Immunorecognition of different ganglioside epitopes on human normal and melanoma tissues

Abstract

There is increasing evidence that cell-surface gangliosides play a role in tumor growth, progression and metastases. In order to determine the frequency of ganglioside GD3 in patients with metastatic malignant melanoma for further therapeutic trials, GD3 ganglioside expression was determined in 119 tissue samples. Of these melanomas, 93% (111/119) were R-24-positive, which indicates the value of this diagnostic marker for melanoma. To study the structural epitopes of gangliosides, 10 ganglioside antibodies with defined specificities and affinities were tested on over 100 fresh-frozen tissue specimens of human normal and melanoma tissues. All the antibodies tested recognize the ganglioside GD3, but vary in their cross-reactivity with other gangliosides. According to their epitope specificity, they can be divided into 5 groups. For example, the antibodies Z-21 and A-4 react like the previously established MAb R-24 with gangliosides GD3 and GQlb, and one MAb (Q-4) detects all gangliosides containing 2 connected sialic acids (GD3, GD2, GDlb, GTlb, GQlb). Specificity on TLC does not always correlate with specificity to melanoma tissues and vice-versa. For example, MAb A-4, which recognizes only GD3 and GQlb on TLC, shows no specific reactivity on tissues. Furthermore, antibodies with the same ganglioside specificity do not have the same staining pattern on human tissues. For example, MAb Z-21, which is directed against the same gangliosides as MAb R-24 on TLC, does not cross-react with as many neuroectodermal tissues as MAb R-24. Because of their distinct properties, some of these antibodies may be even more useful for immunodiagnosis and immunotherapy of malignant melanoma than MAb R-24.

Selection of tumor antigens as targets for immune attack using immunohistochemistry: I. Focus on gangliosides

“…9-O-acetyl-GD3 has been reported to be present in granulocytes and T lymphocytes by Kniep et al (1993) but was not observed in our study in any normal tissues except for weak expression in connective tissue of prostate and ovary, making it a good target for immune attack against melanoma. GD3 and GD2 expression on melanomas, neuroblastomas and sarcomas (Hamilton et al, 1993a,b;Dippold et al, 1985;Cheung et al, 1985), the expression of GD3 in normal brain and connective tissue elements as well as islet cells of the pancreas and the expression of GD2 in brain and connective tissue elements in a variety of organs have all been described before Bernhard et al, 1992;Cheung et al, 1985). However, the presence of GD2 in splenic white pulp, lymph node germinal centers and B cell lymphomas was unexpected and raises the possibility of using GD2 as a target against B cell lymphomas.…”

“…It has been difficult to select optimal antigens and tumor targets based on these previous studies. The distribution of the antigens studied here has been described (Dippold et al, 1985;Bernhard et al, 1992;Cheresh et al, 1984;Brezicka et al, 1989;Bremer et al, 1984;Husmann et al, 1990), but the number and types of tissues studied were generally limited and involved MAbs against one or two antigens without comparison with the expression of other antigens. For this reason, we have begun a large immunohistochemical study on frozen tissue sections of tumor and normal tissues using a panel of well-characterized murine MAbs against a series of these candidate antigens.…”

“…(Bernhard et al, 1992) and GD3-Lactone (in our bands). MAb D1.1 reacts with GD3 O-acylated at the 9, 8 or 7 positions of the proximal or terminal sialic acid (Ritter et al, 1990).…”

Determination of the Localization of Gangliosides Using Anti-ganglioside Antibodies: Comparison of Fixation Methods

“…In summary, whereas previous workers have recognized that the antigenicity and immunogenicity of gangliosides and other glycosphingolipids depends on the density of the antigen on the cell surface, the chain length of the fatty acids and their ␣ -hydroxylation, and the cryptic behavior of glycosphingolipids (Hakomori 1986), the present study demonstrates that the method of fixation is vitally important for unambiguous conclusions to be drawn about the localization of a ganglioside in biological tissues using anti-ganglioside antibodies. This is of importance not only for localizing gangliosides in biological tissues (Schwarz and Futerman 1996) but also for using anti-ganglioside antibodies as diagnostic tools (Bernhard et al 1992;Lloyd et al 1992). …”

Antitumor effects of a novel monoclonal antibody with high binding affinity to ganglioside GD3

“…KM641 was chosen from ten mouse anti-GD3 mAb on these criteria in this study. Several anti-GD3 mAb have been produced so far [2,5,7,14,15,25,27,36,37] and R24 is the only well-studied and clinically used anti-GD3 mAb among them. The binding specificity of KM641 was almost the same as that of R24 in that the both mAb had weak cross-reactivity with GQlb [2,36].…”