Non-specific binding of mouse IgG1 to Heligmosomoides polygyrus: parasite homogenate can affinity purify mouse monoclonal antibodies

Abstract

A characteristic feature of infections with the nematode parasite of mice Heligmosomoides polygyrus, is a marked IgG1 hypergammaglobulinaemia. A possible source for this immunoglobulin has recently been demonstrated, through evidence that H. polygyrus adult worm homogenate (AWH) can induce the in vitro production of non-specific IgG1 from mouse lymphocytes. To determine the interactions between this immunoglobulin and the parasite, the ability of IgG1 to bind to AWH of H. polygyrus was investigated. Protein (Western) blotting indicated that mouse monoclonal antibodies are able to bind non-specifically to selected parasite antigens. Furthermore, by binding H. polygyrus adult worm homogenate to cyanogen bromide (CNBr)-activated Sepharose CL-4B, an affinity column was prepared which could be used to efficiently purify mouse IgG1 monoclonal antibodies. These antibodies were eluted from the affinity column and still retained their original specificity. These results indicate that H. polygyrus not only induces the production of non-specific IgG1 by the host, it can also bind this immunoglobulin to its own specific proteins. Thus, it is possible that IgG1 produced during a primary infection with H. polygyrus may not entirely benefit the host.

Production of a recombinant version of a Heligmosomoides polygyrus antigen that is preferentially recognized by resistant mouse strains

“…We had previously shown that MAb 13B3 binds to proteins found in the adult worm homogenate (AWH) of H. polygyrus (17). We extended these observations to show that the MAb also was able to bind to a combination of proteins found in the E/S of both adult and L4 stages of the parasite (Figure 1).…”

“…(13). The MAb had been initially produced against adult worm homogenate (17), and it was therefore of interest to note that one MAb, called 13B3, was able to bind to a proteins found in both adult and L4 E/S (Figure 1).…”

“…Normal mouse serum was taken from naïve BALB/c mice and stored frozen until required. The production of hyper‐immunized mouse antiserum (HIMS) from BALB/c mice has been described previously (17).…”

“…Immunoglobulin (Ig) G1 monoclonal antibody (MAb) 13B3 was produced in this laboratory according to procedures described elsewhere (16). The binding characteristics of this antibody to H. polygyrus adult worm homogenate have been described previously (17).…”