Detection of IgG3 antibodies specific to the human immunodeficiency virus type 1 (HIV-1) p24 protein as marker for recently acquired infection.

Reducing the risk of human immunodeficiency virus type 1 (HIV-1) transmission is still a public health priority. The development of effective control strategies relies on the quantification of the effects of prophylactic and therapeutic measures in disease incidence. Although several assays can be used to estimate HIV incidence, these estimates are limited by the poor performance of these assays in distinguishing recent from long-standing infections. To address such limitation, we have developed an assay to titrate p24-specific IgG3 antibodies as a marker of recent infection. The assay is based on a recombinant p24 protein capable to detect total IgG antibodies in sera using a liquid micro array and enzyme-linked immunosorbent assay. Subsequently, the assay was optimised to detect and titrate anti-p24 IgG3 responses in a panel of sequential specimens from seroconverters over 24 months. The kinetics of p24-specific IgG3 titres revealed a transient peak in the 4 to 5-month period after seroconversion. It was followed by a sharp decline, allowing infections with less than 6 months to be distinguished from older ones. The developed assay exhibited a mean duration of recent infection of 144 days and a false-recent rate of ca. 14%. Our findings show that HIV-1 p24-specific IgG3 titres can be used as a tool to evaluate HIV incidence in serosurveys and to monitor the efficacy of vaccines and other transmission control strategies.

Development of potent class II transactivator gene delivery systems capable of inducing de novo MHC II expression in human cells, in vitro and ex vivo.

Class II transactivator (CIITA) induces transcription of major histocompatibility complex (MHC) II genes and can potentially be used to improve genetic immunotherapies by converting non-immune cells into cells capable of presenting antigens to CD4+ T cells. However, CIITA expression is tightly controlled and it remains unclear whether distinct non-immune cells differ in this transactivator regulation. Here we describe the development of gene delivery systems capable of promoting the efficient CIITA expression in non-immune cell lines and in primary human cells of an ex vivo skin explant model. Different human cell types undergoing CIITA overexpression presented high-level de novo expression of MHC II, validating the delivery systems as suitable tools for the CIITA evaluation as a molecular adjuvant for gene therapies.

Evaluation of memory immune response to mycobacterium extract among household contact of tuberculosis cases.

The human immune response to tuberculosis (TB) is especially mediated by T CD4(+)lymphocytes. However, more studies are needed in order to understand the exact role of each cytokine in the mechanisms for cures. In this article, our aim was to analyze the production of TNF-alpha, IL-10, and IFN-gamma in peripheral blood mononuclear cells (PBMCs) among the household contacts of common primary TB cases, with or without histories of active TB infection, who were negative to parasitological and HIV tests. In order to characterize the cytokine production, PBMCs from these groups were stimulated with whole-protein extract of M. tuberculosis (WPE) antigen (rAgTb) for 24 and 48 hr. The culture supernatants were collected and IFN-gamma, TNF-alpha, and IL-10 were assayed using capture ELISA. There were no statistical differences between primary TB cases and their household contacts with or without previous histories of lung TB. Our results suggest that T memory cells, T regulatory cells, and the Th1/Th2 dichotomy may be responsible for the results described in this article. Further studies are currently underway.