Abstract
Monoclonal antibodies were generated with a hybridoma technique after immunization of mice with intact human parathyroid cells. Three antibodies of the IgG type reacted in immunohistochemistry and immunofluorescense with parathyroid epithelial cells and proximal tubule cells of the kidney but not with a large number of other human tissues. Adenomatous and hyperplastic parathyroid glands of patients with hyperparathyroidism (HPT) demonstrated a reduced immunohistochemical reactivity with the antibodies as compared to the intense staining of normal human parathyroid tissue. Experiments with dispersed parathyroid cells from the normal and pathological glands showed that 2 of the antibodies blocked the effects of ambient calcium on cytoplasmic calcium and parathyroid hormone release. This indicates that the antibodies interfere with a newly recognized receptor mechanism of parathyroid cells involved in the sensing and gating of calcium and thereby also in the regulation of parathyroid hormone release. A reduced expression of the putative calcium receptor mechanism of abnormal parathyroid cells may, thus, be principally responsible for the hypercalcemia of HPT by causing a decreased sensitivity to extracellular calcium of the parathyroid hormone release from pathological parathyroid glands. The antibodies should improve the parathyroid histological diagnosis by their ability to identify and to distinguish between normal and abnormal parathyroid tissue.
Résumé
Des anticorps monoclonaux ont été fabriqués avec la technique hybridoma après immunisation des souris avec des cellules parathyroïdes humaines intactes. Trois anticorps du type IgG ont réagi en immunohistochimie et en immunofluorescence avec les cellules épithéliales et tubulaires proximales du rein, mais pas avec la plupart des autres tissus humains. Les glandes parathyroïdes adénomateuses et hyperplasiques des patients hyperparathyroïdiens ont présenté une réactivité immunohistoclinique réduite vis-à-vis des anticorps en comparaison avec la coloration intense du tissu parathyroïde humain normal. En utilisant expérimentalement des cellules parathyroïdiennes dispersées provenant des glandes normales et pathologiques, 2 des anticorps bloquaient les effets du calcium ambiant sur le largage du calcium cytoplasmique et de la parathormone. Cela indique que les anticorps interfèrent avec le mécanisme récepteur, récemment reconnu, des cellules parathyroïdiennes, qui détermine la présence du calcium et qui en contrôle les mouvements de l'ion, et par conséquent joue un rôle dans le relargage de la parathormone. La réduction de l'expression du mécanisme suppose des récepteurs de calcium des cellules parathyroïdiennes anormales pourrait donc être responsable de l'hypercalcémie dans l'hyperparathyroïdisme. Le mécanisme serait une baisse de la sensibilité au calcium extracellulaire provoquant une baisse de la sécrétion de parathormone par les glandes parathyroïdiennes pathologiques. Ces anticorps pourraient améliorer le diagnostic histologique par leur capacité d'identifier et de distinguer entre les tissus parathyroïdes normal et pathologique.
Resumen
Anticuerpos monoclonales fueron generados mediante una técnica de hibridoma después de immunizar a ratones con células paratiroideas humanas intactas. Tres anticuerpos del tipo IgG reaccionaron en immunohistoquímica e immunofluorescencia con células paratiroideas epiteliales y con células del túbulo proximal del rinón, pero nó con un gran número de otros tejidos humanos. Las glándulas paratiroides adenomatosas e hiperplásicas de pacientes con hiperparatiroidismo (HPT) mostraron una reducida reactividad immunohistoquímica con los anticuerpos en comparación con la intensa coloración del tejido paratiroideo humano normal. Experimentos con células paratiroideas dispersas provenientes de glándulas normales y patológicas mostraron que 2 de los anticuerpos bloqueaban los efectos del calcio ambiental sobre el calcio citoplásmico y la liberación de hormona paratiroidea. Esto indica que los anticuerpos interfieren con un recientemente descubierto mecanismo receptor de las células paratiroideas involucradas en la percepción del calcio y la regulation de la liberación de hormona paratiroidea. Una expresión reducida del mecanismo receptor putativo de calcio de las células paratiroideas anormales puede ser, por consiguiente, el responsable principal de la hipercalcemia del HPT al causar una disminuida sensibilidad al calcio extracelular en el proceso de la liberación de la hormona por las células paratiroideas patológicas. Estos anticuerpos deben contribuir a mejorar el diagnóstico histológico paratiroideo por su capacidad de identificar y diferenciar el tejido normal del patológico.
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Supported by the Swedish Cancer Society and the Tore Nilsson Fund.
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Juhlin, C., Åkerström, G., Klareskog, L. et al. Monoclonal antiparathyroid antibodies revealing defect expression of a calcium receptor mechanism in hyperparathyroidism. World J. Surg. 12, 552–557 (1988). https://doi.org/10.1007/BF01655449
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DOI: https://doi.org/10.1007/BF01655449