The spirituality of the Order of St. John of Jerusalem is resumed by the motto Tuitio Fidei et Obsequium Pauperum, defense of the Faith and care for the poorest and sickest. The female branch of the Nuns of St. John also uses the motto Pro Fide et Utilitate Hominum, for Faith and in the service of manking.

The roots of the charism of the nuns of the Order of St. John can be found in: St. John the Baptist, with his mission and virtues; the Virgin Mary, in the Mystery of the Nativity; hospitality within the contemplative  life; and the Paschal Mystery of the Lord, consummated in Jerusalem.