Flow cytometry analysis with a new FITC-conjugated monoclonal antibody-3E12 for HLA-B*57:01 rapid screening in prevention of abacavir hypersensitivity in HIV-1-infected patients.

BACKGROUND: Rapid screening for the detection of HLA-B*57:01 in the prevention of abacavir hypersensitivity in HIV-1-infected patients is a hallmark for clinical services. OBJECTIVE: The aim of this work was to analyze the utility of flow cytometry with a new FITC-conjugated B-17 monoclonal antibody (mAb3E12) for HLA-B*57:01 screening in a Spanish cohort of 577 HIV-1+ individuals. METHODS: Cryopreserved peripheral blood mononuclear cell samples from HIV-1+ individuals were analyzed by flow cytometry with the mAb 3E12 that recognizes both HLA-B*57 and HLA-B*58 alleles (members of the group specificity, HLA-B17). Patients' DNA samples had been previously typed for HLA-B*57:01 with PCR-SSO or PCR-SSP and additional DNA sequencing (EPI Study). The results obtained by flow cytometry were compared with the results obtained by the DNA-PCR techniques. RESULTS: By flow cytometry, 46 samples (7.97%) were positive for HLA-B17, 530 (91.86%) were negative, and 1 (0.17%) was undetermined. All samples found negative by flow cytometry were negative for HLA-B*57:01 by DNA-PCR. Of the HLA-B17 positive samples, 31 (67.4%) were positive for HLA-B*57:01, 2 (3.25%) were positive for HLA-B*57:03, 11 (26.1%) were positive for HLA-B*58, and 2 (3.25%) were negative for both HLA-B*57 and HLA-B*58 antigens. The undetermined sample was negative for HLA-B*57 and HLA-B*58 alleles by DNA-PCR. CONCLUSIONS: This study shows that flow cytometry with mAb3E12 is a highly sensitive method (no false negatives) to implement prior to DNA-PCR analysis for rapid screening of HLA-B*57:01. Additional confirmation by molecular HLA typing method would be required in less than 10% of the cohort of HIV-1-infected individuals.

Sublingual therapeutic immunization with a polyvalent bacterial preparation in patients with recurrent respiratory infections: immunomodulatory effect on antigen-specific memory CD4+ T cells and impact on clinical outcome.

Recurrent respiratory tract infections (RRTIs) are common clinical conditions in individuals with alterations of the immune function. A prospective open pilot study in a cohort of patients with RRTIs has been performed to assess whether sublingual immunization with a polyvalent bacterial vaccine could exert an immunomodulatory effect on the antigen-specific immunological responses and have an impact on the clinical outcome. Seventeen patients with RRTIs were recruited. An oral polyvalent bacterial preparation (Bactek®) was administered to all patients daily for 6 months. Immunological assessment was performed at baseline and at the end of immunization. Immunological measurements included: T cell-specific proliferations of CD3+CD4+ and CD3+CD8+ to Bactek® antigens, total immunoglobulin levels, antibodies to pneumococcal polysaccharide and tetanus toxoid and B, T and natural killer (NK) cell subsets. There was a significant increase in the proliferative capacity of CD3+CD4+ T cells specific to Bactek® antigens at month 6 in comparison to baseline (P < 0·0001). A significant increase in total CD3+ T cells was also observed (P < 0·05). No significant differences were observed between baseline and month 6 in levels of total immunoglobulins, specific antibodies and B, T or NK cell subsets. A significant reduction in the patient's rate of RRTIs was observed compared with 1 year prior to initiation of therapy (P < 0·0001). The results demonstrate that long-term administration of a sublingual polyvalent bacterial preparation in patients with RRTIs exerts an immune stimulating effect on CD4+ T helper cell responses to bacterial antigens which could be associated with clinical benefit.

Missense mutation Thr309Lys in the coagulation factor XII gene in a Spanish family with hereditary angioedema type III.

BACKGROUND: A new type of hereditary angioedema (type III) affecting mainly women with normal C1-inhibitor level and function has been described. Exposition to estrogens is an important precipitating factor. Recently, a missense mutation in the gene of the blood coagulation factor XII (Hageman factor) has been reported in a few families with this type of hereditary angioedema. AIM: To study a patient and her family with recurrent swelling attacks during pregnancy. METHODS: Complement factors C3 and C4 as well as C1-inhibitor level and function were determined. Genomic DNA was isolated from venous blood samples and screened for mutations in the coagulation factor XII gene. RESULTS: C3 and C4 levels as well as C1-inhibitor level and function were normal. A missense mutation Thr309Lys was identified in factor XII gene with a heterozygotic pattern. This mutation was also identified in the mother of the patient, her daughter and her son. CONCLUSION: These results support that the mentioned mutation in factor XII gene causes hereditary angioedema type III.

