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Vilnius Cathedral of the Orthodox Monastery of the Holy Spirit is located in the Old Town of Vilnius, near the Gate of Dawn, between Subačiaus, Šv. Dvasios and Aušros Vartų streets, near the Church of St. Theresa. It is a Lithuanian Orthodox centre.

This church is the most important Orthodox Church in Lithuania. Monasteries for men and women located near the church are the only Orthodox monasteries in Lithuania which are still used to this day.

The privilege to build the brick Orthodox Church of the Holy Spirit was granted by the King of Lithuania and Poland Vladislovas Vaza (Władysław IV Vasa).

The interior of the church is decorated with moulding and sculptures from stucco (which is not typical for Orthodox churches of other regions). Stucco is a high quality mixture of plaster and marble, usually either white or slightly yellow. The remains of three Orthodox saints – Anthony, Eustathius, and John rest in the ornate reliquary of the Centre of the Orthodox Church of the Holy Spirit. According to Orthodox tradition, they were killed due to political reasons under the rule of Algirdas in 1346 and 1347. It is said that they were Lithuanians, as well as courtiers of the Grand Duke who secretly accepted the Orthodox faith. Metropolitan Josifas Semaška was buried in the crypt.

The Orthodox Church of the Holy Spirit in Vilnius is the only Orthodox house of worship in Lithuania built according to the Latin cross plan of a Baroque church. There was already a monastery, school and a printing house near the Orthodox Church as early as at the end of the 16th century. The Monastery of the Holy Spirit did not avoid the most significant upheavals of the city in the 18th century: the Northern War and plague, fires, Kościuszko Uprising and the French invasion later on. The Orthodox Church burned down in 1749. It was reconstructed by Jonas Kristupas Glaubicas, one of the most famous Baroque style architects in Vilnius. The church was rebuilt once again in the 19th century.

The church and monastery ensemble is comprised of the Cathedral of the Holy Spirit, a bell tower, and three residential buildings of the monastery. The layout of buildings is free-form. The church is masonry, with three naves, a Latin cross plan, with the style of a basilica, two sacristies, 3 apses, and a tambour near the main western two-tower facade. The exterior of the building has features of the early Baroque period, while the interior has features of the late Baroque and Rococo periods. The facade has two towers (typical for Catholic churches), and three apses typical for Orthodox churches. The western facade is exceptional in that its towers stand near the inner sides of the nave instead of the corners.

The bell tower was built in 1638 (renewed in 1873) and has a square, massive, 3-tier design with the simple style of the early Baroque period. Buildings of the monastery were built in the 16th century (rebuilt in the middle of the 19th century); they have two floors and simple architecture. Men’s monastery is located in the southwestern part of the ensemble.