency virus; HIV-VL: HIV-viral load; ICP: intracranial pressure; IRIS: immune reconstitution inflammatory syndrome; MAC: Mycobacterium avium complex; MRI: magnetic resonance imaging; MSM: man who has sex with men; NR: normal range; OD: omne in die = once daily; OI: opportunistic infection; RAL: raltegravir; TAF: tenofovir alafenamide fumarate.

Effect of anti-interleukin drugs in patients with COVID-19 and signs of cytokine release syndrome (COV-AID): a factorial, randomised, controlled trial.

BACKGROUND: Infections with SARS-CoV-2 continue to cause significant morbidity and mortality. Interleukin (IL)-1 and IL-6 blockade have been proposed as therapeutic strategies in COVID-19, but study outcomes have been conflicting. We sought to study whether blockade of the IL-6 or IL-1 pathway shortened the time to clinical improvement in patients with COVID-19, hypoxic respiratory failure, and signs of systemic cytokine release syndrome. METHODS: We did a prospective, multicentre, open-label, randomised, controlled trial, in hospitalised patients with COVID-19, hypoxia, and signs of a cytokine release syndrome across 16 hospitals in Belgium. Eligible patients had a proven diagnosis of COVID-19 with symptoms between 6 and 16 days, a ratio of the partial pressure of oxygen to the fraction of inspired oxygen (PaO2:FiO2) of less than 350 mm Hg on room air or less than 280 mm Hg on supplemental oxygen, and signs of a cytokine release syndrome in their serum (either a single ferritin measurement of more than 2000 µg/L and immediately requiring high flow oxygen or mechanical ventilation, or a ferritin concentration of more than 1000 µg/L, which had been increasing over the previous 24 h, or lymphopenia below 800/mL with two of the following criteria: an increasing ferritin concentration of more than 700 µg/L, an increasing lactate dehydrogenase concentration of more than 300 international units per L, an increasing C-reactive protein concentration of more than 70 mg/L, or an increasing D-dimers concentration of more than 1000 ng/mL). The COV-AID trial has a 2 × 2 factorial design to evaluate IL-1 blockade versus no IL-1 blockade and IL-6 blockade versus no IL-6 blockade. Patients were randomly assigned by means of permuted block randomisation with varying block size and stratification by centre. In a first randomisation, patients were assigned to receive subcutaneous anakinra once daily (100 mg) for 28 days or until discharge, or to receive no IL-1 blockade (1:2). In a second randomisation step, patients were allocated to receive a single dose of siltuximab (11 mg/kg) intravenously, or a single dose of tocilizumab (8 mg/kg) intravenously, or to receive no IL-6 blockade (1:1:1). The primary outcome was the time to clinical improvement, defined as time from randomisation to an increase of at least two points on a 6-category ordinal scale or to discharge from hospital alive. The primary and supportive efficacy endpoints were assessed in the intention-to-treat population. Safety was assessed in the safety population. This study is registered online with ClinicalTrials.gov (NCT04330638) and EudraCT (2020-001500-41) and is complete. FINDINGS: Between April 4, and Dec 6, 2020, 342 patients were randomly assigned to IL-1 blockade (n=112) or no IL-1 blockade (n=230) and simultaneously randomly assigned to IL-6 blockade (n=227; 114 for tocilizumab and 113 for siltuximab) or no IL-6 blockade (n=115). Most patients were male (265 [77%] of 342), median age was 65 years (IQR 54-73), and median Systematic Organ Failure Assessment (SOFA) score at randomisation was 3 (2-4). All 342 patients were included in the primary intention-to-treat analysis. The estimated median time to clinical improvement was 12 days (95% CI 10-16) in the IL-1 blockade group versus 12 days (10-15) in the no IL-1 blockade group (hazard ratio [HR] 0·94 [95% CI 0·73-1·21]). For the IL-6 blockade group, the estimated median time to clinical improvement was 11 days (95% CI 10-16) versus 12 days (11-16) in the no IL-6 blockade group (HR 1·00 [0·78-1·29]). 55 patients died during the study, but no evidence for differences in mortality between treatment groups was found. The incidence of serious adverse events and serious infections was similar across study groups. INTERPRETATION: Drugs targeting IL-1 or IL-6 did not shorten the time to clinical improvement in this sample of patients with COVID-19, hypoxic respiratory failure, low SOFA score, and low baseline mortality risk. FUNDING: Belgian Health Care Knowledge Center and VIB Grand Challenges program.

