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Immunohistochemical Detection of Salivary Agglutinin/gp-340 in Human Parotid, Submandibular, and Labial Salivary Glands

¹ Department of Dental Basic Sciences, Academic Centre for Dentistry Amsterdam (ACTA), Van der Boechorststraat 7, 1081 BT Amsterdam, the Netherlands;
² Department of Oral & Maxillofacial Surgery/Oral Pathology, University Medical Centre Vrije Universiteit (VUMC), Amsterdam, The Netherlands; and
³ Department of Immunology and Microbiology, Institute for Medical Biology, University of Southern Denmark, 5000 Odense C, Denmark;

View larger version (28K): [in this window] [in a new window]   Figure 1. Human saliva was separated on 7.5% polyacrylamide gels, transferred to nitrocellulose, and immunoblotted with mAb 213-6. Lane 1, parotid saliva; lane 2, submandibular saliva; lane 3, labial saliva. Agglutinin was detected in all samples, but no differences in electrophoretic behavior were observed.

View larger version (30K): [in this window] [in a new window]   Figure 2. Parotid saliva was separated on 7.5% polyacrylamide gels and transferred to nitrocellulose. Subsequently, blots were treated by periodate (P) or neuraminidase (N), columns 2 and 3, respectively. Control blots (C) remained unaffected. - = unreduced; + = reduced. After reduction, the recognition by mAb 213-6 is lost. Periodate and neuraminidase treatment abolished recognition by mAbs 213-1 and 5E9 as well as by lectin MAA. mAb 213-6 was unaffected by these treatments.

View larger version (26K): [in this window] [in a new window]   Figure 3. Parotid and submandibular saliva were separated on 4-15% polyacrylamide gels, transferred to nitrocellulose, and immunoblotted with various mAbs. Parotid saliva, lanes 1, 3, and 5; submandibular saliva, lanes 2, 4, and 6. mAb 213-1 and mAb 213-6 were raised against lung gp-340; CpMG2 was raised against a MUC7 peptide. In contrast to mAb 213-6, mAb 213-1 showed cross-reactivity with MUC7.

View larger version (111K): [in this window] [in a new window]   Figure 4. Immunohistochemical localization of salivary agglutinin/gp-340 in human parotid, submandibular, and labial salivary tissue. (A-D) Serous parotid gland tissue: Duct cells are labeled by mAb 213-1 (A and B) and mAb 213-6 (C and D). The strongest labeling was observed in the striated ducts, but also the excretory and intercalated ducts were stained, albeit less intensely. The serous acini were all negative. (E and F) Seromucous submandibular gland tissue: Serous acini (s) and demilune cells (d) are labeled by mAb 213-6. The mucous acini (m) and duct cells were negative. (G and H) Labial gland tissue: Demilune cells (d, inset), serous acini (s), and the luminal site of the duct cells (l) are labeled by mAb 213-6. Again, all mucous cells (m) were found negative. Magnification A, C, E, and G = 100x; B, D, F, and H = 400x.

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