British Airways, Ryanair and EasyJet have filed a formal legal challenge to the government's quarantine policy.
The airlines say the policy will have "a devastating effect on British tourism and the wider economy" and destroy thousands of jobs.
They have applied for a judicial review at the High Court.
The challenge claims that the quarantine rules for travellers are more stringent than those applied to people who actually have Covid-19.
The new rules came into force this week. They require most inbound travellers to self isolate for 14 days, although there are more than 40 categories of incomers, largely pertaining to certain workers, who are exempt. Rules for those actually infected with the virus require self isolation for seven days.
The airlines state that there was no consultation and no scientific evidence provided to support the policy; that weekly commuters from France or Germany can be exempted; and that the government is preventing people from travelling to and from countries with lower infection rates than the UK.
However, the government has said the quarantine period is a "proportionate and time-limited approach" to protect public health.
In a statement, the three airlines said they had not seen any evidence on how and when so-called "air bridges", allowing quarantine-free travel between the UK and other countries with low infection rates, could be implemented.
They have called on the government instead to re-adopt a previous policy, where quarantine was limited to travellers from high risk countries.
The air industry has been hit particularly hard by the coronavirus outbreak, which has all but stopped their activities. Mass job cuts are under way:
- British Airways is proposing to make 12,000 of its 45,000 staff redundant, with more than 1,000 pilot roles at risk
- Ryanair is set to shed 3,000 jobs - 15% of its workforce - with boss Michael O'Leary saying the planned cuts are "the minimum that we need just to survive the next 12 months"
- EasyJet has said it will cut up to 30% of its workforce - about 4,500 jobs
- Virgin Atlantic, which employs 10,000 people, has said it will cut 3,000 jobs
- Other European airlines cutting back include Germany's Lufthansa, which on Thursday said it would cut 22,000 jobs.
Friday's legal move marks another sign of a breakdown in relations between airlines and the UK government.
Willie Walsh, the boss of IAG, which owns BA, Iberia and Aer Lingus, has called the quarantine policy "irrational", while Ryanair boss Michael O'Leary said the policy was a "stunt" and would not be enforceable.
Industry body Airlines UK has said quarantine "would effectively kill off air travel".
The BBC has approached the Home Office for a comment.