O-acetyl sialic acid binding lectin as a probe for detection of subtle change on cell surface induced during acute lymphoblastic leukemia (ALL) and its clinical application.

A novel probe, a 9-O-acetylated sialic acid binding lectin, namely achatininH (ATNH) has been used for the detection of changes on the cell surface during acute lymphoblastic leukemia (ALL). ATNH does not agglutinate normal human erythrocytes, however it is capable of agglutinating erythrocytes and peripheral blood mononuclear cells (PBMC) of patients suffering from ALL. The differential expression of a key receptor, 9-O-acetylated sialo glyco conjugate (9-O-AcSG), on PBMC was observed using a simple lymphoproliferative assay (LA). The extent of expression of 9-O-AcSG was used as an index to distinguish ALL patients of different clinical stages and assess the probability of relapse. The amount of ATNH needed for maximum stimulation served as a tool to indirectly measure the extent of expression of 9-O-AcSG on PBMC surface. The acetylated sialo glycoconjugate was expressed at a very high concentration during acute phase of the disease. Subsequently it decreased during treatment persisted during maintenance therapy and reappeared with relapse. PBMC of normal human donors required 80 times more ATNH in comparison to the untreated acute phase ALL patients. No cross reactivity was found in non Hodgkin's lymphoma, chronic myelogenous leukemia and thalassaemia patients.

Designing of peptides with immuno-modulatory properties using protein A as a probe.

A series of reports from our laboratory have described the multifarious properties of protein A of Staphylococcus aureus Cowan I, apart from its IgG binding affinity. Original reports regarding its anti-tumor, anti-toxic, anti-carcinogenic and immunomodulatory properties published earlier by the authors have implicated some uniqueness of this bacterial protein. It was conceived that such diversified properties must lie in its specific peptide sequences, rendering it to act and behave as a multipotent "Biological Response Modifier" (BRM). The high resolution X-ray structure of protein A-Fc complex has been delineated earlier, and has been the foundation of many protein engineering studies. This structure along with the amino acid sequence data of its four repetitive domains provided us the basis for designing an octapeptide. This octapeptide was synthesized by solid phase peptide synthesis considering it as the probable site through which PA binds IgG. This octapeptide (NH2-Gln-Asn-Ala-Phe-Tyr-Glu-Ile-Leu-COOH) is present in the first helical segment of B-domain of protein A, and also is a part of domain D, A and C. This octapeptide has been shown to bind IgG by the immunoblotting technique. The binding affinity of the octapeptide appears to be significantly higher than that of intact protein A, as was revealed by calculation of Ka (association constant) and Kd (dissociation constant) values. This octapeptide might serve as a good immunoadsorbant for IgG and/or immune complexes.