Ahly and Esperance fought a 0-0 draw in Alexandria while a Lemponye Tshireletso goal gave surprise qualifiers Rollers a 1-0 win over Kampala Capital City Authority of Uganda in Gaborone.
Rollers top the Group A standings with three points, Ahly and Esperance have one each and KCCA are pointless.
In another match, Mouloudia Alger of Algeria came from behind with 10 men to draw 1-1 at home against Difaa el Jadida of Morocco and share the Group B lead.
TP Mazembe of the Democratic Republic of Congo host Entente Setif of Algeria on Saturday in the other fixture in the mini-league.
Both fixtures in Group C and in Group D will also be staged Saturday with defending champions Wydad Casablanca of Morocco among the clubs in action.
Record eight-time champions Ahly sorely missed two injured strikers, Moroccan Walid Azaro and Nigerian Junior Ajayi, as they failed to score against twice title-holders Esperance.
It was the eighth draw in 17 matches between the teams across various CAF competitions with Ahly winning six and Esperance three.
They clashed in the 2017 Champions League quarter-finals with Ahly winning the second leg near Tunis after being held in Alexandria.
In Gaborone, Tshireletso timed his run perfectly on to a Joel Mogorosi pass and rounded goalkeeper Tom Ikapa to score Rollers' winner on 36 minutes.
Mogorosi wasted a good chance to double the lead almost immediately after the deadlock was broken when he fired wide from inside the six-yard box.
A couple of stoppage-time saves from Rollers goalkeeper Keagile Kgosipula prevented the Ugandan outfit levelling in a clash of group phase debutants.
Rollers shocked Al Merrikh of Sudan and Young Africans of Tanzania to become the first Botswana club to reach the last 16.
Mouloudia had Zidane Mebarakou red-carded early in the second half and fell behind after 77 minutes when Hamid Ahaddad of Jadida scored his seventh Champions League goal this season.
Amir Karaoui equalised with eight minutes remaining in Algiers for Mouloudia, who had scored 15 goals at home in two qualifiers.
Jadida are playing in the group phase for the first time and hope to emulate Moroccan clubs Raja Casablanca, Wydad Casablanca and FAR Rabat by winning the elite African club competition.