Incidence and Risk Factors of COVID-19-Associated Pulmonary Aspergillosis in Intensive Care Unit-A Monocentric Retrospective Observational Study.

Coronavirus disease 2019 (COVID-19)-associated pulmonary aspergillosis (CAPA) is an increasingly recognized complication of COVID-19 and is associated with significant over-mortality. We performed a retrospective monocentric study in patients admitted to the intensive care unit (ICU) for respiratory insufficiency due to COVID-19 from March to December 2020, in order to evaluate the incidence of CAPA and the associated risk factors. We also analysed the diagnostic approach used in our medical centre for CAPA diagnosis. We defined CAPA using recently proposed consensus definitions based on clinical, radiological and microbiological criteria. Probable cases of CAPA occurred in 9 out of 141 patients included in the analysis (6.4%). All cases were diagnosed during the second wave of the pandemic. We observed a significantly higher realization rate of bronchoalveolar lavage (BAL) (51.1% vs. 28.6%, p = 0.01) and Aspergillus testing (through galactomannan, culture, PCR) on BAL samples during the second wave (p < 0.0001). The testing for Aspergillus in patients meeting the clinical and radiological criteria of CAPA increased between the two waves (p < 0.0001). In conclusion, we reported a low but likely underestimated incidence of CAPA in our population. A greater awareness and more systematic testing for Aspergillus are necessary to assess the real incidence and characteristics of CAPA.

A rare case of invasive pulmonary aspergillosis presenting as organizing pneumonia due to Aspergillus niger in an immunocompetent host.

Invasive pulmonary aspergillosis associated with organizing pneumonia is increasingly described and mainly affects the immunocompromised individual. Most of Aspergillus invasive infections in both immunocompetent and immunocompromised patients are attributed to Aspergillus fumigatus. Herein we describe a clinical case of pulmonary Aspergillus niger infection presenting as an organizing pneumonia in an immunocompetent patient. A wedge resection and two successive cures of azoles (voriconazole 6 weeks then itraconazole 6 weeks) were necessary for the patient to be totally recovered. The association of OP and IPA is rare. The involvement of A. niger makes it even rarer, this is the reason why we decided to report on this case.

Infective endocarditis caused by Neisseria mucosa on a prosthetic pulmonary valve with false positive serology for Coxiella burnetii - The first described case.

We present a case of infective endocarditis (IE) on a prosthetic pulmonary valve in a 36-year-old patient with tetralogy of Fallot (TOF). The patient underwent valve replacement surgery and active antibiotic treatment against Gram-negative cocci (Piperacillin Tazobactam then Ceftriaxone) for a total duration of 42 days with a favourable outcome. The causative agent was Neisseria mucosa which was identified on the infected valve by sequencing of 16S ribosomal RNA. To our knowledge, this is the first described case of a N. mucosa infective endocarditis on a pulmonary valve. Initially, serologies performed in clinical settings by immunofluorescence for Coxiella burnetii antibodies showed a major increase in phase I IgG titers at 1024 (normal values <16) corresponding with the diagnostic criteria for Q fever endocarditis. However, this diagnosis could not be confirmed by the National Reference Center, making it the first reported case of a false positive serology for C. burnetii during an infection due to Neisseria spp.

A rare case of voluminous brain abscess due to Actinomyces meyeri and Aggregatibacter aphrophilus: is there any evidence for a prolonged antibiotic oral relay?

