NATO supplies through Pakistan's Torkham border crossing into Afghanistan resumed on Sunday, 11 days after Islamabad closed the point in response to a NATO air attack, officials said.
"The first convoy of more than a dozen vehicles left for Afghanistan this afternoon," customs official Mohammad Nawaz told AFP.
More vehicles loaded with supplies for NATO and US troops were ready to leave, he added.
Pakistan's foreign ministry on Saturday announced the reopening of the main land route for NATO supplies "with immediate effect".
US and NATO forces are fighting a nine-year Taliban insurgency in Afghanistan and the route is vital to the war effort.
The decision came after US ambassador to Pakistan Anne Patterson on Wednesday apologised on behalf of the American people for the "terrible accident".
Pakistan shut the main Khyber Pass border crossing on September 30 after a cross-border NATO helicopter assault killed at least two Pakistani soldiers, straining already tense ties between the anti-terror allies.
The US apology came as Washington was pressing Pakistan to reopen the vital border crossing.
The closure compounded existing difficulties for the Western alliance as NATO supply trucks have come under repeated Taliban attack.
The closure compounded existing difficulties for the Western alliance as NATO supply trucks have come under repeated Taliban attack.
The top US military official, Admiral Mike Mullen, also tried to publicly repair relations damaged by NATO the chopper intrusion, which two Pakistani soldiers mistaken for militants were killed.
"We extend our deepest apology to Pakistan and the families of the Frontier Scouts who were killed and injured," Patterson said late Wednesday. "Pakistan's brave security forces are our allies in a war that threatens both Pakistan and the US."
Separately, in a letter to Pakistan's army chief General Ashfaq Kayani, Mullen on Thursday said the US military took the incident "very seriously" and offered his condolences to the families of the dead.
Mullen said senior commanders would review an investigation into the incident thoroughly in the hope of "avoiding recurrence of a tragedy like this".