Detection of mycobacterial lipoarabinomannan with an antigen-capture ELISA in unprocessed urine of Tanzanian patients with suspected tuberculosis.

A direct antigen-capture ELISA based on the detection of mycobacterial lipoarabinomannan (LAM) in unprocessed urine was evaluated for its usefulness in clinical practice. In Tanzania, 231 patients with suspected pulmonary tuberculosis (TB) and 103 healthy volunteers were screened with standard TB tests and with the new LAM-ELISA. Of 132 patients with confirmed pulmonary mycobacterial disease (positive sputum culture), 106 were positive using the LAM-ELISA (sensitivity 80.3%). In comparison, the sensitivity of acid-fast bacilli (AFB) sputum microscopy was 62.1% (82 of 132 confirmed cases). Of the 231 patients, 17 were both culture- and AFB-negative, but had typical radiographic signs of pulmonary mycobacterial infection and did not respond to antibiotic treatment. Of these 17 patients, 13 (76.5%) had positive LAM-ELISA test results. To define the specificity of the assay, urine samples from 103 healthy volunteers were also screened using LAM-ELISA. All but one had an optical density below the cut-off (specificity 99%). Of interest was a significant correlation between level of microscopic density of mycobacteria in sputum and LAM antigen concentration in urine (chi2=8.44). The LAM-ELISA is a field-adapted tool that can improve screening standards in countries with a high incidence of TB.

Liposomes as carriers of peptide antigens: induction of antibodies and cytotoxic T lymphocytes to conjugated and unconjugated peptides.

In the quest for effective immunization against complex diseases such as cancer, parasitic diseases, AIDS, and other viral infections, numerous peptides and recombinant proteins have been synthesized, examined for the ability to induce antibodies and CTLs, and tested for binding capability and therapeutic or prophylactic efficacy against the original target cell or organism. A liposome formulation, consisting of alum-adsorbed liposomes containing both a potent adjuvant, lipid A, and encapsulated or surface bound antigen, has had a record of safety and strong effectiveness for induction of antibodies in human vaccine trials. These same liposomes can also serve as effective vehicles for delivering conjugated or unconjugated peptides and proteins to antigen presenting cells for presentation via MHC class I and class II pathways for induction of CTLs and antibodies in experimental animal models. Liposomal lipid A appears to be extremely important, and is often a requirement, as an adjuvant for induction of CTLs against liposomal peptide antigens. Computer-generated molecular modelling analysis of small unconjugated or lipid-conjugated peptides strongly suggests that the expression of peptide antigen on the surface of the liposomes can be an important factor both in the induction of antibodies and in determining antibody specificities to small peptides. However, antigenic surface expression of liposomal peptide is not required for induction of CTLs. The data suggest that small synthetic peptides, synthesized with or without a lipid tail, or chemically conjugated to the surface of liposomes, might serve as effective antigenic epitopes, in combination with liposomal lipid A for induction of antibodies and CTLs.