We present the case of a patient with a voluminous cerebral abscess caused by Aggregatibacter aphrophilus and Actinomyces meyeri occurring a week post dental scaling. Both these bacteria are rarely involved in brain abscesses, and so far, cases of cerebral actinomyces have mostly been treated surgically and with intravenous (IV) antibiotics for 3-4 months, then put on oral antibiotic therapy with penicillin or amoxicillin for a further 3-12 months. Our patient underwent drainage through craniotomy and was subsequently put on intravenous ceftriaxone for 3 months accompanied by brain imaging control at the end of this period which showed complete regression of the abscess. Following parenteral treatment, no oral antibiotics were given since pharmacokinetic properties do not allow to attain high tissue concentration in the brain. This treatment gave excellent results.

Aggregatibacter aphrophilus spinal epidural abscess.

A 76-year-old woman with a rare case of spinal epidural abscess (SEA) that had no risk factors for such type of infection, presented symptoms of back pain, progressive neurological deficit of the lower limb and loss of sphincter control. A gadolinium-enhanced MRI confirmed the diagnosis of an SEA. The patient underwent laminectomy with surgical drainage, where cultures showed the presence of Aggregatibacter aphrophilus, a bacterium of the HACEK group (Haemophilus species, Aggregatibacter species, Cardiobacterium hominis, Eikenella corrodens, and Kingella species), rarely involved in SEA. Following surgery, the patient was treated with intravenous ceftriaxone for 6 weeks, and this gave excellent results.

Necrotizing myositis case report and brief literature study.

Necrotizing myositis is an extremely rare soft tissue infection, mainly caused by Group A Streptococci. Although its presentation is nonspecific and seems harmless, it quickly leads to death in almost all cases. Therefore, diagnosis and treatment of necrotizing myositis are considered as medical emergencies. The 27 years old patient we report benefited from early diagnosis and care. Necrotic tissues were surgically removed 24 hours after the appearance of the first clinical signs. Intravenous antibiotherapy as well as immunoglobulin therapy were also given on the first day. Starting from this clinical case, we present a brief explanation of the pathogenesis, the key clinical features and appropriate tools for diagnosis. Then, adequate antibiotherapy, role of immunoglobulin therapy and interest of hyperbaric oxygenotherapy will be discussed.

Disseminated histoplasmosis: case report and review of the literature.

Case report We report the case of a young Cameroonian woman who presented with cough, hyperthermia, weight loss, pancytopenia, and hepatosplenomegaly. A positive HIV serology was discovered and a chest radiography revealed a 'miliary pattern'. Bone marrow aspiration pointed out yeast inclusions within macrophages. Given the morphological aspect, the clinical presentation and immunosuppression, histoplasmosis was retained as a working hypothesis. Antiretroviral and amphotericin B treatments were promptly initiated. Review Given the immigration wave that Europe is currently experiencing, we think it is important to share experience and knowledge, especially in non-endemic areas such as Europe, where clinicians are not used to face this disease. Histoplasmosis is due to Histoplasma capsulatum var. capsulatum, a dimorphic fungus. Infection occurs by inhaling spores contained in soils contaminated by bat or bird droppings. The clinical presentation depends on the immune status of the host and the importance of inoculum, varying from asymptomatic to disseminated forms. AIDS patients are particularly susceptible to develop a severe disease. Antigen detection, molecular biology techniques, and microscopic examination are used to make a rapid diagnosis. However, antigen detection is not available in Europe and diagnosis needs a strong clinical suspicion in non-endemic areas. Because of suggestive imagery, clinicians might focus on tuberculosis. Our case illustrates the need for clinicians to take histoplasmosis in the differential diagnosis, depending on the context and the patient's past history.

Impaired hemoglobin scavenging during an acute HIV-1 retroviral syndrome.

