Pendleton thrashed Meares for the sprint title at the Beijing Games four years ago, but the Australian enters the Hisense Arena holding the world title and is desperate to secure bragging rights ahead of London.
A self-described "emotional" rider, Pendleton nearly quit the sport two years ago but was persuaded to defend her title at London's Velodrome and has added spice to their clash in the lead-up by branding Meares a rider who "pushes the rules".
Meares, who took gold at the 2004 Athens Games in the now defunct 500 metres time trial, signalled a thaw in relations with her 31-year-old rival was unlikely.
"Sometimes with great rivalries, you can't have a great friendship because there is so much riding on the line for those involved that it's an uncomfortable situation to try to switch off and open yourself up to that person when you do need to beat them on the track," 28-year-old Meares said.
The pair, both eight-time world champions, will also face off in the women's team sprint which promises to be another tension-charged affair at the April 4-8 championships.
Meares and Kaarle McCulloch lowered their world record in the qualifying session at the London World Cup, the Olympic test event in February, only to have it wiped by Pendleton and Jessica Varnish in the gold medal decider.
The Meares-Pendleton rivalry is part of a bigger battle for supremacy between Britain and former colony Australia, who are widely tipped to dominate the track cycling medals at London.
Led by Scotsman Chris Hoy's three gold medals, the British riders took seven of the 10 titles on offer in Beijing, leaving Australia with a solitary silver medal and itching to atone for it by humbling the Olympic hosts on home soil.
Australia's "Cyclones" showed their class by storming to eight out of 19 titles at last year's world championships in Apeldoorn, Netherlands, but Britain issued a timely reminder of their pedigree with a dominant World Cup showing at the London velodrome in February.
At that Olympic test event, Hoy, now 36, showed he was far from a spent force by taking the sprint and keirin titles.
Hoy believes he can successfully defend his gold medals at London, which included the sprint, keirin and team sprint.
However, he faces a stiff test just to qualify for the individual sprint at London, with only one berth available per team and compatriot Jason Kenny snapping at his heels.
Kenny was awarded the sprint world title earlier this year after Frenchman Gregory Bauge was stripped of it for failing to comply with the anti-doping "whereabouts rule".
Hoy's path to the keirin title looks easier with Australia's reigning champion Shane Perkins bruised and carrying a sprained wrist after being hit by a car during training over the weekend.
While Bauge should anchor France's bid for the team sprint against a powerful German team that won the London World Cup title, the men's team pursuit appears set to be another showdown between Britain and Australia.
Australia, anchored by individual pursuit world champion Jack Bobridge, edged Russia in the final at Apeldoorn but the British quartet's pace at training in the lead-up has raised eyebrows in Melbourne.
"We are around the same mark as we were before London. I think it is definitely going to take a quicker time here to win than in London. That was a World Cup, this is a world title," Bobridge said.
Britain's world champion women's pursuit team beat Canada and won gold in a world record time at the Olympic test event and are favourites to defend their title in Melbourne.
New Zealand's Shane Archbold will defend his omnium world title ahead of the endurance discipline's Olympic debut, along with Canadian Tara Whitten in the women's event.