AC Milan skipper Calabria, who has not been in the national team since June 2022, was included in the 29-player squad announced on Friday following his recent impressive performances for his club.
Striker Immobile, who earlier this season struggled with a hamstring injury, was not called up despite recent good form for Lazio which included scoring the only goal in Tuesday's Champions League home win against Feyenoord.
Juventus defender Andrea Cambiaso and Monza midfielder Andrea Colpani received their first call-ups.
Spalletti's Italy side are third in Group C with 10 points, three points behind Ukraine but with a game in hand.
Italy host North Macedonia on November 17, a team they haven't beaten in their last three encounters.
They conclude their campaign against Ukraine in Leverkusen on Nov. 20 in the match that will determine who gets the second automatic qualification spot.
England, top of the standings on 16 points, have already qualified for next year's finals in Germany.
Squad:
Goalkeepers: Gianluigi Donnarumma (Paris St Germain), Alex Meret (Napoli), Ivan Provedel (Lazio), Guglielmo Vicario (Tottenham Hotspur)
Defenders: Francesco Acerbi (Inter Milan), Alessandro Bastoni (Inter Milan), Alessandro Buongiorno (Torino), Davide Calabria (AC Milan), Andrea Cambiaso (Juventus), Matteo Darmian (Inter), Giovanni Di Lorenzo (Napoli), Federico Dimarco (Inter), Federico Gatti (Juventus), Rafael Toloi (Atalanta)
Midfielders: Nicolo Barella (Inter), Giacomo Bonaventura (Fiorentina), Andrea Colpani (Monza), Bryan Cristante (AS Roma), Davide Frattesi (Inter), Jorginho (Arsenal), Manuel Locatelli (Juventus)
Forwards: Domenico Berardi (Sassuolo), Federico Chiesa (Juventus), Stephan El Shaarawy (Roma), Moise Kean (Juventus), Matteo Politano (Napoli), Giacomo Raspadori (Napoli), Gianluca Scamacca (Atalanta), Nicolo Zaniolo (Aston Villa).