BACKGROUND: We report a case of temporary impaired hemoglobin scavenging in a patient with an acute HIV-1 retroviral syndrome. The patient was presented at the emergency department in a severe inflammatory state, mimicking bacterial sepsis and/or hemophagocytic syndrome. The serum showed a hemolytic aspect. In contrast, serum haptoglobin concentration was not decreased. METHODS: The hemolysis index was determined and the visual absorbance spectroscopy spectrum of the serum was studied. alpha1 microglobulin and hemopexin concentrations were determined in serum. The presence of circulating hemoglobin:haptoglobin complexes in serum and the saturation of the haptoglobin were investigated using starch gel electrophoresis followed by peroxidase staining. CD163 expression on peripheral blood monocytes was analyzed using flow cytometry. RESULTS: A temporarily impaired hemoglobin scavenging was documented by an increased hemolysis index, absence of decreased haptoglobin levels, presence of circulating hemoglobin:haptoglobin complexes in serum and decreased hemopexin and alpha1 microglobulin concentrations. CONCLUSIONS: A temporarily impaired hemoglobin scavenging was observed due to a transient CD163 pathway impairment following an acute HIV-1 retroviral syndrome. The patient improved clinically and biochemically after initiation of HIV-1 anti-retroviral therapy. The data suggest a transient HIV-1 mediated CD163 impairment, although a latent drug mediated block could not be ruled out completely.

Impact of HIV-1 infection on the hematological recovery after clinical malaria.

BACKGROUND: Anemia is the most frequent cytopenia in HIV-infected individuals and is often associated with malaria. OBJECTIVE: To assess the impact of HIV-1 on the hematological recovery after a clinical malaria episode. METHODS: In Ndola, Zambia, a region with high malaria and HIV prevalence, hemoglobin (Hb) was measured in 634 malaria patients 14 and 45 days after antimalarial treatment. Risk factors for hematological recovery were analyzed in a multivariate linear regression model. RESULTS: At enrollment, HIV-1-infected malaria patients had lower Hb compared with HIV-1 uninfected (122.7 vs 136.0 g/L; P < 0.001). In both groups, mean Hb was significantly lower at day 14 posttreatment than day 0 (P < 0.0001) and significantly higher at day 45 than at day 14 (HIV-1 negative: P = 0.0001; HIV-1 infected: P = 0.005). HIV-1 was a risk factor for a larger Hb decrease until day 14 (P < 0.001) and slower recovery until day 45 (P = 0.048). When considering the whole 45-day follow-up period, mean Hb increased in the HIV-1-negative group (+3.54 g/L; 95% confidence interval: 1.37 to 5.70; P = 0.001) but not in the HIV-1-infected group (-0.72 g/L; 95% confidence interval: -3.85 to +2.40; P = 0.64). HIV-1 infection as such (P < 0.0001), not CD4 cell count (P = 0.46), was an independent risk factor for a slower hematological recovery. CONCLUSIONS: HIV-1-infected malaria patients had a slower hematological recovery after successful parasite clearance. Malaria preventive measures should be targeted to this high-risk group.

Fever after a stay in the tropics: clinical spectrum and outcome in HIV-infected travelers and migrants.

OBJECTIVE: To investigate the epidemiology and clinical spectrum of fever in HIV-infected returning travelers and migrants. METHODS: From April 2000 to December 2006, we explored prospectively, at our referral travel/HIV clinics, the etiology and outcome of febrile illnesses developing within 3 months after a stay in the tropics. For this study, we compared the morbidity profile between HIV-infected individuals and all other cases tested HIV negative. RESULTS: Of the 1850 adults (15 years and older) evaluated for 1921 fever episodes, 93 (5%) had HIV infection, including 5 presenting with primary infection. HIV prevalence was 2% in western travelers or expatriates, 11% in travelers "visiting friends and relatives," and 24% in foreign visitors/migrants. Fever episodes (n = 104) occurring in the HIV-infected individuals were mainly due to opportunistic infections (23%, including tuberculosis), respiratory tract infections (20%), sexually transmitted infections (9%), and noninfectious diseases (7%). All these conditions were more frequently diagnosed than in HIV-negative travelers (1035 fever episodes), although tropical infections (mostly malaria) were proportionally less prevalent. Morbidity (rate and duration of hospitalization) was more considerable in HIV-infected patients than in HIV-negative individuals. CONCLUSIONS: HIV infection was frequent in returning travelers and migrants presenting with fever at our setting and affected strongly the diagnostic spectrum and overall morbidity.