[Introduction to biological drugs]. / Introducción a los fármacos biológicos.

Biological therapies have revolutionized the treatment of chronic systemic diseases in which the immune system disorders form a part of the disease mechanism. In these diseases, the patients follow different drug treatments for long periods of time that causes serious adverse reactions and often obtain unsatisfactory efficacy results. Due to the research conducted in the last 10 years, biological drugs have been introduced into the treatment that are aimed against specific targets, such as inflammatory and immunopathological responses that give rise to tissue injury. The new biological therapies have improved the currently available treatments due to greater efficacy, fast action and greater tolerability. The present work aims to provide a global and up-dated view on the biological agents used most in the usual clinical practice and their importance in the management of the chronic immunologically based inflammatory diseases.

Introducción a los fármacos biológicos / Introduction to biological drugs

Las terapias biológicas han revolucionado el tratamiento de las patologías crónicas sistémicas en las que las alteraciones del sistema inmunológico forman parte de los mecanismos de la enfermedad. En estas enfermedades los pacientes siguen diversos tratamientos farmacológicos durante largo tiempo, lo que produce reacciones adversas serias y, en muchas ocasiones, resultados de eficacia poco satisfactorios. Debido a las investigaciones de los últimos 10 años se han introducido en la clínica fármacos biológicos que están dirigidos contra dianas específicas, como las respuestas inflamatorias e inmunopatológicas que dan lugar a la lesión tisular. Las nuevas terapias biológicas han mejorado los tratamientos actualmente disponibles debido a su mayor eficacia, rapidez de acción y mejor tolerancia. El presente trabajo tiene como objetivo presentar una visión global y actualizada de los agentes biológicos más empleados en la práctica clínica habitual y su importancia en el manejo de las patologías inflamatorias crónicas de base inmunológica (AU)

Biological therapies have revolutionized the treatment of chronic systemic diseases in which the immune system disorders form a part of the disease mechanism. In these diseases, the patients follow different drug treatments for long periods of time that causes serious adverse reactions and often obtain unsatisfactory efficacy results. Due to the research conducted in the last 10 years, biological drugs have been introduced into the treatment that are aimed against specific targets, such as inflammatory and immunopathological responses that give rise to tissue injury. The new biological therapies have improved the currently available treatments due to greater efficacy, fast action and greater tolerability. The present work aims to provide a global and up-dated view on the biological agents used most in the usual clinical practice and their importance in the management of the chronic immunologically based inflammatory diseases (AU)

Immunization with an HIV-1 immunogen induces CD4+ and CD8+ HIV-1-specific polyfunctional responses in patients with chronic HIV-1 infection receiving antiretroviral therapy.

Development of polyfunctional T lymphocyte responses is critical in the immunological response against HIV-1. Fifty-four HIV-1 infected patients receiving antiretroviral treatment (ART) and immunization with an HIV-1 immunogen or placebo, periodically every 3 months throughout a period of 36 months, were evaluated for the purposes of analysing the development of HIV-1-specific CD4+ and CD8+ responses. A significant increase of proliferating and IFN-gamma producing CD8+ HIV-1-specific T cells, of HIV-1-specific precursor frequencies for CD8+ and for CD4+ T cells and of Gag/pol-specific memory CTL precursors (CTLp) was observed in the immunogen group in comparison to placebo. IL-2 intracellular expression and IFN-gamma and TNF-alpha co-expression in HIV-1-specific CD8+ T cells were also substantially increased in the immunized group. A negative correlation between viral load and CD3+CD4+CFSElow HIV-1-specific lymphoproliferative response and frequency of Gag/pol-specific CTLp was solely observed in the HIV-1 immunogen group. Long-term immunization in patients receiving ART helps to develop HIV-1-specific polyfunctional T cell responses.

Therapeutic immunization with an inactivated HIV-1 Immunogen plus antiretrovirals versus antiretroviral therapy alone in asymptomatic HIV-infected subjects.

To determine whether the addition of an inactivated-gp120-depleted HIV-1 Immunogen to antiretrovirals (ARTs) conferred a beneficial effect on delaying time to virologic failure relative to that obtained by ARTs alone, a phase II clinical trial was performed in 243 asymptomatic, ART naïve, HIV-1 seropositive adults. The Cox model showed that HIV-1 Immunogen treatment was associated with a 34% decrease in the risk of virologic failure (P = 0.056). When the analysis incorporated baseline HIV-RNA stratification the risk of virologic failure in the HIV-1 Immunogen Arm was significantly reduced a 37% compared to the IFA placebo Arm (P = 0.034). The data suggest that therapeutic immunization plus ARTs could influence virologic control.