Novel Insights into Lectin Binding Patterns in the Nasopharyngeal Tonsil of Buffaloes (Bubalus bubalis)

Pawan  Kumar

doi:10.6000/1927-520X.2025.14.06

Authors

Pawan Kumar Department of Veterinary Anatomy, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125 004, India

DOI:

https://doi.org/10.6000/1927-520X.2025.14.06

Keywords:

Buffalo, Galactose, Glucose/mannose, Lectin, Nasopharyngeal tonsil, N- acetylgalactosamine, N-acetylglucosamine, Vimentin

Abstract

Background: The present study investigates the specificity of lectin binding in the nasopharyngeal tonsil of six healthy adult buffaloes (Bubalus bubalis), a species not extensively studied regarding its immune system. Lectins, proteins that bind specifically to carbohydrates, are used to identify and characterize different cell types that may have roles in immune responses. This study explores how lectins bind to various cells within the nasopharyngeal tonsil, shedding light on cellular differentiation, interactions, and the potential functional roles of these cells in mucosal immunity.

Methods: A total of 21 biotinylated lectins, grouped into five categories based on their carbohydrate specificity (N-acetylglucosamine, N-acetylgalactosamine, galactose, glucose/ mannose, and fucose), were used to probe the nasopharyngeal tonsil tissue. Lectin histochemistry was applied to identify the binding patterns of these lectins to different cell types within the tissue, including epithelial cells, lymphoid cells, and specialized structures such as M-cells and P-cells. The study also involved the detection of vimentin filaments to explore potential immune responses within the tissue.

Results: Lectin histochemistry revealed a dynamic epithelial composition of the nasopharyngeal tonsil, consisting of pseudostratified columnar ciliated epithelium and lymphoepithelium, with distinct adaptations in the follicle-associated epithelium (FAE). The FAE exhibited M-cells, which are believed to play a role in antigen processing. Additionally, a new class of cells, termed P-cells, was identified based on their lectin-binding patterns, which share similarities with M-cells but are distinct in their function. Lectins targeting N-acetylglucosamine exhibited varying affinities for M- and P-cells, while lectins recognizing N-acetylgalactosamine selectively bound to cilia and goblet cells. Lectins targeting galactose produced complex staining patterns in mucous glands and lymphoid tissues. Specific binding was also observed in lymphoid cells with lectins recognizing glucose/mannose and fucose groups. Vimentin filaments in lymphocytes and specialized epithelial cells suggest an involvement in immune response mechanisms.

Conclusion: This study provides new insights into structural organisation landscape of the buffalo nasopharyngeal tonsil, highlighting the role of lectin-binding patterns in identifying specialized cells and tissues. The M-cells and discovery of P-cells and the detailed lectin-binding profiles may contribute to understanding the cellular dynamics of mucosal immunity. Additionally, the structural details uncovered in this study may serve as a valuable reference for comparative research on mucosal immunity across different species, advancing our understanding of antigen recognition and immune responses at mucosal surfaces.

References

Ibrahim D, Nakamuta N, Taniguchi K, Taniguchi K. Lectin histochemical studies on the vomeronasal organ of the sheep. J Vet Med Sci 2013; 75(8): 1131-7. DOI: https://doi.org/10.1292/jvms.12-0532

Kumar P, Timoney JF, Sheoran AS. M cells and associated lymphoid tissue of the equine nasopharyngeal tonsil. Equine Vet J 2001; 33(2): 224-30. DOI: https://doi.org/10.2746/042516401776249697

Ranjit R, Kumar P, Singh G. Histology, histochemistry, and scanning electron microscopy of nasopharyngeal tonsil of the young pigs. Indian J Vet Anat 2015; 27: 29-32.

Girgiri IA, Kumar P. Histology, histochemistry, and ultrastructure of the nasopharyngeal tonsil of the buffalo (Bubalus bubalis). Anat Histol Embryol 2019; 48(4): 375-83. DOI: https://doi.org/10.1111/ahe.12452

Madara JL, Bye WA, Trier JS. Structural features of and cholesterol distribution in M-cell membranes in guinea pig, rat, and mouse Peyer's patches. Gastroenterology 1984; 87: 1091-103. DOI: https://doi.org/10.1016/S0016-5085(84)80069-4

Smith MW, James PS, Tivey DR. M cell numbers increase after transfer of SPF mice to a normal animal house environment. Am J Pathol 1987; 128(3): 385-9.