Special infectious disease risks of expatriates and long-term travelers in tropical countries. Part I: malaria.

Malaria risk is dependent upon the entomological inoculation rate actually faced by the long-term traveler. Risk is cumulative, increases with duration of exposure, is greatest in rural and periurban areas, and least in urban centers. Risk may be zero in some urban centers, especially during dry seasons. Chemoprophylaxis compliance is hindered by the high adverse event rate often reported by users, is often suboptimal in expatriates, and decreases with duration of stay. Compliance with personal protection measures may also be suboptimal, and use of insecticide-treated nets and effective repellents should be encouraged. Alternative strategies to mitigate risk include seasonal chemoprophylaxis, nonuse of chemoprophylaxis with rapid treatment, self-testing, self-treatment where competent care and quality drugs are unavailable, and vector control. Choice of strategies will depend upon assessment of actual risk and likely compliance, with a combination of measures usually appropriate.

Special infectious disease risks of expatriates and long-term travelers in tropical countries. Part II: infections other than malaria.

A wide range of viral, bacterial, and protozoal diseases pose risk to long-term tropical travelers. Risk varies geographically and with lifestyle. For some infections, risk increases with duration of stay, coming to resemble that of the local population. Risk management strategies include vaccination, chemoprophylaxis, avoidance measures, and screening, where appropriate. Vaccination against hepatitis A and B, typhoid, and rabies is recommended for all long-term travelers to (sub-)tropical areas. Lowering of the vaccination threshold for Japanese encephalitis is suggested. Meningococcal disease is rare in travelers, but vaccination is safe and acceptable. The efficacy of Bacillus Calmette-Guérin (BCG) is uncertain; immunological testing avoids BCG's confounding of tuberculin testing. Diarrhea is common, and self-treatment may be recommended. Sexually transmitted infections including human immunodeficiency virus (HIV) are serious risks; education, screening, and HIV postexposure prophylaxis following involuntary exposure are recommended. Many infections are chronic or asymptomatic, and appropriate screening is recommended on return or after prolonged exposure.

HIV-1 immune suppression and antimalarial treatment outcome in Zambian adults with uncomplicated malaria.

BACKGROUND: Human immunodeficiency virus (HIV)-1 infected adults with low CD4 cell count have a higher risk of malaria infection and clinical malaria. We assessed the influence that HIV-1 immune suppression has on the efficacy of antimalarial treatment in adults with uncomplicated malaria. METHODS: This clinical trial included 971 Zambian adults with uncomplicated malaria. Patients were tested for HIV-1, and, if positive, a CD4 cell count was assessed. The primary outcome was recurrent parasitemia corrected by molecular genotyping within 45 days after treatment. RESULTS: HIV-1 infection was detected in 33% (320/971) of adult patients with malaria. Treatment failure was not associated with HIV-1 infection (relative risk [RR], 1.12 [95% confidence interval {CI}, 0.82-1.53]; P=.45). HIV-1-infected patients with a CD4 cell count <300 cells/microL had an increased risk of recurrent parasitemia, compared with those with a CD4 cell count >or=300 cells/microL (RR, 2.24 [95% CI, 1.20-4.14]; P=.01). After genotyping, the risk of recrudescence was higher in HIV-1-infected patients with a CD4 cell count <300 cells/microL than in the other patients with malaria (RR, 1.67 [95% CI, 1.13-2.47]; P=.02). CONCLUSION: HIV-1-infected patients with malaria with a CD4 cell count <300 cells/microL have a higher risk of experiencing a recrudescent infection, compared with those with a CD4 cell count >or=300 cells/microL or without HIV-1 infection. Trial registered at http://www.clinicaltrials.gov/; reference number NCT00304980.

