Comparison of different monoclonal antibodies against immunosuppressive proteins of Ascaris suum

Abstract

The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract.

PAS-1, an Ascaris suum Protein, Modulates Allergic Airway Inflammation via CD8+ γδTCR+ and CD4+ CD25+ FoxP3+ T Cells

“…Using a monoclonal antibody obtained against the high molecular fraction of the extract, a 200 kDa protein with three polypeptide chains was purified by affinity chromatography and named PAS-1 (protein from Ascaris suum). This protein retained all the immunosuppressive properties of the whole extract [21,22]. In addition, PAS-1 inhibited the inflammatory leucocyte migration induced by bacterial lipopolysaccharide in a murine air pouch model and reduced the synthesis of proinflammatory cytokines.…”

“…Ascaris suum adult worm extract was prepared as described previously [27]. PAS-1 protein was purified from A. suum extract by affinity chromatography using the monoclonal antibody MAIP-1 coupled to Sepharose 4B as previously described [21]. According to the Limulus amebocyte lysate test (Bio Whitaker Inc., Walkersville, MD, USA), endotoxin levels were below the limit of detection in the PAS-1 eluate (<1.25 EU ⁄ mg protein).…”

Monoclonal antibodies in immunodiagnostic assays: a review of recent applications

“…They were shown to react with different antigenic epitopes of high molecular weight proteins. These antibodies have recognized the antigen with different affinity constants (Oshiro et al, 2004). Cryptococcosis is a mycotic disease of major concern and a potentially fatal disease that is the cause of the most common life-threatening meningitis in patients with acquired immunodefficiency syndrome (Andama et al, 2013).…”

Anti-inflammatory activity of PAS-1, a protein component of Ascaris suum

“…While the 29 kDa components induce IgE antibody production (8), the high molecular mass components, like the whole extract, display suppressive effects on the Th1 and Th2 immune responses to unrelated antigens (9). More recently, we isolated from Ascaris suum extract a single protein (PAS-1) that is responsible for the suppressive activity of the whole extract on humoral immune response [10].…”