Owen RL, Bhalla DK. Cytochemical analysis of alkaline phosphatase and esterase activities and of lectin-binding and anionic sites in rat and mouse Peyer's patch M cells. Am J Anat 1983; 168: 199-212. DOI: https://doi.org/10.1002/aja.1001680207

Giannasca PJ, Giannasca KT, Falk P, Gordon JI, Neutra MR. Regional differences in glycoconjugates of intestinal M cells in mice: potential targets for mucosal vaccines. Am J Physiol Gastrointest Liver Physiol 1994; 267: G1108-21. DOI: https://doi.org/10.1152/ajpgi.1994.267.6.G1108

Clark MA, Jepson MA, Simmons NL, Booth TA, Hirst BH. Differential expression of lectin-binding sites defines mouse intestinal M-cells. J Histochem Cytochem 1993; 41: 1679-87. DOI: https://doi.org/10.1177/41.11.7691933

Sharma R, van Damme EJ, Peumans WJ, Sarsfield P, Schumacher U. Lectin binding reveals divergent carbohydrate expression in human and mouse Peyer's patches. Histochem Cell Biol 1996; 105: 459-65. DOI: https://doi.org/10.1007/BF01457659

Murray MC, Bhavanandan VP, Davidson EA, Reinhold V. Modification of sialyl residues of glycoconjugates by reductive amination. Characterization of the modified sialic acids. Carbohydr Res 1989; 186: 255-65. DOI: https://doi.org/10.1016/0008-6215(89)84039-X

Ibrahim D, Nakamuta N, Taniguchi K, Yamamoto Y, Taniguchi K. Histological and lectin histochemical studies on the olfactory and respiratory mucosae of the sheep. J Vet Med Sci 2014; 76: 339-46. DOI: https://doi.org/10.1292/jvms.13-0436

Scocco P, Lepri E, Mercati F, Vitellozzi G, Mechelli L, Ceccarelli P. Glycohistochemical characterization of histologically normal nasal mucosa and enzootic nasal tumor of sheep. Am J Vet Res 2012; 73: 1128-36. DOI: https://doi.org/10.2460/ajvr.73.8.1128

Forstner JF. Intestinal mucins in health and disease. Digestion 1978; 17: 234-63. DOI: https://doi.org/10.1159/000198115

Abraham SN, Beachey EH. Host defenses against adhesion of bacteria to mucosal surfaces. Host Def Mech 1985; 4: 63-88.

Giannasca PJ, Boden JA, Monath TP. Targeted delivery of antigen to hamster nasal lymphoid tissue with M-cell-directed lectins. Infect Immun 1997; 65: 4288-96. DOI: https://doi.org/10.1128/iai.65.10.4288-4298.1997

Castells MT, Ballesta J, Madrid JF, Aviles M, Martinez-Menarguez JA. Characterization of glycoconjugates in developing rat respiratory system by means of conventional and lectin histochemistry. Histochem 1991; 95: 419-26. DOI: https://doi.org/10.1007/BF00266971

Paulsen FP, Tschernig T, Debertin AS, Kleemann WJ, Pabst R, Tillmann BN. Similarities and differences in lectin cytochemistry of laryngeal and tracheal epithelium and subepithelial seromucous glands in cases of sudden infant death and controls. Thorax 2001; 56: 223-7. DOI: https://doi.org/10.1136/thorax.56.3.223

Schroder JM. Epithelial peptide antibiotics. Biochem Pharmacol 1999; 57: 121-34. DOI: https://doi.org/10.1016/S0006-2952(98)00226-3

Gebert A, Hach G, Bartels H. Co-localization of vimentin and cytokeratins in M-cells of rabbit gut-associated lymphoid tissue (GALT). Cell Tissue Res 1992; 269: 331-40. DOI: https://doi.org/10.1007/BF00319625

Gebert A, Rothkötter HJ, Pabst R. M cells in Peyer's patches of the intestine. Int Rev Cytol 1996; 167: 91-159. DOI: https://doi.org/10.1016/S0074-7696(08)61346-7

Lelouard H, Reggio H, Mangeat P, Neutra M, Montcourrier P. Mucin-related epitopes distinguish M cells and enterocytes in rabbit appendix and Peyer's patches. Infect Immun 1999; 67(1): 357-67. DOI: https://doi.org/10.1128/IAI.67.1.357-367.1999

Ramirez C, Gebert A. Vimentin-positive cells in the epithelium of rabbit ileal villi represent cup cells but not M-cells. J Histochem Cytochem 2003; 51: 1533-44. DOI: https://doi.org/10.1177/002215540305101113