Safety and efficacy of lumefantrine-artemether (Coartem) for the treatment of uncomplicated Plasmodium falciparum malaria in Zambian adults.

BACKGROUND: In Zambia, unacceptably high resistance to commonly used antimalarial drugs prompted the choice of artemether-lumefantrine (AL) as first line treatment for uncomplicated Plasmodium falciparum malaria. Although the safety and efficacy of AL have been extensively documented, no clinical trials had been carried out in Zambia. METHODS: Nine hundred seventy one adult patients with uncomplicated malaria were randomized to either sulfadoxine-pyrimethamine (SP)(486) or AL (485) and followed up for 45 days. Outcome of treatment was defined according to the standard WHO classification. Recurrent parasitaemia were genotyped to distinguish between recrudescence and new infection. RESULTS: Fever at day 3 was significantly lower (AL: 0.9%; 4/455; SP: 3,5%; 15/433; p = 0.007) and the mean haemoglobin at day 45 significantly higher (AL: 134 g/l; SP 130 g/l; p = 0.02) in the AL group. Almost all clinical symptoms cleared faster with AL. Early treatment failure was significantly higher in the SP (25/464) than in the AL (2/463) (OR: 13.1 95% CI: 3.08-55.50; P < 0.001). The rate of new infections was similar in both groups (18 with SP and 19 with AL). Late clinical failure (OR: 2.55; 95% CI: 1.34-4.84; P = 0.004) and late parasitological failure (OR:3.18; 95% CI: 1.25-8.09; P = 0.02) were significantly higher in the SP group. Total treatment failure was significantly higher in the SP group (96/393; 19.3%) as compared to the AL (22/403; 5.4%) group (OR: 4.15; 95% CI: 2.52-6.83; P < 0.001). CONCLUSION: In Zambia, the new first line regimen AL is far more efficacious than SP in treating uncomplicated P. falciparum malaria in adults. Data on safety and efficacy of AL in pregnant women are urgently needed.

Efficient stimulation of HIV-1-specific T cells using dendritic cells electroporated with mRNA encoding autologous HIV-1 Gag and Env proteins.

Infection with human immunodeficiency virus type 1 (HIV-1) is characterized by dysfunction of HIV-1-specific T cells. To control the virus, antigen-loaded dendritic cells (DCs) might be useful to boost and broaden HIV-specific T-cell responses. In the present study, monocyte-derived DCs from nontreated HIV-1-seropositive patients were electroporated with codon-optimized ("humanized") mRNA encoding consensus HxB-2 (hHXB-2) Gag protein. These DCs elicited a strong HIV-1 Gag-specific interferon-gamma (IFN-gamma) response by an HLA-A2-restricted CD8+ T-cell line. Moreover, hHXB-2 gag mRNA-electroporated DCs also triggered IFN-gamma secretion by autologous peripheral blood mononuclear cells (PBMCs), CD4+ T cells, and CD8+ T cells from all patients tested. Next, a novel strategy was developed using autologous virus sequences. Significant specific IFN-gamma T-cell responses were induced in all patients tested by DCs electroporated with patients' autologous polymerase chain reaction (PCR)-amplified and in vitro-transcribed proviral and plasma viral mRNA encoding either Gag or Env. The stimulatory effect was seen on PBMCs, CD8+ T cells, and CD4+ T cells, demonstrating both major histocompatibility complex (MHC) class I and MHC class II antigen presentation. Moreover, a significant interleukin-2 (IL-2) T-cell response was induced by DCs electroporated with hHxB-2 or proviral gag mRNA. These findings open a major perspective for the development of patient-specific immunotherapy for HIV-1 disease.